Thermal sterilization of heat-sensitive products using high-temperature short-time sterilization.

Science.gov (United States)

Mann, A; Kiefer, M; Leuenberger, H

2001-03-01

High-temperature short-time (HTST) sterilization with a continuous-flow sterilizer, developed for this study, was evaluated. The evaluation was performed with respect to (a) the chemical degradation of two heat-sensitive drugs in HTST range (140-160 degrees C) and (b) the microbiological effect of HTST sterilization. Degradation kinetics of two heat-sensitive drugs showed that a high peak temperature sterilization process resulted in less chemical degradation for the same microbiological effect than a low peak temperature process. Both drugs investigated could be sterilized with acceptable degradation at HTST conditions. For the evaluation of the microbiological effect, Bacillus stearothermophilus ATCC 7953 spores were used as indicator bacteria. Indicator spore kinetics (D(T), z value, k, and E(a)), were determined in the HTST range. A comparison between the Bigelow model (z value concept) and the Arrhenius model, used to describe the temperature coefficient of the microbial inactivation, demonstrated that the Bigelow model is more accurate in prediction of D(T) values in the HTST range. The temperature coefficient decreased with increasing temperature. The influence of Ca(2+) ions and pH value on the heat resistance of the indicator spores, which is known under typical sterilization conditions, did not change under HTST conditions.

Isolation of temperature-sensitive mutations in murC of Staphylococcus aureus.

Science.gov (United States)

Ishibashi, Mihoko; Kurokawa, Kenji; Nishida, Satoshi; Ueno, Kohji; Matsuo, Miki; Sekimizu, Kazuhisa

2007-09-01

Enzymes in the bacterial peptidoglycan biosynthesis pathway are important targets for novel antibiotics. Of 750 temperature-sensitive (TS) mutants of Gram-positive Staphylococcus aureus, six were complemented by the murC gene, which encodes the UDP-N-acetylmuramic acid:l-alanine ligase. Each mutation resulted in a single amino acid substitution and, in all cases, the TS phenotype was suppressed by high osmotic stress. In mutant strains with the G222E substitution, a decrease in the viable cell number immediately after shift to the restrictive temperature was observed. These results suggest that S. aureus MurC protein is essential for cell growth. The MurC H343Y mutation is located in the putative alanine recognition pocket. Consistent with this, allele-specific suppression was observed of the H343Y mutation by multiple copies of the aapA gene, which encodes an alanine transporter. The results suggest an in vivo role for the H343 residue of S. aureus MurC protein in high-affinity binding to L-alanine.

High-temperature sensitivity and its acclimation for photosynthetic electron reactions of desert succulents

Energy Technology Data Exchange (ETDEWEB)

Chetti, M.B.; Nobel, P.S. (Univ. of California, Los Angeles (USA))

1987-08-01

Photosynthetic electron reactions of succulent plants from hot deserts are able to tolerate extremely high temperatures and to acclimate to seasonal increase in temperature. In this study, we report the influence of relatively long, in vivo, high-temperature treatments on electron transport reactions for two desert succulents, Agave deserti and Opuntia ficus-indica, species which can tolerate 60{degree}C. Whole chain electron transport averaged 3{degree}C more sensitive to a 1-hour high-temperature treatment than did PSII (Photosystem II) which in turn averaged 3{degree}C more sensitive than did PSI. For plants maintained at day/night air temperatures of 30{degree}C/20{degree}C, treatment at 50{degree}C cause these reactions to be inhibited an average of 39% during the first hour, an additional 31% during the next 4 hours, and 100% by 12 hours. Upon shifting the plants from 30{degree}C/20{degree}C to 45{degree}C/35{degree}C, the high temperatures where activity was inhibited 50% increased 3{degree}C to 8{degree}C for the three electron transport reactions, the half-times for acclimation averaging 5 days for A. deserti and 4 days for O. ficus-indica. For the 45{degree}C/35{degree}C plants treated at 60{degree}C for 1 hour, PSI activity was reduced by 54% for A. deserti and 36% for O. ficus-indica. Acclimation leads to a toleration of very high temperatures without substantial disruption of electron transport for these desert succulents, facilitating their survival in hot deserts. Indeed, the electron transport reactions of these species tolerate longer periods at higher temperatures than any other vascular plants so far reported.

The fourth dimension in immunological space: how the struggle for nutrients selects high-affinity lymphocytes.

Science.gov (United States)

Wensveen, Felix M; van Gisbergen, Klaas P J M; Eldering, Eric

2012-09-01

Lymphocyte activation via the antigen receptor is associated with radical shifts in metabolism and changes in requirements for nutrients and cytokines. Concomitantly, drastic changes occur in the expression of pro-and anti-apoptotic proteins that alter the sensitivity of lymphocytes to limiting concentrations of key survival factors. Antigen affinity is a primary determinant for the capacity of activated lymphocytes to access these vital resources. The shift in metabolic needs and the variable access to key survival factors is used by the immune system to eliminate activated low-affinity cells and to generate an optimal high-affinity response. In this review, we focus on the control of apoptosis regulators in activated lymphocytes by nutrients, cytokines, and costimulation. We propose that the struggle among individual clones that leads to the formation of high-affinity effector cell populations is in effect an 'invisible' fourth signal required for effective immune responses. © 2012 John Wiley & Sons A/S.

Temperature uniformity mapping in a high pressure high temperature reactor using a temperature sensitive indicator

NARCIS (Netherlands)

Grauwet, T.; Plancken, van der I.; Vervoort, L.; Matser, A.M.; Hendrickx, M.; Loey, van A.

2011-01-01

Recently, the first prototype ovomucoid-based pressure–temperature–time indicator (pTTI) for high pressure high temperature (HPHT) processing was described. However, for temperature uniformity mapping of high pressure (HP) vessels under HPHT sterilization conditions, this prototype needs to be

Micromolar-Affinity Benzodiazepine Receptors Regulate Voltage-Sensitive Calcium Channels in Nerve Terminal Preparations

Science.gov (United States)

Taft, William C.; Delorenzo, Robert J.

1984-05-01

Benzodiazepines in micromolar concentrations significantly inhibit depolarization-sensitive Ca2+ uptake in intact nerve-terminal preparations. Benzodiazepine inhibition of Ca2+ uptake is concentration dependent and stereospecific. Micromolar-affinity benzodiazepine receptors have been identified and characterized in brain membrane and shown to be distinct from nanomolar-affinity benzodiazepine receptors. Evidence is presented that micromolar, and not nanomolar, benzodiazepine binding sites mediate benzodiazepine inhibition of Ca2+ uptake. Irreversible binding to micromolar benzodiazepine binding sites also irreversibly blocked depolarization-dependent Ca2+ uptake in synaptosomes, indicating that these compounds may represent a useful marker for identifying the molecular components of Ca2+ channels in brain. Characterization of benzodiazepine inhibition of Ca2+ uptake demonstrates that these drugs function as Ca2+ channel antagonists, because benzodiazepines effectively blocked voltage-sensitive Ca2+ uptake inhibited by Mn2+, Co2+, verapamil, nitrendipine, and nimodipine. These results indicate that micromolar benzodiazepine binding sites regulate voltage-sensitive Ca2+ channels in brain membrane and suggest that some of the neuronal stabilizing effects of micromolar benzodiazepine receptors may be mediated by the regulation of Ca2+ conductance.

A mix-and-read drop-based in vitro two-hybrid method for screening high-affinity peptide binders

Science.gov (United States)

Cui, Naiwen; Zhang, Huidan; Schneider, Nils; Tao, Ye; Asahara, Haruichi; Sun, Zhiyi; Cai, Yamei; Koehler, Stephan A.; de Greef, Tom F. A.; Abbaspourrad, Alireza; Weitz, David A.; Chong, Shaorong

2016-01-01

Drop-based microfluidics have recently become a novel tool by providing a stable linkage between phenotype and genotype for high throughput screening. However, use of drop-based microfluidics for screening high-affinity peptide binders has not been demonstrated due to the lack of a sensitive functional assay that can detect single DNA molecules in drops. To address this sensitivity issue, we introduced in vitro two-hybrid system (IVT2H) into microfluidic drops and developed a streamlined mix-and-read drop-IVT2H method to screen a random DNA library. Drop-IVT2H was based on the correlation between the binding affinity of two interacting protein domains and transcriptional activation of a fluorescent reporter. A DNA library encoding potential peptide binders was encapsulated with IVT2H such that single DNA molecules were distributed in individual drops. We validated drop-IVT2H by screening a three-random-residue library derived from a high-affinity MDM2 inhibitor PMI. The current drop-IVT2H platform is ideally suited for affinity screening of small-to-medium-sized libraries (103–106). It can obtain hits within a single day while consuming minimal amounts of reagents. Drop-IVT2H simplifies and accelerates the drop-based microfluidics workflow for screening random DNA libraries, and represents a novel alternative method for protein engineering and in vitro directed protein evolution. PMID:26940078

Electrochemical polarization behavior of sensitized SUS 304 stainless steel in high temperature water

Energy Technology Data Exchange (ETDEWEB)

Kushiya, K [Tohoku Univ., Sendai (Japan); Sugimoto, K; Ejima, T

1978-11-01

Anodic polarization curves for a solution-treated or sensitized SUS 304 stainless steel and solution-treated Fe-Ni-Cr ternary alloys containing 10%Ni and 6 to 14%Cr have been measured in deaerated 0.5 mol/l Na/sub 2/SO/sub 4/ solutions of pH 2.0 to 5.9 at 298, 523 and 553 K. Corrosion potentials for U-bend SCC test specimens of sensitized SUS 304 stainless steel have also been monitored for a long time in the same solutions as those used for the polarization measurements except that they were aerated. It was found that the differences in the current densities in the passive state, i sub(pass), between the solution treated steel and the sensitized one and also between the ternary alloy with higher Cr content and the one with lower Cr content become large with increasing temperature and decreasing pH. This means that the difference in the values of i sub(pass) between grain bodies and Cr-depleted zones along grain boundaries of sensitized steel becomes larger and susceptibility to intergranular corrosion of the sensitized steel in the passivation region becomes higher with increasing temperature and decreasing pH. Since corrosion potentials for the U-bend SCC test specimens in air-satulated solutions lie in the passive region of anodic polarization curves for the sensitized steel in deaerated solutions, the intergranular stress-corrosion cracking of the sensitized steel in high temperature water with dissolved oxygen is considered to be caused by the preferential corrosion in the Cr-depleted zone.

The fabrication of high sensitivity gold nanorod H2S gas sensors utilizing the highly uniform anodic aluminum oxide template

Directory of Open Access Journals (Sweden)

Chien-Yu Li

2016-12-01

Full Text Available Gold nanorod were fabricated using anodic alumina oxide template for H2S gas detection. The nanorod gas sensor exhibits high surface density and contact area, which can increase detection sensitivity. The anodic alumina oxide template contains an array of pores, with a width of 70 nm and a length of 27μm. Au nanorod were obtained through electro-deposition under a pulse bias of −1 V. The resistance of the Au nanorod was recorded upon exposure to various concentrations of H2S. The resistance could be attributed to the high electron affinity between sulfide and Au nanorod. Au–sulfide bonds provide strong bonding, which could alter the conductivity of the sensor. The gas sensor exhibits high sensitivity and short response time for H2S detection at room temperature.

A High-Sensitivity Gas Sensor Toward Methanol Using ZnO Microrods: Effect of Operating Temperature

Science.gov (United States)

Sinha, M.; Mahapatra, R.; Mondal, B.; Ghosh, R.

2017-04-01

In the present work, zinc oxide (ZnO) microrods with the average diameter of 350 nm have been synthesized on fluorine doped tin oxide (FTO) substrate using a hydrothermal reaction process at a low temperature of 90°C. The methanol gas sensing behaviour of as-synthesized ZnO microrods have been studied at different operating temperatures (100-300°C). The gas sensing results show that the ZnO microrods exhibit excellent sensitivity, selectivity, and stability toward methanol gas at 300°C. The as-grown ZnO microrods sensor also shows the good sensitivity for methanol even at a low operating temperature of 100°C. The ultra-high sensitivity of 4.41 × 104% [gas sensitivity, S g = ( I g - I a)/ I a × 100%] and 5.11 × 102% to 100 ppm methanol gas at a temperature of 300°C and 100°C, respectively, has been observed. A fast response time of 200 ms and 270 ms as well as a recovery time of 120 ms and 1330 ms to methanol gas have also been found at an operating temperature of 300°C and 100°C, respectively. The response and recovery time decreases with increasing operation temperature of the sensor.

Influence of pre-deformation, sensitization and oxidation in high temperature water on corrosion resistance of AISI 304 stainless steel

Energy Technology Data Exchange (ETDEWEB)

Lv, Jinlong, E-mail: ljltsinghua@126.com [Beijing Key Laboratory of Fine Ceramics, Institute of Nuclear and New Energy Technology, Tsinghua University, Zhongguancun Street, Haidian District, Beijing 100084 (China); State Key Lab of New Ceramic and Fine Processing, Tsinghua University, Beijing 100084 (China); Liang, Tongxiang [Beijing Key Laboratory of Fine Ceramics, Institute of Nuclear and New Energy Technology, Tsinghua University, Zhongguancun Street, Haidian District, Beijing 100084 (China); State Key Lab of New Ceramic and Fine Processing, Tsinghua University, Beijing 100084 (China); Luo, Hongyun [Key Laboratory of Aerospace Materials and Performance (Ministry of Education), School of Materials Science and Engineering, Beijing University of Aeronautics and Astronautics, Xueyuan Road 37, Beijing 100191 (China)

2016-12-01

Highlights: • The pre-strain accelerated desensitization and sensitization for austenitic stainless steels. • Low temperature sensitization (carbide precipitation) induced α′-martensite. • The sensitization level could affect directly corrosion resistance of the oxide film. - Abstract: The effects of pre-deformation on sensitization of AISI 304 stainless steel were investigated by the double loop electrochemical potentiokinetic reactivation test. The effects of pre-deformation and sensitization on high temperature oxidized film formed in high temperature water were analyzed by a XRD and SEM. The electrochemical impedance spectroscopy at room temperature was used to study corrosion resistance of oxidized film. The point defect density of oxidized film was calculated by Mott–Schottky plots. The results showed that the value of the degree of sensitization first decreased and then slight increased with the increasing of engineering strain. Moreover, low temperature promoted to form sensitization induced “secondary” α′-martensite. The sample with 20% engineering strain had higher impedance value than other samples. The result was supported by further Mott–Schottky experiments. Considering increased α′-martensite with the increasing of strain, the results of the impedance were more consistent with values of the degree of sensitization.

Temperature sensitive surfaces and methods of making same

Science.gov (United States)

Liang, Liang [Richland, WA; Rieke, Peter C [Pasco, WA; Alford, Kentin L [Pasco, WA

2002-09-10

Poly-n-isopropylacrylamide surface coatings demonstrate the useful property of being able to switch charateristics depending upon temperature. More specifically, these coatings switch from being hydrophilic at low temperature to hydrophobic at high temperature. Research has been conducted for many years to better characterize and control the properties of temperature sensitive coatings. The present invention provides novel temperature sensitive coatings on articles and novel methods of making temperature sensitive coatings that are disposed on the surfaces of various articles. These novel coatings contain the reaction products of n-isopropylacrylamide and are characterized by their properties such as advancing contact angles. Numerous other characteristics such as coating thickness, surface roughness, and hydrophilic-to-hydrophobic transition temperatures are also described. The present invention includes articles having temperature-sensitve coatings with improved properties as well as improved methods for forming temperature sensitive coatings.

High-affinity RNA aptamers to C-reactive protein (CRP): newly developed pre-elution methods for aptamer selection

International Nuclear Information System (INIS)

Orito, N; Umekage, S; Sakai, E; Tanaka, T; Kikuchi, Y; Sato, K; Kawauchi, S; Tanaka, H

2012-01-01

We have developed a modified SELEX (systematic evolution of ligands by exponential enrichment) method to obtain RNA aptamers with high affinity to C-reactive protein (CRP). CRP is a clinical biomarker present in plasma, the level of which increases in response to infections and noninfectious inflammation. The CRP level is also an important prognostic indicator in patients with several syndromes. At present, CRP content in blood is measured immunochemically using antibodies. To develop a more sensitive method using RNA aptamers, we have attempted to obtain high-affinity RNA aptamers to CRP. We succeeded in obtaining an RNA aptamer with high affinity to CRP using a CRP-immobilized Sepharose column and pre-elution procedure. Pre-elution is a method that removes the weak binding portion from a selected RNA population by washing for a short time with buffer containing CRP. By surface plasmon-resonance (SPR) analysis, the affinity constant of this aptamer for CRP was calculated to be K D = 2.25x10 -9 (M). The secondary structure, contact sites with CRP protein, and application of this aptamer will be described.

Affinity Crystallography: A New Approach to Extracting High-Affinity Enzyme Inhibitors from Natural Extracts.

Science.gov (United States)

Aguda, Adeleke H; Lavallee, Vincent; Cheng, Ping; Bott, Tina M; Meimetis, Labros G; Law, Simon; Nguyen, Nham T; Williams, David E; Kaleta, Jadwiga; Villanueva, Ivan; Davies, Julian; Andersen, Raymond J; Brayer, Gary D; Brömme, Dieter

2016-08-26

Natural products are an important source of novel drug scaffolds. The highly variable and unpredictable timelines associated with isolating novel compounds and elucidating their structures have led to the demise of exploring natural product extract libraries in drug discovery programs. Here we introduce affinity crystallography as a new methodology that significantly shortens the time of the hit to active structure cycle in bioactive natural product discovery research. This affinity crystallography approach is illustrated by using semipure fractions of an actinomycetes culture extract to isolate and identify a cathepsin K inhibitor and to compare the outcome with the traditional assay-guided purification/structural analysis approach. The traditional approach resulted in the identification of the known inhibitor antipain (1) and its new but lower potency dehydration product 2, while the affinity crystallography approach led to the identification of a new high-affinity inhibitor named lichostatinal (3). The structure and potency of lichostatinal (3) was verified by total synthesis and kinetic characterization. To the best of our knowledge, this is the first example of isolating and characterizing a potent enzyme inhibitor from a partially purified crude natural product extract using a protein crystallographic approach.

Temperature sensitivity of respiration scales with organic matter recalcitrance

Science.gov (United States)

Craine, J. M.; Fierer, N.; McLauchlan, K. K.

2010-12-01

Microbial decomposition of soil organic matter is a key process in determining the carbon sequestration potential of ecosystems and carbon fluxes to the atmosphere. Since microbial decomposition is highly sensitive to short-term changes in temperature, predicting the temperature sensitivity of microbial decomposition is critical to predicting future atmospheric carbon dioxide concentrations and feedbacks to anthropogenic warming. Fundamental principles of enzyme kinetics, embodied in the carbon-quality temperature hypothesis, predict that the temperature sensitivity of microbial decomposition should increase with increasing biochemical recalcitrance of a substrate. To test the generality of this principle, we measured the temperature sensitivity of microbial respiration of soil organic matter with serial short-term temperature manipulations over 365 days for 28 North American soils. When joined with data from similar studies that represent a wide variety of contrasts, we show that the temperature sensitivity of organic matter decomposition scales with biochemical recalcitrance. With physico-chemical protection likely an important covariate for relating plant and soil organic matter decomposition scalars, biochemically recalcitrant organic matter is highly susceptible to short-term increases in temperature, a key link in predicting the effects of warming on carbon cycling.

High Temperature and High Sensitive NOx Gas Sensor with Hetero-Junction Structure using Laser Ablation Method

Science.gov (United States)

Gao, Wei; Shi, Liqin; Hasegawa, Yuki; Katsube, Teruaki

In order to develop a high temperature (200°C˜400°C) and high sensitive NOx gas sensor, we developed a new structure of SiC-based hetero-junction device Pt/SnO2/SiC/Ni, Pt/In2O3/SiC/Ni and Pt/WO3/SiC/Ni using a laser ablation method for the preparation of both metal (Pt) electrode and metal-oxide film. It was found that Pt/In2O3/SiC/Ni sensor shows higher sensitivity to NO2 gas compared with the Pt/SnO2/SiC/Ni and Pt/WO3/SiC/Ni sensor, whereas the Pt/WO3/SiC/Ni sensor had better sensitivity to NO gas. These results suggest that selective detection of NO and NO2 gases may be obtained by choosing different metal oxide films.

High affinity and temperature sensitivity of blood oxygen binding in Pangasianodon hypophthalmus due to lack of chloride-hemoglobin allosteric interaction

DEFF Research Database (Denmark)

Damsgaard, Christian; Phuong, Le My; Huong, Do Thi Thanh

2015-01-01

Air-breathing fishes represent interesting organisms in terms of understanding the physiological changes associated with the terrestrialization of vertebrates, and, further, are of great socio-economic importance for aquaculture in Southeast Asia. To understand how environmental factors......, such as high temperature, affect O2 transport in air-breathing fishes, this study assessed the effects of temperature on O2 binding of blood and Hb in the economically important air-breathing fish Pangasianodon hypophthalmus. To determine blood O2 binding properties, blood was drawn from resting cannulated...

Opioid receptor subtypes mediating the noise-induced decreases in high-affinity choline uptake in the rat brain.

Science.gov (United States)

Lai, H; Carino, M A

1992-07-01

Acute (20 min) exposure to 100-dB white noise elicits a naltrexone-sensitive decrease in sodium-dependent high-affinity choline uptake in the frontal cortex and hippocampus of the rat. In the present study, the subtypes of opioid receptors involved were investigated by pretreating rats with microinjection of specific opioid-receptor antagonists into the lateral cerebroventricle before noise exposure. We found that the noise-induced decrease in high-affinity choline uptake in the hippocampus was blocked by pretreatment with either mu-, delta-, or kappa-opioid-receptor antagonists, whereas the effect of noise on frontal cortical high-affinity choline uptake was blocked by a mu- and delta- but not by a kappa-antagonist. These data further confirm the role of endogenous opioids in mediating the effects of noise on central cholinergic activity and indicate that different neural mechanisms are involved in the effects of noise on the frontal cortical and hippocampal cholinergic systems.

Carbonate-sensitive phytotransferrin controls high-affinity iron uptake in diatoms

Science.gov (United States)

McQuaid, Jeffrey B.; Kustka, Adam B.; Oborník, Miroslav; Horák, Aleš; McCrow, John P.; Karas, Bogumil J.; Zheng, Hong; Kindeberg, Theodor; Andersson, Andreas J.; Barbeau, Katherine A.; Allen, Andrew E.

2018-03-01

In vast areas of the ocean, the scarcity of iron controls the growth and productivity of phytoplankton. Although most dissolved iron in the marine environment is complexed with organic molecules, picomolar amounts of labile inorganic iron species (labile iron) are maintained within the euphotic zone and serve as an important source of iron for eukaryotic phytoplankton and particularly for diatoms. Genome-enabled studies of labile iron utilization by diatoms have previously revealed novel iron-responsive transcripts, including the ferric iron-concentrating protein ISIP2A, but the mechanism behind the acquisition of picomolar labile iron remains unknown. Here we show that ISIP2A is a phytotransferrin that independently and convergently evolved carbonate ion-coordinated ferric iron binding. Deletion of ISIP2A disrupts high-affinity iron uptake in the diatom Phaeodactylum tricornutum, and uptake is restored by complementation with human transferrin. ISIP2A is internalized by endocytosis, and manipulation of the seawater carbonic acid system reveals a second-order dependence on the concentrations of labile iron and carbonate ions. In P. tricornutum, the synergistic interaction of labile iron and carbonate ions occurs at environmentally relevant concentrations, revealing that carbonate availability co-limits iron uptake. Phytotransferrin sequences have a broad taxonomic distribution and are abundant in marine environmental genomic datasets, suggesting that acidification-driven declines in the concentration of seawater carbonate ions will have a negative effect on this globally important eukaryotic iron acquisition mechanism.

Ultrahigh temperature-sensitive silicon MZI with titania cladding

Directory of Open Access Journals (Sweden)

Jong-Moo eLee

2015-05-01

Full Text Available We present a possibility of intensifying temperature sensitivity of a silicon Mach-Zehnder interferometer (MZI by using a highly negative thermo-optic property of titania (TiO2. Temperature sensitivity of an asymmetric silicon MZI with a titania cladding is experimentally measured from +18pm/C to -340 pm/C depending on design parameters of MZI.

Binding Affinity of a Highly Sensitive Au/Ag/Au/Chitosan-Graphene Oxide Sensor Based on Direct Detection of Pb2+ and Hg2+ Ions

Directory of Open Access Journals (Sweden)

Nur Hasiba Kamaruddin

2017-10-01

Full Text Available The study of binding affinity is essential in surface plasmon resonance (SPR sensing because it allows researchers to quantify the affinity between the analyte and immobilised ligands of an SPR sensor. In this study, we demonstrate the derivation of the binding affinity constant, K, for Pb2+ and Hg2+ ions according to their SPR response using a gold/silver/gold/chitosan–graphene oxide (Au/Ag/Au/CS–GO sensor for the concentration range of 0.1–5 ppm. The higher affinity of Pb2+ to binding with the CS–GO sensor explains the outstanding sensitivity of 2.05 °ppm−1 against 1.66 °ppm−1 of Hg2+. The maximum signal-to-noise ratio (SNR upon detection of Pb2+ is 1.53, and exceeds the suggested logical criterion of an SNR. The Au/Ag/Au/CS–GO SPR sensor also exhibits excellent repeatability in Pb2+ due to the strong bond between its functional groups and this cation. The adsorption data of Pb2+ and Hg2+ on the CS–GO sensor fits well with the Langmuir isotherm model where the affinity constant, K, of Pb2+ and Hg2+ ions is computed. The affinity of Pb2+ ions to the Au/Ag/Au/CS–GO sensor is significantly higher than that of Hg2+ based on the value of K, 7 × 105 M−1 and 4 × 105 M−1, respectively. The higher shift in SPR angles due to Pb2+ and Hg2+ compared to Cr3+, Cu2+ and Zn2+ ions also reveals the greater affinity of the CS–GO SPR sensor to them, thus supporting the rationale for obtaining K for these two heavy metals. This study provides a better understanding on the sensing performance of such sensors in detecting heavy metal ions.

Autoradiographic imaging and quantification of the high-affinity GHB binding sites in rodent brain using (3)H-HOCPCA

DEFF Research Database (Denmark)

Klein, A B; Bay, T; Villumsen, I S

2016-01-01

analogue, 3-hydroxycyclopent-1-enecarboxylic acid (HOCPCA) as a tritiated version ((3)H-HOCPCA) to radioactively label the specific GHB high-affinity binding site and gain further insight into the density, distribution and developmental profile of this protein. We show that, in low nanomolar concentrations...... brain development. Due to the high sensitivity of this radioligand, we were able to detect low levels of specific binding already at E15 in mouse brain, which increased progressively until adulthood. Collectively, we show that (3)H-HOCPCA is a highly sensitive radioligand, offering advantages over...

Identification of high-affinity anti-IL-1 α autoantibodies in normal human serum as an interfering substance in a sensitive enzyme-linked immunosorbent assay for IL-1 α

International Nuclear Information System (INIS)

Mae, N.; Liberato, D.J.; Chizzonite, R.; Satoh, H.

1991-01-01

A highly reproducible, sensitive, and specific sandwich enzyme-linked immunosorbent assay (ELISA) for recombinant human IL-1 α (rhIL-1 alpha) has been developed. Results from this ELISA have demonstrated that the concentration of rhIL-1 α added to normal human serum (NHS) decreased by 16.3% after 3 h and 24.9% after 6 h at room temperature. Molecular exclusion column chromatography with Sephacryl S-300 HR revealed that 125I-labeled IL-1 α added to normal human serum rapidly formed higher molecular weight complexes without indication of proteolytic degradation. The observed reduction in immunoreactivity was correlated with this protein complex formation and accounted for the apparent instability of rhIL-1 α in NHS. Immunoblot analysis indicated that the molecular weight of the binding protein was 150-160K, and the IL-1 α binding activity was removed and recovered from NHS by Protein-G affinity chromatography; indicating that the binding protein was IL-1 α-specific IgG. The binding of 125I-labeled IL-1 α to the serum binding proteins could be inhibited by unlabeled IL-1 alpha (IC50 = 7.4 x 10(-11) M) but not by unlabeled IL-1 β. Kinetic analysis with 125I-labeled IL-1 alpha revealed that the average binding affinity of these IL-1 α-specific IgGs was 4.7 x 10(10) M-1. These results suggest that these autoantibodies may interfere with the detection of IL-1 α in human serum by various assay systems and also could be a regulator of circulating IL-1 α

Quantifying high-affinity binding of hydrophobic ligands by isothermal titration calorimetry.

Science.gov (United States)

Krainer, Georg; Broecker, Jana; Vargas, Carolyn; Fanghänel, Jörg; Keller, Sandro

2012-12-18

A fast and reliable quantification of the binding thermodynamics of hydrophobic high-affinity ligands employing a new calorimetric competition experiment is described. Although isothermal titration calorimetry is the method of choice for a quantitative characterization of intermolecular interactions in solution, a reliable determination of a dissociation constant (K(D)) is typically limited to the range 100 μM > K(D) > 1 nM. Interactions displaying higher or lower K(D) values can be assessed indirectly, provided that a suitable competing ligand is available whose K(D) falls within the directly accessible affinity window. This established displacement assay, however, requires the high-affinity ligand to be soluble at high concentrations in aqueous buffer and, consequently, poses serious problems in the study of protein binding involving small-molecule ligands dissolved in organic solvents--a familiar case in many drug-discovery projects relying on compound libraries. The calorimetric competition assay introduced here overcomes this limitation, thus allowing for a detailed thermodynamic description of high-affinity receptor-ligand interactions involving poorly water-soluble compounds. Based on a single titration of receptor into a dilute mixture of the two competing ligands, this competition assay provides accurate and precise values for the dissociation constants and binding enthalpies of both high- and moderate-affinity ligands. We discuss the theoretical background underlying the approach, demonstrate its practical application to metal ion chelation and high-affinity protein-inhibitor interactions, and explore its potential and limitations with the aid of simulations and statistical analyses.

Lectin affinity electrophoresis.

Science.gov (United States)

Kobayashi, Yuka

2014-01-01

An interaction or a binding event typically changes the electrophoretic properties of a molecule. Affinity electrophoresis methods detect changes in the electrophoretic pattern of molecules (mainly macromolecules) that occur as a result of biospecific interactions or complex formation. Lectin affinity electrophoresis is a very effective method for the detection and analysis of trace amounts of glycobiological substances. It is particularly useful for isolating and separating the glycoisomers of target molecules. Here, we describe a sensitive technique for the detection of glycoproteins separated by agarose gel-lectin affinity electrophoresis that uses antibody-affinity blotting. The technique is tested using α-fetoprotein with lectin (Lens culinaris agglutinin and Phaseolus vulgaris agglutinin)-agarose gels.

An Inverse Relationship Links Temperature and Substrate Apparent Affinity in the Ion-Coupled Cotransporters rGAT1 and KAAT1

Directory of Open Access Journals (Sweden)

Antonio Peres

2012-11-01

Full Text Available The effects of temperature on the operation of two ion-coupled cotransporters of the SLC6A family, namely rat GAT1 (SLC6A1 and KAAT1 (SLC6A19 from Manduca sexta, have been studied by electrophysiological means in Xenopus laevis oocytes expressing these proteins. The maximal transport-associated current (Imax and the apparent substrate affinity (K05 were measured. In addition to the expected increase in transport rate (Q10 = 3–6, both transporters showed greater K05 values (i.e., a decrease in apparent affinity at higher temperatures. The transport efficiency, estimated as Imax/K05, increased at negative potentials in both transporters, but did not show statistically significant differences with temperature. The observation that the apparent substrate affinity is inversely related to the transport rate suggests a kinetic regulation of this parameter. Furthermore, the present results indicate that the affinities estimated at room temperature for mammalian cotransporters may not be simply extrapolated to their physiological operating conditions.

Characterization of high affinity [3H]triazolam binding in rat brain

International Nuclear Information System (INIS)

Earle, M.; Concas, A.; Yamamura, H.I.

1986-01-01

The hypnotic Triazolam (TZ), a triazolo (1,4)-benzodiazepine, displays a short physiological half life and has been used for the treatment of insomnia related to anxiety states. Specific binding properties of this recently tritiated TZ were characterized. The authors major objectives were the direct measurement of the temperature dependence and the GABA effect on [ 3 H]TZ binding. Saturation studies showed a shift to lower affinity at 37 0 C (K/sub d/ = 0.25 +/- 0.01 nM at O 0 C; K/sub d/ = 1.46 +/- 0.03 nM at 37 0 C) while the B/sub max/ values remained unchanged (1003 +/- 37 fmoles/mg prot. at 0 0 C and 1001 +/- 43 fmoles/mg prot. at 37 0 C). Inhibition studies showed that [ 3 H]TZ binding displayed no GABA shift at 0 0 C(K/sub i/ 0.37 +/- 0.03 nM/- GABA and K/sub i/ = 0.55 +/- 0.13 nM/+GABA) but a nearly two-fold shift was apparent at 37 0 C (K/sub i/ = 2.92 +/- 0.2 nM/-GABA; K/sub i/ = 1.37 +/- 0.11 mM/+GABA). These results were also confirmed by saturation studies in the presence or absence of GABA showing a shift to higher affinity in the presence of GABA only at 37 0 C. In Ro 15-1788/[ 3 H]TZ competition experiments the presence of GABA did not affect the inhibitory potency of Ro 15-1788 on [ 3 H]TZ binding at both temperatures. In conclusion [ 3 H]TZ binding showed an extremely high affinity for benzodiazepine receptors. In contrast to reported literature, the findings suggest that TZ interacts with benzodiazepine receptors similar to other benzodiazepine agonists

Sensitivity Studies of Advanced Reactors Coupled to High Temperature Electrolysis (HTE) Hydrogen Production Processes

International Nuclear Information System (INIS)

Edwin A. Harvego; Michael G. McKellar; James E. O'Brien; J. Stephen Herring

2007-01-01

High Temperature Electrolysis (HTE), when coupled to an advanced nuclear reactor capable of operating at reactor outlet temperatures of 800 C to 950 C, has the potential to efficiently produce the large quantities of hydrogen needed to meet future energy and transportation needs. To evaluate the potential benefits of nuclear-driven hydrogen production, the UniSim process analysis software was used to evaluate different reactor concepts coupled to a reference HTE process design concept. The reference HTE concept included an Intermediate Heat Exchanger and intermediate helium loop to separate the reactor primary system from the HTE process loops and additional heat exchangers to transfer reactor heat from the intermediate loop to the HTE process loops. The two process loops consisted of the water/steam loop feeding the cathode side of a HTE electrolysis stack, and the steam or air sweep loop used to remove oxygen from the anode side. The UniSim model of the process loops included pumps to circulate the working fluids and heat exchangers to recover heat from the oxygen and hydrogen product streams to improve the overall hydrogen production efficiencies. The reference HTE process loop model was coupled to separate UniSim models developed for three different advanced reactor concepts (a high-temperature helium cooled reactor concept and two different supercritical CO2 reactor concepts). Sensitivity studies were then performed to evaluate the affect of reactor outlet temperature on the power cycle efficiency and overall hydrogen production efficiency for each of the reactor power cycles. The results of these sensitivity studies showed that overall power cycle and hydrogen production efficiencies increased with reactor outlet temperature, but the power cycle producing the highest efficiencies varied depending on the temperature range considered

Identification and properties of very high affinity brain membrane-binding sites for a neurotoxic phospholipase from the taipan venom

International Nuclear Information System (INIS)

Lambeau, G.; Barhanin, J.; Schweitz, H.; Qar, J.; Lazdunski, M.

1989-01-01

Four new monochain phospholipases were purified from the Oxyuranus scutellatus (taipan) venom. Three of them were highly toxic when injected into mice brain. One of these neurotoxic phospholipases, OS2, was iodinated and used in binding experiments to demonstrate the presence of two families of specific binding sites in rat brain synaptic membranes. The affinities were exceptionally high, Kd1 = 1.5 +/- 0.5 pM and Kd2 = 45 +/- 10 pM, and the maximal binding capacities were Bmax 1 = 1 +/- 0.4 and Bmax 2 = 3 +/- 0.5 pmol/mg of protein. Both binding sites were sensitive to proteolysis and demonstrated to be located on proteins of Mr 85,000-88,000 and 36,000-51,000 by cross-linking and photoaffinity labeling techniques. The binding of 125 I-OS2 to synaptic membranes was dependent on Ca2+ ions and enhanced by Zn2+ ions which inhibit phospholipase activity. Competition experiments have shown that, except for beta-bungarotoxin, a number of known toxic snake or bee phospholipases have very high affinities for the newly identified binding sites. A good correlation (r = 0.80) was observed between toxicity and affinity but not between phospholipase activity and affinity

High affinity hemoglobin and Parkinson's disease.

Science.gov (United States)

Graham, Jeffrey; Hobson, Douglas; Ponnampalam, Arjuna

2014-12-01

Parkinson's disease (PD) is a neurodegenerative disorder characterized by the loss of dopaminergic neurons in the substantia nigra (SN) region of the midbrain. Oxidative damage in this region has been shown to play an important role in the pathogenesis of this disease. Human neurons have been discovered to contain hemoglobin, with an increased concentration seen in the neurons of the SN. High affinity hemoglobin is a clinical entity resulting from mutations that create a functional increase in the binding of hemoglobin to oxygen and an inability to efficiently unload it to tissues. This can result in a number of metabolic compensatory changes, including an elevation in circulating hemoglobin and an increase in the molecule 2,3-diphosphoglycerate (2,3-DPG). Population based studies have revealed that patients with PD have elevated hemoglobin as well as 2,3-DPG levels. Based on these observations, we hypothesize that the oxidative damage seen in PD is related to an underlying high affinity hemoglobin subtype. Copyright © 2014 Elsevier Ltd. All rights reserved.

Engineering of bispecific affinity proteins with high affinity for ERBB2 and adaptable binding to albumin.

Directory of Open Access Journals (Sweden)

Johan Nilvebrant

Full Text Available The epidermal growth factor receptor 2, ERBB2, is a well-validated target for cancer diagnostics and therapy. Recent studies suggest that the over-expression of this receptor in various cancers might also be exploited for antibody-based payload delivery, e.g. antibody drug conjugates. In such strategies, the full-length antibody format is probably not required for therapeutic effect and smaller tumor-specific affinity proteins might be an alternative. However, small proteins and peptides generally suffer from fast excretion through the kidneys, and thereby require frequent administration in order to maintain a therapeutic concentration. In an attempt aimed at combining ERBB2-targeting with antibody-like pharmacokinetic properties in a small protein format, we have engineered bispecific ERBB2-binding proteins that are based on a small albumin-binding domain. Phage display selection against ERBB2 was used for identification of a lead candidate, followed by affinity maturation using second-generation libraries. Cell surface display and flow-cytometric sorting allowed stringent selection of top candidates from pools pre-enriched by phage display. Several affinity-matured molecules were shown to bind human ERBB2 with sub-nanomolar affinity while retaining the interaction with human serum albumin. Moreover, parallel selections against ERBB2 in the presence of human serum albumin identified several amino acid substitutions that dramatically modulate the albumin affinity, which could provide a convenient means to control the pharmacokinetics. The new affinity proteins competed for ERBB2-binding with the monoclonal antibody trastuzumab and recognized the native receptor on a human cancer cell line. Hence, high affinity tumor targeting and tunable albumin binding were combined in one small adaptable protein.

Characterization of a high affinity cocaine binding site in rat brain

International Nuclear Information System (INIS)

Calligaro, D.; Eldefrawi, M.

1986-01-01

Binding of [ 3 H]cocaine to synaptic membranes from whole rat brain was reversible and saturable. Nonlinear regression analysis of binding isotherms indicated two binding affinities: one with k/sub d/ = 16 nM, B/sub max/ = 0.65 pmoles/mg protein and the other with K/sub d/ = 660 nM, B/sub max/ = 5.1 pmoles/mg protein. The high-affinity binding of [ 3 H]cocaine was sensitive to the actions of trypsin and chymotrypsin but not carboxypeptidase, and was eliminated by exposure of the membranes to 95 0 C for 5 min. Specific binding at 2 nM was higher at pH 8.8 than at pH 7.0. Binding of [ 3 H]cocaine (15 nM) was inhibited by increasing concentrations of Na + ions. Several cocaine analogues, neurotransmitter uptake inhibitors and local anesthetics displaced specific [ 3 H]cocaine binding at 2 nM with various potencies. The cocaine analogue (-)-norcocaine was the most potent (IC 50 = 10 nM), while the local anesthetic tetracaine was the least potent in inhibiting [ 3 H]cocaine binding. Several biogenic amine uptake inhibitors, including tricyclic antidepressants and phencyclidine, had IC 50 values below μM concentrations

A luminescent Lanthanide-free MOF nanohybrid for highly sensitive ratiometric temperature sensing in physiological range.

Science.gov (United States)

Zhou, You; Zhang, Denan; Zeng, Jin; Gan, Ning; Cuan, Jing

2018-05-01

Luminescent MOF materials with tunable emissions and energy/charge transfer processes have been extensively explored as ratiometric temperature sensors. However, most of the ratiometric MOF thermometers reported thus far are based on the MOFs containing photoactive lanthanides, which are potentially facing cost issue and serious supply shortage. Here, we present a ratiometric luminescent thermometer based on a dual-emitting lanthanide-free MOF hybrid, which is developed by encapsulation of a fluorescent dye into a robust nanocrystalline zirconium-based MOF through a one-pot synthesis approach. The structure and morphology of the hybrid product was characterized by Powder X-ray diffraction (PXRD), N 2 adsorption-desorption measurement and Scanning electron microscopy (SEM). The pore confinement effect well isolates the guest dye molecules and therefore suppresses the nonradiative energy transfer process between dye molecules. The incorporated dye emission is mainly sensitized by the organic linkers within MOF through fluorescence resonance energy transfer. The ratiometric luminescence of the MOF hybrid shows a significant response to temperature due to the thermal-related back energy transfer process from dye molecules and organic linkers, thus can be exploited for self-calibrated temperature sensing. The maximum thermometric sensitivity is 1.19% °C -1 in the physiological temperature range, which is among the highest for the ratiomtric MOF thermometers that operating in 25-45°C. The temperature resolution is better than 0.1°C over the entire operative range (20-60°C). By integrating the advantages of excellent stability, nanoscale nature, and high sensitivity and precision in the physiological temperature range, this dye@MOF hybrid might have potential application in biomedical diagnosis. What' more, this work has expanded the possibility of non-lanthanide luminescent MOF materials for the development of ratiometric temperature sensors. Copyright © 2018

High-throughput fragment screening by affinity LC-MS.

Science.gov (United States)

Duong-Thi, Minh-Dao; Bergström, Maria; Fex, Tomas; Isaksson, Roland; Ohlson, Sten

2013-02-01

Fragment screening, an emerging approach for hit finding in drug discovery, has recently been proven effective by its first approved drug, vemurafenib, for cancer treatment. Techniques such as nuclear magnetic resonance, surface plasmon resonance, and isothemal titration calorimetry, with their own pros and cons, have been employed for screening fragment libraries. As an alternative approach, screening based on high-performance liquid chromatography separation has been developed. In this work, we present weak affinity LC/MS as a method to screen fragments under high-throughput conditions. Affinity-based capillary columns with immobilized thrombin were used to screen a collection of 590 compounds from a fragment library. The collection was divided into 11 mixtures (each containing 35 to 65 fragments) and screened by MS detection. The primary screening was performed in 3500 fragments per day). Thirty hits were defined, which subsequently entered a secondary screening using an active site-blocked thrombin column for confirmation of specificity. One hit showed selective binding to thrombin with an estimated dissociation constant (K (D)) in the 0.1 mM range. This study shows that affinity LC/MS is characterized by high throughput, ease of operation, and low consumption of target and fragments, and therefore it promises to be a valuable method for fragment screening.

The influence of drawing temperature on mechanical properties and organisation of melt spun polyethylene solid-state drawn in the pseudo-affine regime

NARCIS (Netherlands)

Hu, Xin; Alcock, B.; Loos, J.

2006-01-01

Mechanical properties of high density polyethylene (HDPE) solid-state drawn with fixed draw ratio at different temperatures in a fiber/tape spin line were investigated. All drawing experiments were performed in the pseudo-affine regime, i.e. no effective relaxation of the molecules occurs during

Identification and properties of very high affinity brain membrane-binding sites for a neurotoxic phospholipase from the taipan venom

Energy Technology Data Exchange (ETDEWEB)

Lambeau, G.; Barhanin, J.; Schweitz, H.; Qar, J.; Lazdunski, M. (Centre de Biochimie, Nice (France))

1989-07-05

Four new monochain phospholipases were purified from the Oxyuranus scutellatus (taipan) venom. Three of them were highly toxic when injected into mice brain. One of these neurotoxic phospholipases, OS2, was iodinated and used in binding experiments to demonstrate the presence of two families of specific binding sites in rat brain synaptic membranes. The affinities were exceptionally high, Kd1 = 1.5 +/- 0.5 pM and Kd2 = 45 +/- 10 pM, and the maximal binding capacities were Bmax 1 = 1 +/- 0.4 and Bmax 2 = 3 +/- 0.5 pmol/mg of protein. Both binding sites were sensitive to proteolysis and demonstrated to be located on proteins of Mr 85,000-88,000 and 36,000-51,000 by cross-linking and photoaffinity labeling techniques. The binding of {sup 125}I-OS2 to synaptic membranes was dependent on Ca2+ ions and enhanced by Zn2+ ions which inhibit phospholipase activity. Competition experiments have shown that, except for beta-bungarotoxin, a number of known toxic snake or bee phospholipases have very high affinities for the newly identified binding sites. A good correlation (r = 0.80) was observed between toxicity and affinity but not between phospholipase activity and affinity.

Generation and characterization of a human-mouse chimeric high-affinity antibody that detects the DYKDDDDK FLAG peptide.

Science.gov (United States)

Ikeda, Koki; Koga, Tomoaki; Sasaki, Fumiyuki; Ueno, Ayumi; Saeki, Kazuko; Okuno, Toshiaki; Yokomizo, Takehiko

2017-05-13

DYKDDDDK peptide (FLAG) is a useful tool for investigating the function and localization of proteins whose antibodies (Abs) are not available. We recently established a high-affinity monoclonal antibody (mAb) for FLAG (clone 2H8). The 2H8 Ab is highly sensitive for detecting FLAG-tagged proteins by flowcytometry and immunoprecipitation, but it can yield nonspecific signals in immunohistochemistry of mouse tissues because it is of mouse origin. In this study, we reduced nonspecific signals by generating a chimeric 2H8 Ab with Fc fragments derived from human immunoglobulin. We fused a 5' terminal cDNA fragments for the Fab region of 2H8 mAb with 3' terminal cDNA fragments for Fc region of human IgG1. We transfected both chimeric plasmids and purified the resulting human-mouse chimeric 2H8. The chimeric 2H8 Ab successfully detected FLAG-tagged proteins in flowcytometry with anti-human IgG secondary Ab with comparable sensitivity to 2H8 mAb. Importantly, chimeric 2H8 detected specific FLAG peptide signals without nonspecific signals in immunohistochemical analysis with mouse tissues. This human-mouse chimeric high-affinity anti-FLAG Ab will prove useful for future immunohistochemical analysis of mouse tissues. Copyright © 2017 Elsevier Inc. All rights reserved.

Novel trends in affinity biosensors: current challenges and perspectives

International Nuclear Information System (INIS)

Arugula, Mary A; Simonian, Aleksandr

2014-01-01

Molecular biorecognition processes facilitate physical and biochemical interactions between molecules in all crucial metabolic pathways. Perhaps the target analyte and the biorecognition element interactions have the most impactful use in biosensing applications. Traditional analytical sensing systems offer excellent biorecognition elements with the ability to detect and determine the presence of analytes. High affinity antibodies and DNA play an important role in the development of affinity biosensors based on electrochemical, optical and mass sensitive approaches. Advancements in this area routinely employ labels, label free, nanoparticles, multifunctional matrices, carbon nanotubes and other methods to meet the requirements of its own application. However, despite increasing affinity ceilings for conventional biosensors, the field draws back in meeting specifically important demands, such as long-term stability, ultrasensitivity, rapid detection, extreme selectivity, strong biological base, calibration, in vivo measurements, regeneration, satisfactory performance and ease of production. Nevertheless, recent efforts through this line have produced novel high-tech nanosensing systems such as ‘aptamers’ and ‘phages’ which exhibit high-throughput sensing. Aptamers and phages are powerful tools that excel over antibodies in sensibility, stability, multi-detection, in vivo measurements and regeneration. Phages are superior in stability, screening for affinity-based target molecules ranging from small to proteins and even cells, and easy production. In this review, we focus mainly on recent developments in affinity-based biosensors such as immunosensors, DNA sensors, emphasizing aptasensors and phage-based biosensors basing on novel electrochemical, optical and mass sensitive detection techniques. We also address enzyme inhibition-based biosensors and the current problems associated with the above sensors and their future perspectives. (topical review)

High-sensitivity β-Ga_2O_3 solar-blind photodetector on high-temperature pretreated c-plane sapphire substrate

KAUST Repository

Qian, Ling-Xuan

2017-09-20

Recently, monoclinic Ga2O3 (beta-Ga2O3) photodetectors (PDs) have been extensively studied for various commercial and military applications due to the merits of intrinsic solar rejection, high gain, and great compactness. In this work, c-plane sapphire substrates were annealed under different temperatures in a vacuum furnace prior to the molecular beam epitaxy (MBE) of beta-Ga2O3 thin film, which yielded a smoother surface and even a terraceand- step-like morphology on the substrate, resulting in improved crystallinity of the epitaxial film. Accordingly, both the dark and photo currents of beta-Ga2O3 metal-semiconductor-metal (MSM) PDs were increased by the enhanced carrier mobility (mu) of the more crystalline film. However, the substrate-annealing temperature must be sufficiently high to offset the rise of the dark current and thus achieve a remarkable improvement in the photodetection properties. As a result, the PD fabricated on the 1050 degrees C-annealed substrate exhibited extremely high sensitivity, for example, high responsivity (R) of 54.9 A/ W and large specific detectivity (D*) of 3.71 x 10(14) Jones. Both parameters were increased by one order of magnitude because of the combined effects of the dramatic increase in mu and the effective reduction in defect-related recombination centers. Nevertheless, the latter also prolonged the recovery time of the PD. These findings suggest another way to develop beta-Ga2O3 PD with extremely high sensitivity. (C) 2017 Optical Society of America

The effect of label affinity on the sensitivity and specificity of a hapten radioimmunoassay: A comparison of three [125I]diphenylhydantoin radioligands with the 14C-labelled drug

International Nuclear Information System (INIS)

Rowell, F.J.

1979-01-01

The effects on the sensitivity and specificity of a radioimmunoassay for diphenylhydantoin (DPH) has been investigated using three 125 I-labelled tyrosine ester derivatives of DPH having different bridge lengths between the tyrosine moiety and the DPH moeity and 14 C-labelled DPH. The results demonstrate that for a hapten which does not completely fill the antibody binding sites, greatest sensitivity is achieved when the bridge of the iodine label is most dissimilar to that present in the original immunogen, when the hapten and label affinities are nearly equivalent. Greatest specificity is achieved with the label which most resembles the original immunogen. These results illustrate the difficulty of designing satisfactory labels for assays of both high specificity and sensitivity since minimal changes in label structure may produce greatly amplified changes in the subsequent affinity of the label for the antiserum. (Auth.)

Electrocutaneous sensitivity: effects of skin temperature.

Science.gov (United States)

Larkin, W D; Reilly, J P

1986-01-01

The effect of human skin temperature on electrocutaneous sensitivity was examined using brief capacitive discharges. Stimuli were designed to ensure that sensory effects would be independent of skin resistance and would reflect underlying neural excitability as closely as possible. Skin temperature was manipulated by immersing the forearm in circulating hot or cold air. Detection thresholds on the arm and fingertip were raised by cooling, but were not altered by heating. Temperature-related sensitivity shifts were described by the same multiplicative factors for both threshold and suprathreshold levels. The temperature coefficient (Q10) for cutaneous sensitivity under these conditions was approximately 1.3.

Sensitivity of Support Vector Machine Predictions of Passive Microwave Brightness Temperature Over Snow-covered Terrain in High Mountain Asia

Science.gov (United States)

Ahmad, J. A.; Forman, B. A.

2017-12-01

High Mountain Asia (HMA) serves as a water supply source for over 1.3 billion people, primarily in south-east Asia. Most of this water originates as snow (or ice) that melts during the summer months and contributes to the run-off downstream. In spite of its critical role, there is still considerable uncertainty regarding the total amount of snow in HMA and its spatial and temporal variation. In this study, the NASA Land Information Systems (LIS) is used to model the hydrologic cycle over the Indus basin. In addition, the ability of support vector machines (SVM), a machine learning technique, to predict passive microwave brightness temperatures at a specific frequency and polarization as a function of LIS-derived land surface model output is explored in a sensitivity analysis. Multi-frequency, multi-polarization passive microwave brightness temperatures as measured by the Advanced Microwave Scanning Radiometer - Earth Observing System (AMSR-E) over the Indus basin are used as training targets during the SVM training process. Normalized sensitivity coefficients (NSC) are then computed to assess the sensitivity of a well-trained SVM to each LIS-derived state variable. Preliminary results conform with the known first-order physics. For example, input states directly linked to physical temperature like snow temperature, air temperature, and vegetation temperature have positive NSC's whereas input states that increase volume scattering such as snow water equivalent or snow density yield negative NSC's. Air temperature exhibits the largest sensitivity coefficients due to its inherent, high-frequency variability. Adherence of this machine learning algorithm to the first-order physics bodes well for its potential use in LIS as the observation operator within a radiance data assimilation system aimed at improving regional- and continental-scale snow estimates.

New immunogenic form for vasopressin: production of high-affinity antiserum and RIA for plasmatic AVP

International Nuclear Information System (INIS)

Rougon-Rappuzi, G.; Delaage, M.A.; Conte-Devolx, B.; Millet, Y.

1977-01-01

A highly sensitive and specific radioimmunoassay (RIA) for arginine-vasopressin (AVP) was developped and applied to the measurement of AVP in human plasma. High-affinity antivasopressin antibodies with limited association constant heterogeneity have been induced by immunizing rabbits with Lysine-vasopressine (LVP) coupled to a human immunoglobulin (IgA). Replacing air drying of acetone-petroleum ether extracts by lyophilisation increased significantly the yields of AVP. Equilibrium dialysis was used for separating bound and free antigen, thus reducing the total time required for the assay to 48 hours. Only 1 ml of plasma was required for routine determinations due to a sensitivity threshold better than 0.5 pg/ml. Plasma AVP levels of normal subjects and of patients with inappropriate ADH secretion (SIADH) were determined during different hydratation states and following nicotin of ethanol infusions. (orig.) [de

Comparative transcriptome profiling of a thermal resistant vs. sensitive silkworm strain in response to high temperature under stressful humidity condition.

Directory of Open Access Journals (Sweden)

Wenfu Xiao

Full Text Available Thermotolerance is important particularly for poikilotherms such as insects. Understanding the mechanisms by which insects respond to high temperatures can provide insights into their adaptation to the environment. Therefore, in this study, we performed a transcriptome analysis of two silkworm strains with significantly different resistance to heat as well as humidity; the thermo-resistant strain 7532 and the thermos-sensitive strain Knobbed. We identified in total 4,944 differentially expressed genes (DEGs using RNA-Seq. Among these, 4,390 were annotated and 554 were novel. Gene Ontology (GO analysis of 747 DEGs identified between RT_48h (Resistant strain with high-temperature Treatment for 48 hours and ST_48h (Sensitive strain with high-temperature Treatment for 48 hours showed significant enrichment of 12 GO terms including metabolic process, extracellular region and serine-type peptidase activity. Moreover, we discovered 12 DEGs that may contribute to the heat-humidity stress response in the silkworm. Our data clearly showed that 48h post-exposure may be a critical time point for silkworm to respond to high temperature and humidity. These results provide insights into the genes and biological processes involved in high temperature and humidity tolerance in the silkworm, and advance our understanding of thermal tolerance in insects.

Fabrication of ultra-high sensitive and selective CH4 room temperature gas sensing of TiO2nanorods: Detailed study on the annealing temperature

CSIR Research Space (South Africa)

Tshabalala, Zamaswazi P

2016-07-01

Full Text Available Applications of ultra-highly sensitive and selective methane (CH(sub4)) room temperature gas sensors are important for various operations especially in underground mining environment. Therefore, this study is set out to investigate the effect...

Structure of Greyhound hemoglobin: origin of high oxygen affinity.

Science.gov (United States)

Bhatt, Veer S; Zaldívar-López, Sara; Harris, David R; Couto, C Guillermo; Wang, Peng G; Palmer, Andre F

2011-05-01

This study presents the crystal structure of Greyhound hemoglobin (GrHb) determined to 1.9 Å resolution. GrHb was found to crystallize with an α₁β₁ dimer in the asymmetric unit and belongs to the R2 state. Oxygen-affinity measurements combined with the fact that GrHb crystallizes in the R2 state despite the high-salt conditions used for crystallization strongly indicate that GrHb can serve as a model high-oxygen-affinity hemoglobin (Hb) for higher mammals, especially humans. Structural analysis of GrHb and its comparison with the R2-state of human Hb revealed several regions that can potentially contribute to the high oxygen affinity of GrHb and serve to rationalize the additional stability of the R2-state of GrHb. A previously well studied hydrophobic cluster of bar-headed goose Hb near α119 was also incorporated in the comparison between GrHb and human Hb. Finally, a structural comparison with generic dog Hb and maned wolf Hb was conducted, revealing that in contrast to GrHb these structures belong to the R state of Hb and raising the intriguing possibility of an additional allosteric factor co-purifying with GrHb that can modulate its quaternary structure.

Haemoglobin Pierre-Benite--a high affinity variant associated with relative polycythaemia.

Science.gov (United States)

Beard, M E; Potter, H C; Spearing, R L; Brennan, S O

2001-12-01

This is the second reported example of Hb Pierre--Benite (beta90 Glu-->Asp). This mutation is associated with increased oxygen affinity and polycythaemia. No instability was found and there was no charge shift detected by cellulose acetate electrophoresis at pH 8.3. The mutation was however, clearly indicated by electrospray ionization mass spectrometry (ESI MS), which showed an abnormal beta chain with a 14 Da decrease in mass. Blood volume studies documented a relative rather than a true polycythaemia and this finding has been reported in at least two other high affinity haemoglobin variants--Hb Heathrow and Hb Rahere. This finding led to delay in diagnosis because high oxygen affinity variants are conventionally considered to cause a true polycythaemia.

Water Recycling removal using temperature-sensitive hydronen

Energy Technology Data Exchange (ETDEWEB)

Rana B. Gupta

2002-10-30

The overall objective of this project was to study the proposed Water Recycling/Removal Using Temperature-Sensitive Hydrogels. The main element of this technology is the design of a suitable hydrogel that can perform needed water separation for pulp and paper industry. The specific topics studied are to answer following questions: (a) Can water be removed using hydrogel from large molecules such as lignin? (b) Can the rate of separation be made faster? (c) What are the molecular interactions with hydrogel surface? (d) Can a hydrogel be designed for a high ionic strength and high temperature? Summary of the specific results are given.

2017 Guralp Affinity Digitizer Evaluation.

Energy Technology Data Exchange (ETDEWEB)

Merchant, Bion J.

2018-03-01

Sandia National Laboratories has tested and evaluated two Guralp Affinity digitizers. The Affinity digitizers are intended to record sensor output for seismic and infrasound monitoring applications. The purpose of this digitizer evaluation is to measure the performance characteristics in such areas as power consumption, input impedance, sensitivity, full scale, self- noise, dynamic range, system noise, response, passband, and timing. The Affinity digitizers are being evaluated for potential use in the International Monitoring System (IMS) of the Comprehensive Nuclear Test-Ban-Treaty Organization (CTBTO).

Accurate and sensitive quantification of protein-DNA binding affinity.

Science.gov (United States)

Rastogi, Chaitanya; Rube, H Tomas; Kribelbauer, Judith F; Crocker, Justin; Loker, Ryan E; Martini, Gabriella D; Laptenko, Oleg; Freed-Pastor, William A; Prives, Carol; Stern, David L; Mann, Richard S; Bussemaker, Harmen J

2018-04-17

Transcription factors (TFs) control gene expression by binding to genomic DNA in a sequence-specific manner. Mutations in TF binding sites are increasingly found to be associated with human disease, yet we currently lack robust methods to predict these sites. Here, we developed a versatile maximum likelihood framework named No Read Left Behind (NRLB) that infers a biophysical model of protein-DNA recognition across the full affinity range from a library of in vitro selected DNA binding sites. NRLB predicts human Max homodimer binding in near-perfect agreement with existing low-throughput measurements. It can capture the specificity of the p53 tetramer and distinguish multiple binding modes within a single sample. Additionally, we confirm that newly identified low-affinity enhancer binding sites are functional in vivo, and that their contribution to gene expression matches their predicted affinity. Our results establish a powerful paradigm for identifying protein binding sites and interpreting gene regulatory sequences in eukaryotic genomes. Copyright © 2018 the Author(s). Published by PNAS.

Snapshot of methanogen sensitivity to temperature in Zoige wetland from Tibetan plateau

Directory of Open Access Journals (Sweden)

Li eFu

2015-02-01

Full Text Available Zoige wetland in Tibetan plateau represents a cold environment at high altitude where significant methane emission has been observed. However, it remains unknown how the production and emission of CH4 from Zoige wetland will respond to a warming climate. Here we investigated the temperature sensitivity of methanogen community in a Zoige wetland soil under the laboratory incubation conditions. One soil sample was collected and the temperature sensitivity of the methanogenic activity, the structure of methanogen community and the methanogenic pathways were determined. We found that the response of methanogenesis to temperature could be separated into two phases, a high sensitivity in the low temperature range and a modest sensitivity under mesophilic conditions, respectively. The aceticlastic methanogens Methanosarcinaceae were the main methanogens at low temperatures, while hydrogenotrophic Methanobacteriales, Methanomicrobiales and Methanocellales were more abundant at higher temperatures. The total abundance of mcrA genes increased with temperature indicating that the growth of methanogens was stimulated. The growth of hydrogenotrophic methanogens, however, was faster than aceticlastic ones resulting in the shift of methanogen community. Determination of carbon isotopic signatures indicated that methanogenic pathway was also shifted from mainly aceticlastic methanogenesis to a mixture of hydrogenotrophic and aceticlastic methanogenesis with the increase of temperature. Collectively, the shift of temperature responses of methanogenesis was in accordance with the changes in methanogen composition and methanogenic pathway in this Zoige wetland sample. It appears that the aceticlastic methanogenesis dominated at low temperatures is more sensitive than the hydrogenotrophic one at higher temperatures.

High temperature structural silicides

International Nuclear Information System (INIS)

Petrovic, J.J.

1997-01-01

Structural silicides have important high temperature applications in oxidizing and aggressive environments. Most prominent are MoSi 2 -based materials, which are borderline ceramic-intermetallic compounds. MoSi 2 single crystals exhibit macroscopic compressive ductility at temperatures below room temperature in some orientations. Polycrystalline MoSi 2 possesses elevated temperature creep behavior which is highly sensitive to grain size. MoSi 2 -Si 3 N 4 composites show an important combination of oxidation resistance, creep resistance, and low temperature fracture toughness. Current potential applications of MoSi 2 -based materials include furnace heating elements, molten metal lances, industrial gas burners, aerospace turbine engine components, diesel engine glow plugs, and materials for glass processing

High sensitivity and high Q-factor nanoslotted parallel quadrabeam photonic crystal cavity for real-time and label-free sensing

Energy Technology Data Exchange (ETDEWEB)

Yang, Daquan [Rowland Institute at Harvard University, Cambridge, Massachusetts 02142 (United States); State Key Laboratory of Information Photonics and Optical Communications, School of Information and Communication Engineering, Beijing University of Posts and Telecommunications, Beijing 100876 (China); School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts 02138 (United States); Kita, Shota; Wang, Cheng; Lončar, Marko [School of Engineering and Applied Sciences, Harvard University, Cambridge, Massachusetts 02138 (United States); Liang, Feng; Quan, Qimin [Rowland Institute at Harvard University, Cambridge, Massachusetts 02142 (United States); Tian, Huiping; Ji, Yuefeng [State Key Laboratory of Information Photonics and Optical Communications, School of Information and Communication Engineering, Beijing University of Posts and Telecommunications, Beijing 100876 (China)

2014-08-11

We experimentally demonstrate a label-free sensor based on nanoslotted parallel quadrabeam photonic crystal cavity (NPQC). The NPQC possesses both high sensitivity and high Q-factor. We achieved sensitivity (S) of 451 nm/refractive index unit and Q-factor >7000 in water at telecom wavelength range, featuring a sensor figure of merit >2000, an order of magnitude improvement over the previous photonic crystal sensors. In addition, we measured the streptavidin-biotin binding affinity and detected 10 ag/mL concentrated streptavidin in the phosphate buffered saline solution.

Botulinum neurotoxin B recognizes its protein receptor with high affinity and specificity.

Science.gov (United States)

Jin, Rongsheng; Rummel, Andreas; Binz, Thomas; Brunger, Axel T

2006-12-21

Botulinum neurotoxins (BoNTs) are produced by Clostridium botulinum and cause the neuroparalytic syndrome of botulism. With a lethal dose of 1 ng kg(-1), they pose a biological hazard to humans and a serious potential bioweapon threat. BoNTs bind with high specificity at neuromuscular junctions and they impair exocytosis of synaptic vesicles containing acetylcholine through specific proteolysis of SNAREs (soluble N-ethylmaleimide-sensitive fusion protein attachment protein receptors), which constitute part of the synaptic vesicle fusion machinery. The molecular details of the toxin-cell recognition have been elusive. Here we report the structure of a BoNT in complex with its protein receptor: the receptor-binding domain of botulinum neurotoxin serotype B (BoNT/B) bound to the luminal domain of synaptotagmin II, determined at 2.15 A resolution. On binding, a helix is induced in the luminal domain which binds to a saddle-shaped crevice on a distal tip of BoNT/B. This crevice is adjacent to the non-overlapping ganglioside-binding site of BoNT/B. Synaptotagmin II interacts with BoNT/B with nanomolar affinity, at both neutral and acidic endosomal pH. Biochemical and neuronal ex vivo studies of structure-based mutations indicate high specificity and affinity of the interaction, and high selectivity of BoNT/B among synaptotagmin I and II isoforms. Synergistic binding of both synaptotagmin and ganglioside imposes geometric restrictions on the initiation of BoNT/B translocation after endocytosis. Our results provide the basis for the rational development of preventive vaccines or inhibitors against these neurotoxins.

Temperature effects on hemocyanin oxygen binding in an antarctic cephalopod.

Science.gov (United States)

Zielinski, S; Sartoris, F J; Pörtner, H O

2001-02-01

The functional relevance of oxygen transport by hemocyanin of the Antarctic octopod Megaleledone senoi and of the eurythermal cuttlefish Sepia officinalis was analyzed by continuous and simultaneous recordings of changes in pH and hemocyanin oxygen saturation in whole blood at various temperatures. These data were compared to literature data on other temperate and cold-water cephalopods (octopods and giant squid). In S. officinalis, the oxygen affinity of hemocyanin changed at deltaP50/degrees C = 0.12 kPa (pH 7.4) with increasing temperatures; this is similar to observations in temperate octopods. In M. senoi, thermal sensitivity was much smaller (delta log P50/delta pH) increased with increasing temperature in both the cuttlefish and the Antarctic octopod. At low PO2 (1.0 kPa) and pH (7.2), the presence of a large venous oxygen reserve (43% saturation) insensitive to pH reflects reduced pH sensitivity and high oxygen affinity in M. senoi hemocyanin at 0 degrees C. In S. officinalis, this reserve was 19% at pH 7.4, 20 degrees C, and 1.7 kPa O2, a level still higher than in squid. These findings suggest that the lower metabolic rate of octopods and cuttlefish compared to squid is reflected in less pH-dependent oxygen transport. Results of the hemocyanin analysis for the Antarctic octopod were similar to those reported for Vampyroteuthis--an extremely high oxygen affinity supporting a very low metabolic rate. In contrast to findings in cold-adapted giant squid, the minimized thermal sensitivity of oxygen transport in Antarctic octopods will reduce metabolic scope and thereby contribute to their stenothermality.

Effect of oxyanions on the IGSCC inhibition of sensitized 304 stainless steel in high temperature water

International Nuclear Information System (INIS)

Tsuge, Hiroyuki; Murayama, Junichiro; Nagano, Hiroo.

1983-01-01

Effect of oxyanions such as MoO 4 2- , WO 4 2- , and CrO 4 2- on the intergranular stress corrosion cracking (IGSCC) of Type 304 stainless steel in high temperature water was studied. The results obtained are as follows: 1) Addition of such oxyanion as MoO 4 2- , WO 4 2- , and CrO 4 2- suppresses IGSCC of sensitized Type 304 stainless steel in high temperature nondeaerated water. The effectiveness of the inhibitive action by the oxyanion is ranked in the order of MoO 4 2- >WO 4 2- >CrO 4 2- . 2) The mechanism of IGSCC inhibition by MoO 4 2- ion for sentized Type 304 stainless steel in high temperature water is considered as follows, i.e., the presence MoO 4 2- ion decreases the dissolution rate of Cr depleted zone at grain boundaries to the level of matrix by helping the formation of the Cr rich film containing MoO 3 or adsorbed MoO 4 2- ion on the surface of Type 304 stainless steel. (author)

Fundamentals of affinity cell separations.

Science.gov (United States)

Zhang, Ye; Lyons, Veronica; Pappas, Dimitri

2018-03-01

Cell separations using affinity methods continue to be an enabling science for a wide variety of applications. In this review, we discuss the fundamental aspects of affinity separation, including the competing forces for cell capture and elution, cell-surface interactions, and models for cell adhesion. Factors affecting separation performance such as bond affinity, contact area, and temperature are presented. We also discuss and demonstrate the effects of nonspecific binding on separation performance. Metrics for evaluating cell separations are presented, along with methods of comparing separation techniques for cell isolation using affinity capture. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

The monoclonal S9.6 antibody exhibits highly variable binding affinities towards different R-loop sequences.

Directory of Open Access Journals (Sweden)

Fabian König

Full Text Available The monoclonal antibody S9.6 is a widely-used tool to purify, analyse and quantify R-loop structures in cells. A previous study using the surface plasmon resonance technology and a single-chain variable fragment (scFv of S9.6 showed high affinity (0.6 nM for DNA-RNA and also a high affinity (2.7 nM for RNA-RNA hybrids. We used the microscale thermophoresis method allowing surface independent interaction studies and electromobility shift assays to evaluate additional RNA-DNA hybrid sequences and to quantify the binding affinities of the S9.6 antibody with respect to distinct sequences and their GC-content. Our results confirm high affinity binding to previously analysed sequences, but reveals that binding affinities are highly sequence specific. Our study presents R-loop sequences that independent of GC-content and in different sequence variations exhibit either no binding, binding affinities in the micromolar range and as well high affinity binding in the nanomolar range. Our study questions the usefulness of the S9.6 antibody in the quantitative analysis of R-loop sequences in vivo.

Perovskite Nanoparticle-Sensitized Ga2O3 Nanorod Arrays for CO Detection at High Temperature.

Science.gov (United States)

Lin, Hui-Jan; Baltrus, John P; Gao, Haiyong; Ding, Yong; Nam, Chang-Yong; Ohodnicki, Paul; Gao, Pu-Xian

2016-04-13

Noble metal nanoparticles are extensively used for sensitizing metal oxide chemical sensors through the catalytic spillover mechanism. However, due to earth-scarcity and high cost of noble metals, finding replacements presents a great economic benefit. Besides, high temperature and harsh environment sensor applications demand material stability under conditions approaching thermal and chemical stability limits of noble metals. In this study, we employed thermally stable perovskite-type La(0.8)Sr(0.2)FeO3 (LSFO) nanoparticle surface decoration on Ga2O3 nanorod array gas sensors and discovered an order of magnitude enhanced sensitivity to carbon monoxide at 500 °C. The LSFO nanoparticle catalysts was of comparable performance to that achieved by Pt nanoparticles, with a much lower weight loading than Pt. Detailed electron microscopy and X-ray photoelectron spectroscopy studies suggested the LSFO nanoparticle sensitization effect is attributed to a spillover-like effect associated with the gas-LSFO-Ga2O3 triple-interfaces that spread the negatively charged surface oxygen ions from LSFO nanoparticles surfaces over to β-Ga2O3 nanorod surfaces with faster surface CO oxidation reactions.

Highly sensitive and selective room-temperature NO_2 gas sensor based on bilayer transferred chemical vapor deposited graphene

International Nuclear Information System (INIS)

Seekaew, Yotsarayuth; Phokharatkul, Ditsayut; Wisitsoraat, Anurat; Wongchoosuk, Chatchawal

2017-01-01

Highlights: • Simple and low-cost fabrication of bilayer graphene gas sensor was presented. • Layer effects of graphene on NO_2 gas-sensing properties were investigated. • Bilayer graphene sensor exhibited a high linear NO_2 sensitivity of 1.409 ppm"−"1. • The NO_2-sensing mechanisms based on band diagram were highlighted. - Abstract: This work presents a highly sensitive room-temperature gas sensor based on bilayer graphene fabricated by an interfacial transfer of chemical vapor deposited graphene onto nickel interdigitated electrodes. Scanning electron microscopic and Raman spectroscopic characterizations confirm the presence of graphene on interdigitated nickel electrodes with varying numbers of graphene layers. The NO_2 detection performances of bilayer graphene gas sensor have been investigated in comparison with those of monolayer and multilayer graphene gas sensors at room temperature. From results, the bilayer graphene gas sensor exhibits higher response, sensitivity and selectivity to NO_2 than monolayer and multilayer graphene. The sensitivity of bilayer graphene gas sensor is 1.409 ppm"−"1 towards NO_2 over a concentration range of 1–25 ppm, which is more than twice higher than that of monolayer graphene. The NO_2-sensing mechanism of graphene sensing film has been explained based on the direct charge transfer process due to the adsorption of NO_2 molecules.

The role of CH/π interactions in the high affinity binding of streptavidin and biotin.

Science.gov (United States)

Ozawa, Motoyasu; Ozawa, Tomonaga; Nishio, Motohiro; Ueda, Kazuyoshi

2017-08-01

The streptavidin-biotin complex has an extraordinarily high affinity (Ka: 10 15 mol -1 ) and contains one of the strongest non-covalent interactions known. This strong interaction is widely used in biological tools, including for affinity tags, detection, and immobilization of proteins. Although hydrogen bond networks and hydrophobic interactions have been proposed to explain this high affinity, the reasons for it remain poorly understood. Inspired by the deceased affinity of biotin observed for point mutations of streptavidin at tryptophan residues, we hypothesized that a CH/π interaction may also contribute to the strong interaction between streptavidin and biotin. CH/π interactions were explored and analyzed at the biotin-binding site and at the interface of the subunits by the fragment molecular orbital method (FMO) and extended applications: PIEDA and FMO4. The results show that CH/π interactions are involved in the high affinity for biotin at the binding site of streptavidin. We further suggest that the involvement of CH/π interactions at the subunit interfaces and an extended CH/π network play more critical roles in determining the high affinity, rather than involvement at the binding site. Copyright © 2017 Elsevier Inc. All rights reserved.

Identification of high- and low-affinity NGF receptors during development of the chicken central nervous system

International Nuclear Information System (INIS)

Escandon, E.; Chao, M.V.

1990-01-01

In order to study regulation of the nerve growth factor (NGF) receptor during embryogenesis in chick brain, we have used affinity crosslinking of tissues with 125 I-NGF. NGF interacts with high- and low-affinity receptors; high-affinity receptors are required for the majority of NGF's actions. Most measurements of receptor levels do not distinguish between high- and low-affinity forms of the receptor. We have used the lipophilic crosslinking agent HSAB to identify the high-affinity, functional receptor during development of the chicken central nervous system. A peak of expression during Embryonic Days 5-10 was detected in all regions of the chicken central nervous system, but, shortly after birth, only the cerebellar region displays significant levels of NGF receptor protein. The time course of expression confirms the dramatic regulation of the NGF receptor gene during defined embryonic periods. The detection of high-affinity NGF receptors in brain and neural retina provides strong evidence that NGF is involved in essential ontogenetic events in the development of the chicken central nervous system

High affinity calmodulin target sequence in the signalling molecule PI 3-kinase

DEFF Research Database (Denmark)

Fischer, R; Julsgart, J; Berchtold, M W

1998-01-01

M-binding peptide derived from the p110gamma isoform interacts with CaM in a calcium-dependent way. Using gel shift analysis and fluorescence spectrophotometry we discovered that the peptide forms a high affinity complex with CaM. Titration experiments using dansylated CaM gave an affinity constant of 5 n...

Elimination of bus voltage impact on temperature sensitive electrical parameter during turn-on transition for junction temperature estimation of high-power IGBT modules

DEFF Research Database (Denmark)

Luo, Haoze; Iannuzzo, Francesco; Blaabjerg, Frede

2017-01-01

Junction temperature is of great importance to safe operating area of IGBT modules. Various information of the IGBT operating state is reflected on electrical characteristics during turn-on transient. A unified extraction method for internal junction temperature via dynamic thermo......-sensitive electrical parameters (DTSEP) during turn-on transient is proposed. Two DTSEP, turn-on delay time (tdon) and the maximum increasing rate of collector current dic/dt(max), are combined to eliminate the bus voltage impact. Using the inherent emitter-auxiliary inductor LeE in high-power modules, the temperature......-dependent DTSEPs can be converted into a low-voltage and measurable signal. Finally, experiment results are exhibited to verify the effectiveness of proposed method....

Evaluation of high temperature pressure sensors

International Nuclear Information System (INIS)

Choi, In-Mook; Woo, Sam-Yong; Kim, Yong-Kyu

2011-01-01

It is becoming more important to measure the pressure in high temperature environments in many industrial fields. However, there is no appropriate evaluation system and compensation method for high temperature pressure sensors since most pressure standards have been established at room temperature. In order to evaluate the high temperature pressure sensors used in harsh environments, such as high temperatures above 250 deg. C, a specialized system has been constructed and evaluated in this study. The pressure standard established at room temperature is connected to a high temperature pressure sensor through a chiller. The sensor can be evaluated in conditions of changing standard pressures at constant temperatures and of changing temperatures at constant pressures. According to the evaluation conditions, two compensation methods are proposed to eliminate deviation due to sensitivity changes and nonlinear behaviors except thermal hysteresis.

Cold-Adapted Viral Attenuation (CAVA): Highly Temperature Sensitive Polioviruses as Novel Vaccine Strains for a Next Generation Inactivated Poliovirus Vaccine.

Science.gov (United States)

Sanders, Barbara P; de Los Rios Oakes, Isabel; van Hoek, Vladimir; Bockstal, Viki; Kamphuis, Tobias; Uil, Taco G; Song, Yutong; Cooper, Gillian; Crawt, Laura E; Martín, Javier; Zahn, Roland; Lewis, John; Wimmer, Eckard; Custers, Jerome H H V; Schuitemaker, Hanneke; Cello, Jeronimo; Edo-Matas, Diana

2016-03-01

The poliovirus vaccine field is moving towards novel vaccination strategies. Withdrawal of the Oral Poliovirus Vaccine and implementation of the conventional Inactivated Poliovirus Vaccine (cIPV) is imminent. Moreover, replacement of the virulent poliovirus strains currently used for cIPV with attenuated strains is preferred. We generated Cold-Adapted Viral Attenuation (CAVA) poliovirus strains by serial passage at low temperature and subsequent genetic engineering, which contain the capsid sequences of cIPV strains combined with a set of mutations identified during cold-adaptation. These viruses displayed a highly temperature sensitive phenotype with no signs of productive infection at 37°C as visualized by electron microscopy. Furthermore, decreases in infectious titers, viral RNA, and protein levels were measured during infection at 37°C, suggesting a block in the viral replication cycle at RNA replication, protein translation, or earlier. However, at 30°C, they could be propagated to high titers (9.4-9.9 Log10TCID50/ml) on the PER.C6 cell culture platform. We identified 14 mutations in the IRES and non-structural regions, which in combination induced the temperature sensitive phenotype, also when transferred to the genomes of other wild-type and attenuated polioviruses. The temperature sensitivity translated to complete absence of neurovirulence in CD155 transgenic mice. Attenuation was also confirmed after extended in vitro passage at small scale using conditions (MOI, cell density, temperature) anticipated for vaccine production. The inability of CAVA strains to replicate at 37°C makes reversion to a neurovirulent phenotype in vivo highly unlikely, therefore, these strains can be considered safe for the manufacture of IPV. The CAVA strains were immunogenic in the Wistar rat potency model for cIPV, inducing high neutralizing antibody titers in a dose-dependent manner in response to D-antigen doses used for cIPV. In combination with the highly productive

Sensitivity of a soil-plant-atmosphere model to changes in air temperature, dew point temperature, and solar radiation

Energy Technology Data Exchange (ETDEWEB)

Luxmoore, R.J. (Oak Ridge National Lab.,TN); Stolzy, J.L.; Holdeman, J.T.

1981-01-01

Air temperature, dew point temperature and solar radiation were independently varied in an hourly soil-plant-atmosphere model in a sensitivity analysis of these parameters. Results suggested that evapotranspiration in eastern Tennessee is limited more by meteorological conditions that determine the vapor-pressure gradient than by the necessary energy to vaporize water within foliage. Transpiration and soil water drainage were very sensitive to changes in air and dew point temperature and to solar radiation under low atmospheric vapor-pressure deficit conditions associated with reduced air temperature. Leaf water potential and stomatal conductance were reduced under conditions having high evapotranspiration. Representative air and dew point temperature input data for a particular application are necessary for satisfactory results, whereas irradiation may be less well characterized for applications with high atmospheric vapor-pressure deficit. The effects of a general rise in atmospheric temperature on forest water budgets are discussed.

Sensitivity to temperature of nuclear energy generation by hydrogen burning

International Nuclear Information System (INIS)

Mitalas, R.

1981-01-01

The sensitivity to temperature of nuclear energy generation by hydrogen burning is discussed. The complexity of the sensitivity is due to the different equilibration time-scales of the constituents of the p-p chain and CN cycle and the dependence of their abundances and time-scales on temperature. The time-scale of the temperature perturbation, compared to the equilibrium time-scale of a constituent, determines whether the constituent is in equilibrium and affects the sensitivity. The temperature sensitivity of the p-p chain for different values of hydrogen abundance, when different constituents come into equilibrium is presented, as well as its variation with 3 He abundance. The temperature sensitivity is drastically different from n 11 , the temperature sensitivity of the proton-proton reaction, unless the time-scale of temperature perturbation is long enough for 3 He to remain in equilibrium. Even in this case the sensitivity of the p-p chain differs significantly from n 11 , unless the temperature is so low that PP II and PP III chains can be neglected. The variation of the sensitivity of CN energy generation is small for different time-scales of temperature variation, because the temperature sensitivities of individual reactions are so similar. The combined sensitivity to temperature of energy generation by hydrogen burning is presented and shown to have a maximum of 16.4 at T 6 = 24.5. For T 6 > 25 the temperature sensitivity is given by the sensitivity of 14 N + p reaction. (author)

Fiber Fabry-Perot interferometer with controllable temperature sensitivity.

Science.gov (United States)

Zhang, Xinpu; Peng, Wei; Zhang, Yang

2015-12-01

We proposed a fiber taper based on the Fabry-Perot (FP) interferometer structure with controllable temperature sensitivity. The FP interferometer is formed by inserting a segment of tapered fiber tip into the capillary and subsequently splicing the other end of the capillary to a single-mode fiber (SMF), the tapered fiber endface, and the spliced face form the FP cavity. Through controlling the inserted tapered fiber length, a series of FP interferometers were made. Because the inserted taper tip has the degree of freedom along the fiber axial, when the FP interferometer is subjected to temperature variation, the thermal expansion of the fiber taper tip will resist the FP cavity length change caused by the evolution of capillary length, and we can control the temperature sensitivity by adjusting the inserted taper length. In this structure, the equivalent thermal expansion coefficient of the FP interferometer can be defined; it was used to evaluate the temperature sensitivity of the FP interferometer, which provides an effective method to eliminate the temperature effect and to enhance other measurement accuracy. We fabricated the FP interferometers and calibrated their temperature characters by measuring the wavelength shift of the resonance dips in the reflection spectrum. In a temperature range of 50°C to 150°C, the corresponding temperature sensitivities can be controlled between 0 and 1.97 pm/°C when the inserted taper is between 75 and 160 μm. Because of its controllable temperature sensitivity, ease of fabrication, and low cost, this FP interferometer can meet different temperature sensitivity requirements in various application areas, especially in the fields which need temperature insensitivity.

Structural implications of hERG K+ channel block by a high-affinity minimally structured blocker

Science.gov (United States)

Helliwell, Matthew V.; Zhang, Yihong; El Harchi, Aziza; Du, Chunyun; Hancox, Jules C.; Dempsey, Christopher E.

2018-01-01

Cardiac potassium channels encoded by human ether-à-go-go–related gene (hERG) are major targets for structurally diverse drugs associated with acquired long QT syndrome. This study characterized hERG channel inhibition by a minimally structured high-affinity hERG inhibitor, Cavalli-2, composed of three phenyl groups linked by polymethylene spacers around a central amino group, chosen to probe the spatial arrangement of side chain groups in the high-affinity drug-binding site of the hERG pore. hERG current (IhERG) recorded at physiological temperature from HEK293 cells was inhibited with an IC50 of 35.6 nm with time and voltage dependence characteristic of blockade contingent upon channel gating. Potency of Cavalli-2 action was markedly reduced for attenuated inactivation mutants located near (S620T; 54-fold) and remote from (N588K; 15-fold) the channel pore. The S6 Y652A and F656A mutations decreased inhibitory potency 17- and 75-fold, respectively, whereas T623A and S624A at the base of the selectivity filter also decreased potency (16- and 7-fold, respectively). The S5 helix F557L mutation decreased potency 10-fold, and both F557L and Y652A mutations eliminated voltage dependence of inhibition. Computational docking using the recent cryo-EM structure of an open channel hERG construct could only partially recapitulate experimental data, and the high dependence of Cavalli-2 block on Phe-656 is not readily explainable in that structure. A small clockwise rotation of the inner (S6) helix of the hERG pore from its configuration in the cryo-EM structure may be required to optimize Phe-656 side chain orientations compatible with high-affinity block. PMID:29545312

Compound immobilization and drug-affinity chromatography.

Science.gov (United States)

Rix, Uwe; Gridling, Manuela; Superti-Furga, Giulio

2012-01-01

Bioactive small molecules act through modulating a yet unpredictable number of targets. It is therefore of critical importance to define the cellular target proteins of a compound as an entry point to understanding its mechanism of action. Often, this can be achieved in a direct fashion by chemical proteomics. As with any affinity chromatography, immobilization of the bait to a solid support is one of the earliest and most crucial steps in the process. Interfering with structural features that are important for identification of a target protein will be detrimental to binding affinity. Also, many molecules are sensitive to heat or to certain chemicals, such as acid or base, and might be destroyed during the process of immobilization, which therefore needs to be not only efficient, but also mild. The subsequent affinity chromatography step needs to preserve molecular and conformational integrity of both bait compound and proteins in order to result in the desired specific enrichment while ensuring a high level of compatibility with downstream analysis by mass spectrometry. Thus, the right choice of detergent, buffer, and protease inhibitors is also essential. This chapter describes a widely applicable procedure for the immobilization of small molecule drugs and for drug-affinity chromatography with subsequent protein identification by mass spectrometry.

Low temperature fabrication of ZnO compact layer for high performance plastic dye-sensitized ZnO solar cells

International Nuclear Information System (INIS)

Hu Fangyi; Xia Yujing; Guan Zisheng; Yin Xiong; He Tao

2012-01-01

Highlights: ► ZnO compact layer is prepared via simple electrochemical method at low temperature. ► Compact layer can effectively block electron transfer from TCO to electrolyte. ► DSC PCE is improved by 17% when ZnO compact layer is introduced. ► Plastic DSCs with ZnO compact layer show a PCE of 3.29% under AM1.5 100 mW cm −2 . ► The above efficiency is comparable to that with high temperature sintering step. - Abstract: ZnO compact layer has been fabricated on transparent conducting oxide glass and plastic polymer substrates at low temperature via electrodeposition. The results of dark current and cyclic voltammetric measurements demonstrate that the compact layer can effectively reduce the short circuit from transparent conducting oxide to electrolyte in dye-sensitized ZnO solar cells, leading to an increase of open-circuit photovoltage and fill factor of the devices and, thereby, the power conversion efficiency. The resultant plastic dye-sensitized ZnO solar cell presents an efficiency of 3.29% under illumination of 100 mW cm −2 , AM 1.5G. This indicates that electrodeposition is a viable method to fabricate ZnO compact layer for high performance flexible devices.

Temperature-sensitive gating of hCx26: high-resolution Raman spectroscopy sheds light on conformational changes.

Science.gov (United States)

Kniggendorf, Ann-Kathrin; Meinhardt-Wollweber, Merve; Yuan, Xiaogang; Roth, Bernhard; Seifert, Astrid; Fertig, Niels; Zeilinger, Carsten

2014-07-01

The temperature-sensitive gating of human Connexin 26 (hCx26) was analyzed with confocal Raman microscopy. High-resolution Raman spectra covering the spectral range between 400 and 1500 rel. cm(-1) with a spectral resolution of 1 cm(-1) were fully annotated, revealing notable differences between the spectrum recorded from solubilized hCx26 in Ca(2+)-buffered POPC at 10°C and any other set of protein conditions (temperature, Ca(2+) presence, POPC presence). Spectral components originating from specific amino acids show that the TM1/EL1 parahelix and probably the TM4 trans-membrane helix and the plug domain are involved in the gating process responsible for fully closing the hemichannel.

Targeted drug delivery using temperature-sensitive liposomes

International Nuclear Information System (INIS)

Magin, R.L.; Niesman, M.R.

1984-01-01

Liposomes are receiving considerable attention as vehicles for selective drug delivery. One method of targeting liposomal contents involves the combination of local hyperthermia with temperature-sensitive liposomes. Such liposomes have been used to increase the uptake of methotrexate and cis-platinum into locally heated mouse tumors. However, additional information is needed on the mechanism of liposome drug release and the physiologic deposition of liposomes in vivo before clinical trails are begun. Current research is directed at studying the encapsulation and release of water soluble drugs from temperature-sensitive liposomes. The influence of liposome size, structure, and composition on the rapid release in plasma of cytosine arabinoside, cis-platinum, and the radiation sensitizer SR-2508 are described. These results demonstrate potential applications for temperature-sensitive liposomes in selective drug delivery

Molecular basis of a high affinity murine interleukin-5 receptor.

OpenAIRE

Devos, R; Plaetinck, G; Van der Heyden, J; Cornelis, S; Vandekerckhove, J; Fiers, W; Tavernier, J

1991-01-01

The mouse interleukin-5 receptor (mIL-5R) consists of two components one of which, the mIL-5R alpha-chain, binds mIL-5 with low affinity. Recently we demonstrated that monoclonal antibodies (Mabs) recognizing the second mIL-5R beta-chain, immunoprecipitate a p130-140 protein doublet which corresponds to the mIL-3R and the mIL-3R-like protein, the latter chain for which so far no ligand has been identified. In this study we show that a high affinity mIL-5R can be reconstituted on COS1 cells by...

Highly sensitive and selective room-temperature NO{sub 2} gas sensor based on bilayer transferred chemical vapor deposited graphene

Energy Technology Data Exchange (ETDEWEB)

Seekaew, Yotsarayuth [Department of Physics, Faculty of Science, Kasetsart University, Chatuchak, Bangkok 10900 (Thailand); Phokharatkul, Ditsayut; Wisitsoraat, Anurat [Nanoelectronics and MEMS Laboratory, National Electronics and Computer Technology Center, Klong Luang, Pathumthani 12120 (Thailand); Wongchoosuk, Chatchawal, E-mail: chatchawal.w@ku.ac.th [Department of Physics, Faculty of Science, Kasetsart University, Chatuchak, Bangkok 10900 (Thailand)

2017-05-15

Highlights: • Simple and low-cost fabrication of bilayer graphene gas sensor was presented. • Layer effects of graphene on NO{sub 2} gas-sensing properties were investigated. • Bilayer graphene sensor exhibited a high linear NO{sub 2} sensitivity of 1.409 ppm{sup −1}. • The NO{sub 2}-sensing mechanisms based on band diagram were highlighted. - Abstract: This work presents a highly sensitive room-temperature gas sensor based on bilayer graphene fabricated by an interfacial transfer of chemical vapor deposited graphene onto nickel interdigitated electrodes. Scanning electron microscopic and Raman spectroscopic characterizations confirm the presence of graphene on interdigitated nickel electrodes with varying numbers of graphene layers. The NO{sub 2} detection performances of bilayer graphene gas sensor have been investigated in comparison with those of monolayer and multilayer graphene gas sensors at room temperature. From results, the bilayer graphene gas sensor exhibits higher response, sensitivity and selectivity to NO{sub 2} than monolayer and multilayer graphene. The sensitivity of bilayer graphene gas sensor is 1.409 ppm{sup −1} towards NO{sub 2} over a concentration range of 1–25 ppm, which is more than twice higher than that of monolayer graphene. The NO{sub 2}-sensing mechanism of graphene sensing film has been explained based on the direct charge transfer process due to the adsorption of NO{sub 2} molecules.

Novel variable structure control for the temperature of PEM fuel cell stack based on the dynamic thermal affine model

International Nuclear Information System (INIS)

Li Xi; Deng Zhonghua; Wei Dong; Xu Chunshan; Cao Guangyi

2011-01-01

Highlights: → The affine state space control-oriented model is designed and realized for the variant structure control (VSC) strategy. → The VSC with rapid-smooth reaching law and rapid-convergent sliding mode is presented for the PEMFC stack temperature. → Numerical results show that the method can control the operating temperature to reach the target value satisfactorily. - Abstract: Dynamic thermal management of proton exchange membrane fuel cell stack (PEMFC) is a very important aspect, which plays an important role on electro-reaction. Its variation also has a significant influence on the performance and lifespan of PEMFC stack. The temperature of stack should be controlled efficiently, which has great impacts on the performance of PEMFC due to the thermal variation. Based on the control-oriented dynamic thermal affine model identified by optimization algorithm, a novel variable structures control (VSC) with rapid-smooth reaching law (RSRL) and rapid-convergent sliding mode (FCSM) is presented for the temperature control system of PEMFC stack. Numerical test results show that the method can control the operating temperature to reach the target value satisfactorily, which proves the effectiveness and robustness of the algorithm.

Single-experiment displacement assay for quantifying high-affinity binding by isothermal titration calorimetry.

Science.gov (United States)

Krainer, Georg; Keller, Sandro

2015-04-01

Isothermal titration calorimetry (ITC) is the gold standard for dissecting the thermodynamics of a biomolecular binding process within a single experiment. However, reliable determination of the dissociation constant (KD) from a single titration is typically limited to the range 100 μM>KD>1 nM. Interactions characterized by a lower KD can be assessed indirectly by so-called competition or displacement assays, provided that a suitable competitive ligand is available whose KD falls within the directly accessible window. However, this protocol is limited by the fact that it necessitates at least two titrations to characterize one high-affinity inhibitor, resulting in considerable consumption of both sample material and time. Here, we introduce a fast and efficient ITC displacement assay that allows for the simultaneous characterization of both a high-affinity ligand and a moderate-affinity ligand competing for the same binding site on a receptor within a single experiment. The protocol is based on a titration of the high-affinity ligand into a solution containing the moderate-affinity ligand bound to the receptor present in excess. The resulting biphasic binding isotherm enables accurate and precise determination of KD values and binding enthalpies (ΔH) of both ligands. We discuss the theoretical background underlying the approach, demonstrate its practical application to metal ion chelation, explore its potential and limitations with the aid of simulations and statistical analyses, and elaborate on potential applications to protein-inhibitor interactions. Copyright © 2014 Elsevier Inc. All rights reserved.

The probability distribution of intergranular stress corrosion cracking life for sensitized 304 stainless steels in high temperature, high purity water

International Nuclear Information System (INIS)

Akashi, Masatsune; Kenjyo, Takao; Matsukura, Shinji; Kawamoto, Teruaki

1984-01-01

In order to discuss the probability distribution of intergranular stress corrsion carcking life for sensitized 304 stainless steels, a series of the creviced bent beem (CBB) and the uni-axial constant load tests were carried out in oxygenated high temperature, high purity water. The following concludions were resulted; (1) The initiation process of intergranular stress corrosion cracking has been assumed to be approximated by the Poisson stochastic process, based on the CBB test results. (2) The probability distribution of intergranular stress corrosion cracking life may consequently be approximated by the exponential probability distribution. (3) The experimental data could be fitted to the exponential probability distribution. (author)

A high sensitivity nanomaterial based SAW humidity sensor

Energy Technology Data Exchange (ETDEWEB)

Wu, T-T; Chou, T-H [Institute of Applied Mechanics, National Taiwan University, Taipei 106, Taiwan (China); Chen, Y-Y [Department of Mechanical Engineering, Tatung University, Taipei 104, Taiwan (China)], E-mail: wutt@ndt.iam.ntu.edu.tw

2008-04-21

In this paper, a highly sensitive humidity sensor is reported. The humidity sensor is configured by a 128{sup 0}YX-LiNbO{sub 3} based surface acoustic wave (SAW) resonator whose operating frequency is at 145 MHz. A dual delay line configuration is realized to eliminate external temperature fluctuations. Moreover, for nanostructured materials possessing high surface-to-volume ratio, large penetration depth and fast charge diffusion rate, camphor sulfonic acid doped polyaniline (PANI) nanofibres are synthesized by the interfacial polymerization method and further deposited on the SAW resonator as selective coating to enhance sensitivity. The humidity sensor is used to measure various relative humidities in the range 5-90% at room temperature. Results show that the PANI nanofibre based SAW humidity sensor exhibits excellent sensitivity and short-term repeatability.

Temperature sensitivity of soil respiration rates enhanced by microbial community response.

Science.gov (United States)

Karhu, Kristiina; Auffret, Marc D; Dungait, Jennifer A J; Hopkins, David W; Prosser, James I; Singh, Brajesh K; Subke, Jens-Arne; Wookey, Philip A; Agren, Göran I; Sebastià, Maria-Teresa; Gouriveau, Fabrice; Bergkvist, Göran; Meir, Patrick; Nottingham, Andrew T; Salinas, Norma; Hartley, Iain P

2014-09-04

Soils store about four times as much carbon as plant biomass, and soil microbial respiration releases about 60 petagrams of carbon per year to the atmosphere as carbon dioxide. Short-term experiments have shown that soil microbial respiration increases exponentially with temperature. This information has been incorporated into soil carbon and Earth-system models, which suggest that warming-induced increases in carbon dioxide release from soils represent an important positive feedback loop that could influence twenty-first-century climate change. The magnitude of this feedback remains uncertain, however, not least because the response of soil microbial communities to changing temperatures has the potential to either decrease or increase warming-induced carbon losses substantially. Here we collect soils from different ecosystems along a climate gradient from the Arctic to the Amazon and investigate how microbial community-level responses control the temperature sensitivity of soil respiration. We find that the microbial community-level response more often enhances than reduces the mid- to long-term (90 days) temperature sensitivity of respiration. Furthermore, the strongest enhancing responses were observed in soils with high carbon-to-nitrogen ratios and in soils from cold climatic regions. After 90 days, microbial community responses increased the temperature sensitivity of respiration in high-latitude soils by a factor of 1.4 compared to the instantaneous temperature response. This suggests that the substantial carbon stores in Arctic and boreal soils could be more vulnerable to climate warming than currently predicted.

Heterogeneous catalysis in highly sensitive microreactors

DEFF Research Database (Denmark)

Olsen, Jakob Lind

This thesis present a highly sensitive silicon microreactor and examples of its use in studying catalysis. The experimental setup built for gas handling and temperature control for the microreactor is described. The implementation of LabVIEW interfacing for all the experimental parts makes...

Selective induction of high-ouabain-affinity isoform of Na+-K+-ATPase by thyroid hormone

International Nuclear Information System (INIS)

Haber, R.S.; Loeb, J.N.

1988-01-01

The administration of thyroid hormone is known to result in an induction of the Na + -K + -adenosinetriphosphatase (Na + -K + -ATPase) in rat skeletal muscle and other thyroid hormone-responsive tissues. Since the Na + -K + -ATPase in a variety of mammalian tissues has recently been reported to exist in at least two forms distinguishable by differing affinities for the inhibitory cardiac glycoside ouabain. The authors have studied the effects of 3,3',5-triiodo-L-thyronine (T 3 ) treatment on these two forms of the enzyme in rat diaphragm. The inhibition of Na + -K + -ATPase activity in a crude membrane fraction by varying concentrations of ouabain conformed to a biphasic pattern consistent with the presence of two distinct isoforms with inhibition constants (K I s) for ouabain of ∼10 -7 and 10 -4 M, respectively. Measurement of the specific binding of [ 3 H]ouabain to these membranes confirmed the presence of a class of high-affinity ouabain binding sites with a dissociation constant (K d ) of slightly less than 10 -7 M, whose maximal binding capacity was increased by T 3 treatment by 185%. Binding studies in unfractionated homogenates of diaphragm similarly demonstrated the presence of high-affinity sites whose maximal binding capacity was increased by T 3 treatment. Quantitation of both the high- and low-ouabain-affinity forms of the Na + -K + -ATPase by ouabain-dependent phosphorylation from [ 32 P]orthophosphate confirmed that T 3 treatment markedly increased the number of high-affinity sites while having little effect on the number of low-affinity sites. These observations provide, to our knowledge, the first demonstration that these two forms of the Na + -K + -ATPase are subject to selective hormonal induction

Passive Resistor Temperature Compensation for a High-Temperature Piezoresistive Pressure Sensor.

Science.gov (United States)

Yao, Zong; Liang, Ting; Jia, Pinggang; Hong, Yingping; Qi, Lei; Lei, Cheng; Zhang, Bin; Li, Wangwang; Zhang, Diya; Xiong, Jijun

2016-07-22

The main limitation of high-temperature piezoresistive pressure sensors is the variation of output voltage with operating temperature, which seriously reduces their measurement accuracy. This paper presents a passive resistor temperature compensation technique whose parameters are calculated using differential equations. Unlike traditional experiential arithmetic, the differential equations are independent of the parameter deviation among the piezoresistors of the microelectromechanical pressure sensor and the residual stress caused by the fabrication process or a mismatch in the thermal expansion coefficients. The differential equations are solved using calibration data from uncompensated high-temperature piezoresistive pressure sensors. Tests conducted on the calibrated equipment at various temperatures and pressures show that the passive resistor temperature compensation produces a remarkable effect. Additionally, a high-temperature signal-conditioning circuit is used to improve the output sensitivity of the sensor, which can be reduced by the temperature compensation. Compared to traditional experiential arithmetic, the proposed passive resistor temperature compensation technique exhibits less temperature drift and is expected to be highly applicable for pressure measurements in harsh environments with large temperature variations.

High Performance Affinity Chromatography of Antithrombin III Based on Monodisperse Poly (glycidyl methacrylate) Beads

Institute of Scientific and Technical Information of China (English)

无

2001-01-01

A new approach for the separation of antithrombin III with high performance affinity chromatography (HPAC) was described. A novel monodisperse,non-porous,cross-linked poly (glycidyl methacrylate) beads (PGMA) were used as the affinity support. With the water-soluble carbodiimide,heparin was linked covalently to amino-PGMA-beads,which was prepared by amination of PGMA. The adsorbent obtained exhibits high binding activity to antithrombin III (ATIII),good resolution and excellent mechanical properties and can be used under high flow rate.

Role of β/δ101Gln in Regulating the Effect of Temperature and Allosteric Effectors on Oxygen Affinity in Woolly Mammoth Hemoglobin†

Science.gov (United States)

Yuan, Yue; Byrd, Catherine; Shen, Tong-Jian; Simplaceanu, Virgil; Tam, Tsuey Chyi S.; Ho, Chien

2013-01-01

The oxygen affinity of woolly mammoth hemoglobin (rHb WM) is less affected by temperature change than that of Asian elephant hemoglobin (rHb AE) or human adult hemoglobin (Hb A). We report here a biochemical-biophysical study of Hb A, rHb AE, rHb WM and three rHb WM mutants with amino acid substitutions at β/δ101 (β/δ101Gln→Glu, Lys, or Asp) plus a double and a triple mutant, designed to clarify the role of the β/δ101 residue. The β/δ101Gln residue is important for responding to allosteric effectors, such as phosphate, inositol hexaphosphate (IHP), and chloride. The rHb WM mutants studied generally have higher affinity for oxygen under various conditions of pH, temperature, and salt concentration, and in the presence or absence of organic phosphate, than do rHb WM, rHb AE and Hb A. Titrations for the O2 affinity of these mutant rHbs as a function of chloride concentration indicate a lower heterotopic effect of this anion due to the replacement of β/δ101Gln in rHb WM. The alkaline Bohr effect of rHb WM and its mutants is reduced by 20–50% compared to that of Hb A and is independent of changes in temperature, in contrast to what has been observed in the hemoglobins of most mammalian species, including human. The results of our study on the temperature dependence of the O2 affinity of rHb WM and its mutant rHbs illustrate the important role of β/δ101Gln in regulating the functional properties of these hemoglobins. PMID:24228693

Robotic high-throughput purification of affinity-tagged recombinant proteins.

Science.gov (United States)

Wiesler, Simone C; Weinzierl, Robert O J

2015-01-01

Affinity purification of recombinant proteins has become the method of choice to obtain good quantities and qualities of proteins for a variety of downstream biochemical applications. While manual or FPLC-assisted purification techniques are generally time-consuming and labor-intensive, the advent of high-throughput technologies and liquid handling robotics has simplified and accelerated this process significantly. Additionally, without the human factor as a potential source of error, automated purification protocols allow for the generation of large numbers of proteins simultaneously and under directly comparable conditions. The delivered material is ideal for activity comparisons of different variants of the same protein. Here, we present our strategy for the simultaneous purification of up to 24 affinity-tagged proteins for activity measurements in biochemical assays. The protocol described is suitable for the scale typically required in individual research laboratories.

High Temperature, High Power Piezoelectric Composite Transducers

Science.gov (United States)

Lee, Hyeong Jae; Zhang, Shujun; Bar-Cohen, Yoseph; Sherrit, StewarT.

2014-01-01

Piezoelectric composites are a class of functional materials consisting of piezoelectric active materials and non-piezoelectric passive polymers, mechanically attached together to form different connectivities. These composites have several advantages compared to conventional piezoelectric ceramics and polymers, including improved electromechanical properties, mechanical flexibility and the ability to tailor properties by using several different connectivity patterns. These advantages have led to the improvement of overall transducer performance, such as transducer sensitivity and bandwidth, resulting in rapid implementation of piezoelectric composites in medical imaging ultrasounds and other acoustic transducers. Recently, new piezoelectric composite transducers have been developed with optimized composite components that have improved thermal stability and mechanical quality factors, making them promising candidates for high temperature, high power transducer applications, such as therapeutic ultrasound, high power ultrasonic wirebonding, high temperature non-destructive testing, and downhole energy harvesting. This paper will present recent developments of piezoelectric composite technology for high temperature and high power applications. The concerns and limitations of using piezoelectric composites will also be discussed, and the expected future research directions will be outlined. PMID:25111242

Genetically based low oxygen affinities of felid hemoglobins: lack of biochemical adaptation to high-altitude hypoxia in the snow leopard

Science.gov (United States)

Janecka, Jan E.; Nielsen, Simone S. E.; Andersen, Sidsel D.; Hoffmann, Federico G.; Weber, Roy E.; Anderson, Trevor; Storz, Jay F.; Fago, Angela

2015-01-01

ABSTRACT Genetically based modifications of hemoglobin (Hb) function that increase blood–O2 affinity are hallmarks of hypoxia adaptation in vertebrates. Among mammals, felid Hbs are unusual in that they have low intrinsic O2 affinities and reduced sensitivities to the allosteric cofactor 2,3-diphosphoglycerate (DPG). This combination of features compromises the acclimatization capacity of blood–O2 affinity and has led to the hypothesis that felids have a restricted physiological niche breadth relative to other mammals. In seeming defiance of this conjecture, the snow leopard (Panthera uncia) has an extraordinarily broad elevational distribution and occurs at elevations above 6000 m in the Himalayas. Here, we characterized structural and functional variation of big cat Hbs and investigated molecular mechanisms of Hb adaptation and allosteric regulation that may contribute to the extreme hypoxia tolerance of the snow leopard. Experiments revealed that purified Hbs from snow leopard and African lion exhibited equally low O2 affinities and DPG sensitivities. Both properties are primarily attributable to a single amino acid substitution, β2His→Phe, which occurred in the common ancestor of Felidae. Given the low O2 affinity and reduced regulatory capacity of feline Hbs, the extreme hypoxia tolerance of snow leopards must be attributable to compensatory modifications of other steps in the O2-transport pathway. PMID:26246610

A simple highly sensitive and selective aptamer-based colorimetric sensor for environmental toxins microcystin-LR in water samples.

Science.gov (United States)

Li, Xiuyan; Cheng, Ruojie; Shi, Huijie; Tang, Bo; Xiao, Hanshuang; Zhao, Guohua

2016-03-05

A simple and highly sensitive aptamer-based colorimetric sensor was developed for selective detection of Microcystin-LR (MC-LR). The aptamer (ABA) was employed as recognition element which could bind MC-LR with high-affinity, while gold nanoparticles (AuNPs) worked as sensing materials whose plasma resonance absorption peaks red shifted upon binding of the targets at a high concentration of sodium chloride. With the addition of MC-LR, the random coil aptamer adsorbed on Au NPs altered into regulated structure to form MC-LR-aptamer complexes and broke away from the surface of Au NPs, leading to the aggregation of AuNPs, and the color converted from red to blue due to the interparticle plasmon coupling. Results showed that our aptamer-based colorimetric sensor exhibited rapid and sensitive detection performance for MC-LR with linear range from 0.5 nM to 7.5 μM and the detection limit reached 0.37 nM. Meanwhile, the pollutants usually coexisting with MC-LR in pollutant water samples had not demonstrated disturbance for detecting of MC-LR. The mechanism was also proposed suggesting that high affinity interaction between aptamer and MC-LR significantly enhanced the sensitivity and selectivity for MC-LR detection. Besides, the established method was utilized in analyzing real water samples and splendid sensitivity and selectivity were obtained as well. Copyright © 2015 Elsevier B.V. All rights reserved.

Divergent apparent temperature sensitivity of terrestrial ecosystem respiration

Science.gov (United States)

Bing Song; Shuli Niu; Ruise Luo; Yiqi Luo; Jiquan Chen; Guirui Yu; Janusz Olejnik; Georg Wohlfahrt; Gerard Kiely; Ako Noormets; Leonardo Montagnani; Alessandro Cescatti; Vincenzo Magliulo; Beverly Elizabeth Law; Magnus Lund; Andrej Varlagin; Antonio Raschi; Matthias Peichl; Mats B. Nilsson; Lutz Merbold

2014-01-01

Recent studies revealed convergent temperature sensitivity of ecosystem respiration (Re) within aquatic ecosystems and between terrestrial and aquatic ecosystems. We do not know yet whether various terrestrial ecosystems have consistent or divergent temperature sensitivity. Here, we synthesized 163 eddy covariance flux sites across the world and...

Fatty acid and drug binding to a low-affinity component of human serum albumin, purified by affinity chromatography

DEFF Research Database (Denmark)

Vorum, H; Pedersen, A O; Honoré, B

1992-01-01

Binding equilibria for decanoate to a defatted, commercially available human serum albumin preparation were investigated by dialysis exchange rate determinations. The binding isotherm could not be fitted by the general binding equation. It was necessary to assume that the preparation was a mixture...... of two albumin components about 40% of the albumin having high affinity and about 60% having low affinity. By affinity chromatography we succeeded in purifying the low-affinity component from the mixture. The high-affinity component, however, could not be isolated. We further analyzed the fatty acid...... and drug binding abilities of the low-affinity component. The fatty acids decanoate, laurate, myristate and palmitate were bound with higher affinity to the mixture than to the low-affinity component. Diazepam was bound with nearly the same affinity to the low-affinity component as to the albumin mixture...

Characteristics of high affinity and low affinity adenosine binding sites in human cerebral cortex

International Nuclear Information System (INIS)

John, D.; Fox, I.V.

1986-01-01

The binding characteristics of human brain cortical membrane fractions were evaluated to test the hypothesis that there are A 1 and A 2 adenosine binding sites. The ligands used were 2-chloro(8- 3 H) adenosine and N 6 -(adenine-2, 8- 3 H) cyclohexayladenosine. Binding of chloroadenosine to human brain cortical membranes was time dependent, reversible and concentration dependent. The kinetic constant determinations from binding studies of the adenosine receptor are presented. Utilizing tritium-cyclohexyladenosine as ligand the authors observed evidence for a high affinity binding site in human brain cortical membranes with a kd of 5 nM

Temperature sensitivity analysis of polarity controlled electrostatically doped tunnel field-effect transistor

Science.gov (United States)

Nigam, Kaushal; Pandey, Sunil; Kondekar, P. N.; Sharma, Dheeraj

2016-09-01

The conventional tunnel field-effect transistors (TFETs) have shown potential to scale down in sub-22 nm regime due to its lower sub-threshold slope and robustness against short-channel effects (SCEs), however, sensitivity towards temperature variation is a major concern. Therefore, for the first time, we investigate temperature sensitivity analysis of a polarity controlled electrostatically doped tunnel field-effect transistor (ED-TFET). Different performance metrics and analog/RF figure-of-merits were considered and compared for both devices, and simulations were performed using Silvaco ATLAS device tool. We found that the variation in ON-state current in ED-TFET is almost temperature independent due to electrostatically doped mechanism, while, it increases in conventional TFET at higher temperature. Above room temperature, the variation in ION, IOFF, and SS sensitivity in ED-TFET are only 0.11%/K, 2.21%/K, and 0.63%/K, while, in conventional TFET the variations are 0.43%/K, 2.99%/K, and 0.71%/K, respectively. However, below room temperature, the variation in ED-TFET ION is 0.195%/K compared to 0.27%/K of conventional TFET. Moreover, it is analysed that the incomplete ionization effect in conventional TFET severely affects the drive current and the threshold voltage, while, ED-TFET remains unaffected. Hence, the proposed ED-TFET is less sensitive towards temperature variation and can be used for cryogenics as well as for high temperature applications.

Highly Sensitive Temperature Sensors Based on Fiber-Optic PWM and Capacitance Variation Using Thermochromic Sensing Membrane

Directory of Open Access Journals (Sweden)

Md. Rajibur Rahaman Khan

2016-07-01

Full Text Available In this paper, we propose a temperature/thermal sensor that contains a Rhodamine-B sensing membrane. We applied two different sensing methods, namely, fiber-optic pulse width modulation (PWM and an interdigitated capacitor (IDC-based temperature sensor to measure the temperature from 5 °C to 100 °C. To the best of our knowledge, the fiber-optic PWM-based temperature sensor is reported for the first time in this study. The proposed fiber-optic PWM temperature sensor has good sensing ability; its sensitivity is ~3.733 mV/°C. The designed temperature-sensing system offers stable sensing responses over a wide dynamic range, good reproducibility properties with a relative standard deviation (RSD of ~0.021, and the capacity for a linear sensing response with a correlation coefficient of R2 ≈ 0.992 over a wide sensing range. In our study, we also developed an IDC temperature sensor that is based on the capacitance variation principle as the IDC sensing element is heated. We compared the performance of the proposed temperature-sensing systems with different fiber-optic temperature sensors (which are based on the fiber-optic wavelength shift method, the long grating fiber-optic Sagnac loop, and probe type fiber-optics in terms of sensitivity, dynamic range, and linearity. We observed that the proposed sensing systems have better sensing performance than the above-mentioned sensing system.

Multimode fiber tip Fabry-Perot cavity for highly sensitive pressure measurement.

Science.gov (United States)

Chen, W P; Wang, D N; Xu, Ben; Zhao, C L; Chen, H F

2017-03-23

We demonstrate an optical Fabry-Perot interferometer fiber tip sensor based on an etched end of multimode fiber filled with ultraviolet adhesive. The fiber device is miniature (with diameter of less than 60 μm), robust and low cost, in a convenient reflection mode of operation, and has a very high gas pressure sensitivity of -40.94 nm/MPa, a large temperature sensitivity of 213 pm/°C within the range from 55 to 85 °C, and a relatively low temperature cross-sensitivity of 5.2 kPa/°C. This device has a high potential in monitoring environment of high pressure.

Characterization of high-affinity (/sup 3/H)ouabain binding in the rat central nervous system

Energy Technology Data Exchange (ETDEWEB)

Hauger, R.; Luu, H.M.; Meyer, D.K.; Goodwin, F.K.; Paul, S.M.

1985-06-01

The characteristics of (/sup 3/H)ouabain binding were examined in various areas of rat brain. In the striatum, Scatchard analysis revealed a single class of high-affinity binding sites with an apparent binding affinity (KD) of 10.4 +/- 0.9 nM and an estimated binding capacity (Bmax) of 7.6 +/- 1.9 pmol/mg protein. Similar monophasic Scatchard plots were found in the brainstem, cerebellum, hypothalamus, and frontal cerebral cortex. (/sup 3/H)Ouabain binding to rat brain was sodium- and ATP-dependent and strongly inhibited by potassium. Proscillariden A was the most potent cardiac glycoside tested in inhibiting specific (/sup 3/H)ouabain binding to brain membranes, and the rank order of inhibitory potencies for a series of cardiac glycosides was similar to that previously reported for inhibition of heart Na,K-ATPase. To assess whether the high-affinity binding sites for (/sup 3/H)ouabain were localized to neuronal or nonneuronal membranes, the effect of discrete kainic acid lesions on striatal (/sup 3/H)ouabain binding was examined. Kainic acid lesions of the striatum reduced (/sup 3/H)ouabain binding to striatal homogenates by 79.6 +/- 1.6%. This suggests that the high-affinity (/sup 3/H)ouabain binding sites measured in our experiments are localized to neuronal elements. Thus, the high-affinity binding of (/sup 3/H)ouabain to brain membranes may selectively label a neuronal form or conformation of Na,K-ATPase.

Selective high-affinity polydentate ligands and methods of making such

Energy Technology Data Exchange (ETDEWEB)

Denardo, Sally J.; Denardo, Gerald L.; Balhorn, Rodney L.

2018-02-06

This invention provides novel polydentate selective high affinity ligands (SHALs) that can be used in a variety of applications in a manner analogous to the use of antibodies. SHALs typically comprise a multiplicity of ligands that each bind different region son the target molecule. The ligands are joined directly or through a linker thereby forming a polydentate moiety that typically binds the target molecule with high selectivity and avidity.

High-aluminum-affinity silica is a nanoparticle that seeds secondary aluminosilicate formation.

Directory of Open Access Journals (Sweden)

Ravin Jugdaohsingh

Full Text Available Despite the importance and abundance of aluminosilicates throughout our natural surroundings, their formation at neutral pH is, surprisingly, a matter of considerable debate. From our experiments in dilute aluminum and silica containing solutions (pH ~ 7 we previously identified a silica polymer with an extraordinarily high affinity for aluminium ions (high-aluminum-affinity silica polymer, HSP. Here, further characterization shows that HSP is a colloid of approximately 2.4 nm in diameter with a mean specific surface area of about 1,000 m(2 g(-1 and it competes effectively with transferrin for Al(III binding. Aluminum binding to HSP strongly inhibited its decomposition whilst the reaction rate constant for the formation of the β-silicomolybdic acid complex indicated a diameter between 3.6 and 4.1 nm for these aluminum-containing nanoparticles. Similarly, high resolution microscopic analysis of the air dried aluminum-containing silica colloid solution revealed 3.9 ± 1.3 nm sized crystalline Al-rich silica nanoparticles (ASP with an estimated Al:Si ratio of between 2 and 3 which is close to the range of secondary aluminosilicates such as imogolite. Thus the high-aluminum-affinity silica polymer is a nanoparticle that seeds early aluminosilicate formation through highly competitive binding of Al(III ions. In niche environments, especially in vivo, this may serve as an alternative mechanism to polyhydroxy Al(III species binding monomeric silica to form early phase, non-toxic aluminosilicates.

High-Aluminum-Affinity Silica Is a Nanoparticle That Seeds Secondary Aluminosilicate Formation

Science.gov (United States)

Jugdaohsingh, Ravin; Brown, Andy; Dietzel, Martin; Powell, Jonathan J.

2013-01-01

Despite the importance and abundance of aluminosilicates throughout our natural surroundings, their formation at neutral pH is, surprisingly, a matter of considerable debate. From our experiments in dilute aluminum and silica containing solutions (pH ~ 7) we previously identified a silica polymer with an extraordinarily high affinity for aluminium ions (high-aluminum-affinity silica polymer, HSP). Here, further characterization shows that HSP is a colloid of approximately 2.4 nm in diameter with a mean specific surface area of about 1,000 m2 g-1 and it competes effectively with transferrin for Al(III) binding. Aluminum binding to HSP strongly inhibited its decomposition whilst the reaction rate constant for the formation of the β-silicomolybdic acid complex indicated a diameter between 3.6 and 4.1 nm for these aluminum-containing nanoparticles. Similarly, high resolution microscopic analysis of the air dried aluminum-containing silica colloid solution revealed 3.9 ± 1.3 nm sized crystalline Al-rich silica nanoparticles (ASP) with an estimated Al:Si ratio of between 2 and 3 which is close to the range of secondary aluminosilicates such as imogolite. Thus the high-aluminum-affinity silica polymer is a nanoparticle that seeds early aluminosilicate formation through highly competitive binding of Al(III) ions. In niche environments, especially in vivo, this may serve as an alternative mechanism to polyhydroxy Al(III) species binding monomeric silica to form early phase, non-toxic aluminosilicates. PMID:24349573

Biphasic regulation of development of the high-affinity saxitoxin receptor by innervation in rat skeletal muscle

International Nuclear Information System (INIS)

Sherman, S.J.; Catterall, W.A.

1982-01-01

Specific binding of 3 H-saxitoxin (STX) was used to quantitate the density of voltage-sensitive sodium channels in developing rat skeletal muscle. In adult triceps surae, a single class of sites with a KD . 2.9 nM and a density of 21 fmol/mg wet wt was detected. The density of these high-affinity sites increased from 2.0 fmol/mg wet wt to the adult value in linear fashion during days 2-25 after birth. Denervation of the triceps surae at day 11 or 17 reduced final saxitoxin receptor site density to 10.4 or 9.2 fmol/mg wet wt, respectively, without changing KD. Denervation of the triceps surae at day 5 did not alter the subsequent development of saxitoxin receptor sites during days 5-9 and accelerated the increase of saxitoxin receptor sites during days 9-13. After day 13, saxitoxin receptor development abruptly ceased and the density of saxitoxin receptor sites declined to 11 fmol/wg wet wt. These results show that the regulation of high-affinity saxitoxin receptor site density by innervation is biphasic. During the first phase, which is independent of continuing innervation, the saxitoxin receptor density increases to 47-57% of the adult level. After day 11, the second phase of development, which is dependent on continuing innervation, gives rise to the adult density of saxitoxin receptors

Sensitivity analysis of recovery efficiency in high-temperature aquifer thermal energy storage with single well

International Nuclear Information System (INIS)

Jeon, Jun-Seo; Lee, Seung-Rae; Pasquinelli, Lisa; Fabricius, Ida Lykke

2015-01-01

High-temperature aquifer thermal energy storage system usually shows higher performance than other borehole thermal energy storage systems. Although there is a limitation in the widespread use of the HT-ATES system because of several technical problems such as clogging, corrosion, etc., it is getting more attention as these issues are gradually alleviated. In this study, a sensitivity analysis of recovery efficiency in two cases of HT-ATES system with a single well is conducted to select key parameters. For a fractional factorial design used to choose input parameters with uniformity, the optimal Latin hypercube sampling with an enhanced stochastic evolutionary algorithm is considered. Then, the recovery efficiency is obtained using a computer model developed by COMSOL Multiphysics. With input and output variables, the surrogate modeling technique, namely the Gaussian-Kriging method with Smoothly Clopped Absolute Deviation Penalty, is utilized. Finally, the sensitivity analysis is performed based on the variation decomposition. According to the result of sensitivity analysis, the most important input variables are selected and confirmed to consider the interaction effects for each case and it is confirmed that key parameters vary with the experiment domain of hydraulic and thermal properties as well as the number of input variables. - Highlights: • Main and interaction effects on recovery efficiency in HT-ATES was investigated. • Reliability depended on fractional factorial design and interaction effects. • Hydraulic permeability of aquifer had an important impact on recovery efficiency. • Site-specific sensitivity analysis of HT-ATES was recommended.

Differences between high-affinity forskolin binding sites in dopamine-riche and other regions of rat brain

International Nuclear Information System (INIS)

Poat, J.A.; Cripps, H.E.; Iversen, L.L.

1988-01-01

Forskolin labelled with [ 3 H] bound to high- and low-affinity sites in the rat brain. The high-affinity site was discretely located, with highest densities in the striatum, nucleus accumbens, olfactory tubercule, substantia nigra, hippocampus, and the molecular layers of the cerebellum. This site did not correlate well with the distribution of adenylate cyclase. The high-affinity striatal binding site may be associated with a stimulatory guanine nucleotide-binding protein. Thus, the number of sites was increased by the addition of Mg 2+ and guanylyl imidodiphosphate. Cholera toxin stereotaxically injected into rat striatum increased the number of binding sites, and no further increase was noted following the subsequent addition of guanyl nucleotide. High-affinity forskolin binding sites in non-dopamine-rich brain areas (hippocampus and cerebullum) were modulated in a qualitatively different manner by guanyl nucleotides. In these areas the number of binding sites was significantly reduced by the addition of guanyl nucleotide. These results suggest that forskolin may have a potential role in identifying different functional/structural guanine nucleotide-binding proteins

Detection of Waterborne Viruses Using High Affinity Molecularly Imprinted Polymers.

Science.gov (United States)

Altintas, Zeynep; Gittens, Micah; Guerreiro, Antonio; Thompson, Katy-Anne; Walker, Jimmy; Piletsky, Sergey; Tothill, Ibtisam E

2015-07-07

Molecularly imprinted polymers (MIPs) are artificial receptor ligands which can recognize and specifically bind to a target molecule. They are more resistant to chemical and biological damage and inactivation than antibodies. Therefore, target specific-MIP nanoparticles are aimed to develop and implemented to biosensors for the detection of biological toxic agents such as viruses, bacteria, and fungi toxins that cause many diseases and death due to the environmental contamination. For the first time, a molecularly imprinted polymer (MIP) targeting the bacteriophage MS2 as the template was investigated using a novel solid-phase synthesis method to obtain the artificial affinity ligand for the detection and removal of waterborne viruses through optical-based sensors. A high affinity between the artificial ligand and the target was found, and a regenerative MIP-based virus detection assay was successfully developed using a new surface plasmon resonance (SPR)-biosensor which provides an alternative technology for the specific detection and removal of waterborne viruses that lead to high disease and death rates all over the world.

Production and Identification of High Affinity Monoclonal Antibodies Against Pesticide Carbofuran

Institute of Scientific and Technical Information of China (English)

无

2007-01-01

To produce high-affinity monoclonal antibodies against pesticide carbofuran, and the develop immunochemical assays for people's health and environmental protection, the hapten 4-[[(2,3-dihydro-2,2-dimethyl-7-benzofuranyloxy) carbonyl]-amino]-butanoic acid (BFNB) of carbofuran was synthesized and Balb/c mice were immunized by the hapten-carrier (BFNB-bovine serum albumin, BFNB-BSA) conjugates. The splenocytes of immunized mice were fused with Sp2/0 cells and the cultural supernatants of hybridoma cells were screened by the indirect enzyme-linked immunoabsorbent assay (ELISA), based on BFNB-ovoalbumin conjugates (BFNB-OVA). Purified monoclonal antibody (McAb) was obtained from fluids of ascites, deposited by octanoic acid and ammonium sulfate. The affinity and the specificity of McAb were characterized by ELISA or indirect competitive ELISA. A hybridoma cell line (5D3) secreting anti-carbofuran McAb had been established. The titer of culture medium and ascites was up to 1:2.048 × 103 and 1:1.024 × 106, respectively, and the subtype of the McAb was IgG1. The affinity constant of the McAb was about 2.54 × 109 L mol-1, with an IC50 value of 1.18 ng mL-1 and a detection limit of 0.01 ng mL-1. Cross-reactivity studies showed that the McAb was quiet specific for carbofuran, as among the four analogous compounds, they were all hardly recognized (4.59 × 10-4% for 2,3-dihydro-2,2-dimethyl-7-benzofuranol and less than 3.0 × 10-4% for others). The prepared McAb had a very high affinity and specificity,and it could be used to develop ELISA for rapid determination of carbofuran.

Effect of soil moisture on the temperature sensitivity of Northern soils

Science.gov (United States)

Minions, C.; Natali, S.; Ludwig, S.; Risk, D.; Macintyre, C. M.

2017-12-01

Arctic and boreal ecosystems are vast reservoirs of carbon and are particularly sensitive to climate warming. Changes in the temperature and precipitation regimes of these regions could significantly alter soil respiration rates, impacting atmospheric concentrations and affecting climate change feedbacks. Many incubation studies have shown that both temperature and soil moisture are important environmental drivers of soil respiration; this relationship, however, has rarely been demonstrated with in situ data. Here we present the results of a study at six field sites in Alaska from 2016 to 2017. Low-power automated soil gas systems were used to measure soil surface CO2 flux from three forced diffusion chambers and soil profile concentrations from three soil depth chambers at hourly intervals at each site. HOBO Onset dataloggers were used to monitor soil moisture and temperature profiles. Temperature sensitivity (Q10) was determined at each site using inversion analysis applied over different time periods. With highly resolved data sets, we were able to observe the changes in soil respiration in response to changes in temperature and soil moisture. Through regression analysis we confirmed that temperature is the primary driver in soil respiration, but soil moisture becomes dominant beyond a certain threshold, suppressing CO2 flux in soils with high moisture content. This field study supports the conclusions made from previous soil incubation studies and provides valuable insights into the impact of both temperature and soil moisture changes on soil respiration.

Computational modeling and molecular imprinting for the development of acrylic polymers with high affinity for bile salts.

Science.gov (United States)

Yañez, Fernando; Chianella, Iva; Piletsky, Sergey A; Concheiro, Angel; Alvarez-Lorenzo, Carmen

2010-02-05

This work has focused on the rational development of polymers capable of acting as traps of bile salts. Computational modeling was combined with molecular imprinting technology to obtain networks with high affinity for cholate salts in aqueous medium. The screening of a virtual library of 18 monomers, which are commonly used for imprinted networks, identified N-(3-aminopropyl)-methacrylate hydrochloride (APMA.HCl), N,N-diethylamino ethyl methacrylate (DEAEM) and ethyleneglycol methacrylate phosphate (EGMP) as suitable functional monomers with medium-to-high affinity for cholic acid. The polymers were prepared with a fix cholic acid:functional monomer mole ratio of 1:4, but with various cross-linking densities. Compared to polymers prepared without functional monomer, both imprinted and non-imprinted microparticles showed a high capability to remove sodium cholate from aqueous medium. High affinity APMA-based particles even resembled the performance of commercially available cholesterol-lowering granules. The imprinting effect was evident in most of the networks prepared, showing that computational modeling and molecular imprinting can act synergistically to improve the performance of certain polymers. Nevertheless, both the imprinted and non-imprinted networks prepared with the best monomer (APMA.HCl) identified by the modeling demonstrated such high affinity for the template that the imprinting effect was less important. The fitting of adsorption isotherms to the Freundlich model indicated that, in general, imprinting increases the population of high affinity binding sites, except when the affinity of the functional monomer for the target molecule is already very high. The cross-linking density was confirmed as a key parameter that determines the accessibility of the binding points to sodium cholate. Materials prepared with 9% mol APMA and 91% mol cross-linker showed enough affinity to achieve binding levels of up to 0.4 mmol g(-1) (i.e., 170 mg g(-1)) under flow

Optical temperature sensor with enhanced sensitivity by employing hybrid waveguides in a silicon Mach-Zehnder interferometer

DEFF Research Database (Denmark)

Guan, Xiaowei; Wang, Xiaoyan; Frandsen, Lars Hagedorn

2016-01-01

We report on a novel design of an on-chip optical temperature sensor based on a Mach-Zehnder interferometer configuration where the two arms consist of hybrid waveguides providing opposite temperature-dependent phase changes to enhance the temperature sensitivity of the sensor. The sensitivity...... of the fabricated sensor with silicon/polymer hybrid waveguides is measured to be 172 pm/°C, which is two times larger than a conventional all-silicon optical temperature sensor (∼80 pm/°C). Moreover, a design with silicon/titanium dioxide hybrid waveguides is by calculation expected to have a sensitivity as high...

A rhodium(III) complex for high-affinity DNA base-pair mismatch recognition

Science.gov (United States)

Junicke, Henrik; Hart, Jonathan R.; Kisko, Jennifer; Glebov, Oleg; Kirsch, Ilan R.; Barton, Jacqueline K.

2003-01-01

A rhodium(III) complex, rac-[Rh(bpy)2phzi]3+ (bpy, 2,2′-bipyridine; phzi, benzo[a]phenazine-5,6-quinone diimine) has been designed as a sterically demanding intercalator targeted to destabilized mismatched sites in double-helical DNA. The complex is readily synthesized by condensation of the phenazine quinone with the corresponding diammine complex. Upon photoactivation, the complex promotes direct strand scission at single-base mismatch sites within the DNA duplex. As with the parent mismatch-specific reagent, [Rh(bpy)2(chrysi)]3+ [chrysene-5,6-quinone diimine (chrysi)], mismatch selectivity depends on the helix destabilization associated with mispairing. Unlike the parent chrysi complex, the phzi analogue binds and cleaves with high affinity and efficiency. The specific binding constants for CA, CC, and CT mismatches within a 31-mer oligonucleotide duplex are 0.3, 1, and 6 × 107 M−1, respectively; site-specific photocleavage is evident at nanomolar concentrations. Moreover, the specificity, defined as the ratio in binding affinities for mispaired vs. well paired sites, is maintained. The increase in affinity is attributed to greater stability in the mismatched site associated with stacking by the heterocyclic aromatic ligand. The high-affinity complex is also applied in the differential cleavage of DNA obtained from cell lines deficient in mismatch repair vs. those proficient in mismatch repair. Agreement is found between photocleavage by the mismatch-specific probes and deficiency in mismatch repair. This mismatch-specific targeting, therefore, offers a potential strategy for new chemotherapeutic design. PMID:12610209

The RCK1 high-affinity Ca2+ sensor confers carbon monoxide sensitivity to Slo1 BK channels.

Science.gov (United States)

Hou, Shangwei; Xu, Rong; Heinemann, Stefan H; Hoshi, Toshinori

2008-03-11

Carbon monoxide (CO) is a lethal gas, but it is also increasingly recognized as a physiological signaling molecule capable of regulating a variety of proteins. Among them, large-conductance Ca(2+)- and voltage-gated K(+) (Slo1 BK) channels, important in vasodilation and neuronal firing, have been suggested to be directly stimulated by CO. However, the molecular mechanism of the stimulatory action of CO on the Slo1 BK channel has not been clearly elucidated. We report here that CO reliably and repeatedly activates Slo1 BK channels in excised membrane patches in the absence of Ca(2+) in a voltage-sensor-independent manner. The stimulatory action of CO on the Slo1 BK channel requires an aspartic acid and two histidine residues located in the cytoplasmic RCK1 domain, and the effect persists under the conditions known to inhibit the conventional interaction between CO and heme in other proteins. We propose that CO acts as a partial agonist for the high-affinity divalent cation sensor in the RCK1 domain of the Slo1 BK channel.

Application of High Temperature Superconductors to Accelerators

CERN Document Server

Ballarino, A

2000-01-01

Since the discovery of high temperature superconductivity, a large effort has been made by the scientific community to investigate this field towards a possible application of the new oxide superconductors to different devices like SMES, magnetic bearings, flywheels energy storage, magnetic shielding, transmission cables, fault current limiters, etc. However, all present day large scale applications using superconductivity in accelerator technology are based on conventional materials operating at liquid helium temperatures. Poor mechanical properties, low critical current density and sensitivity to the magnetic field at high temperature are the key parameters whose improvement is essential for a large scale application of high temperature superconductors to such devices. Current leads, used for transferring currents from the power converters, working at room temperature, into the liquid helium environment, where the magnets are operating, represent an immediate application of the emerging technology of high t...

High blood oxygen affinity in the air-breathing swamp eel Monopterus albus.

Science.gov (United States)

Damsgaard, Christian; Findorf, Inge; Helbo, Signe; Kocagoz, Yigit; Buchanan, Rasmus; Huong, Do Thi Thanh; Weber, Roy E; Fago, Angela; Bayley, Mark; Wang, Tobias

2014-12-01

The Asian swamp eel (Monopterus albus, Zuiew 1793) is a facultative air-breathing fish with reduced gills. Previous studies have shown that gas exchange seems to occur across the epithelium of the buccopharyngeal cavity, the esophagus and the integument, resulting in substantial diffusion limitations that must be compensated by adaptations in others steps of the O₂ transport system to secure adequate O₂ delivery to the respiring tissues. We therefore investigated O₂ binding properties of whole blood, stripped hemoglobin (Hb), two major isoHb components and the myoglobin (Mb) from M. albus. Whole blood was sampled using indwelling catheters for blood gas analysis and determination of O₂ equilibrium curves. Hb was purified to assess the effects of endogenous allosteric effectors, and Mb was isolated from heart and skeletal muscle to determine its O₂ binding properties. The blood of M. albus has a high O₂ carrying capacity [hematocrit (Hct) of 42.4±4.5%] and binds O₂ with an unusually high affinity (P₅₀=2.8±0.4mmHg at 27°C and pH7.7), correlating with insensitivity of the Hb to the anionic allosteric effectors that normally decrease Hb-O₂ affinity. In addition, Mb is present at high concentrations in both heart and muscle (5.16±0.99 and 1.08±0.19mg ∙ g wet tissue⁻¹, respectively). We suggest that the high Hct and high blood O₂ affinity serve to overcome the low diffusion capacity in the relatively inefficient respiratory surfaces, while high Hct and Mb concentration aid in increasing the O₂ flux from the blood to the muscles. Copyright © 2014 Elsevier Inc. All rights reserved.

Decrease in the red cell cofactor 2,3-diphosphoglycerate increases hemoglobin oxygen affinity in the hibernating brown bear Ursus arctos.

Science.gov (United States)

Revsbech, Inge G; Malte, Hans; Fröbert, Ole; Evans, Alina; Blanc, Stéphane; Josefsson, Johan; Fago, Angela

2013-01-01

During winter hibernation, brown bears (Ursus arctos) reduce basal O(2) consumption rate to ∼25% compared with the active state, while body temperature decreases moderately (to ∼30°C), suggesting a temperature-independent component in their metabolic depression. To establish whether changes in O(2) consumption during hibernation correlate with changes in blood O(2) affinity, we took blood samples from the same six individuals of hibernating and nonhibernating free-ranging brown bears during winter and summer, respectively. A single hemoglobin (Hb) component was detected in all samples, indicating no switch in Hb synthesis. O(2) binding curves measured on red blood cell lysates at 30°C and 37°C showed a less temperature-sensitive O(2) affinity than in other vertebrates. Furthermore, hemolysates from hibernating bears consistently showed lower cooperativity and higher O(2) affinity than their summer counterparts, regardless of the temperature. We found that this increase in O(2) affinity was associated with a significant decrease in the red cell Hb-cofactor 2,3-diphosphoglycerate (DPG) during hibernation to approximately half of the summer value. Experiments performed on purified Hb, to which DPG had been added to match summer and winter levels, confirmed that the low DPG content was the cause of the left shift in the Hb-O(2) equilibrium curve during hibernation. Levels of plasma lactate indicated that glycolysis is not upregulated during hibernation and that metabolism is essentially aerobic. Calculations show that the increase in Hb-O(2) affinity and decrease in cooperativity resulting from decreased red cell DPG may be crucial in maintaining a fairly constant tissue oxygen tension during hibernation in vivo.

Affinity of antibody secreted by a single cell

International Nuclear Information System (INIS)

Doran, D.M.

1978-01-01

It was the intention of this research to measure the affinity of antibody secreted by a single cell, and to describe the spectrum of affinities displayed in response to antigenic stimulation. The single cell secreting specific antibody was isolated by means of the hemolytic plaque assay. The amount of antibody secreted by the cell was to be measured through the use of a solid phase radioimmunoassay. The affinity of the antibody would be estimated by comparing the diameter of the plaque, and the amount of antibody secreted, with a mathematical theory of the formation of a plaque in agar. As a test system, a solid phase radioimmunoassay was developed for human serum albumin using antibody coupled to Sephadex. A sensitivity of 1 nanogram was attained with this assay. A solid phase radioimmunoassay for mouse immunoglobulin M was developed, using antibody coupled to Sepharose. The sensitivity attained with this assay was only on the order of 10 micrograms. The mouse immunoglobulin M radioimmunoassay was not sensitive enough to measure the amount of antibody secreted by a single cell. From a theoretical equation, the relationship between antibody affinity, plaque diameter and antibody secretion rate was calculated for the experimental conditions used in this research. By assuming a constant antibody secretion rate, an effective binding constant for the antibody was estimated from the average plaque diameters. This effective binding constant was observed to increase during the immune response

A Delay Time Measurement of ULTRAS (Ultra-high Temperature Ultrasonic Response Analysis System) for a High Temperature Experiment

International Nuclear Information System (INIS)

Koo, Kil Mo; Kim, Sang Baik

2010-01-01

The temperature measurement of very high temperature core melt is of importance in a high temperature as the molten pool experiment in which gap formation between core melt and the reactor lower head, and the effect of the gap on thermal behavior are to be measured. The existing temperature measurement techniques have some problems, which the thermocouple, one of the contact methods, is restricted to under 2000 .deg. C, and the infrared thermometry, one of the non-contact methods, is unable to measure an internal temperature and very sensitive to the interference from reacted gases. In order to solve these problems, the delay time technique of ultrasonic wavelets due to high temperature has two sorts of stage. As a first stage, a delay time measurement of ULTRAS (Ultra-high Temperature Ultrasonic Response Analysis System) is suggested. As a second stage, a molten material temperature was measured up to 2300 .deg. C. Also, the optimization design of the UTS (ultrasonic temperature sensor) with persistence at the high temperature was suggested in this paper. And the utilization of the theory suggested in this paper and the efficiency of the developed system are performed by special equipment and some experiments supported by KRISS (Korea Research Institute of Standard and Science)

Statistical removal of background signals from high-throughput 1H NMR line-broadening ligand-affinity screens

International Nuclear Information System (INIS)

Worley, Bradley; Sisco, Nicholas J.; Powers, Robert

2015-01-01

NMR ligand-affinity screens are vital to drug discovery, are routinely used to screen fragment-based libraries, and used to verify chemical leads from high-throughput assays and virtual screens. NMR ligand-affinity screens are also a highly informative first step towards identifying functional epitopes of unknown proteins, as well as elucidating the biochemical functions of protein–ligand interaction at their binding interfaces. While simple one-dimensional 1 H NMR experiments are capable of indicating binding through a change in ligand line shape, they are plagued by broad, ill-defined background signals from protein 1 H resonances. We present an uncomplicated method for subtraction of protein background in high-throughput ligand-based affinity screens, and show that its performance is maximized when phase-scatter correction is applied prior to subtraction

Reconstitution of high affinity α2 adrenergic agonist binding by fusion with a pertussis toxin substrate

International Nuclear Information System (INIS)

Kim, M.H.; Neubig, R.R.

1986-01-01

High affinity α 2 adrenergic agonist binding is thought to occur via a coupling of the α 2 receptor with N/sub i/, the inhibitory guanyl nucleotide binding protein. Human platelet membranes pretreated at pH 11.5 exhibit a selective inactivation of agonist binding and N/sub i/. To further study the mechanism of agonist binding, alkali treated membranes (ATM) were mixed with membranes pretreated with 10 μM phenoxybenzamine to block α 2 receptors (POB-M). The combined membrane pellet was incubated in 50% polyethylene glycol (PEG) to promote membrane-membrane fusion and assayed for binding to the α 2 agonist [ 3 H]UK 14,304 (UK) and the antagonist [ 3 H] yohimbine. PEG treatment resulted in a 2-4 fold enhancement of UK binding whereas yohimbine binding was unchanged. No enhancement of UK binding was observed in the absence of PEG treatment. The reconstitution was dependent on the addition of POB-M. They found that a 1:1 ratio of POB-M:ATM was optimal. Reconstituted binding was inhibited by GppNHp. Fusion of rat C6 glioma cell membranes, which do not contain α 2 receptors, also enhanced agonist binding to ATM. Fusion of C6 membranes from cells treated with pertussis toxin did not enhance [ 3 H] UK binding. These data show that a pertussis toxin sensitive membrane component, possibly N/sub i/, can reconstitute high affinity α 2 agonist binding

Selective induction of high-ouabain-affinity isoform of Na sup + -K sup + -ATPase by thyroid hormone

Energy Technology Data Exchange (ETDEWEB)

Haber, R.S.; Loeb, J.N. (Columbia Univ., New York, NY (USA))

1988-12-01

The administration of thyroid hormone is known to result in an induction of the Na{sup +}-K{sup +}-adenosinetriphosphatase (Na{sup +}-K{sup +}-ATPase) in rat skeletal muscle and other thyroid hormone-responsive tissues. Since the Na{sup +}-K{sup +}-ATPase in a variety of mammalian tissues has recently been reported to exist in at least two forms distinguishable by differing affinities for the inhibitory cardiac glycoside ouabain. The authors have studied the effects of 3,3{prime},5-triiodo-L-thyronine (T{sub 3}) treatment on these two forms of the enzyme in rat diaphragm. The inhibition of Na{sup +}-K{sup +}-ATPase activity in a crude membrane fraction by varying concentrations of ouabain conformed to a biphasic pattern consistent with the presence of two distinct isoforms with inhibition constants (K{sub I}s) for ouabain of {approximately}10{sup {minus}7} and 10{sup {minus}4} M, respectively. Measurement of the specific binding of ({sup 3}H)ouabain to these membranes confirmed the presence of a class of high-affinity ouabain binding sites with a dissociation constant (K{sub d}) of slightly less than 10{sup {minus}7}M, whose maximal binding capacity was increased by T{sub 3} treatment by 185%. Binding studies in unfractionated homogenates of diaphragm similarly demonstrated the presence of high-affinity sites whose maximal binding capacity was increased by T{sub 3} treatment. Quantitation of both the high- and low-ouabain-affinity forms of the Na{sup +}-K{sup +}-ATPase by ouabain-dependent phosphorylation from ({sup 32}P)orthophosphate confirmed that T{sub 3} treatment markedly increased the number of high-affinity sites while having little effect on the number of low-affinity sites. These observations provide, to our knowledge, the first demonstration that these two forms of the Na{sup +}-K{sup +}-ATPase are subject to selective hormonal induction.

Selection of DNA aptamers against epidermal growth factor receptor with high affinity and specificity

Energy Technology Data Exchange (ETDEWEB)

Wang, Deng-Liang [The First Clinical Medical College of Fujian Medical University, Fuzhou (China); Department of Neurosurgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou (China); Song, Yan-Ling; Zhu, Zhi; Li, Xi-Lan; Zou, Yuan [State Key Laboratory for Physical Chemistry of Solid Surfaces, Key Laboratory for Chemical Biology of Fujian Province, Key Laboratory of Analytical Chemistry, and Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005 (China); Yang, Hai-Tao; Wang, Jiang-Jie [The First Clinical Medical College of Fujian Medical University, Fuzhou (China); Yao, Pei-Sen [Department of Neurosurgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou (China); Pan, Ru-Jun [The First Clinical Medical College of Fujian Medical University, Fuzhou (China); Yang, Chaoyong James, E-mail: cyyang@xmu.edu.cn [State Key Laboratory for Physical Chemistry of Solid Surfaces, Key Laboratory for Chemical Biology of Fujian Province, Key Laboratory of Analytical Chemistry, and Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005 (China); Kang, De-Zhi, E-mail: kdzy99988@163.com [The First Clinical Medical College of Fujian Medical University, Fuzhou (China); Department of Neurosurgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou (China)

2014-10-31

Highlights: • This is the first report of DNA aptamer against EGFR in vitro. • Aptamer can bind targets with high affinity and selectivity. • DNA aptamers are more stable, cheap and efficient than RNA aptamers. • Our selected DNA aptamer against EGFR has high affinity with K{sub d} 56 ± 7.3 nM. • Our selected DNA aptamer against EGFR has high selectivity. - Abstract: Epidermal growth factor receptor (EGFR/HER1/c-ErbB1), is overexpressed in many solid cancers, such as epidermoid carcinomas, malignant gliomas, etc. EGFR plays roles in proliferation, invasion, angiogenesis and metastasis of malignant cancer cells and is the ideal antigen for clinical applications in cancer detection, imaging and therapy. Aptamers, the output of the systematic evolution of ligands by exponential enrichment (SELEX), are DNA/RNA oligonucleotides which can bind protein and other substances with specificity. RNA aptamers are undesirable due to their instability and high cost of production. Conversely, DNA aptamers have aroused researcher’s attention because they are easily synthesized, stable, selective, have high binding affinity and are cost-effective to produce. In this study, we have successfully identified DNA aptamers with high binding affinity and selectivity to EGFR. The aptamer named TuTu22 with K{sub d} 56 ± 7.3 nM was chosen from the identified DNA aptamers for further study. Flow cytometry analysis results indicated that the TuTu22 aptamer was able to specifically recognize a variety of cancer cells expressing EGFR but did not bind to the EGFR-negative cells. With all of the aforementioned advantages, the DNA aptamers reported here against cancer biomarker EGFR will facilitate the development of novel targeted cancer detection, imaging and therapy.

Selection of DNA aptamers against epidermal growth factor receptor with high affinity and specificity

International Nuclear Information System (INIS)

Wang, Deng-Liang; Song, Yan-Ling; Zhu, Zhi; Li, Xi-Lan; Zou, Yuan; Yang, Hai-Tao; Wang, Jiang-Jie; Yao, Pei-Sen; Pan, Ru-Jun; Yang, Chaoyong James; Kang, De-Zhi

2014-01-01

Highlights: • This is the first report of DNA aptamer against EGFR in vitro. • Aptamer can bind targets with high affinity and selectivity. • DNA aptamers are more stable, cheap and efficient than RNA aptamers. • Our selected DNA aptamer against EGFR has high affinity with K d 56 ± 7.3 nM. • Our selected DNA aptamer against EGFR has high selectivity. - Abstract: Epidermal growth factor receptor (EGFR/HER1/c-ErbB1), is overexpressed in many solid cancers, such as epidermoid carcinomas, malignant gliomas, etc. EGFR plays roles in proliferation, invasion, angiogenesis and metastasis of malignant cancer cells and is the ideal antigen for clinical applications in cancer detection, imaging and therapy. Aptamers, the output of the systematic evolution of ligands by exponential enrichment (SELEX), are DNA/RNA oligonucleotides which can bind protein and other substances with specificity. RNA aptamers are undesirable due to their instability and high cost of production. Conversely, DNA aptamers have aroused researcher’s attention because they are easily synthesized, stable, selective, have high binding affinity and are cost-effective to produce. In this study, we have successfully identified DNA aptamers with high binding affinity and selectivity to EGFR. The aptamer named TuTu22 with K d 56 ± 7.3 nM was chosen from the identified DNA aptamers for further study. Flow cytometry analysis results indicated that the TuTu22 aptamer was able to specifically recognize a variety of cancer cells expressing EGFR but did not bind to the EGFR-negative cells. With all of the aforementioned advantages, the DNA aptamers reported here against cancer biomarker EGFR will facilitate the development of novel targeted cancer detection, imaging and therapy

Two mutations which confer temperature-sensitive radiation sensitivity in the yeast Saccharomyces cerevisiae

International Nuclear Information System (INIS)

Ho, K.S.Y.; Mortimer, R.K.

1975-01-01

X-ray survival curves for two mutations, rad54 and rad55, in the yeast Saccharomyces cerevisiae are presented. These mutations confer temperature sensitive X-ray sensitivity; that is, rad54 and rad55 strains display a wild type X-ray survival response at permissive temperatures and a radiosensitive X-ray survival response at restrictive temperatures. The survival response of cells which were shifted from a permissive to a restrictive temperature or vice versa at various post-irradiation times indicates that repair and fixation of X-ray induced lesions is largely complete three hours after X-irradiation. Experiments to determine the utilization sequence of the rad54 and rad55 gene products in the repair of X-ray induced damage suggest that the two products are required in an interdependent manner

Dopamine modulates metabolic rate and temperature sensitivity in Drosophila melanogaster.

Directory of Open Access Journals (Sweden)

Taro Ueno

Full Text Available Homeothermal animals, such as mammals, maintain their body temperature by heat generation and heat dissipation, while poikilothermal animals, such as insects, accomplish it by relocating to an environment of their favored temperature. Catecholamines are known to regulate thermogenesis and metabolic rate in mammals, but their roles in other animals are poorly understood. The fruit fly, Drosophila melanogaster, has been used as a model system for the genetic studies of temperature preference behavior. Here, we demonstrate that metabolic rate and temperature sensitivity of some temperature sensitive behaviors are regulated by dopamine in Drosophila. Temperature-sensitive molecules like dTrpA1 and shi(ts induce temperature-dependent behavioral changes, and the temperature at which the changes are induced were lowered in the dopamine transporter-defective mutant, fumin. The mutant also displays a preference for lower temperatures. This thermophobic phenotype was rescued by the genetic recovery of the dopamine transporter in dopamine neurons. Flies fed with a dopamine biosynthesis inhibitor (3-iodo-L-tyrosine, which diminishes dopamine signaling, exhibited preference for a higher temperature. Furthermore, we found that the metabolic rate is up-regulated in the fumin mutant. Taken together, dopamine has functions in the temperature sensitivity of behavioral changes and metabolic rate regulation in Drosophila, as well as its previously reported functions in arousal/sleep regulation.

High density and ligand affinity confer ultrasensitive signal detection by a guanylyl cyclase chemoreceptor

Science.gov (United States)

Pichlo, Magdalena; Bungert-Plümke, Stefanie; Weyand, Ingo; Seifert, Reinhard; Bönigk, Wolfgang; Strünker, Timo; Kashikar, Nachiket Dilip; Goodwin, Normann; Müller, Astrid; Körschen, Heinz G.; Collienne, Ursel; Pelzer, Patric; Van, Qui; Enderlein, Jörg; Klemm, Clementine; Krause, Eberhard; Trötschel, Christian; Poetsch, Ansgar; Kremmer, Elisabeth

2014-01-01

Guanylyl cyclases (GCs), which synthesize the messenger cyclic guanosine 3′,5′-monophosphate, control several sensory functions, such as phototransduction, chemosensation, and thermosensation, in many species from worms to mammals. The GC chemoreceptor in sea urchin sperm can decode chemoattractant concentrations with single-molecule sensitivity. The molecular and cellular underpinnings of such ultrasensitivity are not known for any eukaryotic chemoreceptor. In this paper, we show that an exquisitely high density of 3 × 105 GC chemoreceptors and subnanomolar ligand affinity provide a high ligand-capture efficacy and render sperm perfect absorbers. The GC activity is terminated within 150 ms by dephosphorylation steps of the receptor, which provides a means for precise control of the GC lifetime and which reduces “molecule noise.” Compared with other ultrasensitive sensory systems, the 10-fold signal amplification by the GC receptor is surprisingly low. The hallmarks of this signaling mechanism provide a blueprint for chemical sensing in small compartments, such as olfactory cilia, insect antennae, or even synaptic boutons. PMID:25135936

Highly sensitive work function hydrogen gas sensor based on PdNPs/SiO2/Si structure at room temperature

Directory of Open Access Journals (Sweden)

G. Behzadi pour

Full Text Available In this study, fabrication of highly sensitive PdNPs/SiO2/Si hydrogen gas sensor using experimental and theoretical methods has been investigated. Using chemical method the PdNPs are synthesized and characterized by X-ray diffraction (XRD. The average size of PdNPs is 11 nm. The thickness of the oxide film was 20 nm and the surface of oxide film analyzed using Atomic-force microscopy (AFM. The C-V curve for the PdNPs/SiO2/Si hydrogen gas sensor in 1% hydrogen concentration and at the room temperature has been reported. The response time and recovery time for 1% hydrogen concentration at room temperature were 1.2 s and 10 s respectively. The response (R% for PdNPs/SiO2/Si MOS capacitor hydrogen sensor was 96%. The PdNPs/SiO2/Si MOS capacitor hydrogen sensor showed very fast response and recovery times compared to SWCNTs/PdNPs, graphene/PdNPs, nanorod/PdNPs and nanowire/PdNPs hydrogen gas sensors. Keywords: Sensitive, Oxide film, Capacitive, Resistance

Global sensitivity analysis of water age and temperature for informing salmonid disease management

Science.gov (United States)

Javaheri, Amir; Babbar-Sebens, Meghna; Alexander, Julie; Bartholomew, Jerri; Hallett, Sascha

2018-06-01

Many rivers in the Pacific Northwest region of North America are anthropogenically manipulated via dam operations, leading to system-wide impacts on hydrodynamic conditions and aquatic communities. Understanding how dam operations alter abiotic and biotic variables is important for designing management actions. For example, in the Klamath River, dam outflows could be manipulated to alter water age and temperature to reduce risk of parasite infections in salmon by diluting or altering viability of parasite spores. However, sensitivity of water age and temperature to the riverine conditions such as bathymetry can affect outcomes from dam operations. To examine this issue in detail, we conducted a global sensitivity analysis of water age and temperature to a comprehensive set of hydraulics and meteorological parameters in the Klamath River, California, where management of salmonid disease is a high priority. We applied an analysis technique, which combined Latin-hypercube and one-at-a-time sampling methods, and included simulation runs with the hydrodynamic numerical model of the Lower Klamath. We found that flow rate and bottom roughness were the two most important parameters that influence water age. Water temperature was more sensitive to inflow temperature, air temperature, solar radiation, wind speed, flow rate, and wet bulb temperature respectively. Our results are relevant for managers because they provide a framework for predicting how water within 'high infection risk' sections of the river will respond to dam water (low infection risk) input. Moreover, these data will be useful for prioritizing the use of water age (dilution) versus temperature (spore viability) under certain contexts when considering flow manipulation as a method to reduce risk of infection and disease in Klamath River salmon.

Two high-affinity ligand binding states of uterine estrogen receptor distinguished by modulation of hydrophobic environment

International Nuclear Information System (INIS)

Hutchens, T.W.; Li, C.M.; Zamah, N.M.; Besch, P.K.

1987-01-01

The steroid binding function of soluble (cytosolic) estrogen receptors from calf uteri was evaluated under conditions known to modify the extent of hydrophobic interaction with receptor-associated proteins. Receptor preparations were equilibrated into 6 M urea buffers and control buffers by chromatography through small columns of Sephadex G-25 or by dialysis at 0.6 0 C. Equilibrium dissociation constants (K/sub d/) and binding capacities (n) of experimental and control receptor preparations were determined by 13-point Scatchard analyses using concentrations of 17β-[ 3 H]estradiol from 0.05 to 10 nM. Nonspecific binding was determined at each concentration by parallel incubations with a 200-fold molar excess of the receptor-specific competitor diethylstilbestrol. The control receptor population was consistently found to be a single class of binding sites with a high affinity for estradiol which was unaffected by G-25 chromatography, by dialysis, by dilution, or by the presence of 0.4 M KCl. However, equilibration into 6 M urea induced a discrete (10-fold) reduction in receptor affinity to reveal a second, thermodynamically stable, high-affinity binding state. The presence of 0.4 M KCl did not significantly influence the discrete change in receptor affinity induced by urea. The effects of urea on both receptor affinity and binding capacity were reversible, suggesting a lack of covalent modification. These results demonstrate nonenzymatic means by which not only the binding capacity but also the affinity of receptor for estradiol can be reversibly controlled, suggesting that high concentrations of urea might be more effectively utilized during the physicochemical characterization and purification of steroid receptor proteins

Importance of Hypervariable Region 2 for Stability and Affinity of a Shark Single-Domain Antibody Specific for Ebola Virus Nucleoprotein.

Directory of Open Access Journals (Sweden)

George P Anderson

Full Text Available Single-domain antibodies derived from the unique New Antigen Receptor found in sharks have numerous potential applications, ranging from diagnostic reagents to therapeutics. Shark-derived single-domain antibodies possess the same characteristic ability to refold after heat denaturation found in single-domain antibodies derived from camelid heavy-chain-only antibodies. Recently, two shark derived single-domain antibodies specific for the nucleoprotein of Ebola virus were described. Our evaluation confirmed their high affinity for the nucleoprotein, but found their melting temperatures to be low relative to most single-domain antibodies. Our first approach towards improving their stability was grafting antigen-binding regions (complementarity determining regions of one of these single-domain antibodies onto a high melting temperature shark single-domain antibody. This resulted in two variants: one that displayed excellent affinity with a low melting temperature, while the other had poor affinity but a higher melting temperature. These new proteins, however, differed in only 3 amino acids within the complementarity determining region 2 sequence. In shark single-domain antibodies, the complementarity determining region 2 is often referred to as hypervariable region 2, as this segment of the antibody domain is truncated compared to the sequence in camelid single-domain antibodies and conventional heavy chain variable domains. To elucidate which of the three amino acids or combinations thereof were responsible for the affinity and stability we made the 6 double and single point mutants that covered the intermediates between these two clones. We found a single amino acid change that achieved a 10°C higher melting temperature while maintaining sub nM affinity. This research gives insights into the impact of the shark sdAb hypervariable 2 region on both stability and affinity.

Affinity purification of the voltage-sensitive sodium channel from electroplax with resins selective for sialic acid

Energy Technology Data Exchange (ETDEWEB)

James, W.M.; Emerick, M.C.; Agnew, W.S. (Yale Univ. School of medicine, New Haven, CT (USA))

1989-07-11

The voltage-sensitive sodium channel present in the eel (Electrophorus electricus) has an unusually high content of sialic acid, including {alpha}-(2{yields}8)-linked polysialic acid, not found in other electroplax membrane glycopeptides. Lectins from Limax flavus (LFA) and wheat germ (WGA) proved the most effective of 11 lectin resins tried. The most selective resin was prepared from IgM antibodies against Neisseria meningitidis {alpha}-(2{yields}8)-polysialic acid which were affinity purified and coupled to Sepharose 4B. The sodium channel was found to bind to WGA, LFA, and IgM resins and was readily eluted with the appropriate soluble carbohydrates. Experiments with LFA and IgM resins demonstrated binding and unbinding rates and displacement kinetics, which suggest highly specific binding at multiple sites on the sodium channel protein. In preparative-scale purification of protein previously fractionated by anion-exchange chromatography, without stabilizing TTX, high yields were reproducibly obtained. Further, when detergent extracts were prepared from electroplax membranes fractionated by low-speed sedimentation, a single step over the IgM resin provided a 70-fold purification, yielding specific activities of 3,200 pmol of ({sup 3}H)TTX-binding sites/mg of protein and a single polypeptide of {approximately}285,000 Da on SDS-acrylamide gels. No small peptides were observed after this 5-h isolation. The authors describe a cation-dependent stabilization with millimolar levels of monovalent and micromolar levels of divalent species.

The electron affinity of some radiotherapeutic agents used in cancer therapy

International Nuclear Information System (INIS)

Wold, E.; Kaalhus, O.; Johansen, E.S.; Ekse, A.T.

1980-01-01

In order to evaluate whether chemotherapeutic compounds applied in cancer treatment might interact with radiation as anoxic cell sensitizers, the electron-affinic properties of DTIC (5-(3,3-dimethyl-1-triazeno)imidazole-4 carboxamide) AIC 4(5)-aminoimidazole-5(4)-carboxamide, hydroxyurea, busulfan and cyclophosphamide were studied by pulse radiolysis. Reaction rates with hydrated electrons were determined for all these compounds. With the exception of DTIC, they all reacted much more slowly with electrons than do most electron-affinic sensitizers. One-electron reduction potentials were determined for DTIC, AIC and hydroxyurea. The values were all in the region for the onset of sensitization, with hydroxyurea as the most promising (E 7 1 = -0.552 V). For busulfan and cyclophosphamide no value could be determined, but these compounds are probably less electron-affinic than hydroxyurea. A possible application of chemotherapeutic agents as radiosensitizers is discussed. (author)

NK1 receptor fused to beta-arrestin displays a single-component, high-affinity molecular phenotype.

Science.gov (United States)

Martini, Lene; Hastrup, Hanne; Holst, Birgitte; Fraile-Ramos, Alberto; Marsh, Mark; Schwartz, Thue W

2002-07-01

Arrestins are cytosolic proteins that, upon stimulation of seven transmembrane (7TM) receptors, terminate signaling by binding to the receptor, displacing the G protein and targeting the receptor to clathrin-coated pits. Fusion of beta-arrestin1 to the C-terminal end of the neurokinin NK1 receptor resulted in a chimeric protein that was expressed to some extent on the cell surface but also accumulated in transferrin-labeled recycling endosomes independently of agonist stimulation. As expected, the fusion protein was almost totally silenced with respect to agonist-induced signaling through the normal Gq/G11 and Gs pathways. The NK1-beta-arrestin1 fusion construct bound nonpeptide antagonists with increased affinity but surprisingly also bound two types of agonists, substance P and neurokinin A, with high, normal affinity. In the wild-type NK1 receptor, neurokinin A (NKA) competes for binding against substance P and especially against antagonists with up to 1000-fold lower apparent affinity than determined in functional assays and in homologous binding assays. When the NK1 receptor was closely fused to G proteins, this phenomenon was eliminated among agonists, but the agonists still competed with low affinity against antagonists. In contrast, in the NK1-beta-arrestin1 fusion protein, all ligands bound with similar affinity independent of the choice of radioligand and with Hill coefficients near unity. We conclude that the NK1 receptor in complex with arrestin is in a high-affinity, stable, agonist-binding form probably best suited to structural analysis and that the receptor can display binding properties that are nearly theoretically ideal when it is forced to complex with only a single intracellular protein partner.

The high-affinity peptidoglycan binding domain of Pseudomonas phage endolysin KZ144

Energy Technology Data Exchange (ETDEWEB)

Briers, Yves [Division of Gene Technology, Department of Biosystems, Katholieke Universiteit Leuven, Kasteelpark Arenberg 21, B-3001 Leuven (Belgium); Schmelcher, Mathias; Loessner, Martin J. [Institute of Food Science and Nutrition, ETH Zuerich, Schmelzbergstrasse 7, CH-8092 Zuerich (Switzerland); Hendrix, Jelle; Engelborghs, Yves [Laboratory of Biomolecular Dynamics, Department of Chemistry, Katholieke Universiteit Leuven, Celestijnenlaan 200G, B-3001 Leuven (Belgium); Volckaert, Guido [Division of Gene Technology, Department of Biosystems, Katholieke Universiteit Leuven, Kasteelpark Arenberg 21, B-3001 Leuven (Belgium); Lavigne, Rob, E-mail: rob.lavigne@biw.kuleuven.be [Division of Gene Technology, Department of Biosystems, Katholieke Universiteit Leuven, Kasteelpark Arenberg 21, B-3001 Leuven (Belgium)

2009-05-29

The binding affinity of the N-terminal peptidoglycan binding domain of endolysin KZ144 (PBD{sub KZ}), originating from Pseudomonas aeruginosa bacteriophage {phi}KZ, has been examined using a fusion protein of PBD{sub KZ} and green fluorescent protein (PBD{sub KZ}-GFP). A fluorescence recovery after photobleaching analysis of bound PBD{sub KZ}-GFP molecules showed less than 10% fluorescence recovery in the bleached area within 15 min. Surface plasmon resonance analysis confirmed this apparent high binding affinity revealing an equilibrium affinity constant of 2.95 x 10{sup 7} M{sup -1} for the PBD{sub KZ}-peptidoglycan interaction. This unique domain, which binds to the peptidoglycan of all tested Gram-negative species, was harnessed to improve the specific activity of the peptidoglycan hydrolase domain KMV36C. The chimeric peptidoglycan hydrolase (PBD{sub KZ}-KMV36C) exhibits a threefold higher specific activity than the native catalytic domain (KMV36C). These results demonstrate that the modular assembly of functional domains is a rational approach to improve the specific activity of endolysins from phages infecting Gram-negatives.

A high affinity Ca2(+)-ATPase on the surface membrane of Leishmania donovani promastigote

International Nuclear Information System (INIS)

Ghosh, J.; Ray, M.; Sarkar, S.; Bhaduri, A.

1990-01-01

A Ca2(+)-dependent ATP-hydrolytic activity was detected in the crude membrane ghost of the promastigote or vector form of the protozoal parasite Leishmania donovani, the pathogen responsible for kala azar. The Ca2(+)-ATPase was purified to apparent homogeneity after solubilization with deoxycholate. The enzyme consists of two subunits of Mr = 51,000 and 57,000 and has an apparent molecular weight of 215,000 +/- 12,000. The enzyme activity is exclusively dependent on Ca2+, and the pure enzyme can hydrolyze 1.6 mumol of ATP/min/mg of protein. The apparent Km for Ca2+ is 35 nM, which is further reduced to 12 nM in the presence of heterologous calmodulin. The enzyme is sensitive to vanadate, but is insensitive to oligomycin and ouabain. The enzyme is strongly associated with the plasma membrane and has its catalytic site oriented toward the cytoplasmic face. The enzyme spans across the plasma membrane as surface labeling with radioiodine shows considerable radioactivity in the completely purified enzyme. The localization and orientation of this high affinity, calmodulin-sensitive Ca2(+)-ATPase suggest some role of this enzyme in Ca2+ movement in the life cycle of this protozoal parasite

A high affinity Ca2(+)-ATPase on the surface membrane of Leishmania donovani promastigote

Energy Technology Data Exchange (ETDEWEB)

Ghosh, J.; Ray, M.; Sarkar, S.; Bhaduri, A. (Indian Institute of Chemical Biology, Calcutta (India))

1990-07-05

A Ca2(+)-dependent ATP-hydrolytic activity was detected in the crude membrane ghost of the promastigote or vector form of the protozoal parasite Leishmania donovani, the pathogen responsible for kala azar. The Ca2(+)-ATPase was purified to apparent homogeneity after solubilization with deoxycholate. The enzyme consists of two subunits of Mr = 51,000 and 57,000 and has an apparent molecular weight of 215,000 +/- 12,000. The enzyme activity is exclusively dependent on Ca2+, and the pure enzyme can hydrolyze 1.6 mumol of ATP/min/mg of protein. The apparent Km for Ca2+ is 35 nM, which is further reduced to 12 nM in the presence of heterologous calmodulin. The enzyme is sensitive to vanadate, but is insensitive to oligomycin and ouabain. The enzyme is strongly associated with the plasma membrane and has its catalytic site oriented toward the cytoplasmic face. The enzyme spans across the plasma membrane as surface labeling with radioiodine shows considerable radioactivity in the completely purified enzyme. The localization and orientation of this high affinity, calmodulin-sensitive Ca2(+)-ATPase suggest some role of this enzyme in Ca2+ movement in the life cycle of this protozoal parasite.

Ultrasensitive direct competitive FLISA using highly luminescent quantum dot beads for tuning affinity of competing antigens to antibodies

Energy Technology Data Exchange (ETDEWEB)

Xiong, Sicheng; Zhou, Yaofeng; Huang, Xiaolin [State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047 (China); Yu, Ruijin [College of Science, Northwest A& F University, Yangling, Shaanxi 712100 (China); Lai, Weihua [State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047 (China); Xiong, Yonghua, E-mail: yhxiongchen@163.com [State Key Laboratory of Food Science and Technology, Nanchang University, Nanchang 330047 (China)

2017-06-15

Herein, for the first time we report a novel direct competitive fluorescence-linked immunosorbent assay (dcFLISA) for the ultrasensitive detection of ochratoxin A (OTA) by introducing a large size polymer beads loaded with quantum dots (QBs) as carrier of competing antigen for decreasing binding affinity to antibody and enhancing the fluorescent signal intensity. When using 255 nm QBs as carrier of competing antigen, the equilibrium dissociation constant of QB based competing antigen to antibodies can be tuned to 100 times higher than that of the horseradish peroxidase (HRP) based competing antigen by controlling labeled amounts of antigen on the surface of QBs. Various parameters that influenced the sensitivity of dcFLISA were investigated and optimized. Under optimum detection parameters, the dynamic linear range of developed dcFLISA for detecting OTA was established at 0.05 pg/mL to 1.56 pg/mL with a half maximal inhibitory concentration at 0.14 ± 0.04 pg/mL (n = 5), which is three orders of magnitude lower than that of conventional HRP-based dcELISA (0.24 ng/mL). The developed FLISA is also highly accurate, reliable, and shows no cross reaction to other mycotoxins. In summary, the proposed method offers a straightforward approach to improve the sensitivity of direct competitive immunoassay for trace small chemical molecule detection in food quality control, environmental monitoring, and clinical diagnosis. - Highlights: • Highly luminescent QBs were used as a carrier of competing antigen for ultrasensitive detection of OTA. • It is the first time to use a large size QBs as a carrier for tuning affinity of competing antigen to antibodies. • IC{sub 50} value of QB-based dcFLISA is three orders of magnitude lower than that of HRP-based dcELISA.

Ultrasensitive direct competitive FLISA using highly luminescent quantum dot beads for tuning affinity of competing antigens to antibodies

International Nuclear Information System (INIS)

Xiong, Sicheng; Zhou, Yaofeng; Huang, Xiaolin; Yu, Ruijin; Lai, Weihua; Xiong, Yonghua

2017-01-01

Herein, for the first time we report a novel direct competitive fluorescence-linked immunosorbent assay (dcFLISA) for the ultrasensitive detection of ochratoxin A (OTA) by introducing a large size polymer beads loaded with quantum dots (QBs) as carrier of competing antigen for decreasing binding affinity to antibody and enhancing the fluorescent signal intensity. When using 255 nm QBs as carrier of competing antigen, the equilibrium dissociation constant of QB based competing antigen to antibodies can be tuned to 100 times higher than that of the horseradish peroxidase (HRP) based competing antigen by controlling labeled amounts of antigen on the surface of QBs. Various parameters that influenced the sensitivity of dcFLISA were investigated and optimized. Under optimum detection parameters, the dynamic linear range of developed dcFLISA for detecting OTA was established at 0.05 pg/mL to 1.56 pg/mL with a half maximal inhibitory concentration at 0.14 ± 0.04 pg/mL (n = 5), which is three orders of magnitude lower than that of conventional HRP-based dcELISA (0.24 ng/mL). The developed FLISA is also highly accurate, reliable, and shows no cross reaction to other mycotoxins. In summary, the proposed method offers a straightforward approach to improve the sensitivity of direct competitive immunoassay for trace small chemical molecule detection in food quality control, environmental monitoring, and clinical diagnosis. - Highlights: • Highly luminescent QBs were used as a carrier of competing antigen for ultrasensitive detection of OTA. • It is the first time to use a large size QBs as a carrier for tuning affinity of competing antigen to antibodies. • IC_5_0 value of QB-based dcFLISA is three orders of magnitude lower than that of HRP-based dcELISA.

Determination of trace alkaline phosphatase by affinity adsorption solid substrate room temperature phosphorimetry based on wheat germ agglutinin labeled with 8-quinolineboronic acid phosphorescent molecular switch and prediction of diseases

Science.gov (United States)

Liu, Jia-Ming; Gao, Hui; Li, Fei-Ming; Shi, Xiu-Mei; Lin, Chang-Qing; Lin, Li-Ping; Wang, Xin-Xing; Li, Zhi-Ming

2010-09-01

The 8-quinolineboronic acid phosphorescent molecular switch (abbreviated as PMS-8-QBA. Thereinto, 8-QBA is 8-quinolineboronic acid, and PMS is phosphorescent molecular switch) was found for the first time. PMS-8-QBA, which was in the "off" state, could only emit weak room temperature phosphorescence (RTP) on the acetyl cellulose membrane (ACM). However, PMS-8-QBA turned "on" automatically for its changed structure, causing that the RTP of 8-QBA in the system increased, after PMS-8-QBA-WGA (WGA is wheat germ agglutinin) was formed by reaction between -OH of PMS-8-QBA and -COOH of WGA. More interesting is that the -NH 2 of PMS-8-QBA-WGA could react with the -COOH of alkaline phosphatase (AP) to form the affinity adsorption (AA) product WGA-AP-WGA-8-QBA-PMS (containing -NH-CO- bond), which caused RTP of the system to greatly increase. Thus, affinity adsorption solid substrate room temperature phosphorimetry using PMS-8-QBA as labelling reagent (PMS-8-QBA-AA-SSRTP) for the determination of trace AP was established. The method had many advantages, such as high sensitivity (the detection limit (LD) was 2.5 zg spot -1. For sample volume of 0.40 μl spot -1, corresponding concentration was 6.2 × 10 -18 g ml -1), good selectivity (the allowed concentration of coexisting material was higher, when the relative error was ±5%), high accuracy (applied to detection of AP content in serum samples, the result was coincided with those obtained by enzyme-linked immunoassay), which was suitable for the detection of trace AP content in serum samples and the forecast of human diseases. Meanwhile, the mechanism of PMS-8-QBA-AASSRTP was discussed. The new field of analytical application and clinic diagnosis technique of molecule switch are exploited, based on the phosphorescence characteristic of PMS-8-QBA, the AA reaction between WGA and AP, as well as the relation between AP content and human diseases. The research results promote the development and interpenetrate among molecule

Methylation-Sensitive High Resolution Melting (MS-HRM).

Science.gov (United States)

Hussmann, Dianna; Hansen, Lise Lotte

2018-01-01

Methylation-Sensitive High Resolution Melting (MS-HRM) is an in-tube, PCR-based method to detect methylation levels at specific loci of interest. A unique primer design facilitates a high sensitivity of the assays enabling detection of down to 0.1-1% methylated alleles in an unmethylated background.Primers for MS-HRM assays are designed to be complementary to the methylated allele, and a specific annealing temperature enables these primers to anneal both to the methylated and the unmethylated alleles thereby increasing the sensitivity of the assays. Bisulfite treatment of the DNA prior to performing MS-HRM ensures a different base composition between methylated and unmethylated DNA, which is used to separate the resulting amplicons by high resolution melting.The high sensitivity of MS-HRM has proven useful for detecting cancer biomarkers in a noninvasive manner in urine from bladder cancer patients, in stool from colorectal cancer patients, and in buccal mucosa from breast cancer patients. MS-HRM is a fast method to diagnose imprinted diseases and to clinically validate results from whole-epigenome studies. The ability to detect few copies of methylated DNA makes MS-HRM a key player in the quest for establishing links between environmental exposure, epigenetic changes, and disease.

Affinity functions for modeling glass dissolution rates

Energy Technology Data Exchange (ETDEWEB)

Bourcier, W.L. [Lawrence Livermore National Lab., CA (United States)

1997-07-01

Glass dissolution rates decrease dramatically as glass approach ''saturation'' with respect to the leachate solution. Most repository sites are chosen where water fluxes are minimal, and therefore the waste glass is most likely to dissolve under conditions close to ''saturation''. The key term in the rate expression used to predict glass dissolution rates close to ''saturation'' is the affinity term, which accounts for saturation effects on dissolution rates. Interpretations of recent experimental data on the dissolution behaviour of silicate glasses and silicate minerals indicate the following: 1) simple affinity control does not explain the observed dissolution rate for silicate minerals or glasses; 2) dissolution rates can be significantly modified by dissolved cations even under conditions far from saturation where the affinity term is near unity; 3) the effects of dissolved species such as Al and Si on the dissolution rate vary with pH, temperature, and saturation state; and 4) as temperature is increased, the effect of both pH and temperature on glass and mineral dissolution rates decrease, which strongly suggests a switch in rate control from surface reaction-based to diffusion control. Borosilicate glass dissolution models need to be upgraded to account for these recent experimental observations. (A.C.)

Dynamic High-Temperature Characterization of an Iridium Alloy in Compression at High Strain Rates

Energy Technology Data Exchange (ETDEWEB)

Song, Bo [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States). Experimental Environment Simulation Dept.; Nelson, Kevin [Sandia National Lab. (SNL-CA), Livermore, CA (United States). Mechanics of Materials Dept.; Lipinski, Ronald J. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States). Advanced Nuclear Fuel Cycle Technology Dept.; Bignell, John L. [Sandia National Lab. (SNL-NM), Albuquerque, NM (United States). Structural and Thermal Analysis Dept.; Ulrich, G. B. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Radioisotope Power Systems Program; George, E. P. [Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Radioisotope Power Systems Program

2014-06-01

Iridium alloys have superior strength and ductility at elevated temperatures, making them useful as structural materials for certain high-temperature applications. However, experimental data on their high-temperature high-strain-rate performance are needed for understanding high-speed impacts in severe elevated-temperature environments. Kolsky bars (also called split Hopkinson bars) have been extensively employed for high-strain-rate characterization of materials at room temperature, but it has been challenging to adapt them for the measurement of dynamic properties at high temperatures. Current high-temperature Kolsky compression bar techniques are not capable of obtaining satisfactory high-temperature high-strain-rate stress-strain response of thin iridium specimens investigated in this study. We analyzed the difficulties encountered in high-temperature Kolsky compression bar testing of thin iridium alloy specimens. Appropriate modifications were made to the current high-temperature Kolsky compression bar technique to obtain reliable compressive stress-strain response of an iridium alloy at high strain rates (300 – 10000 s^-1) and temperatures (750°C and 1030°C). Uncertainties in such high-temperature high-strain-rate experiments on thin iridium specimens were also analyzed. The compressive stress-strain response of the iridium alloy showed significant sensitivity to strain rate and temperature.

Blood O2 affinity of a large polar elasmobranch, the Greenland shark Somniosus microcephalus

DEFF Research Database (Denmark)

Herbert, N.A.; Skov, Peter Vilhelm; Tirsgaard, B.

2017-01-01

The Greenland shark (Somniosus microcephalus. Bloch & Schneider 1801) is a polar elasmobranch that is hypothesised to possess a unique metabolic physiology due to its extreme large size, the cold waters it inhabits and its slow swimming lifestyle. Our results therefore provide the first insight...... into the metabolic physiology of this unique shark, with a focus on blood O2 affinity. An evaluation of blood O2 affinity at 2 °C using tonometry revealed a P50 of 11.7 mmHg at a PCO2 of 2.25 mmHg and a Bohr effect (binding sensitivity of blood to pH, ϕ = Δlog P50/ΔpH) of −0.26. A comparative evaluation of blood O2...... affinity across elasmobranch fishes suggests that S. microcephalus has a high blood O2 affinity (i.e., low P50) and a small Bohr effect but these are common traits in sluggish elasmobranch fishes, with little evidence for any relationship of blood O2 affinity to the low metabolic rates, low environmental...

Influence of temperature on the formation and encapsulation of gold nanoparticles using a temperature-sensitive template

Directory of Open Access Journals (Sweden)

Noel Peter Bengzon Tan

2015-12-01

Full Text Available This data article describes the synthesis of temperature-sensitive and amine-rich microgel particle as a dual reductant and template to generate smart gold/polymer nanocomposite particle. TEM images illustrate the influence of reaction temperature on the formation and in-site encapsulation of gold nanoparticles using the temperature-sensitive microgel template. Thermal stability of the resultant gold/polymer composite particles was also examined.

Isolation of Anti-Ricin Protective Antibodies Exhibiting High Affinity from Immunized Non-Human Primates

Directory of Open Access Journals (Sweden)

Tal Noy-Porat

2016-03-01

Full Text Available Ricin, derived from the castor bean plant Ricinus communis, is one of the most potent and lethal toxins known, against which there is no available antidote. To date, the use of neutralizing antibodies is the most promising post-exposure treatment for ricin intoxication. The aim of this study was to isolate high affinity anti-ricin antibodies that possess potent toxin-neutralization capabilities. Two non-human primates were immunized with either a ricin-holotoxin- or subunit-based vaccine, to ensure the elicitation of diverse high affinity antibodies. By using a comprehensive set of primers, immune scFv phage-displayed libraries were constructed and panned. A panel of 10 antibodies (five directed against the A subunit of ricin and five against the B subunit was isolated and reformatted into a full-length chimeric IgG. All of these antibodies were found to neutralize ricin in vitro, and several conferred full protection to ricin-intoxicated mice when given six hours after exposure. Six antibodies were found to possess exceptionally high affinity toward the toxin, with KD values below pM (koff < 1 × 10−7 s−1 that were well correlated with their ability to neutralize ricin. These antibodies, alone or in combination, could be used for the development of a highly-effective therapeutic preparation for post-exposure treatment of ricin intoxication.

Mutational analysis of affinity and selectivity of kringle-tetranectin interaction. Grafting novel kringle affinity ontp the trtranectin lectin scaffold

DEFF Research Database (Denmark)

Graversen, Jonas Heilskov; Jacobsen, C; Sigurskjold, B W

2000-01-01

-type lectin-like domain of tetranectin, involving Lys-148, Glu-150, and Asp-165, which mediates calcium-sensitive binding to plasminogen kringle 4. Here, we investigate the effect of conservative substitutions of these and a neighboring amino acid residue. Substitution of Thr-149 in tetranectin...... with a tyrosine residue considerably increases the affinity for plasminogen kringle 4, and, in addition, confers affinity for plasminogen kringle 2. As shown by isothermal titration calorimetry analysis, this new interaction is stronger than the binding of wild-type tetranectin to plasminogen kringle 4...

Electrochemical affinity biosensors for detection of mycotoxins: A review.

Science.gov (United States)

Vidal, Juan C; Bonel, Laura; Ezquerra, Alba; Hernández, Susana; Bertolín, Juan R; Cubel, Carlota; Castillo, Juan R

2013-11-15

This review discusses the current state of electrochemical biosensors in the determination of mycotoxins in foods. Mycotoxins are highly toxic secondary metabolites produced by molds. The acute toxicity of these results in serious human and animal health problems, although it has been only since early 1960s when the first studied aflatoxins were found to be carcinogenic. Mycotoxins affect a broad range of agricultural products, most important cereals and cereal-based foods. A majority of countries, mentioning especially the European Union, have established preventive programs to control contamination and strict laws of the permitted levels in foods. Official methods of analysis of mycotoxins normally requires sophisticated instrumentation, e.g. liquid chromatography with fluorescence or mass detectors, combined with extraction procedures for sample preparation. For about sixteen years, the use of simpler and faster analytical procedures based on affinity biosensors has emerged in scientific literature as a very promising alternative, particularly electrochemical (i.e., amperometric, impedance, potentiometric or conductimetric) affinity biosensors due to their simplicity and sensitivity. Typically, electrochemical biosensors for mycotoxins use specific antibodies or aptamers as affinity ligands, although recombinant antibodies, artificial receptors and molecular imprinted polymers show potential utility. This article deals with recent advances in electrochemical affinity biosensors for mycotoxins and covers complete literature from the first reports about sixteen years ago. Copyright © 2013 Elsevier B.V. All rights reserved.

Greater temperature sensitivity of plant phenology at colder sites

DEFF Research Database (Denmark)

Prevey, Janet; Vellend, Mark; Ruger, Nadja

2017-01-01

Warmer temperatures are accelerating the phenology of organisms around the world. Temperature sensitivity of phenology might be greater in colder, higher latitude sites than in warmer regions, in part because small changes in temperature constitute greater relative changes in thermal balance...

Paper-based immune-affinity arrays for detection of multiple mycotoxins in cereals.

Science.gov (United States)

Li, Li; Chen, Hongpu; Lv, Xiaolan; Wang, Min; Jiang, Xizhi; Jiang, Yifei; Wang, Heye; Zhao, Yongfu; Xia, Liru

2018-03-01

Mycotoxins produced by different species of fungi may coexist in cereals and feedstuffs, and could be highly toxic for humans and animals. For quantification of multiple mycotoxins in cereals, we developed a paper-based mycotoxin immune-affinity array. First, paper-based microzone arrays were fabricated by photolithography. Then, monoclonal mycotoxin antibodies were added in a copolymerization reaction with a cross-linker to form an immune-affinity monolith on the paper-based microzone array. With use of a competitive immune-response format, paper-based mycotoxin immune-affinity arrays were successfully applied to detect mycotoxins in samples. The detection limits for deoxynivalenol, zearalenone, T-2 toxin, and HT-2 toxin were 62.7, 10.8, 0.36, and 0.23 μg·kg -1 , respectively, which meet relevant requirements for these compounds in food. The recovery rates were 81-86% for deoxynivalenol, 89-117% for zearalenone, 79-86% for T-2 toxin, and 78-83% for HT-2 toxin, and showed the paper-based immune-affinity arrays had good reproducibility. In summary, the paper-based mycotoxin immune-affinity array provides a sensitive, rapid, accurate, stable, and convenient platform for detection of multiple mycotoxins in agro-foods. Graphical abstract Paper-based immune-affinity monolithic array. DON deoxynivalenol, HT-2 HT-2 toxin, T-2 T-2 toxin, PEGDA polyethylene glycol diacrylate, ZEN zearalenone.

Effect of Propellant Composition to the Temperature Sensitivity of Composite Propellant

International Nuclear Information System (INIS)

Aziz, Amir; Mamat, Rizalman; Amin, Makeen; Wan Ali, Wan Khairuddin

2012-01-01

The propellant composition is one of several parameter that influencing the temperature sensitivity of composite propellant. In this paper, experimental investigation of temperature sensitivity in burning rate of composite propellant was conducted. Four sets of different propellant compositions had been prepared with the combination of ammonium perchlorate (AP) as an oxidizer, aluminum (Al) as fuel and hydroxy-terminated polybutadiene (HTPB) as fuel and binder. For each mixture, HTPB binder was fixed at 15% and cured with isophorone diisocyanate (IPDI). By varying AP and Al, the effect of oxidizer- fuel mixture ratio (O/F) on the whole propellant can be determined. The propellant strands were manufactured using compression molded method and burnt in a strand burner using wire technique over a range of pressure from 1 atm to 31 atm. The results obtained shows that the temperature sensitivity, a, increases with increasing O/F. Propellant p80 which has O/F ratio of 80/20 gives the highest value of temperature sensitivity which is 1.687. The results shows that the propellant composition has significant effect on the temperature sensitivity of composite propellant

SENSITIVITY TEMPERATURE DEPENDENCE RESEARCH OF TV-CAMERAS BASED ON SILICON MATRIXES

Directory of Open Access Journals (Sweden)

Alexey N. Starchenko

2017-07-01

Full Text Available Subject of Research. The research is dedicated to the analysis of sensitivity change patterns of the cameras based on silicon CMOS-matrixes in various ambient temperatures. This information is necessary for the correct camera application for photometric measurements in-situ. The paper deals with studies of sensitivity variations of two digital cameras with different silicon CMOS matrixes in visible and near IR regions of the spectrum at temperature change. Method. Due to practical restrictions the temperature changes were recorded in separate spectral intervals important for practical use of the cameras. The experiments were carried out with the use of a climatic chamber, providing change and keeping the temperature range from minus 40 to plus 50 °C at a pitch of 10 о С. Two cameras were chosen for research: VAC-135-IP with OmniVision OV9121 matrix and VAC-248-IP with OnSemiconductor VITA2000 matrix. The two tested devices were placed in a climatic chamber at the same time and illuminated by one radiation source with a color temperature about 3000 K in order to eliminate a number of methodological errors. Main Results. The temperature dependence of the signals was shown to be linear and the matrixes sensitivities were determined. The results obtained are consistent with theoretical views, in general. The coefficients of thermal sensitivity were computed by these dependencies. It is shown that the greatest affect of temperature on the sensitivity occurs in the area (0.7–1.1 mkm. Temperature coefficients of sensitivity increase with the downward radiation wavelength increase. The experiments carried out have shown that it is necessary to take into account the changes in temperature sensitivity of silicon matrixes in the red and near in IR regions of the spectrum. The effect reveals itself in a clearly negative way in cameras with an amplitude resolution of 10-12 bits used for aerospace and space spectrozonal photography. Practical Relevance

Temperature sensitivity of extracellular enzyme kinetics in subtropical wetland soils under different nutrient and water level conditions

Science.gov (United States)

Goswami, S.; Inglett, K.; Inglett, P.

2012-12-01

Microbial extracellular enzymes play an important role in the initial steps of soil organic matter decomposition and are involved in regulating nutrient cycle processes. Moreover, with the recent concern of climate change, microbial extracellular enzymes may affect the functioning (C losses, C sequestration, greenhouse gas emissions, vegetation changes) of different ecosystems. Hence, it is imperative to understand the biogeochemical processes that may be climate change sensitive. Here, we have measured the Michaelis Menten Kinetics [maximal rate of velocity (Vmax) and half-saturation constant (Km)] of 6 enzymes involved in soil organic matter decomposition (phosphatase, phosphodiesterase, β-D-glucosidase, cellobiohydrolase, leucine aminopeptidase, N-Acetyl-β-D glucosaminidase) in different nutrient(P) concentration both aerobically and anaerobically in Everglade water conservation area 2A (F1, F4-slough and U3-slough). Temperature sensitivity of different enzymes is assessed within soil of different P concentrations. We hypothesized that the temperature sensitivity of the enzyme changes with the biogeochemical conditions including water level and nutrient condition. Furthermore, we have tested specific hypothesis that higher P concentration will initiate more C demand for microbes leading to higher Vmax value for carbon processing enzymes in high P site. We found temperature sensitivity of all enzymes for Vmax and Km under both aerobic and anaerobic condition ranges from 0.6 to 3.2 for Vmax and 0.5 to 2.5 for Km. Q10 values of Km for glucosidase indicate more temperature sensitivity under anaerobic condition. Under aerobic condition higher temperature showed significant effect on Vmax for bisphosphatase between high P and low P site. Decreasing P concentration from F1 site to U3-S site had showed significant effect in all temperature on carbon processing enzyme. This suggests that in high P site, microbes will use more carbon-processing enzyme to get more carbon

Thermal-depth matching in dynamic scene based on affine projection and feature registration

Science.gov (United States)

Wang, Hongyu; Jia, Tong; Wu, Chengdong; Li, Yongqiang

2018-03-01

This paper aims to study the construction of 3D temperature distribution reconstruction system based on depth and thermal infrared information. Initially, a traditional calibration method cannot be directly used, because the depth and thermal infrared camera is not sensitive to the color calibration board. Therefore, this paper aims to design a depth and thermal infrared camera calibration board to complete the calibration of the depth and thermal infrared camera. Meanwhile a local feature descriptors in thermal and depth images is proposed. The belief propagation matching algorithm is also investigated based on the space affine transformation matching and local feature matching. The 3D temperature distribution model is built based on the matching of 3D point cloud and 2D thermal infrared information. Experimental results show that the method can accurately construct the 3D temperature distribution model, and has strong robustness.

A high affinity monoclonal antibody recognizing the light chain of human coagulating factor VII.

Science.gov (United States)

Sarial, Sheila; Asadi, Farzad; Jeddi-Tehrani, Mahmood; Hadavi, Reza; Bayat, Ali Ahmad; Mahmoudian, Jafar; Taghizadeh-Jahed, Masoud; Shokri, Fazel; Rabbani, Hodjattallah

2012-12-01

Factor VII (FVII) is a serine protease-coagulating element responsible for the initiation of an extrinsic pathway of clot formation. Here we generated and characterized a high affinity monoclonal antibody that specifically recognizes human FVII. Recombinant human FVII (rh-FVII) was used for the production of a monoclonal antibody using BALB/c mice. The specificity of the antibody was determined by Western blot using plasma samples from human, mouse, sheep, goat, bovine, rabbit, and rat. Furthermore, the antibody was used to detect transiently expressed rh-FVII in BHK21 cell line using Western blot and sandwich ELISA. A mouse IgG1 (kappa chain) monoclonal antibody clone 1F1-B11 was produced against rh-FVII. The affinity constant (K(aff)) of the antibody was calculated to be 6.4×10(10) M(-1). The antibody could specifically recognize an epitope on the light chain of hFVII, with no reactivity with factor VII from several other animals. In addition, transiently expressed rh-FVII in BHK21 cells was recognized by 1F1-B11. The high affinity as well as the specificity of 1F1-B11 for hFVII will facilitate the affinity purification of hFVII and also production of FVII deficient plasma and minimizes the risk of bovine FVII contamination when fetal bovine serum-supplemented media are used for production and subsequent purification of rh-FVII.

Affine and quasi-affine frames for rational dilations

DEFF Research Database (Denmark)

Bownik, Marcin; Lemvig, Jakob

2011-01-01

In this paper we extend the investigation of quasi-affine systems, which were originally introduced by Ron and Shen [J. Funct. Anal. 148 (1997), 408-447] for integer, expansive dilations, to the class of rational, expansive dilations. We show that an affine system is a frame if, and only if......, the corresponding family of quasi-affine systems are frames with uniform frame bounds. We also prove a similar equivalence result between pairs of dual affine frames and dual quasi-affine frames. Finally, we uncover some fundamental differences between the integer and rational settings by exhibiting an example...

Arabidopsis ZED1-related kinases mediate the temperature-sensitive intersection of immune response and growth homeostasis.

Science.gov (United States)

Wang, Zhicai; Cui, Dayong; Liu, Jing; Zhao, Jingbo; Liu, Cheng; Xin, Wei; Li, Yuan; Liu, Na; Ren, Dongtao; Tang, Dingzhong; Hu, Yuxin

2017-07-01

Activation of the immune response in plants antagonizes growth and development in the absence of pathogens, and such an autoimmune phenotype is often suppressed by the elevation of ambient temperature. However, molecular regulation of the ambient temperature-sensitive intersection of immune response and growth is largely elusive. A genetic screen identified an Arabidopsis mutant, zed1-D, by its high temperature-dependent growth retardation. A combination of molecular, cytological and genetic approaches was used to investigate the molecular basis behind the temperature-sensitive growth and immune response in zed1-D. A dominant mutation in HOPZ-ETI-DEFICIENT 1 (ZED1) is responsible for a high temperature-dependent autoimmunity and growth retardation in zed1-D. The autoimmune phenotype in zed1-D is dependent on the HOPZ-ACTIVATED RESISTANCE 1 (ZAR1). ZED1 and some ZED1-related kinases (ZRKs) are induced by elevated temperature and function cooperatively to suppress the immune response by modulating the transcription of SUPPRESSOR OF NPR1-1 CONSTITUTIVE 1 (SNC1) in the absence of pathogens. Our data reveal a previously unidentified role of ZRKs in the ambient temperature-sensitive immune response in the absence of pathogens, and thus reveals a possible molecular mechanism underlying the temperature-mediated intersection of immune response and growth in plants. © 2017 The Authors. New Phytologist © 2017 New Phytologist Trust.

High-temperature sensor based on an abrupt-taper Michelson interferometer in single-mode fiber.

Science.gov (United States)

Xu, Le; Jiang, Lan; Wang, Sumei; Li, Benye; Lu, Yongfeng

2013-04-01

This study proposes a high-temperature sensor based on an abrupt fiber-taper Michelson interferometer (FTMI) in single-mode fiber fabricated by a fiber-taper machine and electric-arc discharge. The proposed FTMI is applied to measure temperature and refractive index (RI). A high temperature sensitivity of 118.6 pm/°C is obtained in the temperature range of 500°C-800°C. The wavelength variation is only -0.335 nm for the maximum attenuation peak, with the external RI changed from 1.333 to 1.3902, which is desirable for high-temperature sensing to eliminate the cross sensitivity to RI.

High transition temperature superconducting integrated circuit

International Nuclear Information System (INIS)

DiIorio, M.S.

1985-01-01

This thesis describes the design and fabrication of the first superconducting integrated circuit capable of operating at over 10K. The primary component of the circuit is a dc SQUID (Superconducting QUantum Interference Device) which is extremely sensitive to magnetic fields. The dc SQUID consists of two superconductor-normal metal-superconductor (SNS) Josephson microbridges that are fabricated using a novel step-edge process which permits the use of high transition temperature superconductors. By utilizing electron-beam lithography in conjunction with ion-beam etching, very small microbridges can be produced. Such microbridges lead to high performance dc SQUIDs with products of the critical current and normal resistance reaching 1 mV at 4.2 K. These SQUIDs have been extensively characterized, and exhibit excellent electrical characteristics over a wide temperature range. In order to couple electrical signals into the SQUID in a practical fashion, a planar input coil was integrated for efficient coupling. A process was developed to incorporate the technologically important high transition temperature superconducting materials, Nb-Sn and Nb-Ge, using integrated circuit techniques. The primary obstacles were presented by the metallurgical idiosyncrasies of the various materials, such as the need to deposit the superconductors at elevated temperatures, 800-900 0 C, in order to achieve a high transition temperature

Ultra-high sensitive hydrazine chemical sensor based on low-temperature grown ZnO nanoparticles

International Nuclear Information System (INIS)

Mehta, S.K.; Singh, Kulvinder; Umar, Ahmad; Chaudhary, G.R.; Singh, Sukhjinder

2012-01-01

Graphical abstract: Systematic representation of the fabricated amperometric hydrazine chemical sensor based on ZnO NPs/Au modified electrode. Highlights: ► Synthesis of well-crystalline ZnO NPs has been achieved in aqueous solution. ► ZnO NPs act as efficient electron mediators for hydrazine sensor. ► Extremely high sensitivity and low-detection limit have been obtained. - Abstract: Using well-crystalline ZnO nanoparticles (NPs), an ultra high sensitive hydrazine amperometric sensor has been fabricated and reported in this paper. The ZnO NPs have been synthesized by very simple aqueous solution process at 90 °C and characterized in detail in terms of their morphological, compositional, structural and optical properties. The detailed investigations reveal that the synthesized products are well-crystalline NPs, possessing wurtzite hexagonal phase and exhibit good optical properties. The fabricated amperometric hydrazine sensor exhibits ultra-high sensitivity of ∼97.133 μA cm −2 μM −1 and very low-detection limit of 147.54 nM. To the best of our knowledge, this is the first report in which an ultra-high sensitivity and low-detection limit have been obtained for the hydrazine chemical sensor based on ZnO nanostructures.

Borehole Stability in High-Temperature Formations

Science.gov (United States)

Yan, Chuanliang; Deng, Jingen; Yu, Baohua; Li, Wenliang; Chen, Zijian; Hu, Lianbo; Li, Yang

2014-11-01

In oil and gas drilling or geothermal well drilling, the temperature difference between the drilling fluid and formation will lead to an apparent temperature change around the borehole, which will influence the stress state around the borehole and tend to cause borehole instability in high geothermal gradient formations. The thermal effect is usually not considered as a factor in most of the conventional borehole stability models. In this research, in order to solve the borehole instability in high-temperature formations, a calculation model of the temperature field around the borehole during drilling is established. The effects of drilling fluid circulation, drilling fluid density, and mud displacement on the temperature field are analyzed. Besides these effects, the effect of temperature change on the stress around the borehole is analyzed based on thermoelasticity theory. In addition, the relationships between temperature and strength of four types of rocks are respectively established based on experimental results, and thermal expansion coefficients are also tested. On this basis, a borehole stability model is established considering thermal effects and the effect of temperature change on borehole stability is also analyzed. The results show that the fracture pressure and collapse pressure will both increase as the temperature of borehole rises, and vice versa. The fracture pressure is more sensitive to temperature. Temperature has different effects on collapse pressures due to different lithological characters; however, the variation of fracture pressure is unrelated to lithology. The research results can provide a reference for the design of drilling fluid density in high-temperature wells.

Structure-based engineering to restore high affinity binding of an isoform-selective anti-TGFβ1 antibody

Science.gov (United States)

Honey, Denise M.; Best, Annie; Qiu, Huawei

2018-01-01

ABSTRACT Metelimumab (CAT192) is a human IgG4 monoclonal antibody developed as a TGFβ1-specific antagonist. It was tested in clinical trials for the treatment of scleroderma but later terminated due to lack of efficacy. Subsequent characterization of CAT192 indicated that its TGFβ1 binding affinity was reduced by ∼50-fold upon conversion from the parental single-chain variable fragment (scFv) to IgG4. We hypothesized this result was due to decreased conformational flexibility of the IgG that could be altered via engineering. Therefore, we designed insertion mutants in the elbow region and screened for binding and potency. Our results indicated that increasing the elbow region linker length in each chain successfully restored the isoform-specific and high affinity binding of CAT192 to TGFβ1. The crystal structure of the high binding affinity mutant displays large conformational rearrangements of the variable domains compared to the wild-type antigen-binding fragment (Fab) and the low binding affinity mutants. Insertion of two glycines in both the heavy and light chain elbow regions provided sufficient flexibility for the variable domains to extend further apart than the wild-type Fab, and allow the CDR3s to make additional interactions not seen in the wild-type Fab structure. These interactions coupled with the dramatic conformational changes provide a possible explanation of how the scFv and elbow-engineered Fabs bind TGFβ1 with high affinity. This study demonstrates the benefits of re-examining both structure and function when converting scFv to IgG molecules, and highlights the potential of structure-based engineering to produce fully functional antibodies. PMID:29333938

Energy-dependent dissociation of ATP from high affinity catalytic sites of beef heart mitochondrial adenosine triphosphatase

International Nuclear Information System (INIS)

Penefsky, H.S.

1985-01-01

Incubation of [gamma- 32 P]ATP with a molar excess of the membrane-bound form of mitochondrial ATPase (F1) results in binding of the bulk of the radioactive nucleotide in high affinity catalytic sites (Ka = 10(12) M-1). Subsequent initiation of respiration by addition of succinate or NADH is accompanied by a profound decrease in the affinity for ATP. About one-third of the bound radioactive ATP appears to dissociate, that is, the [gamma- 32 P]ATP becomes accessible to hexokinase. The NADH-stimulated dissociation of [gamma- 32 P]ATP is energy-dependent since the stimulation is inhibited by uncouplers of oxidative phosphorylation and is prevented by respiratory chain inhibitors. The rate of the energy-dependent dissociation of ATP that occurs in the presence of NADH, ADP, and Pi is commensurate with the measured initial rate of ATP synthesis in NADH-supported oxidative phosphorylation catalyzed by the same submitochondrial particles. Thus, the rate of dissociation of ATP from the high affinity catalytic site of submitochondrial particles meets the criterion of kinetic competency under the conditions of oxidative phosphorylation. These experiments provide evidence in support of the argument that energy conserved during the oxidation of substrates by the respiratory chain can be utilized to reduce the very tight binding of product ATP in high affinity catalytic sites and to promote dissociation of the nucleotide

Simple and Efficient Purification of Recombinant Proteins Using the Heparin-Binding Affinity Tag.

Science.gov (United States)

Jayanthi, Srinivas; Gundampati, Ravi Kumar; Kumar, Thallapuranam Krishnaswamy Suresh

2017-11-01

Heparin, a member of the glycosaminoglycan family, is known to interact with more than 400 different types of proteins. For the past few decades, significant progress has been made to understand the molecular details involved in heparin-protein interactions. Based on the structural knowledge available from the FGF1-heparin interaction studies, we have designed a novel heparin-binding peptide (HBP) affinity tag that can be used for the simple, efficient, and cost-effective purification of recombinant proteins of interest. HBP-tagged fusion proteins can be purified by heparin Sepharose affinity chromatography using a simple sodium chloride gradient to elute the bound fusion protein. In addition, owing to the high density of positive charges on the HBP tag, recombinant target proteins are preferably expressed in their soluble forms. The purification of HBP-fusion proteins can also be achieved in the presence of chemical denaturants, including urea. Additionally, polyclonal antibodies raised against the affinity tag can be used to detect HBP-fused target proteins with high sensitivity. © 2017 by John Wiley & Sons, Inc. Copyright © 2017 John Wiley & Sons, Inc.

Stress corrosion cracking of austenitic stainless steel in high temperature and high pressure water

International Nuclear Information System (INIS)

Uragami, Ken

1977-01-01

Austenitic stainless steels used in for equipment in chemical plants have failed owing to stress corrosion cracking (SCC). These failures brought about great problems in some cases. The failures were caused by chloride, sulfide and alkali solution environment, in particular, by chloride solution environment. It was known that SCC was caused not only by high content chloride solution such as 42% MgCl 2 solution but also by high temperature water containing Cl - ions as NaCl. In order to estimate quantitatively the effects of some factors on SCC in high temperature water environment, the effects of Cl - ion contents, oxygen partial pressure (increasing in proportion to dissolved oxygen), pH and temperature were investigated. Moreover SCC sensitivity owing to the difference of materials and heat treatments was also investigated. The experimental results obtained are summarized as follows: (1) Regarding the effect of contaminant Cl - ions in proportion as Cl - ion contents increased, the material life extremely decreased owing to SCC. The tendency of decreasing was affected by the level of oxygen partial pressure. (2) Three regions of SCC sensitivity existed and they depended upon oxygen partial pressure. These were a region that did not show SCC sensitivity, a region of the highest SCC sensitivity and a region of somewhat lower SCC sensitivity. (3) In the case of SUS304 steel and 500 ppm Cl - ion contents SCC did not occur at 150 0 C, but it occurred and caused failures at 200 0 C and 250 0 C. (auth.)

High temperature sensors for exhaust diagnosis

Energy Technology Data Exchange (ETDEWEB)

Svenningstorp, Henrik

2000-07-01

One of the largest problems that we will have to deal with on this planet this millennium is to stop the pollution of our environment. In many of the ongoing works to reduce toxic emissions, gas sensors capable of enduring rough environments and high temperatures, would be a great tool. The different applications where sensors like this would be useful vary between everything from online measurement in the paper industry and food industry to measurement in the exhaust pipe of a car. In my project we have tested Schottky diodes and MlSiCFET sensor as gas sensors operating at high temperatures. The measurement condition in the exhaust pipe of a car is extremely tough, not only is the temperature high and the different gases quite harmful, there are also a lot of particles that can affect the sensors in an undesirable way. In my project we have been testing Schottky diodes and MlSiCFET sensors based on SiC as high temperature sensors, both in the laboratory with simulated exhaust and after a real engine. In this thesis we conclude that these sensors can work in the hostile environment of an engines exhaust. It is shown that when measuring in a gas mixture with a fixed I below one, where the I-value is controlled by the O{sub 2} concentration, a sensor with a catalytic gate metal as sensitive material respond more to the increased O{sub 2} concentration than the increased HC concentration when varying the two correspondingly. A number of different sensors have been tested in simulated exhaust towards NO{sub x}. It was shown that resistivity changes in the thin gate metal influenced the gas response. Tests have been performed where sensors were a part of a SCR system with promising results concerning NH{sub 3} sensitivity. With a working temperature of 300 deg C there is no contamination of the metal surface.

New Synthesis and Tritium Labeling of a Selective Ligand for Studying High-affinity γ-Hydroxybutyrate (GHB) Binding Sites

Science.gov (United States)

Vogensen, Stine B.; Marek, Aleš; Bay, Tina; Wellendorph, Petrine; Kehler, Jan; Bundgaard, Christoffer; Frølund, Bente; Pedersen, Martin H.F.; Clausen, Rasmus P.

2013-01-01

3-Hydroxycyclopent-1-enecarboxylic acid (HOCPCA, 1) is a potent ligand for the high-affinity GHB binding sites in the CNS. An improved synthesis of 1 together with a very efficient synthesis of [3H]-1 is described. The radiosynthesis employs in situ generated lithium trimethoxyborotritide. Screening of 1 against different CNS targets establishes a high selectivity and we demonstrate in vivo brain penetration. In vitro characterization of [3H]-1 binding shows high specificity to the high-affinity GHB binding sites. PMID:24053696

Affinity selection-mass spectrometry and its emerging application to the high throughput screening of G protein-coupled receptors.

Science.gov (United States)

Whitehurst, Charles E; Annis, D Allen

2008-07-01

Advances in combinatorial chemistry and genomics have inspired the development of novel affinity selection-based screening techniques that rely on mass spectrometry to identify compounds that preferentially bind to a protein target. Of the many affinity selection-mass spectrometry techniques so far documented, only a few solution-based implementations that separate target-ligand complexes away from unbound ligands persist today as routine high throughput screening platforms. Because affinity selection-mass spectrometry techniques do not rely on radioactive or fluorescent reporters or enzyme activities, they can complement traditional biochemical and cell-based screening assays and enable scientists to screen targets that may not be easily amenable to other methods. In addition, by employing mass spectrometry for ligand detection, these techniques enable high throughput screening of massive library collections of pooled compound mixtures, vastly increasing the chemical space that a target can encounter during screening. Of all drug targets, G protein coupled receptors yield the highest percentage of therapeutically effective drugs. In this manuscript, we present the emerging application of affinity selection-mass spectrometry to the high throughput screening of G protein coupled receptors. We also review how affinity selection-mass spectrometry can be used as an analytical tool to guide receptor purification, and further used after screening to characterize target-ligand binding interactions, enabling the classification of orthosteric and allosteric binders.

Temperature sensitive riboflavin mutants of Penicillium vermiculatum Dangeard

International Nuclear Information System (INIS)

Mitra, J.; Chaudhari, K.L.

1974-01-01

Two temperature sensitive UV induced riboflavin mutants rib 1 and rib 6 have been physiologically and genetically characterized. The two mutants behave differently with regard to their temperature sensitivity. The rib 1 mutant exhibits a leaky growth in minimal medium between 15 0 C and 30 0 C but grows well when the medium is supplemented with riboflavin. At 35 0 C the growth response of the mutant is at its max. and at 40 0 C and below 15 0 C it ceases to grow. The rib 6 mutant which is red brown in colour shows wild type character at temp. below 25 0 C in minimal medium but requires riboflavin at 30 0 C and above. Heterokaryotic analysis revealed the nonallelic nature of the two temperature mutants. Genetic tests of allelic relationship between riboflavin markers by crossing were also done. (author)

Temperature-sensitive elastin-mimetic dendrimers: Effect of peptide length and dendrimer generation to temperature sensitivity.

Science.gov (United States)

Kojima, Chie; Irie, Kotaro; Tada, Tomoko; Tanaka, Naoki

2014-06-01

Dendrimers are synthetic macromolecules with unique structure, which are a potential scaffold for peptides. Elastin is one of the main components of extracellular matrix and a temperature-sensitive biomacromolecule. Previously, Val-Pro-Gly-Val-Gly peptides have been conjugated to a dendrimer for designing an elastin-mimetic dendrimer. In this study, various elastin-mimetic dendrimers using different length peptides and different dendrimer generations were synthesized to control the temperature dependency. The elastin-mimetic dendrimers formed β-turn structure by heating, which was similar to the elastin-like peptides. The elastin-mimetic dendrimers exhibited an inverse phase transition, largely depending on the peptide length and slightly depending on the dendrimer generation. The elastin-mimetic dendrimers formed aggregates after the phase transition. The endothermal peak was observed in elastin-mimetic dendrimers with long peptides, but not with short ones. The peptide length and the dendrimer generation are important factors to tune the temperature dependency on the elastin-mimetic dendrimer. Copyright © 2013 Wiley Periodicals, Inc.

Amine-functionalized PVA-co-PE nanofibrous membrane as affinity membrane with high adsorption capacity for bilirubin.

Science.gov (United States)

Wang, Wenwen; Zhang, Hao; Zhang, Zhifeng; Luo, Mengying; Wang, Yuedan; Liu, Qiongzhen; Chen, Yuanli; Li, Mufang; Wang, Dong

2017-02-01

In this study, poly(vinyl alcohol-co-ethylene) (PVA-co-PE) nanofibrous membrane was activated by sodium hydroxide and cyanuric chloride, and then the activated membranes were functionalized by 1,3-propanediamine, hexamethylenediamine and diethylenetriamine to be affinity membranes for bilirubin removal, respectively. The chemical structures and morphologies of membranes were investigated by SEM, FTIR and XPS. And the adsorption ability of different amine-functionalized nanofibrous membranes for bilirubin was characterized. Furthermore, the effects of temperature, initial concentration of bilirubin, NaCl concentration and BSA concentration on the adsorption capacity for bilirubin of diethylenetriamine-functionalized nanofibrous membrane were studied. Results indicated that the adsorption capacity for bilirubin of diethylenetriamine-functionalized nanofibrous membrane could reach 85mg/g membrane when the initial bilirubin concentration was 200mg/L while the adsorption capacity could be increased to 110mg/g membrane if the initial bilirubin concentration was more than 400mg/L. The dynamic adsorption of diethylenetriamine-functionalized nanofibrous membrane showed that the ligands of amine groups on the membrane surface could be used as far as possible by recirculating the plasma with certain flow rates. Therefore, the diethylenetriamine-functionalized PVA-co-PE nanofibrous membrane possessed high adsorption capacity for bilirubin and it can be candidate as affinity membrane for bilirubin removal. Copyright © 2016. Published by Elsevier B.V.

High-temperature bulk acoustic wave sensors

International Nuclear Information System (INIS)

Fritze, Holger

2011-01-01

Piezoelectric crystals like langasite (La 3 Ga 5 SiO 14 , LGS) and gallium orthophosphate (GaPO 4 ) exhibit piezoelectrically excited bulk acoustic waves at temperatures of up to at least 1450 °C and 900 °C, respectively. Consequently, resonant sensors based on those materials enable new sensing approaches. Thereby, resonant high-temperature microbalances are of particular interest. They correlate very small mass changes during film deposition onto resonators or gas composition-dependent stoichiometry changes of thin films already deposited onto the resonators with the resonance frequency shift of such devices. Consequently, the objective of the work is to review the high-temperature properties, the operation limits and the measurement principles of such resonators. The electromechanical properties of high-temperature bulk acoustic wave resonators such as mechanical stiffness, piezoelectric and dielectric constant, effective viscosity and electrical conductivity are described using a one-dimensional physical model and determined accurately up to temperatures as close as possible to their ultimate limit. Insights from defect chemical models are correlated with the electromechanical properties of the resonators. Thereby, crucial properties for stable operation as a sensor under harsh conditions are identified to be the formation of oxygen vacancies and the bulk conductivity. Operation limits concerning temperature, oxygen partial pressure and water vapor pressure are given. Further, application-relevant aspects such as temperature coefficients, temperature compensation and mass sensitivity are evaluated. In addition, approximations are introduced which make the exact model handy for routine data evaluation. An equivalent electrical circuit for high-temperature resonator devices is derived based on the one-dimensional physical model. Low- and high-temperature approximations are introduced. Thereby, the structure of the equivalent circuit corresponds to the

High-temperature bulk acoustic wave sensors

Science.gov (United States)

Fritze, Holger

2011-01-01

Piezoelectric crystals like langasite (La3Ga5SiO14, LGS) and gallium orthophosphate (GaPO4) exhibit piezoelectrically excited bulk acoustic waves at temperatures of up to at least 1450 °C and 900 °C, respectively. Consequently, resonant sensors based on those materials enable new sensing approaches. Thereby, resonant high-temperature microbalances are of particular interest. They correlate very small mass changes during film deposition onto resonators or gas composition-dependent stoichiometry changes of thin films already deposited onto the resonators with the resonance frequency shift of such devices. Consequently, the objective of the work is to review the high-temperature properties, the operation limits and the measurement principles of such resonators. The electromechanical properties of high-temperature bulk acoustic wave resonators such as mechanical stiffness, piezoelectric and dielectric constant, effective viscosity and electrical conductivity are described using a one-dimensional physical model and determined accurately up to temperatures as close as possible to their ultimate limit. Insights from defect chemical models are correlated with the electromechanical properties of the resonators. Thereby, crucial properties for stable operation as a sensor under harsh conditions are identified to be the formation of oxygen vacancies and the bulk conductivity. Operation limits concerning temperature, oxygen partial pressure and water vapor pressure are given. Further, application-relevant aspects such as temperature coefficients, temperature compensation and mass sensitivity are evaluated. In addition, approximations are introduced which make the exact model handy for routine data evaluation. An equivalent electrical circuit for high-temperature resonator devices is derived based on the one-dimensional physical model. Low- and high-temperature approximations are introduced. Thereby, the structure of the equivalent circuit corresponds to the Butterworth

Characterization of an AtCCX5 gene from Arabidopsis thaliana that involves in high-affinity K+ uptake and Na+ transport in yeast

International Nuclear Information System (INIS)

Zhang, Xinxin; Zhang, Min; Takano, Tetsuo; Liu, Shenkui

2011-01-01

Highlights: → The AtCCX5 protein coding a putative cation calcium exchanger was characterized. → AtCCX5 expressed in yeast was localized in the plasma membrane and nuclear periphery. → AtCCX5 protein did not show the same transport properties as the CAXs. → AtCCX5 protein involves in mediating high-affinity K + uptake in yeast. → AtCCX5 protein also involves in Na + transport in yeast. -- Abstract: The gene for a putative cation calcium exchanger (CCX) from Arabidopsis thaliana, AtCCX5, was cloned and its function was analyzed in yeast. Green fluorescent protein-tagged AtCCX5 expressed in yeast was localized in the plasma membrane and nuclear periphery. The yeast transformants expressing AtCCX5 were created and their growth in the presence of various cations (K + , Na + , Ca 2+ , Mg 2+ , Fe 2+ , Cu 2+ , Co 2+ , Cd 2+ , Mn 2+ , Ba 2+ , Ni 2+ , Zn 2+ , and Li + ) were analyzed. AtCCX5 expression was found to affect the response to K + and Na + in yeast. The AtCCX5 transformant also showed a little better growth to Zn 2+ . The yeast mutant 9.3 expressing AtCCX5 restored growth of the mutant on medium with low K + (0.5 mM), and also suppressed its Na + sensitivity. Ion uptake experiments showed that AtCCX5 mediated relatively high-affinity K + uptake and was also involved in Na + transport in yeast. Taken together, these findings suggest that the AtCCX5 is a novel transport protein involves in mediating high-affinity K + uptake and Na + transport in yeast.

Highly sensitive microcalorimeters for radiation research

International Nuclear Information System (INIS)

Avaev, V.N.; Demchuk, B.N.; Ioffe, L.A.; Efimov, E.P.

1984-01-01

Calorimetry is used in research at various types of nuclear-physics installations to obtain information on the quantitative and qualitative composition of ionizing radiation in a reactor core and in the surrounding layers of the biological shield. In this paper, the authors examine the characteristics of highly sensitive microcalorimeters with modular semiconductor heat pickups designed for operation in reactor channels. The microcalorimeters have a thin-walled aluminum housing on whose inner surface modular heat pickups are placed radially as shown here. The results of measurements of the temperature dependence of the sensitivity of the microcalorimeters are shown. The results of measuring the sensitivity of a PMK-2 microcalorimeter assembly as a function of integrated neutron flux for three energy intervals and the adsorbed gamma energy are shown. In order to study specimens with different shapes and sizes, microcalorimeters with chambers in the form of cylinders and a parallelepiped were built and tested

High sensitivity thermal sensors on insulating diamond

Energy Technology Data Exchange (ETDEWEB)

Job, R. [Fernuniversitaet Hagen (Gesamthochschule) (Germany). Electron. Devices; Denisenko, A.V. [Fernuniversitaet Hagen (Gesamthochschule) (Germany). Electron. Devices; Zaitsev, A.M. [Fernuniversitaet Hagen (Gesamthochschule) (Germany). Electron. Devices; Melnikov, A.A. [Belarussian State Univ., Minsk (Belarus). HEII and FD; Werner, M. [VDI/VDE-IT, Teltow (Germany); Fahrner, W.R. [Fernuniversitaet Hagen (Gesamthochschule) (Germany). Electron. Devices

1996-12-15

Diamond is a promising material to develop sensors for applications in harsh environments. To increase the sensitivity of diamond temperature sensors the effect of thermionic hole emission (TE) over an energetic barrier formed in the interface between highly boron-doped p-type and intrinsic insulating diamond areas has been suggested. To study the TE of holes a p-i-p diode has been fabricated and analyzed by electrical measurements in the temperature range between 300 K and 700 K. The experimental results have been compared with numerical simulations of its electrical characteristics. Based on a model of the thermionic emission of carriers into an insulator it has been suggested that the temperature sensitivity of the p-i-p diode on diamond is strongly affected by the re-emission of holes from a group of donor-like traps located at a level of 0.7-1.0 eV above the valence band. The mechanism of thermal activation of the current includes a spatial redistribution of the potential, which results in the TE regime from a decrease of the immobilized charge of the ionized traps within the i-zone of the diode and the correspondent lowering of the forward biased barrier. The characteristics of the p-i-p diode were studied with regard to temperature sensor applications. The temperature coefficient of resistance (TCR=-0.05 K{sup -1}) for temperatures above 600 K is about four times larger than the maximal attainable TCR for conventional boron-doped diamond resistors. (orig.)

Rapid Diagnostic Assay for Intact Influenza Virus Using a High Affinity Hemagglutinin Binding Protein.

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Anderson, Caitlin E; Holstein, Carly A; Strauch, Eva-Maria; Bennett, Steven; Chevalier, Aaron; Nelson, Jorgen; Fu, Elain; Baker, David; Yager, Paul

2017-06-20

Influenza is a ubiquitous and recurring infection that results in approximately 500 000 deaths globally each year. Commercially available rapid diagnostic tests are based upon detection of the influenza nucleoprotein, which are limited in that they are unable to differentiate by species and require an additional viral lysis step. Sample preprocessing can be minimized or eliminated by targeting the intact influenza virus, thereby reducing assay complexity and leveraging the large number of hemagglutinin proteins on the surface of each virus. Here, we report the development of a paper-based influenza assay that targets the hemagglutinin protein; the assay employs a combination of antibodies and novel computationally designed, recombinant affinity proteins as the capture and detection agents. This system leverages the customizability of recombinant protein design to target the conserved receptor-binding pocket of the hemagglutinin protein and to match the trimeric nature of hemagglutinin for improved avidity. Using this assay, we demonstrate the first instance of intact influenza virus detection using a combination of antibody and affinity proteins within a porous network. The recombinant head region binder based assays yield superior analytical sensitivity as compared to the antibody based assay, with lower limits of detection of 3.54 × 10 7 and 1.34 × 10 7 CEID 50 /mL for the mixed and all binder stacks, respectively. Not only does this work describe the development of a novel influenza assay, it also demonstrates the power of recombinant affinity proteins for use in rapid diagnostic assays.

A high-temperature silicon-on-insulator stress sensor

International Nuclear Information System (INIS)

Wang Zheyao; Tian Kuo; Zhou Youzheng; Pan Liyang; Liu Litian; Hu Chaohong

2008-01-01

A piezoresistive stress sensor is developed using silicon-on-insulator (SOI) wafers and calibrated for stress measurement for high-temperature applications. The stress sensor consists of 'silicon-island-like' piezoresistor rosettes that are etched on the SOI layer. This eliminates leakage current and enables excellent electrical insulation at high temperature. To compensate for the measurement errors caused by the misalignment of the piezoresistor rosettes with respect to the crystallographic axes, an anisotropic micromachining technique, tetramethylammonium hydroxide etching, is employed to alleviate the misalignment issue. To realize temperature-compensated stress measurement, a planar diode is fabricated as a temperature sensor to decouple the temperature information from the piezoresistors, which are sensitive to both stress and temperature. Design, fabrication and calibration of the piezoresistors are given. SOI-related characteristics such as piezoresistive coefficients and temperature coefficients as well as the influence of the buried oxide layer are discussed in detail

Irreversible blockade of the high and low affinity (3H) naloxone binding sites by C-6 derivatives of morphinane-6-ones

International Nuclear Information System (INIS)

Krizsan, D.; Varga, E.; Benyhe, S.; Szucs, M.; Borsodi, A.; Hosztafi, S.

1991-01-01

C-6 derivatives-hydrazones, phenylhydrazones, dinitrophenylhydrazones, oximes and semicarbazones - of morphinane-6-ones were synthesized and their binding characteristics were studied on rat brain membranes. The dihydromorphinone and oxymorphone derivatives compete for the ( 3 H)naloxone binding sites with high affinity, while the dihydrocodeinone and oxycodone derivatives are less potent. The affinity of the new compounds is decreased for the delta sites as compared to the parent ligands. The ligands bearing bulky substituents also bind with low affinity to the kappa sites. The modification decreased the Na + -index of compounds indicating their mixed agonist-antagonist character. The dihydromorphinone derivatives are all capable to block irreversibly the high affinity binding site of ( 3 H)naloxone, whereas the dihydrocodeinone derivatives block irreversibly the low affinity site. A possible mechanism for the inhibition is suggested

A novel lentiviral scFv display library for rapid optimization and selection of high affinity antibodies.

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Qudsia, Sehar; Merugu, Siva B; Mangukiya, Hitesh B; Hema, Negi; Wu, Zhenghua; Li, Dawei

2018-04-30

Antibody display libraries have become a popular technique to screen monoclonal antibodies for therapeutic purposes. An important aspect of display technology is to generate an optimization library by changing antibody affinity to antigen through mutagenesis and screening the high affinity antibody. In this study, we report a novel lentivirus display based optimization library antibody in which Agtuzumab scFv is displayed on cell membrane of HEK-293T cells. To generate an optimization library, hotspot mutagenesis was performed to achieve diverse antibody library. Based on sequence analysis of randomly selected clones, library size was estimated approximately to be 1.6 × 10 6 . Lentivirus display vector was used to display scFv antibody on cell surface and flow cytometery was performed to check the antibody affinity to antigen. Membrane bound scFv antibodies were then converted to secreted antibody through cre/loxP recombination. One of the mutant clones, M8 showed higher affinity to antigen in flow cytometery analysis. Further characterization of cellular and secreted scFv through western blot showed that antibody affinity was increased by three fold after mutagenesis. This study shows successful construction of a novel antibody library and suggests that hotspot mutagenesis could prove a useful and rapid optimization tool to generate similar libraries with various degree of antigen affinity. Copyright © 2018 Elsevier Inc. All rights reserved.

Interactions of dopaminergic agonists and antagonists with dopaminergic D3 binding sites in rat striatum. Evidence that [3H]dopamine can label a high affinity agonist-binding state of the D1 dopamine receptor

International Nuclear Information System (INIS)

Leff, S.E.; Creese, I.

1985-01-01

The interactions of dopaminergic agonists and antagonists with 3 H-agonist labeled D3 dopaminergic binding sites of rat striatum have been characterized by radioligand-binding techniques. When the binding of [ 3 H]dopamine and [ 3 H]apomorphine to D2 dopamine receptors is blocked by the inclusion of D2 selective concentrations of unlabeled spiroperidol or domperidone, these ligands appear to label selectively the previously termed D3 binding site. Antagonist/[ 3 H]dopamine competition curves are of uniformly steep slope (nH . 1.0), suggesting the presence of a single D3 binding site. The relative potencies of antagonists to inhibit D3 specific [ 3 H]dopamine binding are significantly correlated with their potencies to block D1 dopamine receptors as measured by the inhibition of both dopamine-stimulated adenylate cyclase and [ 3 H]flupentixol-binding activities. The affinities of agonists to inhibit D3 specific [ 3 H]dopamine binding are also correlated with estimates of these agonists affinities for the high affinity binding component of agonist/[ 3 H]flupentixol competition curves. Both D3 specific [ 3 H] dopamine binding and the high affinity agonist-binding component of dopamine/[ 3 H]flupentixol competition curves show a similar sensitivity to guanine nucleotides. Taken together, these data strongly suggest that the D3 binding site is related to a high affinity agonist-binding state of the D1 dopamine receptor

On the sensitivity of annual streamflow to air temperature

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Milly, Paul C.D.; Kam, Jonghun; Dunne, Krista A.

2018-01-01

Although interannual streamflow variability is primarily a result of precipitation variability, temperature also plays a role. The relative weakness of the temperature effect at the annual time scale hinders understanding, but may belie substantial importance on climatic time scales. Here we develop and evaluate a simple theory relating variations of streamflow and evapotranspiration (E) to those of precipitation (P) and temperature. The theory is based on extensions of the Budyko water‐balance hypothesis, the Priestley‐Taylor theory for potential evapotranspiration ( ), and a linear model of interannual basin storage. The theory implies that the temperature affects streamflow by modifying evapotranspiration through a Clausius‐Clapeyron‐like relation and through the sensitivity of net radiation to temperature. We apply and test (1) a previously introduced “strong” extension of the Budyko hypothesis, which requires that the function linking temporal variations of the evapotranspiration ratio (E/P) and the index of dryness ( /P) at an annual time scale is identical to that linking interbasin variations of the corresponding long‐term means, and (2) a “weak” extension, which requires only that the annual evapotranspiration ratio depends uniquely on the annual index of dryness, and that the form of that dependence need not be known a priori nor be identical across basins. In application of the weak extension, the readily observed sensitivity of streamflow to precipitation contains crucial information about the sensitivity to potential evapotranspiration and, thence, to temperature. Implementation of the strong extension is problematic, whereas the weak extension appears to capture essential controls of the temperature effect efficiently.

Drought-induced weakening of growth-temperature associations in high-elevation Iberian pines

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Diego Galván, J.; Büntgen, Ulf; Ginzler, Christian; Grudd, Håkan; Gutiérrez, Emilia; Labuhn, Inga; Julio Camarero, J.

2015-01-01

The growth/climate relationship of theoretically temperature-controlled high-elevation forests has been demonstrated to weaken over recent decades. This is likely due to new tree growth limiting factors, such as an increasing drought risk for ecosystem functioning and productivity across the Mediterranean Basin. In addition, declining tree growth sensitivity to spring temperature may emerge in response to increasing drought stress. Here, we evaluate these ideas by assessing the growth/climate sensitivity of 1500 tree-ring width (TRW) and 102 maximum density (MXD) measurement series from 711 and 74 Pinus uncinata trees, respectively, sampled at 28 high-elevation forest sites across the Pyrenees and two relict populations of the Iberian System. Different dendroclimatological standardization and split period approaches were used to assess the high- to low-frequency behavior of 20th century tree growth in response to temperature means, precipitation totals and drought indices. Long-term variations in TRW track summer temperatures until about 1970 but diverge afterwards, whereas MXD captures the recent temperature increase in the low-frequency domain fairly well. On the other hand summer drought has increasingly driven TRW along the 20th century. Our results suggest fading temperature sensitivity of Iberian high-elevation P. uncinata forest growth, and reveal the importance of summer drought that is becoming the emergent limiting factor of tree ring width formation in many parts of the Mediterranean Basin.

Hemoglobin affinity in Andean rodents

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HRVOJ OSTOJIC

2002-01-01

Full Text Available Blood hemoglobin oxygen affinity (P50 was measured in three Andean species and in the laboratory rat (control, all raised near sea level. Chinchilla lanigera (Molina, 1792 has an altitudinal habitat range from low Andean slopes up to 3000 m., while Chinchilla brevicaudata (Waterhouse, 1848 has an altitudinal range from 3000 to 5000 m. The laboratory type guinea pig, wild type guinea pig (Cavia porcellus, (Waterhouse, 1748, and laboratory rat (Rattus norvegicus were also raised at sea level. The Andean species had high hemoglobin oxygen affinities (low P50 compared with the rat. Chinchilla brevicaudata had a higher affinity than Chinchilla lanigera. The wild type guinea pig had a higher affinity than the laboratory type. As has been shown in other species, this is another example of an inverse correlation between the altitude level and the P50 values. This is the first hemoglobin oxygen affinity study in Chinchilla brevicaudata.

Silicon Carbide-Based Hydrogen Gas Sensors for High-Temperature Applications

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Sangchoel Kim

2013-10-01

Full Text Available We investigated SiC-based hydrogen gas sensors with metal-insulator-semiconductor (MIS structure for high temperature process monitoring and leak detection applications in fields such as the automotive, chemical and petroleum industries. In this work, a thin tantalum oxide (Ta2O5 layer was exploited with the purpose of sensitivity improvement, because tantalum oxide has good stability at high temperature with high permeability for hydrogen gas. Silicon carbide (SiC was used as a substrate for high-temperature applications. We fabricated Pd/Ta2O5/SiC-based hydrogen gas sensors, and the dependence of their I-V characteristics and capacitance response properties on hydrogen concentrations were analyzed in the temperature range from room temperature to 500 °C. According to the results, our sensor shows promising performance for hydrogen gas detection at high temperatures.

High-affinity receptors for bombesin-like peptides in normal guinea pig lung membranes

International Nuclear Information System (INIS)

Lach, E.; Trifilieff, A.; Landry, Y.; Gies, J.P.

1991-01-01

The binding of the radiolabeled bombesin analogue [ 125 I-Tyr 4 ]bombesin to guinea-pig lung membranes was investigated. Binding of [ 125 I-Tyr 4 ]bombesin was specific, saturable, reversible and linearly related to the protein concentration. Scatchard analysis of equilibrium binding data at 25C indicated the presence of a single class of non-interacting binding sites for bombesin (B max = 7.7 fmol/mg protein). The value of the equilibrium dissociation constant (K D = 90 pM) agrees with a high-affinity binding site. Bombesin and structurally related peptides such as [ 125 I-Tyr 4 ]bombesin, neuromedin B and neuromedin C inhibited the binding of [ 125 I-Tyr 4 ]bombesin in an order of potencies as follows: [ 125 I-Tyr 4 ]bombesin > bombesin ≥ neuromedin C much-gt neuromedin B. These results indicate that guinea-pig lung membranes possess a single class of bombesin receptors with a high affinity for bombesin and a lower one for neuromedin B

High-throughput bioscreening system utilizing high-performance affinity magnetic carriers exhibiting minimal non-specific protein binding

International Nuclear Information System (INIS)

Hanyu, Naohiro; Nishio, Kosuke; Hatakeyama, Mamoru; Yasuno, Hiroshi; Tanaka, Toshiyuki; Tada, Masaru; Nakagawa, Takashi; Sandhu, Adarsh; Abe, Masanori; Handa, Hiroshi

2009-01-01

For affinity purification of drug target protein we have developed magnetic carriers, narrow in size distribution (184±9 nm), which exhibit minimal non-specific binding of unwanted proteins. The carriers were highly dispersed in aqueous solutions and highly resistant to organic solvents, which enabled immobilization of various hydrophobic chemicals as probes on the carrier surfaces. Utilizing the carriers we have automated the process of separation and purification of the target proteins that had been done by manual operation previously.

The relationship between virtual body ownership and temperature sensitivity

Science.gov (United States)

Llobera, Joan; Sanchez-Vives, M. V.; Slater, Mel

2013-01-01

In the rubber hand illusion, tactile stimulation seen on a rubber hand, that is synchronous with tactile stimulation felt on the hidden real hand, can lead to an illusion of ownership over the rubber hand. This illusion has been shown to produce a temperature decrease in the hidden hand, suggesting that such illusory ownership produces disownership of the real hand. Here, we apply immersive virtual reality (VR) to experimentally investigate this with respect to sensitivity to temperature change. Forty participants experienced immersion in a VR with a virtual body (VB) seen from a first-person perspective. For half the participants, the VB was consistent in posture and movement with their own body, and in the other half, there was inconsistency. Temperature sensitivity on the palm of the hand was measured before and during the virtual experience. The results show that temperature sensitivity decreased in the consistent compared with the inconsistent condition. Moreover, the change in sensitivity was significantly correlated with the subjective illusion of virtual arm ownership but modulated by the illusion of ownership over the full VB. This suggests that a full body ownership illusion results in a unification of the virtual and real bodies into one overall entity—with proprioception and tactile sensations on the real body integrated with the visual presence of the VB. The results are interpreted in the framework of a ‘body matrix’ recently introduced into the literature. PMID:23720537

Time-temperature-sensitization and time-temperature-precipitation behavior of alloy 625

International Nuclear Information System (INIS)

Koehler, M.; Heubner, U.

1996-01-01

Time-Temperature-Sensitization diagrams have been established for a low-carbon version of alloy 625 (UNS N06625). Sensitization in terms of a 50 microm (2 mils) intergranular penetration criterion starts after about 3 h aging time at 750 C (soft annealed condition) or after less than 1 h aging time at 800 C (solution annealed condition) when tested according to ASTM-G 28 method A. Grain boundary precipitation of carbides occurs during aging of both the soft annealed and the solution annealed material, but the soft annealed material exhibits a more pronounced general precipitation of Ni 3 (Nb,Mo) phase giving rise to more distinct loss of ductility. Sensitization of alloy 625 may be retarded by lowering its iron content

Sensitivity of glaciation in the arid subtropical Andes to changes in temperature, precipitation, and solar radiation

Science.gov (United States)

Vargo, L. J.; Galewsky, J.; Rupper, S.; Ward, D. J.

2018-04-01

The subtropical Andes (18.5-27 °S) have been glaciated in the past, but are presently glacier-free. We use idealized model experiments to quantify glacier sensitivity to changes in climate in order to investigate the climatic drivers of past glaciations. We quantify the equilibrium line altitude (ELA) sensitivity (the change in ELA per change in climate) to temperature, precipitation, and shortwave radiation for three distinct climatic regions in the subtropical Andes. We find that in the western cordillera, where conditions are hyper-arid with the highest solar radiation on Earth, ELA sensitivity is as high as 34 m per % increase in precipitation, and 70 m per % decrease in shortwave radiation. This is compared with the eastern cordillera, where precipitation is the highest of the three regions, and ELA sensitivity is only 10 m per % increase in precipitation, and 25 m per % decrease in shortwave radiation. The high ELA sensitivity to shortwave radiation highlights the influence of radiation on mass balance of high elevation and low-latitude glaciers. We also consider these quantified ELA sensitivities in context of previously dated glacial deposits from the regions. Our results suggest that glaciation of the humid eastern cordillera was driven primarily by lower temperatures, while glaciations of the arid Altiplano and western cordillera were also influenced by increases in precipitation and decreases in shortwave radiation. Using paleoclimate records from the timing of glaciation, we find that glaciation of the hyper-arid western cordillera can be explained by precipitation increases of 90-160% (1.9-2.6× higher than modern), in conjunction with associated decreases in shortwave radiation of 7-12% and in temperature of 3.5 °C.

Enhancement of the cytotoxicity of radiosensitizers by modest hyperthermia: the electron-affinity relationship

Energy Technology Data Exchange (ETDEWEB)

Rajaratnam, S.; Adams, G.E.; Stratford, I.J.; Clarke, C.

1982-01-01

The cytotoxicity of 3 electron-affinic radiosensitizers has been studied in Chinese hamster V-79 cells as a function of pH and modest hyperthermia. When equitoxic concentrations were used and temperature was increased from 34 to 41/sup 0/C metronidazole, the compound with the lowest electron affinity showed the greatest enhancement of hypoxic-cell toxicity, and nitrofurantoin, the compound with the highest electron affinity, the least. The results can be explained if the mechanisms of toxicity involves a redox reaction, since it would be expected that the least toxic compound (lowest electron affinity) would have the largest activation energy and hence the greatest temperature effect. This appears to hold for these 3 compounds. Experiments also showed that nitrofurantoin which exhibits no increase in toxicity when the temperature was increased from 37 to 41/sup 0/C at pH 7.4, showed an increase in toxicity for the same temperature change at the pH of 7.0 and 6.6. Under aerobic conditions only metronidazole showed significant toxicity at 41/sup 0/C, where the differential between aerobic and hypoxic cell toxicity was minimal, both at pH 7.4, and at the low pH values of 7.0 and 6.6. In the clinical setting there is evidence that tumor cells are at a lower pH than their surrounding normal tissues. Hypoxic-cell cytotoxicity is enhanced at low pH, and even further enhanced at low pH in combination with a temperature of 41/sup 0/C. However, this finding correlates conversely with electron affinity.

Sensitivity calculation of the coolant temperature regarding the thermohydraulic parameters

International Nuclear Information System (INIS)

Andrade Lima, F.R. de; Silva, F.C. da; Thome Filho, Z.D.; Alvim, A.C.M.; Oliveira Barroso, A.C. de.

1985-01-01

It's studied the application of the Generalized Perturbation Theory (GPT) in the sensitivity calculation of thermalhydraulic problems, aiming at verifying the viability of the extension of the method. For this, the axial distribution, transient, of the coolant temperature in a PWR channel are considered. Perturbation expressions are developed using the GPT formalism, and a computer code (Tempera) is written, to calculate the channel temperature distribution and the associated importance function, as well as the effect of the thermalhydraulic parameters variations in the coolant temperature (sensitivity calculation). The results are compared with those from the direct calculation. (E.G.) [pt

Visual and Plasmon Resonance Absorption Sensor for Adenosine Triphosphate Based on the High Affinity between Phosphate and Zr(IV).

Science.gov (United States)

Qi, Wenjing; Liu, Zhongyuan; Zhang, Wei; Halawa, Mohamed Ibrahim; Xu, Guobao

2016-10-12

Zr(IV) can form phosphate and Zr(IV) (-PO₃ 2- -Zr 4+ -) complex owing to the high affinity between Zr(IV) with phosphate. Zr(IV) can induce the aggregation of gold nanoparticles (AuNPs), while adenosine triphosphate(ATP) can prevent Zr(IV)-induced aggregation of AuNPs. Herein, a visual and plasmon resonance absorption (PRA)sensor for ATP have been developed using AuNPs based on the high affinity between Zr(IV)with ATP. AuNPs get aggregated in the presence of certain concentrations of Zr(IV). After the addition of ATP, ATP reacts with Zr(IV) and prevents AuNPs from aggregation, enabling the detection of ATP. Because of the fast interaction of ATP with Zr(IV), ATP can be detected with a detection limit of 0.5 μM within 2 min by the naked eye. Moreover, ATP can be detected by the PRA technique with higher sensitivity. The A 520nm / A 650nm values in PRA spectra increase linearly with the concentrations of ATP from 0.1 μM to 15 μM (r = 0.9945) with a detection limit of 28 nM. The proposed visual and PRA sensor exhibit good selectivity against adenosine, adenosine monophosphate, guanosine triphosphate, cytidine triphosphate and uridine triphosphate. The recoveries for the analysis of ATP in synthetic samples range from 95.3% to 102.0%. Therefore, the proposed novel sensor for ATP is promising for real-time or on-site detection of ATP.

Low-Temperature Synthesis and Gas Sensitivity of Perovskite-Type LaCoO3 Nanoparticles

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Lorenzo Gildo Ortiz

2014-01-01

Full Text Available LaCoO3 nanoparticles with perovskite-type structure were prepared by a microwave-assisted colloidal method. Lanthanum nitrate, cobalt nitrate, and ethylenediamine were used as precursors and ethyl alcohol as solvent. The thermal decomposition of the precursors leads to the formation of LaCoO3 from a temperature of 500°C. The structural, morphological, and compositional properties of LaCoO3 nanoparticles were studied in this work by X-ray diffraction (XRD, scanning electron microscopy (SEM, transmission electron microscopy (TEM, and atomic force microscopy (AFM. Pellets were manufactured in order to test the gas sensing properties of LaCoO3 powders in carbon monoxide (CO and propane (C3H8 atmospheres. Agglomerates of nanoparticles with high connectivity, forming a porous structure, were observed from SEM and TEM analysis. LaCoO3 pellets presented a high sensitivity in both CO and C3H8 at different concentrations and operating temperatures. As was expected, sensitivity increased with the gas concentration and operation temperature increase.

Microfabrication of a Novel Ceramic Pressure Sensor with High Sensitivity Based on Low-Temperature Co-Fired Ceramic (LTCC Technology

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Chen Li

2014-06-01

Full Text Available In this paper, a novel capacitance pressure sensor based on Low-Temperature Co-Fired Ceramic (LTCC technology is proposed for pressure measurement. This approach differs from the traditional fabrication process for a LTCC pressure sensor because a 4J33 iron-nickel-cobalt alloy is applied to avoid the collapse of the cavity and to improve the performance of the sensor. Unlike the traditional LTCC sensor, the sensitive membrane of the proposed sensor is very flat, and the deformation of the sensitivity membrane is smaller. The proposed sensor also demonstrates a greater responsivity, which reaches as high as 13 kHz/kPa in range of 0–100 kPa. During experiments, the newly fabricated sensor, which is only about 6.5 cm2, demonstrated very good performance: the repeatability error, hysteresis error, and nonlinearity of the sensor are about 4.25%, 2.13%, and 1.77%, respectively.

Synthesis of site-heterologous haptens for high-affinity anti-pyraclostrobin antibody generation.

Science.gov (United States)

Mercader, Josep V; Agulló, Consuelo; Abad-Somovilla, Antonio; Abad-Fuentes, Antonio

2011-03-07

The design and synthesis of functional chemical derivatives of small organic molecules is usually a key step for the intricate production of a variety of bioconjugates. In this respect, the derivatization site at which the spacer arm is introduced in immunizing conjugates constitutes a highly critical parameter for the generation of high-affinity and selective antibodies. However, due to the usual complexity of the required synthetic procedures, the appropriate comparison of alternative tethering positions has often been neglected. In the present study, meticulous strategies were followed to prepare synthetic derivatives of pyraclostrobin with the same linkers located at diverse rationally-chosen sites. Activity appraisal of antibodies and bioconjugates was carried out by bidimensional competitive direct and indirect immunoassays, and a superior performance of two of the three synthesized haptens was found. Finally, a detailed analysis of the conformations of the target molecule and the synthesized haptens in aqueous solution was done using computer assisted molecular modeling techniques. This study suggested that the lower titers and affinities of one set of antibodies are most probably due to conformational effects of the spacer arm in the immunizing bioconjugate.

Temperature- and pH-sensitive wearable materials for monitoring foot ulcers

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Salvo P

2017-01-01

Full Text Available Pietro Salvo,1,2 Nicola Calisi,1 Bernardo Melai,1 Valentina Dini,3 Clara Paoletti,1 Tommaso Lomonaco,1 Andrea Pucci,1 Fabio Di Francesco,1 Alberto Piaggesi,4 Marco Romanelli3 1Department of Chemistry and Industrial Chemistry, University of Pisa, 2Institute of Clinical Physiology, National Council of Research, 3Wound Healing Research Unit, Department of Dermatology, University of Pisa, 4Diabetic Foot Section, Department of Medicine, University of Pisa, Pisa, Italy Abstract: Foot ulcers account for 15% of comorbidities associated with diabetes. Presently, no device allows the status of foot ulcers to be continuously monitored when patients are not hospitalized. In this study, we describe a temperature and a pH sensor capable of monitoring diabetic foot and venous leg ulcers developed in the frame of the seventh framework program European Union project SWAN-iCare (smart wearable and autonomous negative pressure device for wound monitoring and therapy. Temperature is measured by exploiting the variations in the electrical resistance of a nanocomposite consisting of multiwalled carbon nanotubes and poly(styrene-b-(ethylene-co-butylene-b-styrene. The pH sensor used a graphene oxide (GO layer that changes its electrical potential when pH changes. The temperature sensor has a sensitivity of ~85 Ω/°C in the range 25°C–50°C and a high repeatability (maximum standard deviation of 0.1% over seven repeated measurements. For a GO concentration of 4 mg/mL, the pH sensor has a sensitivity of ~42 mV/pH and high linearity (R2=0.99. Keywords: diabetic foot ulcer, wearable sensors, wound temperature, wound pH

Affinity imaging mass spectrometry (AIMS): high-throughput screening for specific small molecule interactions with frozen tissue sections.

Science.gov (United States)

Yoshimi, T; Kawabata, S; Taira, S; Okuno, A; Mikawa, R; Murayama, S; Tanaka, K; Takikawa, O

2015-11-07

A novel screening system, using affinity imaging mass spectrometry (AIMS), has been developed to identify protein aggregates or organ structures in unfixed human tissue. Frozen tissue sections are positioned on small (millimetre-scale) stainless steel chips and incubated with an extensive library of small molecules. Candidate molecules showing specific affinity for the tissue section are identified by imaging mass spectrometry (IMS). As an example application, we screened over a thousand compounds against Alzheimer's disease (AD) brain tissue and identified several compounds with high affinity for AD brain sections containing tau deposits compared to age-matched controls. It should also be possible to use AIMS to isolate chemical compounds with affinity for tissue structures or components that have been extensively modified by events such as oxidation, phosphorylation, acetylation, aggregation, racemization or truncation, for example, due to aging. It may also be applicable to biomarker screening programs.

Validation of temperature-sensitive radio transmitters for measurement of body temperature in small animals

NARCIS (Netherlands)

Williams, Joseph B.; Tieleman, B. I.; Shobrak, Mohammed

2009-01-01

As part of a study on the core body temperature (T(b)) of desert birds, we purposed to use temperature-sensitive implantable radio transmitters. Because of the difficulty in recapturing these birds, we needed to know if these electronic devices held their calibration over the duration of normal

Temperature Crosstalk Sensitivity of the Kummerow Rainfall Algorithm

Science.gov (United States)

Spencer, Roy W.; Petrenko, Boris

1999-01-01

Even though the signal source for passive microwave retrievals is thermal emission, retrievals of non-temperature geophysical parameters typically do not explicitly take into account the effects of temperature change on the retrievals. For global change research, changes in geophysical parameters (e.g. water vapor, rainfall, etc.) are referenced to the accompanying changes in temperature. If the retrieval of a certain parameter has a cross-talk response from temperature change alone, the retrievals might not be very useful for climate research. We investigated the sensitivity of the Kummerow rainfall retrieval algorithm to changes in air temperature. It was found that there was little net change in total rainfall with air temperature change. However, there were non-negligible changes within individual rain rate categories.

Molecular electron affinities

International Nuclear Information System (INIS)

Fukuda, E.K.

1983-01-01

Molecular electron affinities have historically been difficult quantities to measure accurately. These difficulties arise from differences in structure between the ion and neutral as well as the existence of excited negative ion states. To circumvent these problems, relative electron affinities were determined in this dissertation by studying equilibrium electron transfer reactions using a pulsed ion cyclotron resonance (ICR) spectrometer. Direct measurement of ion and neutral concentrations for reactions of the general type, A - + B = B - + A, allow calculation of the equilibrium constant and, therefore, the free energy change. The free energy difference is related to the difference in electron affinities between A and B. A relative electron affinity scale covering a range of about 45 kcal/mol was constructed with various substituted p-benzoquinones, nitrobenzenes, anhydrides, and benzophenones. To assign absolute electron affinities, various species with accurately known electron affinities are tied to the scale via ion-cyclotron double resonance bracketing techniques. After the relative scale is anchored to these species with well-known electron affinities, the scale is then used as a check on other electron affinity values as well as generating new electron affinity values. Many discrepancies were found between the electron affinities measured using the ICR technique and previous literature determinations

Highly sensitive voltammetric biosensor for nitric oxide based on its high affinity with hemoglobin

International Nuclear Information System (INIS)

Fan Chunhai; Liu Xinjian; Pang Jiantao; Li Genxi; Scheer, Hugo

2004-01-01

Although heme protein-based, amperometric nitric oxide (NO) biosensors have been well documented in previous studies, most have been conducted in anaerobic conditions. Herein we report a novel hemoglobin-based NO biosensor that is not only very sensitive but also usable in air. The heme protein was entrapped in a sodium montmorillonite film, which was immobilized at a pyrolytic graphite electrode surface. Film-entrapped hemoglobin can directly exchange electrons with the electrode, and this process has proven to favor the catalytic reduction of oxygen. In addition, NO induced a cathodic potential shift of the catalytic reduction peak of oxygen. This potential shift was proportional to the logarithm of NO concentration ranging from 4.0 x 10 -11 to 5.0 x 10 -6 mol/L. The detection limit has been estimated to be 20 pM, approximately four orders lower than previously reported amperometric detectors

Potentialities in electronics of new high critical temperature superconductors

International Nuclear Information System (INIS)

Hartemann, P.

1989-01-01

The main electronic applications of superconductors involve the signal processing, the electromagnetic wave detection and the magnetometry. Characteristics of devices based on conventional superconductors cooled by liquid helium are given and the changes induced by incorporating high-temperature superconductors are estimated. After a survey of new superconductor properties, the superconducting devices for analog or digital signal processing are reviewed. The gains predicted for high-temperature superconducting analog devices are considered in greater detail. Different sections deal with the infrared or (sub)millimeter wave detection. The most sensitive apparatuses for magnetic measurements are based on SQUIDs. Features of SQUIDs made of granular high-temperature superconducting material samples (grain boundaries behave as barriers of intrinsic junctions) are discussed [fr

Surface Effects and Challenges for Application of Piezoelectric Langasite Substrates in Surface Acoustic Wave Devices Caused by High Temperature Annealing under High Vacuum.

Science.gov (United States)

Seifert, Marietta; Rane, Gayatri K; Kirbus, Benjamin; Menzel, Siegfried B; Gemming, Thomas

2015-12-19

Substrate materials that are high-temperature stable are essential for sensor devices which are applied at high temperatures. Although langasite is suggested as such a material, severe O and Ga diffusion into an O-affine deposited film was observed during annealing at high temperatures under vacuum conditions, leading to a damage of the metallization as well as a change of the properties of the substrate and finally to a failure of the device. Therefore, annealing of bare LGS (La 3 Ga 5 SiO 14 ) substrates at 800 ∘ C under high vacuum conditions is performed to analyze whether this pretreatment improves the suitability and stability of this material for high temperature applications in vacuum. To reveal the influence of the pretreatment on the subsequently deposited metallization, RuAl thin films are used as they are known to oxidize on LGS at high temperatures. A local study of the pretreated and metallized substrates using transmission electron microscopy reveals strong modification of the substrate surface. Micro cracks are visible. The composition of the substrate is strongly altered at those regions. Severe challenges for the application of LGS substrates under high-temperature vacuum conditions arise from these substrate damages, revealing that the pretreatment does not improve the applicability.

Surface Effects and Challenges for Application of Piezoelectric Langasite Substrates in Surface Acoustic Wave Devices Caused by High Temperature Annealing under High Vacuum

Directory of Open Access Journals (Sweden)

Marietta Seifert

2015-12-01

Full Text Available Substrate materials that are high-temperature stable are essential for sensor devices which are applied at high temperatures. Although langasite is suggested as such a material, severe O and Ga diffusion into an O-affine deposited film was observed during annealing at high temperatures under vacuum conditions, leading to a damage of the metallization as well as a change of the properties of the substrate and finally to a failure of the device. Therefore, annealing of bare LGS (La 3 Ga 5 SiO 14 substrates at 800 ∘ C under high vacuum conditions is performed to analyze whether this pretreatment improves the suitability and stability of this material for high temperature applications in vacuum. To reveal the influence of the pretreatment on the subsequently deposited metallization, RuAl thin films are used as they are known to oxidize on LGS at high temperatures. A local study of the pretreated and metallized substrates using transmission electron microscopy reveals strong modification of the substrate surface. Micro cracks are visible. The composition of the substrate is strongly altered at those regions. Severe challenges for the application of LGS substrates under high-temperature vacuum conditions arise from these substrate damages, revealing that the pretreatment does not improve the applicability.

Modeling DNA affinity landscape through two-round support vector regression with weighted degree kernels

KAUST Repository

Wang, Xiaolei

2014-12-12

Background: A quantitative understanding of interactions between transcription factors (TFs) and their DNA binding sites is key to the rational design of gene regulatory networks. Recent advances in high-throughput technologies have enabled high-resolution measurements of protein-DNA binding affinity. Importantly, such experiments revealed the complex nature of TF-DNA interactions, whereby the effects of nucleotide changes on the binding affinity were observed to be context dependent. A systematic method to give high-quality estimates of such complex affinity landscapes is, thus, essential to the control of gene expression and the advance of synthetic biology. Results: Here, we propose a two-round prediction method that is based on support vector regression (SVR) with weighted degree (WD) kernels. In the first round, a WD kernel with shifts and mismatches is used with SVR to detect the importance of subsequences with different lengths at different positions. The subsequences identified as important in the first round are then fed into a second WD kernel to fit the experimentally measured affinities. To our knowledge, this is the first attempt to increase the accuracy of the affinity prediction by applying two rounds of string kernels and by identifying a small number of crucial k-mers. The proposed method was tested by predicting the binding affinity landscape of Gcn4p in Saccharomyces cerevisiae using datasets from HiTS-FLIP. Our method explicitly identified important subsequences and showed significant performance improvements when compared with other state-of-the-art methods. Based on the identified important subsequences, we discovered two surprisingly stable 10-mers and one sensitive 10-mer which were not reported before. Further test on four other TFs in S. cerevisiae demonstrated the generality of our method. Conclusion: We proposed in this paper a two-round method to quantitatively model the DNA binding affinity landscape. Since the ability to modify

High affinity, ligand specific uptake of complexed copper-67 by brain tissue incubated in vitro

International Nuclear Information System (INIS)

Barnea, A.; Hartter, D.E.

1987-01-01

Copper is an essential metal that is highly concentrated in the brain. The blood, the sole source of tissue Cu, contains 16-20 μM Cu, of which >95% is complexed to proteins and 2 was 10 times greater than that of CuAlbumin or Cu(II). Within the range of 0.2-150μM Cu, multiple uptake sites for CuHis were apparent. Increasing the molar ratio of His:Cu had a differential effect on Cu uptake: enhancing uptake at [Cu] 1 μM. Thus, using a His:Cu ratio of 1000, they observed a high affinity process exhibiting saturating and half saturating values of 5 μM and 1.5 μM Cu, respectively; using a His:Cu ratio of 2, they observed a low affinity process exhibiting saturating and half-saturating values of 100 μM and 40 μM Cu, respectively. Both processes required thermic but not metabolic energy, suggestive of facilitated diffusion. Considering the blood brain barrier for proteins, CuHis appears to be the major substrate for Cu uptake by neuronal tissue. They demonstrate the existence of a ligand specific, high affinity (apparent Km about 1.5 μM Cu) uptake process for CuHis in the brain, operative at the physiological concentration range of CuHis and histidine

Amyloid-beta binds catalase with high affinity and inhibits hydrogen peroxide breakdown.

OpenAIRE

Milton, N G

1999-01-01

Amyloid-beta (Abeta) specifically bound purified catalase with high affinity and inhibited catalase breakdown of H(2)O(2). The Abeta-induced catalase inhibition involved formation of the inactive catalase Compound II and was reversible. CatalaseAbeta interactions provide rapid functional assays for the cytotoxic domain of Abeta and suggest a mechanism for some of the observed actions of Abeta plus catalase in vitro.

Mobile Technology Affinity in Renal Transplant Recipients.

Science.gov (United States)

Reber, S; Scheel, J; Stoessel, L; Schieber, K; Jank, S; Lüker, C; Vitinius, F; Grundmann, F; Eckardt, K-U; Prokosch, H-U; Erim, Y

Medication nonadherence is a common problem in renal transplant recipients (RTRs). Mobile health approaches to improve medication adherence are a current trend, and several medication adherence apps are available. However, it is unknown whether RTRs use these technologies and to what extent. In the present study, the mobile technology affinity of RTRs was analyzed. We hypothesized significant age differences in mobile technology affinity and that mobile technology affinity is associated with better cognitive functioning as well as higher educational level. A total of 109 RTRs (63% male) participated in the cross-sectional study, with an overall mean age of 51.8 ± 14.2 years. The study included the Technology Experience Questionnaire (TEQ) for the assessment of mobile technology affinity, a cognitive test battery, and sociodemographic data. Overall, 57.4% of the patients used a smartphone or tablet and almost 45% used apps. The TEQ sum score was 20.9 in a possible range from 6 (no affinity to technology) to 30 (very high affinity). Younger patients had significantly higher scores in mobile technology affinity. The only significant gender difference was found in having fun with using electronic devices: Men enjoyed technology more than women did. Mobile technology affinity was positively associated with cognitive functioning and educational level. Young adult patients might profit most from mobile health approaches. Furthermore, high educational level and normal cognitive functioning promote mobile technology affinity. This should be kept in mind when designing mobile technology health (mHealth) interventions for RTRs. For beneficial mHealth interventions, further research on potential barriers and desired technologic features is necessary to adapt apps to patients' needs. Copyright © 2017 Elsevier Inc. All rights reserved.

Identification of high-affinity calmodulin-binding proteins in rat liver

International Nuclear Information System (INIS)

Hanley, R.M.; Dedman, J.R.; Shenolikar, S.

1987-01-01

The Ca 2+ -dependent binding of [ 125 I] calmodulin (CaM) to hepatic proteins separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was utilized to identify CaM binding or acceptor proteins or CAPs. Two proteins of apparent molecular weight of 60,000 (CAP-60) and 45,000 (CAP-45) comprised > 80% of the Ca 2+ -dependent CaM binding in rat liver cytosol. CAP-60 and CAP-45 were partially purified by a variety of chromatographic steps, including affinity chromatography on CaM Sepharose. CAP-60 possessed a native molecular size of 400,000, indicating it to be the CaM-binding subunit of a larger oligomeric complex. In contrast, CAP-45 was monomeric as judged by gel filtration. Neither CAP-60 nor CAP-45 possessed chromatographic properties consistent with known CaM-dependent enzymes reported in the literature. Two-dimensional peptide mapping provided convincing evidence that CAP-60 and CAP-45 were unrelated to other well-characterized CAPs, namely Ca 2+ (CaM)-dependent protein kinase II, calcineurin, or the CaM-dependent cyclic nucleotide phosphodiesterase. The relative abundance and high affinity for CaM could suggest that these novel target proteins, CAP-60 and CAP-45, represent a dominant pathway for CaM action in the mammalian liver

High night temperature strongly impacts TCA cycle, amino acid and polyamine biosynthetic pathways in rice in a sensitivity-dependent manner.

Science.gov (United States)

Glaubitz, Ulrike; Erban, Alexander; Kopka, Joachim; Hincha, Dirk K; Zuther, Ellen

2015-10-01

Global climate change combined with asymmetric warming can have detrimental effects on the yield of crop plants such as rice (Oryza sativa L.). Little is known about metabolic responses of rice to high night temperature (HNT) conditions. Twelve cultivars with different HNT sensitivity were used to investigate metabolic changes in the vegetative stage under HNT compared to control conditions. Central metabolism, especially TCA cycle and amino acid biosynthesis, were strongly affected particularly in sensitive cultivars. Levels of several metabolites were correlated with HNT sensitivity. Furthermore, pool sizes of some metabolites negatively correlated with HNT sensitivity under control conditions, indicating metabolic pre-adaptation in tolerant cultivars. The polyamines putrescine, spermidine and spermine showed increased abundance in sensitive cultivars under HNT conditions. Correlations between the content of polyamines and 75 other metabolites indicated metabolic shifts from correlations with sugar-phosphates and 1-kestose under control to correlations with sugars and amino and organic acids under HNT conditions. Increased expression levels of ADC2 and ODC1, genes encoding enzymes catalysing the first committed steps of putrescine biosynthesis, were restricted to sensitive cultivars under HNT. Additionally, transcript levels of eight polyamine biosynthesis genes were correlated with HNT sensitivity. Responses to HNT in the vegetative stage result in distinct differences between differently responding cultivars with a dysregulation of central metabolism and an increase of polyamine biosynthesis restricted to sensitive cultivars under HNT conditions and a pre-adaptation of tolerant cultivars already under control conditions with higher levels of potentially protective compatible solutes. © The Author 2015. Published by Oxford University Press on behalf of the Society for Experimental Biology.

Sensors based on mesoporous SnO{sub 2}-CuWO{sub 4} with high selective sensitivity to H{sub 2}S at low operating temperature

Energy Technology Data Exchange (ETDEWEB)

Stanoiu, Adelina; Simion, Cristian E. [National Institute of Materials Physics, Atomistilor 405A, P.O. Box MG-7, 077125 Bucharest, Măgurele (Romania); Calderon-Moreno, Jose Maria; Osiceanu, Petre [“Ilie Murgulescu” Institute of Physical Chemistry, Romanian Academy, Surface Chemistry and Catalysis Laboratory, Spl. Independentei 202, 060021, Bucharest (Romania); Florea, Mihaela [University of Bucharest, Faculty of Chemistry, Department of Organic Chemistry, Biochemistry and Catalysis, B-dul Regina Elisabeta 4-12, Bucharest (Romania); National Institute of Materials Physics, Atomistilor 405A, P.O. Box MG-7, 077125 Bucharest, Măgurele (Romania); Teodorescu, Valentin S. [National Institute of Materials Physics, Atomistilor 405A, P.O. Box MG-7, 077125 Bucharest, Măgurele (Romania); Somacescu, Simona, E-mail: somacescu.simona@gmail.com [“Ilie Murgulescu” Institute of Physical Chemistry, Romanian Academy, Surface Chemistry and Catalysis Laboratory, Spl. Independentei 202, 060021, Bucharest (Romania)

2017-06-05

Highlights: • Mesoporous SnO{sub 2}-CuWO{sub 4} obtained by an inexpensive synthesis route. • Powders characterization performed by a variety of complementary techniques. • SnO{sub 2}-CuWO{sub 4} layers with high selective sensitivity to H{sub 2}S. • Low operating temperature and relative humidity influences. - Abstract: Development of new sensitive materials by different synthesis routes in order to emphasize the sensing properties for hazardous H{sub 2}S detection is one of a nowadays challenge in the field of gas sensors. In this study we obtained mesoporous SnO{sub 2}-CuWO{sub 4} with selective sensitivity to H{sub 2}S by an inexpensive synthesis route with low environmental pollution level, using tripropylamine (TPA) as template and polyvinylpyrrolidone (PVP) as dispersant/stabilizer. In order to bring insights about the intrinsic properties, the powders were characterized by means of a variety of complementary techniques such as: X-Ray Diffraction, XRD; Transmission Electron Microscopy, TEM; High Resolution TEM, HRTEM; Raman Spectroscopy, RS; Porosity Analysis by N{sub 2} adsorption/desorption, BET; Scanning Electron Microscopy, SEM and X-ray Photoelectron Spectroscopy, XPS. The sensors were fabricated by powders deposition via screen-printing technique onto planar commercial Al{sub 2}O{sub 3} substrates. The sensor signals towards H{sub 2}S exposure at low operating temperature (100 °C) reaches values from 10{sup 5} (for SnWCu600) to 10{sup 6} (for SnWCu800) over the full range of concentrations (5–30 ppm). The recovery processes were induced by a short temperature trigger of 500 °C. The selective sensitivity was underlined with respect to the H{sub 2}S, relative to other potential pollutants and relative humidity (10–70% RH).

The effect of molybdenum addition on SCC susceptibility of stainless steels in oxygenated high temperature water

International Nuclear Information System (INIS)

Akashi, Masatsune; Kawamoto, Teruaki

1978-01-01

The effect of molybdenum addition on the SCC susceptibility of sensitized stainless steel in oxygenated high temperature water has been studied through the creviced bent beam SCC test (CBB test) and A262E intergranular corrosion test. The molybdenum addition improved the SCC susceptibility of sensitized stainless steels in oxygenated high temperature water not only by delaying the sensitization at lower temperatures but also by increasing the material resistance to the SCC under a given degree of sensitization. These laboratory test results reveal that the molybdenum addition is quite beneficial for improving the SCC susceptibility of stainless steel pipe weld joints in boiling water reactor environment. (auth.)

Antibody Binding Selectivity: Alternative Sets of Antigen Residues Entail High-Affinity Recognition.

Directory of Open Access Journals (Sweden)

Yves Nominé

Full Text Available Understanding the relationship between protein sequence and molecular recognition selectivity remains a major challenge. The antibody fragment scFv1F4 recognizes with sub nM affinity a decapeptide (sequence 6TAMFQDPQER15 derived from the N-terminal end of human papilloma virus E6 oncoprotein. Using this decapeptide as antigen, we had previously shown that only the wild type amino-acid or conservative replacements were allowed at positions 9 to 12 and 15 of the peptide, indicating a strong binding selectivity. Nevertheless phenylalanine (F was equally well tolerated as the wild type glutamine (Q at position 13, while all other amino acids led to weaker scFv binding. The interfaces of complexes involving either Q or F are expected to diverge, due to the different physico-chemistry of these residues. This would imply that high-affinity binding can be achieved through distinct interfacial geometries. In order to investigate this point, we disrupted the scFv-peptide interface by modifying one or several peptide positions. We then analyzed the effect on binding of amino acid changes at the remaining positions, an altered susceptibility being indicative of an altered role in complex formation. The 23 starting variants analyzed contained replacements whose effects on scFv1F4 binding ranged from minor to drastic. A permutation analysis (effect of replacing each peptide position by all other amino acids except cysteine was carried out on the 23 variants using the PEPperCHIP® Platform technology. A comparison of their permutation patterns with that of the wild type peptide indicated that starting replacements at position 11, 12 or 13 modified the tolerance to amino-acid changes at the other two positions. The interdependence between the three positions was confirmed by SPR (Biacore® technology. Our data demonstrate that binding selectivity does not preclude the existence of alternative high-affinity recognition modes.

3D printed high performance strain sensors for high temperature applications

Science.gov (United States)

Rahman, Md Taibur; Moser, Russell; Zbib, Hussein M.; Ramana, C. V.; Panat, Rahul

2018-01-01

Realization of high temperature physical measurement sensors, which are needed in many of the current and emerging technologies, is challenging due to the degradation of their electrical stability by drift currents, material oxidation, thermal strain, and creep. In this paper, for the first time, we demonstrate that 3D printed sensors show a metamaterial-like behavior, resulting in superior performance such as high sensitivity, low thermal strain, and enhanced thermal stability. The sensors were fabricated using silver (Ag) nanoparticles (NPs), using an advanced Aerosol Jet based additive printing method followed by thermal sintering. The sensors were tested under cyclic strain up to a temperature of 500 °C and showed a gauge factor of 3.15 ± 0.086, which is about 57% higher than that of those available commercially. The sensor thermal strain was also an order of magnitude lower than that of commercial gages for operation up to a temperature of 500 °C. An analytical model was developed to account for the enhanced performance of such printed sensors based on enhanced lateral contraction of the NP films due to the porosity, a behavior akin to cellular metamaterials. The results demonstrate the potential of 3D printing technology as a pathway to realize highly stable and high-performance sensors for high temperature applications.

The utility of affine variables and affine coherent states

International Nuclear Information System (INIS)

Klauder, John R

2012-01-01

Affine coherent states are generated by affine kinematical variables much like canonical coherent states are generated by canonical kinematical variables. Although all classical and quantum formalisms normally entail canonical variables, it is shown that affine variables can serve equally well for many classical and quantum studies. This general purpose analysis provides tools to discuss two major applications: (1) the completely successful quantization of a nonrenormalizable scalar quantum field theory by affine techniques, in complete contrast to canonical techniques which only offer triviality; and (2) a formulation of the kinematical portion of quantum gravity that favors affine kinematical variables over canonical kinematical variables, and which generates a framework in which a favorable analysis of the constrained dynamical issues can take place. All this is possible because of the close connection between the affine and the canonical stories, while the few distinctions can be used to advantage when appropriate. This article is part of a special issue of Journal of Physics A: Mathematical and Theoretical devoted to ‘Coherent states: mathematical and physical aspects’. (review)

Heparin-binding peptide as a novel affinity tag for purification of recombinant proteins.

Science.gov (United States)

Morris, Jacqueline; Jayanthi, Srinivas; Langston, Rebekah; Daily, Anna; Kight, Alicia; McNabb, David S; Henry, Ralph; Kumar, Thallapuranam Krishnaswamy Suresh

2016-10-01

Purification of recombinant proteins constitutes a significant part of the downstream processing in biopharmaceutical industries. Major costs involved in the production of bio-therapeutics mainly depend on the number of purification steps used during the downstream process. Affinity chromatography is a widely used method for the purification of recombinant proteins expressed in different expression host platforms. Recombinant protein purification is achieved by fusing appropriate affinity tags to either N- or C- terminus of the target recombinant proteins. Currently available protein/peptide affinity tags have proved quite useful in the purification of recombinant proteins. However, these affinity tags suffer from specific limitations in their use under different conditions of purification. In this study, we have designed a novel 34-amino acid heparin-binding affinity tag (HB-tag) for the purification of recombinant proteins expressed in Escherichia coli (E. coli) cells. HB-tag fused recombinant proteins were overexpressed in E. coli in high yields. A one-step heparin-Sepharose-based affinity chromatography protocol was developed to purify HB-fused recombinant proteins to homogeneity using a simple sodium chloride step gradient elution. The HB-tag has also been shown to facilitate the purification of target recombinant proteins from their 8 M urea denatured state(s). The HB-tag has been demonstrated to be successfully released from the fusion protein by an appropriate protease treatment to obtain the recombinant target protein(s) in high yields. Results of the two-dimensional NMR spectroscopy experiments indicate that the purified recombinant target protein(s) exist in the native conformation. Polyclonal antibodies raised against the HB-peptide sequence, exhibited high binding specificity and sensitivity to the HB-fused recombinant proteins (∼10 ng) in different crude cell extracts obtained from diverse expression hosts. In our opinion, the HB-tag provides a

Selection of imprinted nanoparticles by affinity chromatography.

Science.gov (United States)

Guerreiro, António R; Chianella, Iva; Piletska, Elena; Whitcombe, Michael J; Piletsky, Sergey A

2009-04-15

Soluble molecularly imprinted nanoparticles were synthesised via iniferter initiated polymerisation and separated by size via gel permeation chromatography. Subsequent fractionation of these particles by affinity chromatography allowed the separation of high affinity fractions from the mixture of nanoparticles. Fractions selected this way possess affinity similar to that of natural antibodies (K(d) 6.6x10(-8)) M and were also able to discriminate between related functional analogues of the template.

Modeling FBG sensors sensitivity from cryogenic temperatures to room temperature as a function of metal coating thickness

Science.gov (United States)

Vendittozzi, Cristian; Felli, Ferdinando; Lupi, Carla

2018-05-01

Fiber optics with photo-imprinted Bragg grating have been studied in order to be used as temperature sensors in cryogenic applications. The main disadvantage presented by Fiber Bragg Grating (FBG) sensors is the significant drop in sensitivity as temperature decreases, mainly due to the critical lowering of the thermo-optic coefficient of the fiber and the very low thermal expansion coefficient (CTE) of fused silica at cryogenic temperatures. Thus, especially for the latter, it is important to enhance sensitivity to temperature by depositing a metal coating presenting higher CTE. In this work the thermal sensitivity of metal-coated FBG sensors has been evaluated by considering their elongation within temperature variations in the cryogenic range, as compared to bare fiber sensors. To this purpose, a theoretical model simulating elongation of metal-coated sensors has been developed. The model has been used to evaluate the behaviour of different metals which can be used as coating (Ni, Cu, Al, Zn, Pb and In). The optimal coating thickness has been calculated at different fixed temperature (from 5 K to 100 K) for each metal. It has been found that the metal coating effectiveness depends on thickness and operating temperature in accordance to our previous experimental work and theory suggest.

Structural insights into a high affinity nanobody:antigen complex by homology modelling

DEFF Research Database (Denmark)

Skottrup, Peter Durand

2017-01-01

Porphyromonas gingivalis is a major periodontitis-causing pathogens. P. gingivalis secrete a cysteine protease termed RgpB, which is specific for Arg-Xaa bonds in substrates. Recently, a nanobody-based assay was used to demonstrate that RgpB could represent a novel diagnostic target, thereby...... simplifying. P. gingivalis detection. The nanobody, VHH7, had a high binding affinity and was specific for RgpB, when tested towards the highly identical RgpA. In this study a homology model of VHH7 was build. The complementarity determining regions (CDR) comprising the paratope residues responsible for Rgp...

Fragment screening of cyclin G-associated kinase by weak affinity chromatography.

Science.gov (United States)

Meiby, Elinor; Knapp, Stefan; Elkins, Jonathan M; Ohlson, Sten

2012-11-01

Fragment-based drug discovery (FBDD) has become a new strategy for drug discovery where lead compounds are evolved from small molecules. These fragments form low affinity interactions (dissociation constant (K(D)) = mM - μM) with protein targets, which require fragment screening methods of sufficient sensitivity. Weak affinity chromatography (WAC) is a promising new technology for fragment screening based on selective retention of fragments by a drug target. Kinases are a major pharmaceutical target, and FBDD has been successfully applied to several of these targets. In this work, we have demonstrated the potential to use WAC in combination with mass spectrometry (MS) detection for fragment screening of a kinase target-cyclin G-associated kinase (GAK). One hundred seventy fragments were selected for WAC screening by virtual screening of a commercial fragment library against the ATP-binding site of five different proteins. GAK protein was immobilized on a capillary HPLC column, and compound binding was characterized by frontal affinity chromatography. Compounds were screened in sets of 13 or 14, in combination with MS detection for enhanced throughput. Seventy-eight fragments (46 %) with K(D) < 200 μM were detected, including a few highly efficient GAK binders (K(D) of 2 μM; ligand efficiency = 0.51). Of special interest is that chiral screening by WAC may be possible, as two stereoisomeric fragments, which both contained one chiral center, demonstrated twin peaks. This ability, in combination with the robustness, sensitivity, and simplicity of WAC makes it a new method for fragment screening of considerable potential.

Alternative affinity tools: more attractive than antibodies?

NARCIS (Netherlands)

Ruigrok, V.J.B.; Levisson, M.; Eppink, M.H.M.; Smidt, H.; Oost, van der J.

2011-01-01

Antibodies are the most successful affinity tools used today, in both fundamental and applied research (diagnostics, purification and therapeutics). Nonetheless, antibodies do have their limitations, including high production costs and low stability. Alternative affinity tools based on nucleic acids

Signature of electron-phonon interaction in high temperature superconductors

Directory of Open Access Journals (Sweden)

Vinod Ashokan

2011-09-01

Full Text Available The theory of thermal conductivity of high temperature superconductors (HTS based on electron and phonon line width (life times formulation is developed with Quantum dynamical approach of Green's function. The frequency line width is observed as an extremely sensitive quantity in the transport phenomena of HTS as a collection of large number of scattering processes. The role of resonance scattering and electron-phonon interaction processes is found to be most prominent near critical temperature. The theory successfully explains the spectacular behaviour of high Tc superconductors in the vicinity of transition temperature. A successful agreement between theory and experiment has been obtained by analyzing the thermal conductivity data for the sample La1.8Sr0.2CuO4 in the temperature range 0 − 200K. The theory is equally and successfully applicable to all other high Tc superconductors.

Temperature sensitivity of soil respiration is dependent on readily decomposable C substrate concentration

Science.gov (United States)

Larionova, A. A.; Yevdokimov, I. V.; Bykhovets, S. S.

2007-06-01

Temperature acclimation of soil organic matter (SOM) decomposition is one of the major uncertainties in predicting soil CO2 efflux by the increase in global mean temperature. A reasonable explanation for an apparent acclimation proposed by Davidson and colleagues (2006) based on Michaelis-Menten kinetics suggests that temperature sensitivity decreases when both maximal activity of respiratory enzymes (Vmax) and half- saturation constant (Ks) cancel each other upon temperature increase. We tested the hypothesis of the canceling effect by the mathematical simulation of the data obtained in the incubation experiments with forest and arable soils. Our data confirm the hypothesis and suggest that concentration of readily decomposable C substrate as glucose equivalent is an important factor controlling temperature sensitivity. The highest temperature sensitivity was observed when C substrate concentration was much lower than Ks. Increase of substrate content to the half-saturation constant resulted in temperature acclimation associated with the canceling effect. Addition of the substrate to the level providing respiration at a maximal rate Vmax leads to the acclimation of the whole microbial community as such. However, growing microbial biomass was more sensitive to the temperature alterations. This study improves our understanding of the instability of temperature sensitivity of soil respiration under field conditions, explaining this phenomenon by changes in concentration of readily decomposable C substrate. It is worth noting that this pattern works regardless of the origin of C substrate: production by SOM decomposition, release into the soil by rhizodeposition, litter fall or drying-rewetting events.

Further studies on a temperature-sensitive mutant of Escherichia coli with defective repair capacity

International Nuclear Information System (INIS)

Morfiadakis, I.; Geissler, E.; Akademie der Wissenschaften der DDR, Berlin. Zentralinstitut fuer Molekularbiologie)

1981-01-01

A temperature-sensitive mutant of E. coli, WG24, was studied with respect to its sensitivity to photodynamic action, its capacity to perform host controlled reactivation, and its sensitivity to transduction at elevated temperatures. Mutant cells are much more sensitive than wild type cells to photodynamic action by thiopyronine and visible light at elevated temperatures. As well defined rec mutants, WG24 cells are less able to reactivate UV irradiated lambdac phages at elevated temperatures, while their ability to repair T1 phages is less impaired. Mutant cells cannot be transduced to T6 resistance at a detectable rate at elevated temperature. It is concluded, therefore, that some rec gene carries a ts mutation in this mutant. (author)

Trace element affinities in two high-Ge coals from China

Energy Technology Data Exchange (ETDEWEB)

Jing Li; Xinguo Zhuang; Xavier Querol [China University of Geosciences, Wuhan (China). Faculty of Earth Resources

2011-01-15

The Lincang (Yunnan Province, Southwest China) and Wulantuga (Inner Mongolia, Northeast China) coal deposits are known because of the high-Ge content. These coals have also a high concentration of a number of other elements. To determine the mode of occurrence of the enriched elements in both coals, six density fractions from {lt} 1.43 to {gt} 2.8 g/cm{sup 3} were obtained from two representative samples using heavy-liquids. A number of peculiar geochemical patterns characterize these high-Ge coals. Thus, the results of the chemical analysis of these density fractions showed that both coals (very distant and of a different geological age) are highly enriched (compared with the usual worldwide coal concentration ranges) in Ge, As, Sb, W, Be, and Tl. This may be due to similar geochemistry of hydrothermal fluids influencing the Earth Crust in these regions of China. Moreover, Wulantuga coal (Early Cretaceous subbituminous coal) is also enriched in Ca, Mg, and Na, and Lincang coal (Neogene subbituminous coal) in K, Rb, Nb, Mo, Sn, Cs, and U. A group of elements consisting of Ge, W, B, Nb, and Sb mostly occur with an organic affinity in both coals. Additionally, Be, U, and Mo (and partially Mn and Zn) in Lincang, and Na and Mg in Wulantuga occur also with a major organic affinity. Both coals have sulfide-arsenide mineral assemblages (Fe, S, As, Sn, and Pb, and in addition to Tl, Ta, and Cs in the Lincang coal). The occurrence of Al, P, Li, Sc, Ti, V, Cr, and Zr in both coals, and Ba in Lincang, are associated with the mineral assemblage of silico-aluminates and minor heavy minerals. Furthermore, P, Na, Li, Sc, Ti, Ga, Rb, Zr, Cr, Ba, Th, and LREE (La, Ce, Pr, Nd, and Gd) in Lincang are associated with mineral assemblages of phosphates and minor heavy minerals. The two later mineral assemblages are derived from the occurrence of detrital minerals. 34 refs., 7 figs., 3 tabs.

How does pressure overload cause cardiac hypertrophy and dysfunction? High-ouabain affinity cardiac Na+ pumps are crucial.

Science.gov (United States)

Blaustein, Mordecai P

2017-11-01

Left ventricular hypertrophy is frequently observed in hypertensive patients and is believed to be due to the pressure overload and cardiomyocyte stretch. Three recent reports on mice with genetically engineered Na + pumps, however, have demonstrated that cardiac ouabain-sensitive α 2 -Na + pumps play a key role in the pathogenesis of transaortic constriction-induced hypertrophy. Hypertrophy was delayed/attenuated in mice with mutant, ouabain-resistant α 2 -Na + pumps and in mice with cardiac-selective knockout or transgenic overexpression of α 2 -Na + pumps. The latter, seemingly paradoxical, findings can be explained by comparing the numbers of available (ouabain-free) high-affinity (α 2 ) ouabain-binding sites in wild-type, knockout, and transgenic hearts. Conversely, hypertrophy was accelerated in α 2 -ouabain-resistant (R) mice in which the normally ouabain-resistant α 1 -Na + pumps were mutated to an ouabain-sensitive (S) form (α 1 S/S α 2 R/R or "SWAP" vs. wild-type or α 1 R/R α 2 S/S mice). Furthermore, transaortic constriction-induced hypertrophy in SWAP mice was prevented/reversed by immunoneutralizing circulating endogenous ouabain (EO). These findings show that EO and its receptor, ouabain-sensitive α 2 , are critical factors in pressure overload-induced cardiac hypertrophy. This complements reports linking elevated plasma EO to hypertension, cardiac hypertrophy, and failure in humans and elucidates the underappreciated role of the EO-Na + pump pathway in cardiovascular disease. Copyright © 2017 the American Physiological Society.

Report: Affinity Chromatography.

Science.gov (United States)

Walters, Rodney R.

1985-01-01

Supports, affinity ligands, immobilization, elution methods, and a number of applications are among the topics considered in this discussion of affinity chromatography. An outline of the basic principles of affinity chromatography is included. (JN)

Low soil moisture during hot periods drives apparent negative temperature sensitivity of soil respiration in a dryland ecosystem: A multi-model comparison

Science.gov (United States)

Tucker, Colin; Reed, Sasha C.

2016-01-01

Arid and semiarid ecosystems (drylands) may dominate the trajectory of biosphere-to-atmosphere carbon (C) flux over the coming century. Accordingly, understanding dryland CO2 efflux controls is important for understanding C cycling at the global-scale: key unknowns regarding how temperature and moisture interact to regulate dryland C cycling remain. Further, the patchiness of dryland vegetation can create ‘islands of fertility’, with spatially heterogeneous rates of soil respiration (Rs). At our study site in southeastern Utah, USA we added or removed litter (0 to 650% of control) in paired plots that were either associated with a shrub or with interspaces between vascular plants. We measured Rs, soil temperature, and water content (θ) on eight sampling dates between October 2013 and November 2014. Rs was highest following monsoon rains in late summer when soil temperature was ~30°C. During mid-summer, Rs was low, associated with high soil temperatures (>40°C), resulting in an apparent negative temperature sensitivity of Rs at high temperatures, and positive temperature sensitivity at low-moderate temperatures. We used Bayesian statistical methods to compare multiple competing models capturing a wide range of hypothesized relationships between temperature, moisture, and Rs. The best fit model indicates apparent negative temperature sensitivity of soil respiration at high temperatures reflects the control of soil moisture – not high temperatures – in limiting Rs. The modeled Q10 ranged from 2.7 at 5°C to 1.4 at 45°C. Litter addition had no effect on temperature sensitivity or reference respiration (Rref = Rs at 20°C and optimum moisture) beneath shrubs, and little effect on Rref in interspaces, yet Rref was 1.5 times higher beneath shrubs than in interspaces. Together, these results suggest reduced Rs often observed at high temperatures in drylands is dominated by the control of moisture, and that variable litter inputs – at least over the short

The high temperature out-of-pile test of LVDT for elongation measurement of fuel pellet

Energy Technology Data Exchange (ETDEWEB)

Son, J. M.; Kim, B. K.; Jo, M. S.; Joo, K. N.; Park, S. J.; Gang, Y. H.; Kim, Y. J. [KAERI, Taejon (Korea, Republic of)

2003-10-01

As a part of the development of instrumentation technologies for the nuclear fuel irradiation test in HANARO(High-flux Advanced Nuclear Application Reactor), the elongation measurement technique of the fuel pellet is being developed using LVDT(Linear Variable Differential Transformer). The well qualified out-of-pile test were needed to understand the LVDT's detail characteristics at high temperature for the detail design of the fuel irradiation instrumented capsule, because LVDT is very sensitive to variation of temperature. Therefore, the high temperature out-of-pile test system for fuel pellet elongation was developed, and this test was performed under the temperature condition between room temperature and 300 .deg. C with increasing the elongation from 0 to 5 mm. The LVDT's high temperature characteristics and temperature sensitivity of LVDT were analyzed through this experiment. Based on the result of this test, the method for the application of LVDT and elongation detector at high temperature was introduced. It is known that the results will be used to predict accurately the elongation of fuel pellet during irradiation test.

Palladium Gate All Around - Hetero Dielectric -Tunnel FET based highly sensitive Hydrogen Gas Sensor

Science.gov (United States)

Madan, Jaya; Chaujar, Rishu

2016-12-01

The paper presents a novel highly sensitive Hetero-Dielectric-Gate All Around Tunneling FET (HD-GAA-TFET) based Hydrogen Gas Sensor, incorporating the advantages of band to band tunneling (BTBT) mechanism. Here, the Palladium supported silicon dioxide is used as a sensing media and sensing relies on the interaction of hydrogen with Palladium-SiO2-Si. The high surface to volume ratio in the case of cylindrical GAA structure enhances the fortuities for surface reactions between H2 gas and Pd, and thus improves the sensitivity and stability of the sensor. Behaviour of the sensor in presence of hydrogen and at elevated temperatures is discussed. The conduction path of the sensor which is dependent on sensors radius has also been varied for the optimized sensitivity and static performance analysis of the sensor where the proposed design exhibits a superior performance in terms of threshold voltage, subthreshold swing, and band to band tunneling rate. Stability of the sensor with respect to temperature affectability has also been studied, and it is found that the device is reasonably stable and highly sensitive over the bearable temperature range. The successful utilization of HD-GAA-TFET in gas sensors may open a new door for the development of novel nanostructure gas sensing devices.

Discovery of PF-06928215 as a high affinity inhibitor of cGAS enabled by a novel fluorescence polarization assay.

Science.gov (United States)

Hall, Justin; Brault, Amy; Vincent, Fabien; Weng, Shawn; Wang, Hong; Dumlao, Darren; Aulabaugh, Ann; Aivazian, Dikran; Castro, Dana; Chen, Ming; Culp, Jeffrey; Dower, Ken; Gardner, Joseph; Hawrylik, Steven; Golenbock, Douglas; Hepworth, David; Horn, Mark; Jones, Lyn; Jones, Peter; Latz, Eicke; Li, Jing; Lin, Lih-Ling; Lin, Wen; Lin, David; Lovering, Frank; Niljanskul, Nootaree; Nistler, Ryan; Pierce, Betsy; Plotnikova, Olga; Schmitt, Daniel; Shanker, Suman; Smith, James; Snyder, William; Subashi, Timothy; Trujillo, John; Tyminski, Edyta; Wang, Guoxing; Wong, Jimson; Lefker, Bruce; Dakin, Leslie; Leach, Karen

2017-01-01

Cyclic GMP-AMP synthase (cGAS) initiates the innate immune system in response to cytosolic dsDNA. After binding and activation from dsDNA, cGAS uses ATP and GTP to synthesize 2', 3' -cGAMP (cGAMP), a cyclic dinucleotide second messenger with mixed 2'-5' and 3'-5' phosphodiester bonds. Inappropriate stimulation of cGAS has been implicated in autoimmune disease such as systemic lupus erythematosus, thus inhibition of cGAS may be of therapeutic benefit in some diseases; however, the size and polarity of the cGAS active site makes it a challenging target for the development of conventional substrate-competitive inhibitors. We report here the development of a high affinity (KD = 200 nM) inhibitor from a low affinity fragment hit with supporting biochemical and structural data showing these molecules bind to the cGAS active site. We also report a new high throughput cGAS fluorescence polarization (FP)-based assay to enable the rapid identification and optimization of cGAS inhibitors. This FP assay uses Cy5-labelled cGAMP in combination with a novel high affinity monoclonal antibody that specifically recognizes cGAMP with no cross reactivity to cAMP, cGMP, ATP, or GTP. Given its role in the innate immune response, cGAS is a promising therapeutic target for autoinflammatory disease. Our results demonstrate its druggability, provide a high affinity tool compound, and establish a high throughput assay for the identification of next generation cGAS inhibitors.

Visual and Plasmon Resonance Absorption Sensor for Adenosine Triphosphate Based on the High Affinity between Phosphate and Zr(IV

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Wenjing Qi

2016-10-01

Full Text Available Zr(IV can form phosphate and Zr(IV (–PO32−–Zr4+– complex owing to the high affinity between Zr(IV with phosphate. Zr(IV can induce the aggregation of gold nanoparticles (AuNPs, while adenosine triphosphate(ATP can prevent Zr(IV-induced aggregation of AuNPs. Herein, a visual and plasmon resonance absorption (PRAsensor for ATP have been developed using AuNPs based on the high affinity between Zr(IVwith ATP. AuNPs get aggregated in the presence of certain concentrations of Zr(IV. After the addition of ATP, ATP reacts with Zr(IV and prevents AuNPs from aggregation, enabling the detection of ATP. Because of the fast interaction of ATP with Zr(IV, ATP can be detected with a detection limit of 0.5 μM within 2 min by the naked eye. Moreover, ATP can be detected by the PRA technique with higher sensitivity. The A520nm/A650nm values in PRA spectra increase linearly with the concentrations of ATP from 0.1 μM to 15 μM (r = 0.9945 with a detection limit of 28 nM. The proposed visual and PRA sensor exhibit good selectivity against adenosine, adenosine monophosphate, guanosine triphosphate, cytidine triphosphate and uridine triphosphate. The recoveries for the analysis of ATP in synthetic samples range from 95.3% to 102.0%. Therefore, the proposed novel sensor for ATP is promising for real-time or on-site detection of ATP.

High-Affinity Methanotrophy Informed by Genome-Wide Analysis of Upland Soil Cluster Alpha (USCα) from Axel Heiberg Island, Canadian High Arctic

Science.gov (United States)

Rusley, C.; Onstott, T. C.; Lau, M.

2017-12-01

Methane (CH4) is a potent greenhouse gas whose proper budgeting is vital to climate predictions. Recent studies have identified upland Arctic mineral cryosols as consistent CH4 sinks, drawing CH4 from both the atmosphere and underlying anaerobic soil layers. Global atmospheric CH4 uptake is proposed to be mediated by high-affinity methanotrophs based on the detection of the marker gene pmoA (particulate methane monooxygenase beta subunit). However, a lack of pure cultures and scarcity of genomic information have hindered our understanding of their metabolic capabilities and versatility. Together with the missing genetic linkage between its pmoA and 16S ribosomal RNA (rRNA) gene, the factors that control the distribution and magnitude of high-affinity methanotrophy in the Arctic permafrost-affected region have remained elusive. Using 21 metagenomic datasets of surface soils obtained from long-term core incubation experiments,1 this bioinformatics study aimed to reconstruct the draft genome of the Upland Soil Cluster α-proteobacteria (USCα), the high-affinity methanotroph previously detected in the samples,2 and to determine its phylogeny and metabolic requirements. We obtained a genome bin containing the high-affinity form of the USCα-like pmoA gene. The 3.03 Mbp assembly is 91.6% complete with a unique set of single-copy marker genes. The 16S rRNA gene fragment of USCα belongs to the α-proteobacterial family Beijerinckiaceae. Genome annotation indicates possible formaldehyde oxidation via tetrahydromethanopterin-linked C1 transfer pathways, acetate utilization, carbon fixation via the Calvin-Benson-Bassham cycle, and glycogen production. Notably, the key enzymes for formaldehyde assimilation via the serine and ribulose monophosphate pathways are missing. The presence of genes encoding nitrate reductase and hemoglobin suggests adaptation to low O2 under water-logged conditions. Since USCα has versatile carbon metabolisms, it may not be an obligate methanotroph

Ion channel regulation by phosphoinositides analyzed with VSPs – PI(4,5P2 affinity, phosphoinositide selectivity, and PI(4,5P2 pool accessibility

Directory of Open Access Journals (Sweden)

Alexandra eRjasanow

2015-06-01

Full Text Available The activity of many proteins depends on the phosphoinositide (PI content of the membrane. E.g., dynamic changes of the concentration of PI(4,5P2 are cellular signals that regulate ion channels. The susceptibility of a channel to such dynamics depends on its affinity for PI(4,5P2. Yet, measuring affinities for endogenous PIs has not been possible directly, but has relied largely on the response to soluble analogs, which may not quantitatively reflect binding to native lipids.Voltage-sensitive phosphatases (VSPs turn over PI(4,5P2 to PI(4P when activated by depolarization. In combination with voltage-clamp electrophysiology VSPs are useful tools for rapid and reversible depletion of PI(4,5P2. Because cellular PI(4,5P2 is resynthesized rapidly, steady state PI(4,5P2 changes with the degree of VSP activation and thus depends on membrane potential.Here we show that titration of endogenous PI(4,5P2 with Ci-VSP allows for the quantification of relative PI(4,5P2 affinities of ion channels. The sensitivity of inward rectifier and voltage-gated K+ channels to Ci-VSP allowed for comparison of PI(4,5P2 affinities within and across channel subfamilies and detected changes of affinity in mutant channels. The results also reveal that VSPs are useful only for PI effectors with high binding specificity among PI isoforms, because PI(4,5P2 depletion occurs at constant overall PI level. Thus, Kir6.2, a channel activated by PI(4,5P2 and PI(4P was insensitive to VSP.Surprisingly, despite comparable PI(4,5P2 affinity as determined by Ci-VSP, the Kv7 and Kir channel families strongly differed in their sensitivity to receptor-mediated depletion of PI(4,5P2. While Kv7 members were highly sensitive to activation of PLC by Gq-coupled receptors, Kir channels were insensitive even when PI(4,5P2 affinity was lowered by mutation. We hypothesize that different channels may be associated with distinct pools of PI(4,5P2 that differ in their accessibility to PLC and VSPs.

Sensitive element of multifunctional sensor for measuring temperature, strain and magnetic field induction

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Druzhinin A. A.

2017-12-01

Full Text Available Sensitive element of multifunctional sensor for measuring temperature, strain and magnetic field induction has been developed based on the studies of electrical conductivity and magnetoresistance of silicon and germanium microcrystals in the temperature range 4.2—70 K, strain ±1.5*10–3 rel.un. and magnetic fields of 0—14 T. The feature of the sensitive element is the using of the p- and n-type conductivity germanium microcrystals as mechanical and magnetic field sensors, respectively, and the p-type silicon microcrystal — as temperature sensor. That allows providing the compensation of temperature influence on piezoresistance and on sensitivity to the magnetic field.

Spectral affinity in protein networks.

Science.gov (United States)

Voevodski, Konstantin; Teng, Shang-Hua; Xia, Yu

2009-11-29

Protein-protein interaction (PPI) networks enable us to better understand the functional organization of the proteome. We can learn a lot about a particular protein by querying its neighborhood in a PPI network to find proteins with similar function. A spectral approach that considers random walks between nodes of interest is particularly useful in evaluating closeness in PPI networks. Spectral measures of closeness are more robust to noise in the data and are more precise than simpler methods based on edge density and shortest path length. We develop a novel affinity measure for pairs of proteins in PPI networks, which uses personalized PageRank, a random walk based method used in context-sensitive search on the Web. Our measure of closeness, which we call PageRank Affinity, is proportional to the number of times the smaller-degree protein is visited in a random walk that restarts at the larger-degree protein. PageRank considers paths of all lengths in a network, therefore PageRank Affinity is a precise measure that is robust to noise in the data. PageRank Affinity is also provably related to cluster co-membership, making it a meaningful measure. In our experiments on protein networks we find that our measure is better at predicting co-complex membership and finding functionally related proteins than other commonly used measures of closeness. Moreover, our experiments indicate that PageRank Affinity is very resilient to noise in the network. In addition, based on our method we build a tool that quickly finds nodes closest to a queried protein in any protein network, and easily scales to much larger biological networks. We define a meaningful way to assess the closeness of two proteins in a PPI network, and show that our closeness measure is more biologically significant than other commonly used methods. We also develop a tool, accessible at http://xialab.bu.edu/resources/pnns, that allows the user to quickly find nodes closest to a queried vertex in any protein

Structural insights into a high affinity nanobody:antigen complex by homology modelling.

Science.gov (United States)

Skottrup, Peter Durand

2017-09-01

Porphyromonas gingivalis is a major periodontitis-causing pathogens. P. gingivalis secrete a cysteine protease termed RgpB, which is specific for Arg-Xaa bonds in substrates. Recently, a nanobody-based assay was used to demonstrate that RgpB could represent a novel diagnostic target, thereby simplifying. P. gingivalis detection. The nanobody, VHH7, had a high binding affinity and was specific for RgpB, when tested towards the highly identical RgpA. In this study a homology model of VHH7 was build. The complementarity determining regions (CDR) comprising the paratope residues responsible for RgpB binding were identified and used as input to the docking. Furthermore, residues likely involved in the RgpB epitope was identified based upon RgpB:RgpA alignment and analysis of residue surface accessibility. CDR residues and putitative RgpB epitope residues were used as input to an information-driven flexible docking approach using the HADDOCK server. Analysis of the VHH7:RgpB model demonstrated that the epitope was found in the immunoglobulin-like domain and residue pairs located at the molecular paratope:epitope interface important for complex stability was identified. Collectively, the VHH7 homology model and VHH7:RgpB docking supplies knowledge of the residues involved in the high affinity interaction. This information could prove valuable in the design of an antibody-drug conjugate for specific RgpB targeting. Copyright © 2017 Elsevier Inc. All rights reserved.

Current advances in screening for bioactive components from medicinal plants by affinity ultrafiltration mass spectrometry.

Science.gov (United States)

Chen, Guilin; Huang, Bill X; Guo, Mingquan

2018-05-21

Medicinal plants have played an important role in maintaining human health for thousands of years. However, the interactions between the active components in medicinal plants and some certain biological targets during a disease are still unclear in most cases. To conduct the high-throughput screening for small active molecules that can interact with biological targets, which is of great theoretical significance and practical value. The ultrafiltration mass spectrometry (UF-LC/MS) is a powerful bio-analytical method by combining affinity ultrafiltration and liquid chromatography-mass spectrometry (LC/MS), which could rapidly screen and identify small active molecules that bind to biological targets of interest at the same time. Compared with other analytical methods, affinity UF-LC/MS has the characteristics of fast, sensitive and high throughput, and is especially suitable for the complicated extracts of medicinal plants. In this review, the basic principle, characteristics and some most recent challenges in UF-LC/MS have been demonstrated. Meanwhile, the progress and applications of affinity UF-LC/MS in the discovery of the active components from natural medicinal plants and the interactions between small molecules and biological target proteins are also briefly summarised. In addition, the future directions for UF-LC/MS are also prospected. Affinity UF-LC/MS is a powerful tool in studies on the interactions between small active molecules and biological protein targets, especially in the high-throughput screening of active components from the natural medicinal plants. Copyright © 2018 John Wiley & Sons, Ltd.

MEMS fiber-optic Fabry-Perot pressure sensor for high temperature application

Science.gov (United States)

Fang, G. C.; Jia, P. G.; Cao, Q.; Xiong, J. J.

2016-10-01

We design and demonstrate a fiber-optic Fabry-Perot pressure sensor (FOFPPS) for high-temperature sensing by employing micro-electro-mechanical system (MEMS) technology. The FOFPPS is fabricated by anodically bonding the silicon wafer and the Pyrex glass together and fixing the facet of the optical fiber in parallel with the silicon surface by glass frit and organic adhesive. The silicon wafer can be reduced through dry etching technology to construct the sensitive diaphragm. The length of the cavity changes with the deformation of the diaphragm due to the loaded pressure, which leads to a wavelength shift of the interference spectrum. The pressure can be gauged by measuring the wavelength shift. The pressure experimental results show that the sensor has linear pressure sensitivities ranging from 0 kPa to 600 kPa at temperature range between 20°C to 300°C. The pressure sensitivity at 300°C is approximately 27.63 pm/kPa. The pressure sensitivities gradually decrease with increasing the temperature. The sensor also has a linear thermal drift when temperature changes from 20°C - 300°C.

Responses and mechanisms of positive electron affinity molecules in the N2 mode of the thermionic ionization detector and the electron-capture detector

International Nuclear Information System (INIS)

Jones, C.S.

1989-01-01

Very little knowledge has been acquired in the past on the mechanistic pathway by which molecules respond in the N 2 mode of the thermionic ionization detector. An attempt is made here to elucidate the response mechanism of the detector. The basic response mechanisms are known for the electron capture detector, and an attempt is made to identify the certain mechanism by which selected molecules respond. The resonance electron capture rate constant has been believed to be temperature independent, and investigations of the temperature dependence of electron capture responses are presented. Mechanisms for the N 2 mode of the thermionic ionization detector have been proposed by examining the detector response to positive electron affinity molecules and by measurement of the ions produced by the detector. Electron capture mechanisms for selected molecules have been proposed by examining their temperature dependent responses in the electron capture detector and negative ion mass spectra of the samples. In studies of the resonance electron capture rate constant, the relative responses of selected positive electron affinity molecules and their temperature dependent responses were investigated. Positive electron affinity did not guarantee large responses in the N 2 mode thermionic ionization detector. High mass ions were measured following ionization of samples in the detector. Responses in the electron capture detector varied with temperature and electron affinity

Quality and sensitivity of high-resolution numerical simulation of urban heat islands

International Nuclear Information System (INIS)

Li, Dan; Bou-Zeid, Elie

2014-01-01

High-resolution numerical simulations of the urban heat island (UHI) effect with the widely-used Weather Research and Forecasting (WRF) model are assessed. Both the sensitivity of the results to the simulation setup, and the quality of the simulated fields as representations of the real world, are investigated. Results indicate that the WRF-simulated surface temperatures are more sensitive to the planetary boundary layer (PBL) scheme choice during nighttime, and more sensitive to the surface thermal roughness length parameterization during daytime. The urban surface temperatures simulated by WRF are also highly sensitive to the urban canopy model (UCM) used. The implementation in this study of an improved UCM (the Princeton UCM or PUCM) that allows the simulation of heterogeneous urban facets and of key hydrological processes, together with the so-called CZ09 parameterization for the thermal roughness length, significantly reduce the bias (<1.5 °C) in the surface temperature fields as compared to satellite observations during daytime. The boundary layer potential temperature profiles are captured by WRF reasonable well at both urban and rural sites; the biases in these profiles relative to aircraft-mounted senor measurements are on the order of 1.5 °C. Changing UCMs and PBL schemes does not alter the performance of WRF in reproducing bulk boundary layer temperature profiles significantly. The results illustrate the wide range of urban environmental conditions that various configurations of WRF can produce, and the significant biases that should be assessed before inferences are made based on WRF outputs. The optimal set-up of WRF-PUCM developed in this paper also paves the way for a confident exploration of the city-scale impacts of UHI mitigation strategies in the companion paper (Li et al 2014). (letter)

A new real-time method for investigation of affinity properties and binding kinetics of magnetic nanoparticles

Energy Technology Data Exchange (ETDEWEB)

Orlov, Alexey V. [Prokhorov General Physics Institute, Russian Academy of Sciences, 38 Vavilov St, 119991 Moscow (Russian Federation); Nikitin, Maxim P. [Prokhorov General Physics Institute, Russian Academy of Sciences, 38 Vavilov St, 119991 Moscow (Russian Federation); Shemyakin–Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, 16/10 Miklukho-Maklaya St, 117997 Moscow (Russian Federation); Moscow Institute of Physics and Technology, 9 Institutskii per., Dolgoprudny, Moscow Region 141700 (Russian Federation); Bragina, Vera A.; Znoyko, Sergey L.; Zaikina, Marina N.; Ksenevich, Tatiana I.; Gorshkov, Boris G. [Prokhorov General Physics Institute, Russian Academy of Sciences, 38 Vavilov St, 119991 Moscow (Russian Federation); Nikitin, Petr I., E-mail: nikitin@kapella.gpi.ru [Prokhorov General Physics Institute, Russian Academy of Sciences, 38 Vavilov St, 119991 Moscow (Russian Federation); National Research Nuclear University MEPhI (Moscow Engineering Physics Institute), 31 Kashirskoe shosse, 115409 Moscow (Russian Federation)

2015-04-15

A method for quantitative investigation of affinity constants of receptors immobilized on magnetic nanoparticles (MP) is developed based on spectral correlation interferometry (SCI). The SCI records with a picometer resolution the thickness changes of a layer of molecules or nanoparticles due to a biochemical reaction on a cover slip, averaged over the sensing area. The method is compatible with other types of sensing surfaces employed in biosensing. The measured values of kinetic association constants of magnetic nanoparticles are 4 orders of magnitude higher than those of molecular antibody association with antigen. The developed method also suggests highly sensitive detection of antigens in a wide dynamic range. The limit of detection of 92 pg/ml has been demonstrated for prostate-specific antigen (PSA) with 50-nm MP employed as labels, which produce 3-order amplification of the SCI signals. The calibration curve features high sensitivity (slope) of 3-fold signal raise per 10-fold increase of PSA concentration within 4-order dynamic range, which is an attractive compromise for precise quantitative and highly sensitive immunoassay. The proposed biosensing technique offers inexpensive disposable sensor chips of cover slips and represents an economically sound alternative to traditional immunoassays for disease diagnostics, detection of pathogens in food and environmental monitoring. - Highlights: • Method for study of affinity constants of magnetic nanoparticles with receptors is proposed. • Association constants of such particles are 4 orders higher than for biomolecules. • Method is compatible with widely used biosensor surfaces and affordable consumables. • It has high sensitivity: 3-fold signal increasing per 10-fold of PSA concentration. • Limit of detection for PSA is 92 pg/ml, dynamic range – 4 orders of concentration.

Sensitivities and uncertainties of modeled ground temperatures in mountain environments

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S. Gubler

2013-08-01

Full Text Available Model evaluation is often performed at few locations due to the lack of spatially distributed data. Since the quantification of model sensitivities and uncertainties can be performed independently from ground truth measurements, these analyses are suitable to test the influence of environmental variability on model evaluation. In this study, the sensitivities and uncertainties of a physically based mountain permafrost model are quantified within an artificial topography. The setting consists of different elevations and exposures combined with six ground types characterized by porosity and hydraulic properties. The analyses are performed for a combination of all factors, that allows for quantification of the variability of model sensitivities and uncertainties within a whole modeling domain. We found that model sensitivities and uncertainties vary strongly depending on different input factors such as topography or different soil types. The analysis shows that model evaluation performed at single locations may not be representative for the whole modeling domain. For example, the sensitivity of modeled mean annual ground temperature to ground albedo ranges between 0.5 and 4 °C depending on elevation, aspect and the ground type. South-exposed inclined locations are more sensitive to changes in ground albedo than north-exposed slopes since they receive more solar radiation. The sensitivity to ground albedo increases with decreasing elevation due to shorter duration of the snow cover. The sensitivity in the hydraulic properties changes considerably for different ground types: rock or clay, for instance, are not sensitive to uncertainties in the hydraulic properties, while for gravel or peat, accurate estimates of the hydraulic properties significantly improve modeled ground temperatures. The discretization of ground, snow and time have an impact on modeled mean annual ground temperature (MAGT that cannot be neglected (more than 1 °C for several

Glucose uptake and growth of glucose-limited chemostat cultures of Aspergillus niger and a disruptant lacking MstA, a high-affinity glucose transporter

DEFF Research Database (Denmark)

Jørgensen, Thomas R; vanKuyk, Patricia A; Poulsen, Bjarne R

2007-01-01

This is a study of high-affinity glucose uptake in Aspergillus niger and the effect of disruption of a high-affinity monosaccharide-transporter gene, mstA. The substrate saturation constant (K(s)) of a reference strain was about 15 microM in glucose-limited chemostat culture. Disruption of mst......-affinity uptake system of A. niger. The mstA disruptant and a reference strain were cultivated in glucose-limited chemostat cultures at low, intermediate and high dilution rate (D=0.07 h(-1), 0.14 h(-1) and 0.20 h(-1)). Mycelium harvested from steady-state cultures was subjected to glucose uptake assays...

Temperature dependence of heat sensitization and thermotolerance induction with ethanol

International Nuclear Information System (INIS)

Henle, K.J.; Nagle, W.A.; Moss, A.J.

1987-01-01

Cytoxicity of 1 M ethanol was strongly temperature dependent; survival curves between 34 0 and 39 0 C were similar to heat survival curves between 40 and 45 0 without ethanol. Ethanol was non-toxic at 22 0 ; at 34.5 0 and 35.5 0 ethanol survival curves were biphasic. The major effect of 1 M ethanol was an effective temperature shift of 6.4 Celsius degrees, although temperatures between 34 0 and 36 0 caused additional sensitization reminiscent of the stepdown heating phenomenon. Induction of thermotolerance with equitoxic ethanol exposures at 35.5 0 and 37 0 or with heat alone (10 min, 45 0 ) resulted in tolerance development with similar kinetics; in contrast, ethanol exposures at 22 0 did not induce any tolerance development with similar kinetics; in contrast, ethanol exposures at 22 0 did not induce any tolerance to hyperthermia. These data provide a rationale for conflicting reports in the literature regarding thermotolerance induction by ethanol and suggest that ethanol causes ''heat'' stress at temperatures that are generally considered to be physiological. This interpretation predicts that the use of ethanol and other organic solvents in high concentrations will cause effects at 37 0 that normally occur only at hyperthermic temperatures, including membrane perturbations and HSP synthesis, and that ''physiological'' temperatures must be precisely controlled under those conditions

The selectively bred high alcohol sensitivity (HAS) and low alcohol sensitivity (LAS) rats differ in sensitivity to nicotine.

Science.gov (United States)

de Fiebre, NancyEllen C; Dawson, Ralph; de Fiebre, Christopher M

2002-06-01

Studies in rodents selectively bred to differ in alcohol sensitivity have suggested that nicotine and ethanol sensitivities may cosegregate during selective breeding. This suggests that ethanol and nicotine sensitivities may in part be genetically correlated. Male and female high alcohol sensitivity (HAS), control alcohol sensitivity, and low alcohol sensitivity (LAS) rats were tested for nicotine-induced alterations in locomotor activity, body temperature, and seizure activity. Plasma and brain levels of nicotine and its primary metabolite, cotinine, were measured in these animals, as was the binding of [3H]cytisine, [3H]epibatidine, and [125I]alpha-bungarotoxin in eight brain regions. Both replicate HAS lines were more sensitive to nicotine-induced locomotor activity depression than the replicate LAS lines. No consistent HAS/LAS differences were seen on other measures of nicotine sensitivity; however, females were more susceptible to nicotine-induced seizures than males. No HAS/LAS differences in nicotine or cotinine levels were seen, nor were differences seen in the binding of nicotinic ligands. Females had higher levels of plasma cotinine and brain nicotine than males but had lower brain cotinine levels than males. Sensitivity to a specific action of nicotine cosegregates during selective breeding for differential sensitivity to a specific action of ethanol. The differential sensitivity of the HAS/LAS rats is due to differences in central nervous system sensitivity and not to pharmacokinetic differences. The differential central nervous system sensitivity cannot be explained by differences in the numbers of nicotinic receptors labeled in ligand-binding experiments. The apparent genetic correlation between ethanol and nicotine sensitivities suggests that common genes modulate, in part, the actions of both ethanol and nicotine and may explain the frequent coabuse of these agents.

A Novel Recombinant DNA System for High Efficiency Affinity Purification of Proteins in Saccharomyces cerevisiae

Directory of Open Access Journals (Sweden)

Brian H. Carrick

2016-03-01

Full Text Available Isolation of endogenous proteins from Saccharomyces cerevisiae has been facilitated by inserting encoding polypeptide affinity tags at the C-termini of chromosomal open reading frames (ORFs using homologous recombination of DNA fragments. Tagged protein isolation is limited by a number of factors, including high cost of affinity resins for bulk isolation and low concentration of ligands on the resin surface, leading to low isolation efficiencies and trapping of contaminants. To address this, we have created a recombinant “CelTag” DNA construct from which PCR fragments can be created to easily tag C-termini of S. cerevisiae ORFs using selection for a nat1 marker. The tag has a C-terminal cellulose binding module to be used in the first affinity step. Microgranular cellulose is very inexpensive and has an effectively continuous ligand on its surface, allowing rapid, highly efficient purification with minimal background. Cellulose-bound proteins are released by specific cleavage of an included site for TEV protease, giving nearly pure product. The tag can be lifted from the recombinant DNA construct either with or without a 13x myc epitope tag between the target ORF and the TEV protease site. Binding of CelTag protein fusions to cellulose is stable to high salt, nonionic detergents, and 1 M urea, allowing stringent washing conditions to remove loosely associated components, as needed, before specific elution. It is anticipated that this reagent could allow isolation of protein complexes from large quantities of yeast extract, including soluble, membrane-bound, or nucleic acid-associated assemblies.

Discovery of PF-06928215 as a high affinity inhibitor of cGAS enabled by a novel fluorescence polarization assay

Energy Technology Data Exchange (ETDEWEB)

Hall, Justin; Brault, Amy; Vincent, Fabien; Weng, Shawn; Wang, Hong; Dumlao, Darren; Aulabaugh, Ann; Aivazian, Dikran; Castro, Dana; Chen, Ming; Culp, Jeffrey; Dower, Ken; Gardner, Joseph; Hawrylik, Steven; Golenbock, Douglas; Hepworth, David; Horn, Mark; Jones, Lyn; Jones, Peter; Latz, Eicke; Li, Jing; Lin, Lih-Ling; Lin, Wen; Lin, David; Lovering, Frank; Niljanskul, Nootaree; Nistler, Ryan; Pierce, Betsy; Plotnikova, Olga; Schmitt, Daniel; Shanker, Suman; Smith, James; Snyder, William; Subashi, Timothy; Trujillo, John; Tyminski, Edyta; Wang, Guoxing; Wong, Jimson; Lefker, Bruce; Dakin, Leslie; Leach, Karen (UMASS, MED); (Pfizer)

2017-09-21

Cyclic GMP-AMP synthase (cGAS) initiates the innate immune system in response to cytosolic dsDNA. After binding and activation from dsDNA, cGAS uses ATP and GTP to synthesize 2', 3' -cGAMP (cGAMP), a cyclic dinucleotide second messenger with mixed 2'-5' and 3'-5' phosphodiester bonds. Inappropriate stimulation of cGAS has been implicated in autoimmune disease such as systemic lupus erythematosus, thus inhibition of cGAS may be of therapeutic benefit in some diseases; however, the size and polarity of the cGAS active site makes it a challenging target for the development of conventional substrate-competitive inhibitors. We report here the development of a high affinity (K_D = 200 nM) inhibitor from a low affinity fragment hit with supporting biochemical and structural data showing these molecules bind to the cGAS active site. We also report a new high throughput cGAS fluorescence polarization (FP)-based assay to enable the rapid identification and optimization of cGAS inhibitors. This FP assay uses Cy5-labelled cGAMP in combination with a novel high affinity monoclonal antibody that specifically recognizes cGAMP with no cross reactivity to cAMP, cGMP, ATP, or GTP. Given its role in the innate immune response, cGAS is a promising therapeutic target for autoinflammatory disease. Our results demonstrate its druggability, provide a high affinity tool compound, and establish a high throughput assay for the identification of next generation cGAS inhibitors.

Boundary integral method to calculate the sensitivity temperature error of microstructured fibre plasmonic sensors

International Nuclear Information System (INIS)

Esmaeilzadeh, Hamid; Arzi, Ezatollah; Légaré, François; Hassani, Alireza

2013-01-01

In this paper, using the boundary integral method (BIM), we simulate the effect of temperature fluctuation on the sensitivity of microstructured optical fibre (MOF) surface plasmon resonance (SPR) sensors. The final results indicate that, as the temperature increases, the refractometry sensitivity of our sensor decreases from 1300 nm/RIU at 0 °C to 1200 nm/RIU at 50 °C, leading to ∼7.7% sensitivity reduction and the sensitivity temperature error of 0.15% °C −1 for this case. These results can be used for biosensing temperature-error adjustment in MOF SPR sensors, since biomaterials detection usually happens in this temperature range. Moreover, the signal-to-noise ratio (SNR) of our sensor decreases from 0.265 at 0 °C to 0.154 at 100 °C with the average reduction rate of ∼0.42% °C −1 . The results suggest that at lower temperatures the sensor has a higher SNR. (paper)

Topography of the high-affinity lysine binding site of plasminogen as defined with a specific antibody probe

International Nuclear Information System (INIS)

Miles, L.A.; Plow, E.F.

1986-01-01

An antibody population that reacted with the high-affinity lysine binding site of human plasminogen was elicited by immunizing rabbits with an elastase degradation product containing kringles 1-3 (EDP I). This antibody was immunopurified by affinity chromatography on plasminogen-Sepharose and elution with 0.2 M 6-aminohexanoic acid. The eluted antibodies bound [ 125 I]EDP I, [ 125 I]Glu-plasminogen, and [ 125 I]Lys-plasminogen in radioimmunoassays, and binding of each ligand was at least 99% inhibited by 0.2 M 6-aminohexanoic acid. The concentrations for 50% inhibition of [ 125 I]EDP I binding by tranexamic acid, 6-aminohexanoic acid, and lysine were 2.6, 46, and l730 μM, respectively. Similar values were obtained with plasminogen and suggested that an unoccupied high-affinity lysine binding site was required for antibody recognition. The antiserum reacted exclusively with plasminogen derivatives containing the EDP I region and did not react with those lacking an EDP I region, or with tissue plasminogen activator or prothrombin, which also contains kringles. By immunoblotting analyses, a chymotryptic degradation product of M/sub r/ 20,000 was derived from EDP I that retained reactivity with the antibody. α 2 -Antiplasmin inhibited the binding of radiolabeled EDP I, Glu-plasminogen, or Lys-plasminogen by the antiserum, suggesting that the recognized site is involved in the noncovalent interaction of the inhibitor with plasminogen. The binding of [ 125 I]EDP I to fibrin was also inhibited by the antiserum. The observations provide independent evidence for the role of the high-affinity lysine binding site in the functional interactions of plasminogen with its primary substrate and inhibitor

A High-Temperature Piezoresistive Pressure Sensor with an Integrated Signal-Conditioning Circuit

Directory of Open Access Journals (Sweden)

Zong Yao

2016-06-01

Full Text Available This paper focuses on the design and fabrication of a high-temperature piezoresistive pressure sensor with an integrated signal-conditioning circuit, which consists of an encapsulated pressure-sensitive chip, a temperature compensation circuit and a signal-conditioning circuit. A silicon on insulation (SOI material and a standard MEMS process are used in the pressure-sensitive chip fabrication, and high-temperature electronic components are adopted in the temperature-compensation and signal-conditioning circuits. The entire pressure sensor achieves a hermetic seal and can be operated long-term in the range of −50 °C to 220 °C. Unlike traditional pressure sensor output voltage ranges (in the dozens to hundreds of millivolts, the output voltage of this sensor is from 0 V to 5 V, which can significantly improve the signal-to-noise ratio and measurement accuracy in practical applications of long-term transmission based on experimental verification. Furthermore, because this flexible sensor’s output voltage is adjustable, general follow-up pressure transmitter devices for voltage converters need not be used, which greatly reduces the cost of the test system. Thus, the proposed high-temperature piezoresistive pressure sensor with an integrated signal-conditioning circuit is expected to be highly applicable to pressure measurements in harsh environments.

Titanium dioxide as chemo-affinity chromatographic sorbent of biomolecular compounds - Applications in acidic modification-specific proteomics

DEFF Research Database (Denmark)

Engholm-Keller, Kasper; Larsen, Martin R

2011-01-01

biomolecules due to its unique ion and ligand exchange properties and high stability towards pH and temperature. Recently, titanium dioxide chromatography was introduced in proteomics as a highly specific method for enriching phosphorylated peptides - a method, which has been widely adapted by the field...... matrices for further characterization is affinity chromatography, which relies on the specific interaction between an analyte in solution and a solid adsorbent. Titanium dioxide-based affinity chromatography has proven to be a versatile tool in enrichment of various compounds such as phosphorylated....... The development of TiO(2)-based chromatographic strategies for separation of various biomolecules from its introduction for small molecules more than 20years ago until recent proteomics applications today will be reviewed here....

Visual and Plasmon Resonance Absorption Sensor for Adenosine Triphosphate Based on the High Affinity between Phosphate and Zr(IV)

OpenAIRE

Qi, Wenjing; Liu, Zhongyuan; Zhang, Wei; Halawa, Mohamed Ibrahim; Xu, Guobao

2016-01-01

Zr(IV) can form phosphate and Zr(IV) (?PO3 2??Zr4+?) complex owing to the high affinity between Zr(IV) with phosphate. Zr(IV) can induce the aggregation of gold nanoparticles (AuNPs), while adenosine triphosphate(ATP) can prevent Zr(IV)-induced aggregation of AuNPs. Herein, a visual and plasmon resonance absorption (PRA)sensor for ATP have been developed using AuNPs based on the high affinity between Zr(IV)with ATP. AuNPs get aggregated in the presence of certain concentrations of Zr(IV). Aft...

Quality and sensitivity of high-resolution numerical simulation of urban heat islands

Science.gov (United States)

Li, Dan; Bou-Zeid, Elie

2014-05-01

High-resolution numerical simulations of the urban heat island (UHI) effect with the widely-used Weather Research and Forecasting (WRF) model are assessed. Both the sensitivity of the results to the simulation setup, and the quality of the simulated fields as representations of the real world, are investigated. Results indicate that the WRF-simulated surface temperatures are more sensitive to the planetary boundary layer (PBL) scheme choice during nighttime, and more sensitive to the surface thermal roughness length parameterization during daytime. The urban surface temperatures simulated by WRF are also highly sensitive to the urban canopy model (UCM) used. The implementation in this study of an improved UCM (the Princeton UCM or PUCM) that allows the simulation of heterogeneous urban facets and of key hydrological processes, together with the so-called CZ09 parameterization for the thermal roughness length, significantly reduce the bias (Changing UCMs and PBL schemes does not alter the performance of WRF in reproducing bulk boundary layer temperature profiles significantly. The results illustrate the wide range of urban environmental conditions that various configurations of WRF can produce, and the significant biases that should be assessed before inferences are made based on WRF outputs. The optimal set-up of WRF-PUCM developed in this paper also paves the way for a confident exploration of the city-scale impacts of UHI mitigation strategies in the companion paper (Li et al 2014).

A highly sensitive and selective aptamer-based colorimetric sensor for the rapid detection of PCB 77.

Science.gov (United States)

Cheng, Ruojie; Liu, Siyao; Shi, Huijie; Zhao, Guohua

2018-01-05

A highly sensitive, specific and simple colorimetric sensor based on aptamer was established for the detection of polychlorinated biphenyls (PCB 77). The use of unmodified gold nanoparticles as a colorimetric probe for aptamer sensors enabled the highly sensitive and selective detection of polychlorinated biphenyls (PCB 77). A linear range of 0.5nM to 900nM was obtained for the colorimetric assay with a minimum detection limit of 0.05nM. In addition, by the methods of circular dichroism, UV and naked eyes, we found that the 35 base fragments retained after cutting 5 bases from the 5 'end of aptamer plays the most significant role in the PCB 77 specific recognition process. We found a novel way to truncated nucleotides to optimize the detection of PCB 77, and the selected nucleotides also could achieve high affinity with PCB 77. At the same time, the efficient detection of the PCB 77 by our colorimetric sensor in the complex environmental water samples was realized, which shows a good application prospect. Copyright © 2017 Elsevier B.V. All rights reserved.

Study on temperature sensitivity of topological insulators based on long-period fiber grating

Science.gov (United States)

Luo, Jianhua; Zhao, Chenghai; Li, Jianbo; He, Mengdong

2017-06-01

Based on a long-period fiber grating, we conducted experimental research on the temperature sensitivity of topological insulators. The long-period fiber grating and topological insulators solution were encapsulated in a capillary tube using UV glue, and the temperature response was measured. Within a range of 35 to 75 centigrade, one resonance dip of a long-period fiber grating exhibits a redshift of 1.536 nm. The temperature sensitivity is about 7.7 times of an ordinary long-period fiber grating's sensitivity (0.005 nm/°C). A numerical simulation is also performed on the basis of the experiments.

14-O-Methylmorphine: A Novel Selective Mu-Opioid Receptor Agonist with High Efficacy and Affinity.

Science.gov (United States)

Zádor, Ferenc; Balogh, Mihály; Váradi, András; Zádori, Zoltán S; Király, Kornél; Szűcs, Edina; Varga, Bence; Lázár, Bernadette; Hosztafi, Sándor; Riba, Pál; Benyhe, Sándor; Fürst, Susanna; Al-Khrasani, Mahmoud

2017-11-05

14-O-methyl (14-O-Me) group in morphine-6-O-sulfate (M6SU) or oxymorphone has been reported to be essential for enhanced affinity, potency and antinociceptive effect of these opioids. Herein we report on the pharmacological properties (potency, affinity and efficacy) of the new compound, 14-O-methylmorphine (14-O-MeM) in in vitro. Additionally, we also investigated the antinociceptive effect of the novel compound, as well as its inhibitory action on gastrointestinal transit in in vivo. The potency and efficacy of test compound were measured by [ 35 S]GTPγS binding, isolated mouse vas deferens (MVD) and rat vas deferens (RVD) assays. The affinity of 14-O-MeM for opioid receptors was assessed by radioligand binding and MVD assays. The antinociceptive and gastrointestinal effects of the novel compound were evaluated in the rat tail-flick test and charcoal meal test, respectively. Morphine, DAMGO, Ile 5,6 deltorphin II, deltorphin II and U-69593 were used as reference compounds. 14-O-MeM showed higher efficacy (E max ) and potency (EC 50 ) than morphine in MVD, RVD or [ 35 S]GTPγS binding. In addition, 14-O-MeM compared to morphine showed higher affinity for μ-opioid receptor (MOR). In vivo, in rat tail-flick test 14-O-MeM proved to be stronger antinociceptive agent than morphine after peripheral or central administration. Additionally, both compounds inhibited the gastrointestinal peristalsis. However, when the antinociceptive and antitransit doses for each test compound are compared, 14-O-MeM proved to have slightly more favorable pharmacological profile. Our results affirm that 14-O-MeM, an opioid of high efficacy and affinity for MOR can be considered as a novel analgesic agent of potential clinical value. Copyright © 2017 Elsevier B.V. All rights reserved.

Novel high-resolution temperature probe for radiofrequency dosimetry

Energy Technology Data Exchange (ETDEWEB)

Schuderer, Juergen [Foundation for Research on Information Technologies in Society (IT' IS), Integrated Systems Laboratory IIS, Swiss Federal Institute of Technology (ETH), CH-8092 Zurich (Switzerland); Schmid, Thomas [Schmid and Partner Engineering AG, 8004 Zurich (Switzerland); Urban, Gerald [IMTEK, Albert-Ludwigs University Freiburg, 79110 Freiburg (Germany); Samaras, Theodoros [Department of Physics, Aristotle University of Thessaloniki, 54124 Thessaloniki (Greece); Kuster, Niels [Foundation for Research on Information Technologies in Society (IT' IS), Integrated Systems Laboratory IIS, Swiss Federal Institute of Technology (ETH), CH-8092 Zurich (Switzerland)

2004-03-21

A novel integrated thermistor probe for temperature evaluations in radiofrequency-heated environments was realized. The probe's sensitive area is based on a highly resistive 50 {mu}m x 100 {mu}m layer of amorphous germanium processed on a glass tip. The small dimensions allow measurements with a distance as close as 150 {mu}m from solid boundaries. Due to its high temperature resolution of 4 mK and its short response time of the order of 10 ms, the sensor is very well suited for dosimetric measurements in strong absorption gradients. The influence of radiofrequency (RF) electric fields on the signal is minimized due to the high resistance of the sensor and the leads. The probe was successfully used to determine the highly nonuniform absorption distribution resulting from the RF exposure of cell cultures placed in Petri dishes. (note)

Novel high-resolution temperature probe for radiofrequency dosimetry

International Nuclear Information System (INIS)

Schuderer, Juergen; Schmid, Thomas; Urban, Gerald; Samaras, Theodoros; Kuster, Niels

2004-01-01

A novel integrated thermistor probe for temperature evaluations in radiofrequency-heated environments was realized. The probe's sensitive area is based on a highly resistive 50 μm x 100 μm layer of amorphous germanium processed on a glass tip. The small dimensions allow measurements with a distance as close as 150 μm from solid boundaries. Due to its high temperature resolution of 4 mK and its short response time of the order of 10 ms, the sensor is very well suited for dosimetric measurements in strong absorption gradients. The influence of radiofrequency (RF) electric fields on the signal is minimized due to the high resistance of the sensor and the leads. The probe was successfully used to determine the highly nonuniform absorption distribution resulting from the RF exposure of cell cultures placed in Petri dishes. (note)

Affinity-tuning leukocyte integrin for development of safe therapeutics

Science.gov (United States)

Park, Spencer

Much attention has been given to the molecular and cellular pathways linking inflammation with cancer and the local tumor environment to identify new target molecules that could lead to improved diagnosis and treatment. Among the many molecular players involved in the complex response, central to the induction of inflammation is intercellular adhesion molecule (ICAM)-1, which is of particular interest for its highly sensitive and localized expression in response to inflammatory signals. ICAM-1, which has been implicated to play a critical role in tumor progression in various types of cancer, has also been linked to cancer metastases, where ICAM-1 facilitates the spread of metastatic cancer cells to secondary sites. This unique expression profile of ICAM-1 throughout solid tumor microenvironment makes ICAM-1 an intriguing molecular target, which holds great potential as an important diagnostic and therapeutic tool. Herein, we have engineered the ligand binding domain, or the inserted (I) domain of a leukocyte integrin, to exhibit a wide range of monovalent affinities to the natural ligand, ICAM-1. Using the resulting I domain variants, we have created drug and gene delivery nanoparticles, as well as targeted immunotherapeutics that have the ability to bind and migrate to inflammatory sites prevalent in tumors and the associated microenvironment. Through the delivery of diagnostic agents, chemotherapeutics, and immunotherapeutics, the following chapters demonstrate that the affinity enhancements achieved by directed evolution bring the affinity of I domains into the range optimal for numerous applications.

High-sensitivity β-Ga_2O_3 solar-blind photodetector on high-temperature pretreated c-plane sapphire substrate

KAUST Repository

Qian, Ling-Xuan; Zhang, Hua-Fan; Lai, P. T.; Wu, Ze-Han; Liu, Xing-Zhao

2017-01-01

-annealing temperature must be sufficiently high to offset the rise of the dark current and thus achieve a remarkable improvement in the photodetection properties. As a result, the PD fabricated on the 1050 degrees C-annealed substrate exhibited extremely high

Development and application of a species sensitivity distribution for temperature-induced mortality in the aquatic environment

NARCIS (Netherlands)

Vries, de P.; Tamis, J.E.; Murk, A.J.; Smit, M.G.D.

2008-01-01

Current European legislation has static water quality objectives for temperature effects, based on the most sensitive species. In the present study a species sensitivity distribution (SSD) for elevated temperatures is developed on the basis of temperature sensitivity data (mortality) of 50 aquatic

Acylated heptapeptide binds albumin with high affinity and application as tag furnishes long-acting peptides.

Science.gov (United States)

Zorzi, Alessandro; Middendorp, Simon J; Wilbs, Jonas; Deyle, Kaycie; Heinis, Christian

2017-07-17

The rapid renal clearance of peptides in vivo limits this attractive platform for the treatment of a broad range of diseases that require prolonged drug half-lives. An intriguing approach for extending peptide circulation times works through a 'piggy-back' strategy in which peptides bind via a ligand to the long-lived serum protein albumin. In accordance with this strategy, we developed an easily synthesized albumin-binding ligand based on a peptide-fatty acid chimera that has a high affinity for human albumin (K d =39 nM). This ligand prolongs the elimination half-life of cyclic peptides in rats 25-fold to over seven hours. Conjugation to a peptide factor XII inhibitor developed for anti-thrombotic therapy extends the half-life from 13 minutes to over five hours, inhibiting coagulation for eight hours in rabbits. This high-affinity albumin ligand could potentially extend the half-life of peptides in human to several days, substantially broadening the application range of peptides as therapeutics.

Acylated heptapeptide binds albumin with high affinity and application as tag furnishes long-acting peptides

Science.gov (United States)

Zorzi, Alessandro; Middendorp, Simon J.; Wilbs, Jonas; Deyle, Kaycie; Heinis, Christian

2017-07-01

The rapid renal clearance of peptides in vivo limits this attractive platform for the treatment of a broad range of diseases that require prolonged drug half-lives. An intriguing approach for extending peptide circulation times works through a `piggy-back' strategy in which peptides bind via a ligand to the long-lived serum protein albumin. In accordance with this strategy, we developed an easily synthesized albumin-binding ligand based on a peptide-fatty acid chimera that has a high affinity for human albumin (Kd=39 nM). This ligand prolongs the elimination half-life of cyclic peptides in rats 25-fold to over seven hours. Conjugation to a peptide factor XII inhibitor developed for anti-thrombotic therapy extends the half-life from 13 minutes to over five hours, inhibiting coagulation for eight hours in rabbits. This high-affinity albumin ligand could potentially extend the half-life of peptides in human to several days, substantially broadening the application range of peptides as therapeutics.

Multiple temperature effects on up-conversion fluorescences of Er3+-Y b3+-Mo6+ codoped TiO2 and high thermal sensitivity

Directory of Open Access Journals (Sweden)

B. S. Cao

2015-08-01

Full Text Available We report multiple temperature effects on green and red up-conversion emissions in Er3+-Y b3+-Mo6+ codoped TiO2 phosphors. With increasing temperature, the decrease of the red emission from 4F9/2→4I15/2, the increase of green emission from 2H11/2→4I15/2 and another unchanged green emission from 4S3/2→4I15/2 were simultaneously observed, which are explained by steady-state rate equations analysis. Due to different evolution with temperature of the two green emissions, higher thermal sensitivity of optical thermal sensor was obtained based on the transitions with the largest fluorescence intensity ratio. Two parameters, maximum theoretical sensitivity (Smax and optimum operating temperature (Tmax are given to describe thermal sensing properties of the produced sensors. The intensity ratio and energy difference ΔE of a pair of energy levels are two main factors for the sensitivity and accuracy of sensors, which should be referred to design sensors with optimized sensing properties.

Physiological epidermal growth factor concentrations activate high affinity receptors to elicit calcium oscillations.

Directory of Open Access Journals (Sweden)

Béatrice Marquèze-Pouey

Full Text Available Signaling mediated by the epidermal growth factor (EGF is crucial in tissue development, homeostasis and tumorigenesis. EGF is mitogenic at picomolar concentrations and is known to bind its receptor on high affinity binding sites depending of the oligomerization state of the receptor (monomer or dimer. In spite of these observations, the cellular response induced by EGF has been mainly characterized for nanomolar concentrations of the growth factor, and a clear definition of the cellular response to circulating (picomolar concentrations is still lacking. We investigated Ca2+ signaling, an early event in EGF responses, in response to picomolar doses in COS-7 cells where the monomer/dimer equilibrium is unaltered by the synthesis of exogenous EGFR. Using the fluo5F Ca2+ indicator, we found that picomolar concentrations of EGF induced in 50% of the cells a robust oscillatory Ca2+ signal quantitatively similar to the Ca2+ signal induced by nanomolar concentrations. However, responses to nanomolar and picomolar concentrations differed in their underlying mechanisms as the picomolar EGF response involved essentially plasma membrane Ca2+ channels that are not activated by internal Ca2+ store depletion, while the nanomolar EGF response involved internal Ca2+ release. Moreover, while the picomolar EGF response was modulated by charybdotoxin-sensitive K+ channels, the nanomolar response was insensitive to the blockade of these ion channels.

Physiological epidermal growth factor concentrations activate high affinity receptors to elicit calcium oscillations.

Science.gov (United States)

Marquèze-Pouey, Béatrice; Mailfert, Sébastien; Rouger, Vincent; Goaillard, Jean-Marc; Marguet, Didier

2014-01-01

Signaling mediated by the epidermal growth factor (EGF) is crucial in tissue development, homeostasis and tumorigenesis. EGF is mitogenic at picomolar concentrations and is known to bind its receptor on high affinity binding sites depending of the oligomerization state of the receptor (monomer or dimer). In spite of these observations, the cellular response induced by EGF has been mainly characterized for nanomolar concentrations of the growth factor, and a clear definition of the cellular response to circulating (picomolar) concentrations is still lacking. We investigated Ca2+ signaling, an early event in EGF responses, in response to picomolar doses in COS-7 cells where the monomer/dimer equilibrium is unaltered by the synthesis of exogenous EGFR. Using the fluo5F Ca2+ indicator, we found that picomolar concentrations of EGF induced in 50% of the cells a robust oscillatory Ca2+ signal quantitatively similar to the Ca2+ signal induced by nanomolar concentrations. However, responses to nanomolar and picomolar concentrations differed in their underlying mechanisms as the picomolar EGF response involved essentially plasma membrane Ca2+ channels that are not activated by internal Ca2+ store depletion, while the nanomolar EGF response involved internal Ca2+ release. Moreover, while the picomolar EGF response was modulated by charybdotoxin-sensitive K+ channels, the nanomolar response was insensitive to the blockade of these ion channels.

Temperature-sensitive host range mutants of herpes simplex virus type 2

International Nuclear Information System (INIS)

Koment, R.W.; Rapp, F.

1975-01-01

Herpesviruses are capable of several types of infection of a host cell. To investigate the early events which ultimately determine the nature of the virus-host cell interaction, a system was established utilizing temperature-sensitive mutants of herpes simplex virus type 2. Four mutants have been isolated which fail to induce cytopathic effects and do not replicate at 39 C in hamster embryo fibroblast cells. At least one mutant is virus DNA negative. Since intracellular complementation is detectable between pairs of mutants, a virus function is known to be temperature sensitive. However, all four mutants induce cytopathic effects and replicate to parental virus levels in rabbit kidney cells at 39 C. This suggests that a host cell function, lacking or nonfunctional in HEF cells but present in rabbit kidney cells at 39 C, is required for the replication of these mutants in hamster embryo fibroblast cells at 39 C. Therefore, we conclude that these mutants are both temperature sensitive and exhibit host range properties

Temperature sensitivity of the penicillin-induced autolysis mechanism in nongrowing cultures of Escherichia coli.

OpenAIRE

Kusser, W; Ishiguro, E E

1987-01-01

The effect of incubation temperature on the ampicillin-induced autolysis of nongrowing Escherichia coli was determined. The autolysis mechanisms in amino acid-deprived relA mutant cells treated with chloramphenicol were temperature sensitive. This temperature-sensitive autolysis was demonstrated in three independent ways: turbidimetric determinations, viable cell counts, and solubilization of radiolabeled peptidoglycan.

Hirota's solitons in the affine and the conformal affine Toda models

International Nuclear Information System (INIS)

Aratyn, H.; Constantinidis, C.P.; Ferreira, L.A.; Gomes, J.F.; Zimerman, A.H.

1993-01-01

We use Hirota's method formulated as a recursive scheme to construct a complete set of soliton solutions for the affine Toda field theory based on an arbitrary Lie algebra. Our solutions include a new class of solitons connected with two different types of degeneracies encountered in Hirota's perturbation approach. We also derive an universal mass formula for all Hirota's solutions to the affine Toda model valid for all underlying Lie groups. Embedding of the affine Toda model in the conformal affine Toda model plays a crucial role in this analysis. (orig.)

Improving image segmentation by learning region affinities

Energy Technology Data Exchange (ETDEWEB)

Prasad, Lakshman [Los Alamos National Laboratory; Yang, Xingwei [TEMPLE UNIV.; Latecki, Longin J [TEMPLE UNIV.

2010-11-03

We utilize the context information of other regions in hierarchical image segmentation to learn new regions affinities. It is well known that a single choice of quantization of an image space is highly unlikely to be a common optimal quantization level for all categories. Each level of quantization has its own benefits. Therefore, we utilize the hierarchical information among different quantizations as well as spatial proximity of their regions. The proposed affinity learning takes into account higher order relations among image regions, both local and long range relations, making it robust to instabilities and errors of the original, pairwise region affinities. Once the learnt affinities are obtained, we use a standard image segmentation algorithm to get the final segmentation. Moreover, the learnt affinities can be naturally unutilized in interactive segmentation. Experimental results on Berkeley Segmentation Dataset and MSRC Object Recognition Dataset are comparable and in some aspects better than the state-of-art methods.

Selection of antigenic markers on a GFP-Cκ fusion scaffold with high sensitivity by eukaryotic ribosome display

International Nuclear Information System (INIS)

Yang Yongmin; Barankiewicz, Teresa J.; He Mingyue; Taussig, Michael J.; Chen, Swey-Shen

2007-01-01

Ribosome display is a cell-free system permitting gene selection through the physical association of genetic material (mRNA) and its phenotypic (protein) product. While often used to select single-chain antibodies from large libraries by panning against immobilized antigens, we have adapted ribosome display for use in the 'reverse' format in order to select high affinity antigenic determinants against solid-phase antibody. To create an antigenic scaffold, DNA encoding green fluorescent protein (GFP) was fused to a light chain constant domain (Cκ) with stop codon deleted, and with 5' signals (T7 promoter, Kozak) enabling coupled transcription/translation in a eukaryotic cell-free system. Epitopes on either GFP (5') or Cκ (3') were selected by anti-GFP or anti-Cκ antibodies, respectively, coupled to magnetic beads. After selection, mRNA was amplified directly from protein-ribosome-mRNA (PRM) complexes by in situ PCR followed by internal amplification and reassembly PCR. As little as 10 fg of the 1 kb DNA construct, i.e. approximately 7500 molecules, could be recovered following a single round of interaction with solid-phase anti-GFP antibody. This platform is highly specific and sensitive for the antigen-antibody interaction and may permit selection and reshaping of high affinity antigenic variants of scaffold proteins

Influence of Temperature Variation on Optical Receiver Sensitivity and its Compensation

Directory of Open Access Journals (Sweden)

A. Prokes

2007-09-01

Full Text Available In the paper, the influence of temperature variation on the sensitivity of an avalanche-photodiode-based optical receiver applied in the free space optical communication link is discussed. Communication systems of this type are exposed to a wide range of operating temperatures, which markedly affect many photodiode and preamplifier parameters. The paper presents a receiver sensitivity calculation, taking into consideration the temperature dependence of avalanche photodiode gain, excess noise factor, dark current and thermal noise of preamplifier resistances, and describes the compensation of temperature effects on photodiode gain based on a corresponding change in the reverse voltage applied to the diode. The calculations are demonstrated on the connection of a small-area silicon APD operating in the wavelength range from 820 to 1150 nm with a transimpedance preamplifier using a bipolar junction transistor.

The solutions of affine and conformal affine Toda field theory

International Nuclear Information System (INIS)

Papadopoulos, G.; Spence, B.

1994-02-01

We give new formulations of the solutions of the field equations of the affine Toda and conformal affine Toda theories on a cylinder and two-dimensional Minkowski space-time. These solutions are parameterised in terms of initial data and the resulting covariant phase spaces are diffeomorphic to the Hamiltonian ones. We derive the fundamental Poisson brackets of the parameters of the solutions and give the general static solutions for the affine theory. (authors). 10 refs

Low-Temperature Crystalline Titanium Dioxide by Atomic Layer Deposition for Dye-Sensitized Solar Cells

KAUST Repository

Chandiran, Aravind Kumar

2013-04-24

Low-temperature processing of dye-sensitized solar cells (DSCs) is crucial to enable commercialization with low-cost, plastic substrates. Prior studies have focused on mechanical compression of premade particles on plastic or glass substrates; however, this did not yield sufficient interconnections for good carrier transport. Furthermore, such compression can lead to more heterogeneous porosity. To circumvent these problems, we have developed a low-temperature processing route for photoanodes where crystalline TiO2 is deposited onto well-defined, mesoporous templates. The TiO2 is grown by atomic layer deposition (ALD), and the crystalline films are achieved at a growth temperature of 200 C. The ALD TiO2 thickness was systematically studied in terms of charge transport and performance to lead to optimized photovoltaic performance. We found that a 15 nm TiO2 overlayer on an 8 μm thick SiO2 film leads to a high power conversion efficiency of 7.1% with the state-of-the-art zinc porphyrin sensitizer and cobalt bipyridine redox mediator. © 2013 American Chemical Society.

Analysis of fatigue reliability for high temperature and high pressure multi-stage decompression control valve

Science.gov (United States)

Yu, Long; Xu, Juanjuan; Zhang, Lifang; Xu, Xiaogang

2018-03-01

Based on stress-strength interference theory to establish the reliability mathematical model for high temperature and high pressure multi-stage decompression control valve (HMDCV), and introduced to the temperature correction coefficient for revising material fatigue limit at high temperature. Reliability of key dangerous components and fatigue sensitivity curve of each component are calculated and analyzed by the means, which are analyzed the fatigue life of control valve and combined with reliability theory of control valve model. The impact proportion of each component on the control valve system fatigue failure was obtained. The results is shown that temperature correction factor makes the theoretical calculations of reliability more accurate, prediction life expectancy of main pressure parts accords with the technical requirements, and valve body and the sleeve have obvious influence on control system reliability, the stress concentration in key part of control valve can be reduced in the design process by improving structure.

Bimodular high temperature planar oxygen gas sensor

Directory of Open Access Journals (Sweden)

Xiangcheng eSun

2014-08-01

Full Text Available A bimodular planar O2 sensor was fabricated using NiO nanoparticles (NPs thin film coated yttria-stabilized zirconia (YSZ substrate. The thin film was prepared by radio frequency (r.f. magnetron sputtering of NiO on YSZ substrate, followed by high temperature sintering. The surface morphology of NiO nanoparticles film was characterized by atomic force microscopy (AFM and scanning electron microscopy (SEM. X-ray diffraction (XRD patterns of NiO NPs thin film before and after high temperature O2 sensing demonstrated that the sensing material possesses a good chemical and structure stability. The oxygen detection experiments were performed at 500 °C, 600 °C and 800 °C using the as-prepared bimodular O2 sensor under both potentiometric and resistance modules. For the potentiometric module, a linear relationship between electromotive force (EMF output of the sensor and the logarithm of O2 concentration was observed at each operating temperature, following the Nernst law. For the resistance module, the logarithm of electrical conductivity was proportional to the logarithm of oxygen concentration at each operating temperature, in good agreement with literature report. In addition, this bimodular sensor shows sensitive, reproducible and reversible response to oxygen under both sensing modules. Integration of two sensing modules into one sensor could greatly enrich the information output and would open a new venue in the development of high temperature gas sensors.

High-Temperature Piezoelectric Sensing

Directory of Open Access Journals (Sweden)

Xiaoning Jiang

2013-12-01

Full Text Available Piezoelectric sensing is of increasing interest for high-temperature applications in aerospace, automotive, power plants and material processing due to its low cost, compact sensor size and simple signal conditioning, in comparison with other high-temperature sensing techniques. This paper presented an overview of high-temperature piezoelectric sensing techniques. Firstly, different types of high-temperature piezoelectric single crystals, electrode materials, and their pros and cons are discussed. Secondly, recent work on high-temperature piezoelectric sensors including accelerometer, surface acoustic wave sensor, ultrasound transducer, acoustic emission sensor, gas sensor, and pressure sensor for temperatures up to 1,250 °C were reviewed. Finally, discussions of existing challenges and future work for high-temperature piezoelectric sensing are presented.

Influence of self-affine roughness on the friction coefficient of rubber at high sliding velocity

NARCIS (Netherlands)

Palasantzas, G

2004-01-01

In this work we investigate the influence of self-affine roughness on the friction coefficient of a rubber body onto a solid surface at high speeds. The roughness is characterized by the rms amplitude w, the correlation length xi, and the roughness exponent H. It is shown that the friction

Spectral affinity in protein networks

Directory of Open Access Journals (Sweden)

Teng Shang-Hua

2009-11-01

Full Text Available Abstract Background Protein-protein interaction (PPI networks enable us to better understand the functional organization of the proteome. We can learn a lot about a particular protein by querying its neighborhood in a PPI network to find proteins with similar function. A spectral approach that considers random walks between nodes of interest is particularly useful in evaluating closeness in PPI networks. Spectral measures of closeness are more robust to noise in the data and are more precise than simpler methods based on edge density and shortest path length. Results We develop a novel affinity measure for pairs of proteins in PPI networks, which uses personalized PageRank, a random walk based method used in context-sensitive search on the Web. Our measure of closeness, which we call PageRank Affinity, is proportional to the number of times the smaller-degree protein is visited in a random walk that restarts at the larger-degree protein. PageRank considers paths of all lengths in a network, therefore PageRank Affinity is a precise measure that is robust to noise in the data. PageRank Affinity is also provably related to cluster co-membership, making it a meaningful measure. In our experiments on protein networks we find that our measure is better at predicting co-complex membership and finding functionally related proteins than other commonly used measures of closeness. Moreover, our experiments indicate that PageRank Affinity is very resilient to noise in the network. In addition, based on our method we build a tool that quickly finds nodes closest to a queried protein in any protein network, and easily scales to much larger biological networks. Conclusion We define a meaningful way to assess the closeness of two proteins in a PPI network, and show that our closeness measure is more biologically significant than other commonly used methods. We also develop a tool, accessible at http://xialab.bu.edu/resources/pnns, that allows the user to

Different endothelin receptor affinities in dog tissues

International Nuclear Information System (INIS)

Loeffler, B.M.L.; Loehrer, W.

1991-01-01

Endothelin (ET) is a long-lasting potent vasoconstrictor-peptide. Here the authors report different binding affinities of endothelin-1 (ET-1) to ET-receptors of various dog tissues. Crude microsomal fractions were prepared after homogenisation of dog tissues in 50 mM Tris/HCl, 20 mM MnCl2, 1 mM EDTA, pH 7.4 by differential centrifugation. Aliquots of microsomal fractions (70 micrograms of protein) were incubated at 25 degrees C for 180 min in the presence of 20 pM 125I-ET-1 and various concentrations of cold ET-1. Four different ET-1 receptor binding affinities were found: adrenals, cerebrum, liver, heart, skeletal muscle and stomach microsomal membranes contained high affinity binding sites (Kd 50 - 80 pM, Bmax 60 - 250 fmol/mg). In cerebellum and spleen medium affinity ET-1 receptors (Kd 350 pM, Bmax 880 and 1200 fmol/mg respectively) were present. In comparison lung and kidney microsomes contained a low affinity ET-1 receptor (Kd 800 and 880 pM, Bmax 1600 and 350 fmol/mg). Receptors of even lower affinity were present in heart, intestine and liver microsomes with Kd values of 3 - 6 nM

Low affinity uniporter carrier proteins can increase net substrate uptake rate by reducing efflux

NARCIS (Netherlands)

Bosdriesz, Evert; Wortel, Meike T.; Haanstra, Jurgen R.; Wagner, Marijke J.; De La Torre Cortés, Pilar; Teusink, Bas

2018-01-01

Many organisms have several similar transporters with different affinities for the same substrate. Typically, high-affinity transporters are expressed when substrate is scarce and low-affinity ones when it is abundant. The benefit of using low instead of high-affinity transporters remains unclear,

Low affinity uniporter carrier proteins can increase net substrate uptake rate by reducing efflux

NARCIS (Netherlands)

Bosdriesz, Evert; Wortel, M.T.; Haanstra, Jurgen R.; Wagner, Marijke J.; De La Torre, P.; Teusink, Bas

2018-01-01

Many organisms have several similar transporters with different affinities for the same substrate. Typically, high-affinity transporters are expressed when substrate is scarce and low-affinity ones when it is abundant. The benefit of using low instead of high-affinity transporters remains

Photo-sensitive Ge nanocrystal based films controlled by substrate deposition temperature

KAUST Repository

Stavarache, Ionel

2017-07-21

Lowering the temperature of crystallization by deposition of thin films on a heated substrate represents the easiest way to find new means to develop and improve new working devices based on nanocrystals embedded in thin films. The improvements are strongly related with the increasing of operation speed, substantially decreasing the energy consumption and reducing unit fabrication costs of the respective semiconductor devices. This approach avoids major problems, such as those related to diffusion or difficulties in controlling of nanocrystallites size, which appear during thermal treatments at high temperatures after deposition. It is reported here the significant progress introduced by synthesis procedure to the in-situ structuring of Ge nanocrystallites in SiO₂ thin films by heating the substrate at low temperature, 400 °C during co-deposition of Ge and SiO₂ by magnetron sputtering. As a proof-of-concept, a Si/Ge-NCs:SiO₂ photo-sensitive structure was fabricated thereof and characterized. The structure shows superior performance on broad operation bandwidth from visible to near-infrared, as strong rectification properties in dark, significant current rise in the inversion mode when illuminated, high responsivity, high photo-detectivity of 10¹⁴ Jones, quick response and significant conversion efficiency of 850 %. This simple preparation approach brings an important contribution to the efort of structuring Ge nanocrystallites in SiO₂ thin films at a lower temperature for the purpose of using these materials for devices in optoelectronics, solar cells and electronics on flexible substrates.

Cu-modified carbon spheres/reduced graphene oxide as a high sensitivity of gas sensor for NO2 detection at room temperature

Science.gov (United States)

Su, Zhibin; Tan, Li; Yang, Ruiqiang; Zhang, Yu; Tao, Jin; Zhang, Nan; Wen, Fusheng

2018-03-01

Nitrogen dioxide (NO2) as one of the most serious air pollution is harmful to people's health, therefore high-performance gas sensors is critically needed. Here, Cu-modified carbon spheres/reduced graphene oxide (Cu@CS/RGO) composite have been prepared as NO2 gas sensor material. Carbon sphere in the interlayer of RGO can increase the specific surface area of RGO. Copper nanoparticles decorated on the surface of CS can effectively enhance the adsorption activity of RGO as supplier of free electrons. The experimental results showed that its particular structure improved the gas sensitivity of RGO at different NO2 concentrations at room temperature.

Temperature Sensitivity of an Atomic Vapor Cell-Based Dispersion-Enhanced Optical Cavity

Science.gov (United States)

Myneni, K.; Smith, D. D.; Chang, H.; Luckay, H. A.

2015-01-01

Enhancement of the response of an optical cavity to a change in optical path length, through the use of an intracavity fast-light medium, has previously been demonstrated experimentally and described theoretically for an atomic vapor cell as the intracavity resonant absorber. This phenomenon may be used to enhance both the scale factor and sensitivity of an optical cavity mode to the change in path length, e.g. in gyroscopic applications. We study the temperature sensitivity of the on-resonant scale factor enhancement, S(sub o), due to the thermal sensitivity of the lower-level atom density in an atomic vapor cell, specifically for the case of the Rb-87 D(sub 2) transition. A semi-empirical model of the temperature-dependence of the absorption profile, characterized by two parameters, a(sub o)(T) and gamma(sub a)(T) allows the temperature-dependence of the cavity response, S(sub o)(T) and dS(sub o)/dT to be predicted over a range of temperature. We compare the predictions to experiment. Our model will be useful in determining the useful range for S(sub o), given the practical constraints on temperature stability for an atomic vapor cell.

High-Affinity Accumulation of Chloroquine by Mouse Erythrocytes Infected with Plasmodium berghei

Science.gov (United States)

Fitch, Coy D.; Yunis, Norman G.; Chevli, Rekha; Gonzalez, Yolanda

1974-01-01

Washed erythrocytes infected with chloroquine-susceptible (CS) or with chloroquine-resistant (CR) P. berghei were used in model systems in vitro to study the accumulation of chloroquine with high affinity. The CS model could achieve distribution ratios (chloroquine in cells: chloroquine in medium) of 100 in the absence of substrate. 200—300 in the presence of 10 mM pyruvate or lactate, and over 600 in the presence of 1 mM glucose or glycerol. In comparable studies of the CR model, the distribution ratios were 100 in the absence of substrate and 300 or less in the presence of glucose or glycerol. The presence of lactate stimulated chloroquine accumulation in the CR model, whereas the presence of pyruvate did not. Lactate production from glucose and glycerol was undiminished in the CR model, and ATP concentrations were higher than in the CS model. Cold, iodoacetate, 2,4-dinitrophenol, or decreasing pH inhibited chloroquine accumulation in both models. These findings demonstrate substrate involvement in the accumulation of chloroquine with high affinity. In studies of the CS model, certain compounds competitively inhibited chloroquine accumulation, while others did not. This finding is attributable to a specific receptor that imposes structural constraints on the process of accumulation. For chloroquine analogues, the position and length of the side chain, the terminal nitrogen atom of the side chain, and the nitrogen atom in the quinoline ring are important determinants of binding to this receptor. PMID:4600044

Na+-Dependent High-Affinity Nitrate, Phosphate and Amino Acids Transport in Leaf Cells of the Seagrass Posidonia oceanica (L. Delile

Directory of Open Access Journals (Sweden)

Lourdes Rubio

2018-05-01

Full Text Available Posidonia oceanica (L. Delile is a seagrass, the only group of vascular plants to colonize the marine environment. Seawater is an extreme yet stable environment characterized by high salinity, alkaline pH and low availability of essential nutrients, such as nitrate and phosphate. Classical depletion experiments, membrane potential and cytosolic sodium measurements were used to characterize the high-affinity NO3−, Pi and amino acids uptake mechanisms in this species. Net uptake rates of both NO3− and Pi were reduced by more than 70% in the absence of Na+. Micromolar concentrations of NO3− depolarized mesophyll leaf cells plasma membrane. Depolarizations showed saturation kinetics (Km = 8.7 ± 1 μM NO3−, which were not observed in the absence of Na+. NO3− induced depolarizations at increasing Na+ also showed saturation kinetics (Km = 7.2 ± 2 mM Na+. Cytosolic Na+ measured in P. oceanica leaf cells (17 ± 2 mM Na+ increased by 0.4 ± 0.2 mM Na+ upon the addition of 100 μM NO3−. Na+-dependence was also observed for high-affinity l-ala and l-cys uptake and high-affinity Pi transport. All together, these results strongly suggest that NO3−, amino acids and Pi uptake in P. oceanica leaf cells are mediated by high-affinity Na+-dependent transport systems. This mechanism seems to be a key step in the process of adaptation of seagrasses to the marine environment.

High Affinity IgE-Fc Receptor alpha and gamma Subunit Interactions

International Nuclear Information System (INIS)

Rashid, A.; Housden, J. E. M.; Sabban, S.; Helm, B.

2014-01-01

Objective: To explore the relationships between the subunits (alpha, beta and gamma) of the high affinity IgE receptor (Fc and RI) and its ability to mediate transmembrane signaling. Study Design: Experimental study. Place and Duration of Study: Department of Molecular Biology and Biotechnology, University of Sheffield, UK, from 2008 to 2009. Methodology: The approach employed was to create a chimera (human alpha-gamma-gamma) using the extracellular (EC) domain of the human high affinity IgE receptor. The alpha subunit (huFc and RIalpha) of IgE receptor was spliced onto the rodent gamma TM and cytoplasmic domain (CD). This was transfected into the Rat Basophilic Leukemia cell line in order to assess the possibility of selectively activating cells transfected with this single pass construct for antigen induced mediator release. Results: The RBLs cell lines transfected with the huFc and RIalpha/gamma/gamma cDNA constructs were assessed for the cell surface expression of the huFc and RIalpha subunit and the response to the antigenic stimulus by looking for degranulation and intracellular Ca2+ mobilisation. The results obtained showed the absence of huFc and RIalpha subunit expression on the surface of transfected cells as seen by flowcytometric studies, beta-hexosaminidase assays and intracellular calcium mobilisation studies. Conclusion: In the present study the grounds for non-expression of huFc and RIalpha/gamma/gamma cDNA remains elusive but may be due to the fact that the human-rodent chimeric receptors are assembled differently than the endogenous rodent receptors as seen in study in which COS 7 cells were transfected with human/rat chimeric complexes. (author)

Low temperature sensitization of austenitic stainless steel: an ageing effect during BWR service

International Nuclear Information System (INIS)

Shah, B.K.; Sinha, A.K.; Rastogi, P.K.; Kulkarni, P.G.

1994-01-01

Sensitization in austenitic stainless steel refers to chromium carbide precipitation at the grain boundaries with concomitant depletion of chromium below 12% near grain boundaries. This makes the material susceptible to either intergranular corrosion (IGC) or intergranular stress corrosion cracking (IGSCC). This effect is predominant whenever austenitic stainless steel is subjected to thermal exposure in the temperature range 723-1073K either during welding or during heat treatment. Low temperature sensitization (LTS) refers to sensitization at temperature below the typical range of sensitization i.e. 723-1073K. A prerequisite for LTS phenomenon is reported to be the presence of chromium carbide nuclei at the grain boundaries which can grow during boiling water reactor service even at a relatively lower temperature of around 560K. LTS can lead to failure of BWR pipe due to IGSCC. The paper reviews the phenomenological and mechanistic aspects of LTS. Studies carried out regarding effect of prior cold work on LTS are reported. Summary of the studies reported in literature to examine the occurrence of LTS during BWR service has also been included. (author). 10 refs., 3 figs

N-Oxide analogs of WAY-100635 : new high affinity 5-HT (1A) receptor antagonists

NARCIS (Netherlands)

Oberwinkler - Marchais, Sandrine; Nowicki, B; Pike, VW; Halldin, C; Sandell, J; Chou, YH; Gulyas, B; Brennum, LT; Farde, L; Wikstrom, H V

2005-01-01

WAY-100635 [N-(2-(1-(4-(2-methoxyphenyl)piperazinyl)ethyl))-N-(2-pyridinyl)cyclohexanecarboxamide] 1 and its O-des-methyl derivative DWAY 2 are well-known high affinity 5-HT1A receptor antagonists. which when labeled with carbon-II (beta(+): t(1/2) 20.4min) in the carbonyl group are effective

Affinity chromatography with pseudobiospecific ligands on high-performance supports for purification of proteins of biotechnological interest

Directory of Open Access Journals (Sweden)

N.B. Iannucci

2003-03-01

Full Text Available High-performance affinity matrices were obtained by attaching pseudobiospecific ligands to hollow-fibre membranes. The neutral protease contained in FlavourzymeTM was purified to homogeneity with Yellow 4R-HE affinity hollow-fibre membranes. Immobilisation of Red HE-3B allowed purification of a milk-clotting enzyme obtained by solid-state culture of Mucor bacilliformis. Copper immobilisation through iminodiacetic acid allowed fractionation of Biocon Bioconcentrated PlusTM to separate the pectinesterase-containing fraction. The productivity of the developed processes - 1900, 94 and 750 U/ml.min, respectively - was 10- to 15-fold higher than that achieved with the same ligands immobilised on agarose-based soft gels, mainly due to the shortening of the purification processes.

Fabrication of surface plasmon resonance sensor surface with control of the nonspecific adsorption and affinity for the detection of 2,4,6-trinitrotoluene using an antifouling copolymer

Directory of Open Access Journals (Sweden)

Rui eYatabe

2014-04-01

Full Text Available We fabricated a surface plasmon resonance (SPR sensor using a hydrophilic polymer for the highly sensitive detection of 2,4,6-trinitrotoluene (TNT. The hydrophilic polymer was made from mono-2-(methacryloyloxyethylsuccinate (MES and 2-hydroxyethylmethacrylate (HEMA by surface-initiated atom transfer radical polymerization (SI-ATRP. The detection of TNT was carried out by displacement assay with the SPR measurement. In displacement assay, the affinity between anti-TNT antibody and the sensor surface, affects to the sensitivity. In the SPR measurement, nonspecific adsorption should be controlled because SPR sensor cannot discriminate between specific and nonspecific adsorption. Therefore, the affinity and nonspecific adsorption were controlled by changing the ratio of HEMA to MES. A detection limit of 0.4 ng/ml (ppb for TNT was achieved using a sensor surface with the lowest affinity without nonspecific adsorption.

Design and Fabrication of a Piezoresistive Pressure Sensor for Ultra High Temperature Environment

International Nuclear Information System (INIS)

Zhao, L B; Zhao, Y L; Jiang, Z D

2006-01-01

In order to solve the pressure measurement problem in the harsh environment, a piezoresistive pressure sensor has been developed, which can be used under high temperature above 200 deg. C and is able to endure instantaneous ultra high temperature (2000deg. C, duration≤2s) impact. Based on the MEMS (Micro Electro-Mechanical System) and integrated circuit technology, the piezoresistive pressure sensor's sensitive element was fabricated and constituted by silicon substrate, a thin buried silicon dioxide layer, four p-type resistors in the measuring circuit layer by boron ion implantation and photolithography, the top SiO2 layer by oxidation, stress matching Si3N4 layer, and a Ti-Pt-Au beam lead layer for connecting p-type resistors by sputtering. In order to decrease the leak-current influence to sensor in high temperature above 200deg. C, the buried SiO2 layer with the thickness 367 nm was fabricated by the SIMOX (Separation by Implantation of Oxygen) technology, which was instead of p-n junction to isolate the upper measuring circuit layer from Si substrate. In order to endure instantaneous ultra high temperature impact, the mechanical structure with cantilever and diaphragm and transmitting beam was designed. By laser welding and high temperature packaging technology, the high temperature piezoresistive pressure sensor was fabricated with range of 120MPa. After the thermal compensation, the sensor's thermal zero drift k 0 and thermal sensitivity drift k s were easy to be less than 3x10 -4 FS/deg. C. The experimental results show that the developed piezoresistive pressure sensor has good performances under high temperature and is able to endure instantaneous ultra high temperature impact, which meets the requirements of modern industry, such as aviation, oil, engine, etc

pH and temperature dual-sensitive liposome gel based on novel cleavable mPEG-Hz-CHEMS polymeric vaginal delivery system

Science.gov (United States)

Chen, Daquan; Sun, Kaoxiang; Mu, Hongjie; Tang, Mingtan; Liang, Rongcai; Wang, Aiping; Zhou, Shasha; Sun, Haijun; Zhao, Feng; Yao, Jianwen; Liu, Wanhui

2012-01-01

Background In this study, a pH and temperature dual-sensitive liposome gel based on a novel cleavable hydrazone-based pH-sensitive methoxy polyethylene glycol 2000-hydrazone-cholesteryl hemisuccinate (mPEG-Hz-CHEMS) polymer was used for vaginal administration. Methods The pH-sensitive, cleavable mPEG-Hz-CHEMS was designed as a modified pH-sensitive liposome that would selectively degrade under locally acidic vaginal conditions. The novel pH-sensitive liposome was engineered to form a thermogel at body temperature and to degrade in an acidic environment. Results A dual-sensitive liposome gel with a high encapsulation efficiency of arctigenin was formed and improved the solubility of arctigenin characterized by Fourier transform infrared spectroscopy and differential scanning calorimetry. The dual-sensitive liposome gel with a sol-gel transition at body temperature was degraded in a pH-dependent manner, and was stable for a long period of time at neutral and basic pH, but cleavable under acidic conditions (pH 5.0). Arctigenin encapsulated in a dual-sensitive liposome gel was more stable and less toxic than arctigenin loaded into pH-sensitive liposomes. In vitro drug release results indicated that dual-sensitive liposome gels showed constant release of arctigenin over 3 days, but showed sustained release of arctigenin in buffers at pH 7.4 and pH 9.0. Conclusion This research has shed some light on a pH and temperature dual-sensitive liposome gel using a cleavable mPEG-Hz-CHEMS polymer for vaginal delivery. PMID:22679372

Fenobody: A Ferritin-Displayed Nanobody with High Apparent Affinity and Half-Life Extension.

Science.gov (United States)

Fan, Kelong; Jiang, Bing; Guan, Zhe; He, Jiuyang; Yang, Dongling; Xie, Ni; Nie, Guohui; Xie, Can; Yan, Xiyun

2018-04-10

Nanobodies consist of a single domain variable fragment of a camelid heavy-chain antibody. Nanobodies have potential applications in biomedical fields because of their simple production procedures and low cost. Occasionally, nanobody clones of interest exhibit low affinities for their target antigens, which, together with their short half-life limit bioanalytical or therapeutic applications. Here, we developed a novel platform we named fenobody, in which a nanobody developed against H5N1 virus is displayed on the surface of ferritin in the form of a 24mer. We constructed a fenobody by substituting the fifth helix of ferritin with the nanobody. TEM analysis showed that nanobodies were displayed on the surface of ferritin in the form of 6 × 4 bundles, and that these clustered nanobodies are flexible for antigen binding in spatial structure. Comparing fenobodies with conventional nanobodies currently used revealed that the antigen binding apparent affinity of anti-H5N1 fenobody was dramatically increased (∼360-fold). Crucially, their half-life extension in a murine model was 10-fold longer than anti-H5N1 nanobody. In addition, we found that our fenobodies are highly expressed in Escherichia coli, and are both soluble and thermo-stable nanocages that self-assemble as 24-polymers. In conclusion, our results demonstrate that fenobodies have unique advantages over currently available systems for apparent affinity enhancement and half-life extension of nanobodies. Our fenobody system presents a suitable platform for various large-scale biotechnological processes and should greatly facilitate the application of nanobody technology in these areas.

Calculation of coolant temperature sensitivity related to thermohydraulic parameters

International Nuclear Information System (INIS)

Silva, F.C. da; Andrade Lima, F.R. de

1985-01-01

It is verified the viability to apply the generalized Perturbation Theory (GPT) in the calculation of sensitivity for thermal-hydraulic problems. It was developed the TEMPERA code in FORTRAN-IV to transient calculations in the axial temperature distribution in a channel of PWR reactor and the associated importance function, as well as effects of variations of thermalhydraulic parameters in the coolant temperature. The results are compared with one which were obtained by direct calculation. (M.C.K.) [pt

A highly sensitive electrochemical glucose sensor structuring with nickel hydroxide and enzyme glucose oxidase

International Nuclear Information System (INIS)

Mathew, Manjusha; Sandhyarani, N.

2013-01-01

Graphical abstract: A combination of Ni 2+ /Ni 3+ redox couple and glucose oxidase has successfully been exploited for the realization of a highly sensitive glucose sensor for the first time. -- Highlights: • A multilayered glucose biosensor with enhanced sensitivity was fabricated. • Combination of Ni 2+ /Ni 3+ redox couple and glucose oxidase has been exploited for the first time. • Exhibits a lower detection limit of 100 nM with a high sensitivity of 16,840 μA mM −1 cm −2 . • The surface shows a low Michaelis–Menten constant value of 2.4 μM. • Detailed mechanism of sensing was proposed and justified. -- Abstract: A multilayered glucose biosensor with enhanced electron transport was fabricated via the sequential electrodeposition of chitosan gold nanocomposite (CGNC) and nickel hydroxide (Ni(OH) 2 ) on a bare gold electrode and subsequent immobilization of glucose oxidase. A thin film of Ni(OH) 2 deposited on CGNC modified gold electrode serves as an electrochemical redox probe as well as a matrix for the immobilization of glucose oxidase retaining its activity. Electron transport property of CGNC has been exploited to enhance the electron transport between the analyte and electrode. Electrochemical characteristics of the biosensor were studied by cyclic voltammetry and chronoamperometry. Under optimal conditions the biosensor exhibits a linear range from 1 μM to 100 μM with a limit of detection (lod) down to 100 nM. The sensor shows a low Michaelis-Menten constant value of 2.4 μM indicates the high affinity of enzyme to the analyte points to the retained activity of enzyme after immobilization. The present glucose sensor with the high selectivity, sensitivity and stability is promising for practical clinical applications

Potentialities in electronics of new high critical temperature superconductors. Potentialites en electronique des nouveaux supraconducteurs a haute temperature critique

Energy Technology Data Exchange (ETDEWEB)

Hartemann, P [Thomson-CSF, 75 - Paris (FR)

1989-09-01

The main electronic applications of superconductors involve the signal processing, the electromagnetic wave detection and the magnetometry. Characteristics of devices based on conventional superconductors cooled by liquid helium are given and the changes induced by incorporating high-temperature superconductors are estimated. After a survey of new superconductor properties, the superconducting devices for analog or digital signal processing are reviewed. The gains predicted for high-temperature superconducting analog devices are considered in greater detail. Different sections deal with the infrared or (sub)millimeter wave detection. The most sensitive apparatuses for magnetic measurements are based on SQUIDs. Features of SQUIDs made of granular high-temperature superconducting material samples (grain boundaries behave as barriers of intrinsic junctions) are discussed.

Highly efficient high temperature electrolysis

DEFF Research Database (Denmark)

Hauch, Anne; Ebbesen, Sune; Jensen, Søren Højgaard

2008-01-01

High temperature electrolysis of water and steam may provide an efficient, cost effective and environmentally friendly production of H-2 Using electricity produced from sustainable, non-fossil energy sources. To achieve cost competitive electrolysis cells that are both high performing i.e. minimum...... internal resistance of the cell, and long-term stable, it is critical to develop electrode materials that are optimal for steam electrolysis. In this article electrolysis cells for electrolysis of water or steam at temperatures above 200 degrees C for production of H-2 are reviewed. High temperature...... electrolysis is favourable from a thermodynamic point of view, because a part of the required energy can be supplied as thermal heat, and the activation barrier is lowered increasing the H-2 production rate. Only two types of cells operating at high temperature (above 200 degrees C) have been described...

In Vivo Neutralization of α-Cobratoxin with High-Affinity Llama Single-Domain Antibodies (VHHs) and a VHH-Fc Antibody

Science.gov (United States)

Richard, Gabrielle; Meyers, Ashley J.; McLean, Michael D.; Arbabi-Ghahroudi, Mehdi; MacKenzie, Roger; Hall, J. Christopher

2013-01-01

Small recombinant antibody fragments (e.g. scFvs and VHHs), which are highly tissue permeable, are being investigated for antivenom production as conventional antivenoms consisting of IgG or F(ab’)2 antibody fragments do not effectively neutralize venom toxins located in deep tissues. However, antivenoms composed entirely of small antibody fragments may have poor therapeutic efficacy due to their short serum half-lives. To increase serum persistence and maintain tissue penetration, we prepared low and high molecular mass antivenom antibodies. Four llama VHHs were isolated from an immune VHH-displayed phage library and were shown to have high affinity, in the low nM range, for α-cobratoxin (α–Cbtx), the most lethal component of Naja kaouthia venom. Subsequently, our highest affinity VHH (C2) was fused to a human Fc fragment to create a VHH2-Fc antibody that would offer prolonged serum persistence. After in planta (Nicotiana benthamiana) expression and purification, we show that our VHH2-Fc antibody retained high affinity binding to α–Cbtx. Mouse α–Cbtx challenge studies showed that our highest affinity VHHs (C2 and C20) and the VHH2-Fc antibody effectively neutralized lethality induced by α–Cbtx at an antibody:toxin molar ratio as low as ca. 0.75×:1. Further research towards the development of an antivenom therapeutic involving these anti-α-Cbtx VHHs and VHH2-Fc antibody molecules should involve testing them as a combination, to determine whether they maintain tissue penetration capability and low immunogenicity, and whether they exhibit improved serum persistence and therapeutic efficacy. PMID:23894495

Unconventional route to encapsulated ultrasmall gold nanoparticles for high-temperature catalysis.

Science.gov (United States)

Zhang, Tingting; Zhao, Hongyu; He, Shengnan; Liu, Kai; Liu, Hongyang; Yin, Yadong; Gao, Chuanbo

2014-07-22

Ultrasmall gold nanoparticles (us-AuNPs, gold hydroxide nanoparticles, which have excellent affinity to silica, then carrying out controllable silica coating in reverse micelles, and finally converting gold hydroxide particles into well-protected us-AuNPs. With a single-core/shell configuration that prevents sintering of nearby us-AuNPs and amino group modification of the Au/SiO2 interface that provides additional coordinating interactions, the resulting us-AuNP@SiO2 nanospheres are highly stable at high temperatures and show high activity in catalytic CO oxidation reactions. A dramatic and continuous increase in the catalytic activity has been observed when the size of the us-AuNPs decreases from 2.3 to 1.5 nm, which reflects the intrinsic size effect of the Au nanoparticles on an inert support. The synthesis scheme described in this work is believed to be extendable to many other ultrasmall metal@oxide nanostructures for much broader catalytic applications.

Human body temperature and new approaches to constructing temperature-sensitive bacterial vaccines.

Science.gov (United States)

White, Matthew D; Bosio, Catharine M; Duplantis, Barry N; Nano, Francis E

2011-09-01

Many of the live human and animal vaccines that are currently in use are attenuated by virtue of their temperature-sensitive (TS) replication. These vaccines are able to function because they can take advantage of sites in mammalian bodies that are cooler than the core temperature, where TS vaccines fail to replicate. In this article, we discuss the distribution of temperature in the human body, and relate how the temperature differential can be exploited for designing and using TS vaccines. We also examine how one of the coolest organs of the body, the skin, contains antigen-processing cells that can be targeted to provoke the desired immune response from a TS vaccine. We describe traditional approaches to making TS vaccines, and highlight new information and technologies that are being used to create a new generation of engineered TS vaccines. We pay particular attention to the recently described technology of substituting essential genes from Arctic bacteria for their homologues in mammalian pathogens as a way of creating TS vaccines.

Lp-dual affine surface area

Science.gov (United States)

Wei, Wang; Binwu, He

2008-12-01

According to the notion of Lp-affine surface area by Lutwak, in this paper, we introduce the concept of Lp-dual affine surface area. Further, we establish the affine isoperimetric inequality and the Blaschke-Santaló inequality for Lp-dual affine surface area. Besides, the dual Brunn-Minkowski inequality for Lp-dual affine surface area is presented.

Shrub encroachment alters sensitivity of soil respiration to temperature and moisture

Science.gov (United States)

Cable, Jessica M.; Barron-Gafford, Greg A.; Ogle, Kiona; Pavao-Zuckerman, Mitchell; Scott, Russell L.; Williams, David G.; Huxman, Travis E.

2012-03-01

A greater abundance of shrubs in semiarid grasslands affects the spatial patterns of soil temperature, moisture, and litter, resulting in fertile islands with potentially enhanced soil metabolic activity. The goal of this study was to quantify the microsite specificity of soil respiration in a semiarid riparian ecosystem experiencing shrub encroachment. We quantified the response of soil respiration to different microsite conditions created by big mesquite shrubs (near the trunk and the canopy edge), medium-sized mesquite, sacaton bunchgrasses, and open spaces. We hypothesized that soil respiration would be more temperature sensitive and less moisture sensitive and have a greater magnitude in shrub microsites compared with grass and open microsites. Field and incubation soil respiration data were simultaneously analyzed in a Bayesian framework to quantify the microsite-specific temperature and moisture sensitivities and magnitude of respiration. The analysis showed that shrub expansion increases the heterogeneity of respiration. Respiration has greater temperature sensitivity near the shrub canopy edge, and respiration rates are higher overall under big mesquite compared with those of the other microsites. Respiration in the microsites beneath medium-sized mesquites does not behave like a downscaled version of big mesquite microsites. The grass microsites show more similarity to big mesquite microsites than medium-sized shrubs. This study shows there can be a great deal of fine-scale spatial heterogeneity that accompanies shifts in vegetation structure. Such complexity presents a challenge in scaling soil respiration fluxes to the landscape for systems experiencing shrub encroachment, but quantifying this complexity is significantly important in determining overall ecosystem metabolic behavior.

Highly sensitive, colorimetric detection of mercury(II) in aqueous media by quaternary ammonium group-capped gold nanoparticles at room temperature.

Science.gov (United States)

Liu, Dingbin; Qu, Weisi; Chen, Wenwen; Zhang, Wei; Wang, Zhuo; Jiang, Xingyu

2010-12-01

We provide a highly sensitive and selective assay to detect Hg(2+) in aqueous solutions using gold nanoparticles modified with quaternary ammonium group-terminated thiols at room temperature. The mechanism is the abstraction of thiols by Hg(2+) that led to the aggregation of nanoparticles. With the assistance of solar light irradiation, the detection limit can be as low as 30 nM, which satisfies the guideline concentration of Hg(2+) in drinking water set by the WHO. In addition, the dynamic range of detection is wide (3 × 10(-8)-1 × 10(-2) M). This range, to our best knowledge, is the widest one that has been reported so far in gold nanoparticle (AuNP)-based assays for Hg(2+).

High Temperature Radio Frequency Loads

CERN Document Server

Federmann, S; Grudiev, A; Montesinos, E; Syratchev, I

2011-01-01

In the context of energy saving and recovery requirements the design of reliable and robust RF power loads which permit a high outlet temperature and high pressure of the cooling water is desirable. Cooling water arriving at the outlet withmore than 150 ◦C and high pressure has a higher value than water with 50 ◦C under low pressure. Conventional RF power loads containing dielectric and magnetic materials as well as sensitive ceramic windows usually do not permit going much higher than 90 ◦C. Here we present and discuss several design concepts for "metal only" RF high power loads. One concept is the application of magnetic steel corrugated waveguides near cutoff – this concept could find practical use above several GHz. Another solution are resonant structures made of steel to be installed in large waveguides for frequencies of 500 MHz or lower. Similar resonant structures above 100 MHz taking advantage of the rather high losses of normal steel may also be used in coaxial line geometries with large di...

Temperature sheets and aspect sensitive radar echoes

Directory of Open Access Journals (Sweden)

H. Luce

Full Text Available here have been years of discussion and controversy about the existence of very thin and stable temperature sheets and their relationship to the VHF radar aspect sensitivity. It is only recently that very high-resolution in situ temperature observations have brought credence to the reality and ubiquity of these structures in the free atmosphere and to their contribution to radar echo enhancements along the vertical. Indeed, measurements with very high-resolution sensors are still extremely rare and rather difficult to obtain outside of the planetary boundary layer. They have only been carried out up to the lower stratosphere by Service d’A´ eronomie (CNRS, France for about 10 years. The controversy also persisted due to the volume resolution of the (Mesosphere-Stratosphere-Troposphere VHF radars which is coarse with respect to sheet thickness, although widely sufficient for meteorological or mesoscale investigations. The contribution within the range gate of many of these structures, which are advected by the wind, and decay and grow at different instants and could be distorted either by internal gravity waves or turbulence fields, could lead to radar echoes with statistical properties similar to those produced by anisotropic turbulence. Some questions thus remain regarding the manner in which temperature sheets contribute to VHF radar echoes. In particular, the zenithal and azimuthal angular dependence of the echo power may not only be produced by diffuse reflection on stable distorted or corrugated sheets, but also by extra contributions from anisotropic turbulence occurring in the stratified atmosphere. Thus, for several years, efforts have been put forth to improve the radar height resolution in order to better describe thin structures. Frequency interferometric techniques are widely used and have been recently further developed with the implementation of high-resolution data processings. We begin by reviewing briefly some characteristics

Temperature sheets and aspect sensitive radar echoes

Directory of Open Access Journals (Sweden)

H. Luce

2001-08-01

Full Text Available here have been years of discussion and controversy about the existence of very thin and stable temperature sheets and their relationship to the VHF radar aspect sensitivity. It is only recently that very high-resolution in situ temperature observations have brought credence to the reality and ubiquity of these structures in the free atmosphere and to their contribution to radar echo enhancements along the vertical. Indeed, measurements with very high-resolution sensors are still extremely rare and rather difficult to obtain outside of the planetary boundary layer. They have only been carried out up to the lower stratosphere by Service d’A´ eronomie (CNRS, France for about 10 years. The controversy also persisted due to the volume resolution of the (Mesosphere-Stratosphere-Troposphere VHF radars which is coarse with respect to sheet thickness, although widely sufficient for meteorological or mesoscale investigations. The contribution within the range gate of many of these structures, which are advected by the wind, and decay and grow at different instants and could be distorted either by internal gravity waves or turbulence fields, could lead to radar echoes with statistical properties similar to those produced by anisotropic turbulence. Some questions thus remain regarding the manner in which temperature sheets contribute to VHF radar echoes. In particular, the zenithal and azimuthal angular dependence of the echo power may not only be produced by diffuse reflection on stable distorted or corrugated sheets, but also by extra contributions from anisotropic turbulence occurring in the stratified atmosphere. Thus, for several years, efforts have been put forth to improve the radar height resolution in order to better describe thin structures. Frequency interferometric techniques are widely used and have been recently further developed with the implementation of high-resolution data processings. We begin by reviewing briefly some characteristics

A fundamental study on stress corrosion cracking of SUS 304 steel in high temperature water

International Nuclear Information System (INIS)

Mukai, Yoshihiko; Murata, Masato

1985-01-01

SCC susceptibility of sensitized SUS 304 stainless steel in high temperature water was studied. The results obtained are as follows. SCC susceptibility was increased by adding crevices to the tensile specimen surface, for the corrodent became acidified by hydrolysis in crevices. SCC susceptibility was best fit to TTS curve obtained by EPR test, not by other corrosion tests such as Strauss test or the grain boundary corrosion test in high temperature water. In addition, by giving a simulated weld thermal cycle before the sensitizing heat treatment, the sensitization was clearly promoted. This seemed to be caused by the reason that nucleation of carbide occured in the simulated weld thermal cycle process and it promoted the carbide growth and the formation of Cr poor layer around carbide in the subsequent sensitization process. (author)

High temperature materials and mechanisms

CERN Document Server

2014-01-01

The use of high-temperature materials in current and future applications, including silicone materials for handling hot foods and metal alloys for developing high-speed aircraft and spacecraft systems, has generated a growing interest in high-temperature technologies. High Temperature Materials and Mechanisms explores a broad range of issues related to high-temperature materials and mechanisms that operate in harsh conditions. While some applications involve the use of materials at high temperatures, others require materials processed at high temperatures for use at room temperature. High-temperature materials must also be resistant to related causes of damage, such as oxidation and corrosion, which are accelerated with increased temperatures. This book examines high-temperature materials and mechanisms from many angles. It covers the topics of processes, materials characterization methods, and the nondestructive evaluation and health monitoring of high-temperature materials and structures. It describes the ...

Specific recognition of the C-terminal end of A beta 42 by a high affinity monoclonal antibody

DEFF Research Database (Denmark)

Axelsen, Trine Veje; Holm, Arne; Birkelund, Svend

2009-01-01

The neurotoxic peptide A beta(42) is derived from the amyloid precursor protein by proteolytic cleavage and is deposited in the brain of patients suffering from Alzheimer's disease (AD). In this study we generate a high affinity monoclonal antibody that targets the C-terminal end of A beta(42......) with high specificity. By this is meant that the paratope of the antibody must enclose the C-terminal end of A beta(42) including the carboxy-group of amino acid 42, and not just recognize a linear epitope in the C-terminal part of A beta. This has been accomplished by using a unique antigen construct made...... by the Ligand Presenting Assembly technology (LPA technology). This strategy results in dimeric presentation of the free C-terminal end of A beta(42). The generated Mab A beta1.1 is indeed specific for the C-terminal end of A beta(42) to which it binds with high affinity. Mab A beta1.1 recognizes the epitope...

Selection of antigenic markers on a GFP-C{kappa} fusion scaffold with high sensitivity by eukaryotic ribosome display

Energy Technology Data Exchange (ETDEWEB)

Yongmin, Yang [Institute of Genetics, San Diego, CA 92121-2233 (United States); IgE Therapeutics, Inc., San Diego, CA 92121-2233 (United States); Barankiewicz, Teresa J [Institute of Genetics, San Diego, CA 92121-2233 (United States); IgE Therapeutics, Inc., San Diego, CA 92121-2233 (United States); Mingyue, He [Babraham Institute, Cambridge CB2 4AT (United Kingdom); Taussig, Michael J [Babraham Institute, Cambridge CB2 4AT (United Kingdom); Chen, Swey-Shen [Institute of Genetics, San Diego, CA 92121-2233 (United States) and IgE Therapeutics, Inc., San Diego, CA 92121-2233 (United States)

2007-07-27

Ribosome display is a cell-free system permitting gene selection through the physical association of genetic material (mRNA) and its phenotypic (protein) product. While often used to select single-chain antibodies from large libraries by panning against immobilized antigens, we have adapted ribosome display for use in the 'reverse' format in order to select high affinity antigenic determinants against solid-phase antibody. To create an antigenic scaffold, DNA encoding green fluorescent protein (GFP) was fused to a light chain constant domain (C{kappa}) with stop codon deleted, and with 5' signals (T7 promoter, Kozak) enabling coupled transcription/translation in a eukaryotic cell-free system. Epitopes on either GFP (5') or C{kappa} (3') were selected by anti-GFP or anti-C{kappa} antibodies, respectively, coupled to magnetic beads. After selection, mRNA was amplified directly from protein-ribosome-mRNA (PRM) complexes by in situ PCR followed by internal amplification and reassembly PCR. As little as 10 fg of the 1 kb DNA construct, i.e. approximately 7500 molecules, could be recovered following a single round of interaction with solid-phase anti-GFP antibody. This platform is highly specific and sensitive for the antigen-antibody interaction and may permit selection and reshaping of high affinity antigenic variants of scaffold proteins.

Novel High Temperature Materials for In-Situ Sensing Devices

Energy Technology Data Exchange (ETDEWEB)

Florian Solzbacher; Anil Virkar; Loren Rieth; Srinivasan Kannan; Xiaoxin Chen; Hannwelm Steinebach

2009-12-31

The overriding goal of this project was to develop gas sensor materials and systems compatible with operation at temperatures from 500 to 700 C. Gas sensors operating at these temperatures would be compatible with placement in fossil-energy exhaust streams close to the combustion chamber, and therefore have advantages for process regulation, and feedback for emissions controls. The three thrusts of our work included investigating thin film gas sensor materials based on metal oxide materials and electroceramic materials, and also development of microhotplate devices to support the gas sensing films. The metal oxide materials NiO, In{sub 2}O{sub 3}, and Ga{sub 2}O{sub 3} were investigated for their sensitivity to H{sub 2}, NO{sub x}, and CO{sub 2}, respectively, at high temperatures (T > 500 C), where the sensing properties of these materials have received little attention. New ground was broken in achieving excellent gas sensor responses (>10) for temperatures up to 600 C for NiO and In{sub 2}O{sub 3} materials. The gas sensitivity of these materials was decreasing as temperatures increased above 500 C, which indicates that achieving strong sensitivities with these materials at very high temperatures (T {ge} 650 C) will be a further challenge. The sensitivity, selectivity, stability, and reliability of these materials were investigated across a wide range of deposition conditions, temperatures, film thickness, as using surface active promoter materials. We also proposed to study the electroceramic materials BaZr{sub (1-x)}Y{sub x}O{sub (3-x/2)} and BaCe{sub (2-x)}Ca{sub x}S{sub (4-x/2)} for their ability to detect H{sub 2}O and H{sub 2}S, respectively. This report focuses on the properties and gas sensing characteristics of BaZr{sub (1-x)}Y{sub x}O{sub (3-x/2)} (Y-doped BaZrO{sub 3}), as significant difficulties were encounter in generating BaCe{sub (2-x)}Ca{sub x}S{sub (4-x/2)} sensors. Significant new results were achieved for Y-doped BaZrO{sub 3}, including

Selection of DNA aptamers against epidermal growth factor receptor with high affinity and specificity.

Science.gov (United States)

Wang, Deng-Liang; Song, Yan-Ling; Zhu, Zhi; Li, Xi-Lan; Zou, Yuan; Yang, Hai-Tao; Wang, Jiang-Jie; Yao, Pei-Sen; Pan, Ru-Jun; Yang, Chaoyong James; Kang, De-Zhi

2014-10-31

Epidermal growth factor receptor (EGFR/HER1/c-ErbB1), is overexpressed in many solid cancers, such as epidermoid carcinomas, malignant gliomas, etc. EGFR plays roles in proliferation, invasion, angiogenesis and metastasis of malignant cancer cells and is the ideal antigen for clinical applications in cancer detection, imaging and therapy. Aptamers, the output of the systematic evolution of ligands by exponential enrichment (SELEX), are DNA/RNA oligonucleotides which can bind protein and other substances with specificity. RNA aptamers are undesirable due to their instability and high cost of production. Conversely, DNA aptamers have aroused researcher's attention because they are easily synthesized, stable, selective, have high binding affinity and are cost-effective to produce. In this study, we have successfully identified DNA aptamers with high binding affinity and selectivity to EGFR. The aptamer named TuTu22 with Kd 56±7.3nM was chosen from the identified DNA aptamers for further study. Flow cytometry analysis results indicated that the TuTu22 aptamer was able to specifically recognize a variety of cancer cells expressing EGFR but did not bind to the EGFR-negative cells. With all of the aforementioned advantages, the DNA aptamers reported here against cancer biomarker EGFR will facilitate the development of novel targeted cancer detection, imaging and therapy. Copyright © 2014 Elsevier Inc. All rights reserved.

High temperature refrigerator

International Nuclear Information System (INIS)

Steyert, W.A. Jr.

1978-01-01

A high temperature magnetic refrigerator is described which uses a Stirling-like cycle in which rotating magnetic working material is heated in zero field and adiabatically magnetized, cooled in high field, then adiabatically demagnetized. During this cycle the working material is in heat exchange with a pumped fluid which absorbs heat from a low temperature heat source and deposits heat in a high temperature reservoir. The magnetic refrigeration cycle operates at an efficiency 70% of Carnot

Peptides in headlock--a novel high-affinity and versatile peptide-binding nanobody for proteomics and microscopy.

Science.gov (United States)

Braun, Michael B; Traenkle, Bjoern; Koch, Philipp A; Emele, Felix; Weiss, Frederik; Poetz, Oliver; Stehle, Thilo; Rothbauer, Ulrich

2016-01-21

Nanobodies are highly valuable tools for numerous bioanalytical and biotechnical applications. Here, we report the characterization of a nanobody that binds a short peptide epitope with extraordinary affinity. Structural analysis reveals an unusual binding mode where the extended peptide becomes part of a β-sheet structure in the nanobody. This interaction relies on sequence-independent backbone interactions augmented by a small number of specificity-determining side chain contacts. Once bound, the peptide is fastened by two nanobody side chains that clamp it in a headlock fashion. Exploiting this unusual binding mode, we generated a novel nanobody-derived capture and detection system. Matrix-coupled nanobody enables the fast and efficient isolation of epitope-tagged proteins from prokaryotic and eukaryotic expression systems. Additionally, the fluorescently labeled nanobody visualizes subcellular structures in different cellular compartments. The high-affinity-binding and modifiable peptide tag of this system renders it a versatile and robust tool to combine biochemical analysis with microscopic studies.

Highly sensitive room temperature organic vapor sensor based on polybenzoxazine-derived carbon aerogel thin film composite

International Nuclear Information System (INIS)

Thubsuang, Uthen; Sukanan, Darunee; Sahasithiwat, Somboon; Wongkasemjit, Sujitra; Chaisuwan, Thanyalak

2015-01-01

Graphical abstract: - Highlights: • Activated carbon aerogel with high surface area can be prepared from polybenzoxazine. • Activated carbon aerogel enhances the adsorption capacity of gas sensor. • Organic vapors with very low concentration can be detected by the as-prepared sensor. • The as-prepared sensor shows impressive short exposure and recovery time. • The response to different organic vapors can be tailored by changing polymer matrix. - Abstract: Gas sensing composites were fabricated using polybenzoxazine-based activated carbon aerogel as a conductive filler. The activated carbon aerogel is a nano-porous material, which has high pore volume of 0.57 cm 3 /g and surface area of 917 m 2 /g. The activated carbon aerogel/polybutadiene composite displayed good response of 11.2 and 6.7 to toluene and n-hexane, respectively, compared to those of graphite/polybutadiene composite. The activated carbon aerogel/polybutadiene composite also showed high sensitivity of 3.09 × 10 2 ppm −1 to toluene. However, the sensitivity of activated carbon aerogel/polybutadiene composite drastically decreased to 1.99 ppm −1 and zero when exposed to acetone and water, respectively. Contrarily, when polyvinyl alcohol was used as a matrix, the sensitivity was about 4.19 ppm −1 to water. While the composite was found to be not sensitive to toluene. The activated carbon aerogel/polybutadiene composite also showed good recovery as the electrical resistance came back to the original value within minutes when exposed to nitrogen gas

The high temperature out-of-pile test of LVDT for internal pressure measurement of nuclear fuel rod

Energy Technology Data Exchange (ETDEWEB)

Son, J. M.; Kim, B. K.; Kim, D. S.; Yoon, K. B.; Sin, Y. T.; Park, S. J.; Kang, Y. H. [KAERI, Taejon (Korea, Republic of)

2002-10-01

As a part of the development of instrumentation technologies for the nuclear fuel irradiation test in HANARO(High-flux Advanced Nuclear Application Reactor), the internal pressure measurement technique of the nuclear fuel rod is being developed using LVDT(Linear Variable Differential Transformer). As the results of out-of-pile test at room temperature, it was concluded that the well qualified out-of-pile tests were needed to understand the LVDT's detail characteristics at high temperature for the detail design of the fuel irradiation capsule, because LVDT is very sensitive to variation of temperature. Therefore, the high temperature out-of-pile test system for pressure measurement was developed, and this test was performed under the temperature condition between room temperature and 300 .deg. C increasing the pressure from 0 bar to 30 bar. The LVDT's high temperature characteristics and temperature sensitivity of LVDT were analyzed through this experiment. Based on the result of this test, the method for the application of LVDT at high temperature was introduced. It is known that the results will be used to predict accurately the internal pressure of fuel rod during irradiation test.

Characteristics of the interaction of calcium with casein submicelles as determined by analytical affinity chromatography

International Nuclear Information System (INIS)

Jang, H.D.; Swaisgood, H.E.

1990-01-01

Interaction of calcium with casein submicelles was investigated in CaCl2 and calcium phosphate buffers and with synthetic milk salt solutions using the technique of analytical affinity chromatography. Micelles that had been prepared by size exclusion chromatography with glycerolpropyl controlled-pore glass from fresh raw skim milk that had never been cooled, were dialyzed at room temperature against calcium-free imidazole buffer, pH 6.7. Resulting submicelles were covalently immobilized on succinamidopropyl controlled-pore glass (300-nm pore size). Using 45Ca to monitor the elution retardation, the affinity of free Ca2+ and calcium salt species was determined at temperatures of 20 to 40 degrees C and pH 6.0 to 7.5. Increasing the pH in this range or increasing the temperature strengthened the binding of calcium to submicelles, similar to previous observations with individual caseins. However, the enthalpy change obtained from the temperature dependence was considerably greater than that reported for alpha s1- and beta-caseins. Furthermore, the elution profiles for 45Ca in milk salt solutions were decidedly different from those in CaCl2 or calcium phosphate buffers and the affinities were also greater. For example, at pH 6.7 and 30 degrees C the average dissociation constant for the submicelle-calcium complex is 0.074 mM for CaCl2 and calcium phosphate buffers, vs 0.016 mM for the milk salt solution. The asymmetric frontal boundaries and higher average affinities observed with milk salts may be due to binding of calcium salts with greater affinity in addition to the binding of free Ca2+ in these solutions

Blockage of High-Affinity Choline Transporter Increases Visceral Hypersensitivity in Rats with Chronic Stress

Science.gov (United States)

2018-01-01

Background Visceral hypersensitivity is a common feature of irritable bowel syndrome. Cholinergic system involves in the development of visceral hypersensitivity, and high-affinity choline transporter (CHT1) is of crucial importance in choline uptake system. However, involvement of CHT1 in visceral hypersensitivity remains unknown. The research aimed to study the CHT1 expression in dorsal root ganglions (DRGs) and the role of CHT1 in visceral hypersensitivity. Methods Repetitive water avoidance stress (WAS) was used to induce visceral hypersensitivity in rats. Colorectal distension (CRD) was determined, and the abdominal withdrawal reflex (AWR) and threshold intensity data were recorded to measure the visceral sensitivity. After intraperitoneal injection of hemicholinium-3 (HC-3), the specific inhibitor of CHT1, CRD data were also recorded. The CHT1 expression of DRGs was investigated by Western blotting, immunohistochemistry, and quantitative RT-PCR. Acetylcholine levels in the DRGs were detected by the assay kit. Results Repetitive WAS increased the AWR score of CRD at high distension pressure and decreased the mean threshold of rats. The CHT1 expression and acetylcholine concentration of DRG were significantly increased in WAS rats. After the administration of HC-3, the AWR score in WAS group was significantly increased at higher distension pressure while the threshold intensity was significantly reduced compared to the normal saline group. Acetylcholine concentration was significantly lower than the normal saline rats. Conclusion Our research firstly reports that CHT1 is overexpressed in noninflammatory visceral hypersensitivity, and blockage of CHT1 can enhance the visceral hypersensitivity. CHT1 may play an inhibitory role in visceral hypersensitivity. PMID:29849603

N-Acetyl-2-Aminofluorene (AAF) Processing in Adult Rat Hepatocytes in Primary Culture Occurs by High-Affinity Low-Velocity and Low-Affinity High-Velocity AAF Metabolite-Forming Systems.

Science.gov (United States)

Koch, Katherine S; Moran, Tom; Shier, W Thomas; Leffert, Hyam L

2018-05-01

N-acetyl-2-aminofluorene (AAF) is a procarcinogen used widely in physiological investigations of chemical hepatocarcinogenesis. Its metabolic pathways have been described extensively, yet little is known about its biochemical processing, growth cycle expression, and pharmacological properties inside living hepatocytes-the principal cellular targets of this hepatocarcinogen. In this report, primary monolayer adult rat hepatocyte cultures and high specific-activity [ring G-3 H]-N-acetyl-2-aminofluorene were used to extend previous observations of metabolic activation of AAF by highly differentiated, proliferation-competent hepatocytes in long-term cultures. AAF metabolism proceeded by zero-order kinetics. Hepatocytes processed significant amounts of procarcinogen (≈12 μg AAF/106 cells/day). Five ring-hydroxylated and one deacetylated species of AAF were secreted into the culture media. Extracellular metabolite levels varied during the growth cycle (days 0-13), but their rank quantitative order was time invariant: 5-OH-AAF > 7-OH-AAF > 3-OH-AAF > N-OH-AAF > aminofluorene (AF) > 1-OH-AAF. Lineweaver-Burk analyses revealed two principal classes of metabolism: System I (high-affinity and low-velocity), Km[APPARENT] = 1.64 × 10-7  M and VMAX[APPARENT] = 0.1 nmol/106 cells/day and System II (low-affinity and high-velocity), Km[APPARENT] = 3.25 × 10-5  M and VMAX[APPARENT] = 1000 nmol/106 cells/day. A third system of metabolism of AAF to AF, with Km[APPARENT] and VMAX[APPARENT] constants of 9.6 × 10-5  M and 4.7 nmol/106 cells/day, was also observed. Evidence provided in this report and its companion paper suggests selective roles and intracellular locations for System I- and System II-mediated AAF metabolite formation during hepatocarcinogenesis, although some of the molecules and mechanisms responsible for multi-system processing remain to be fully defined.

Novel Methods of Tritium Sequestration: High Temperature Gettering and Separation Membrane Materials Discovery for Nuclear Energy Systems

Energy Technology Data Exchange (ETDEWEB)

Chen, Franglin [Univ. of South Carolina, Columbia, SC (United States); Sholl, David [Georgia Inst. of Technology, Atlanta, GA (United States); Brinkman, Kyle [Savannah River Site (SRS), Aiken, SC (United States). Savannah River National Lab. (SRNL); Lyer, Ratnasabapathy [Claflin Univ., Orangeburg, SC (United States); Iyer, Ratnasabapathy [Claflin Univ., Orangeburg, SC (United States); Reifsnider, Kenneth [Univ. of South Carolina, Columbia, SC (United States)

2015-01-22

This project is aimed at addressing critical issues related to tritium sequestration in next generation nuclear energy systems. A technical hurdle to the use of high temperature heat from the exhaust produced in the next generation nuclear processes in commercial applications such as nuclear hydrogen production is the trace level of tritium present in the exhaust gas streams. This presents a significant challenge since the removal of tritium from the high temperature gas stream must be accomplished at elevated temperatures in order to subsequently make use of this heat in downstream processing. One aspect of the current project is to extend the techniques and knowledge base for metal hydride materials being developed for the ''hydrogen economy'' based on low temperature absorption/desorption of hydrogen to develop materials with adequate thermal stability and an affinity for hydrogen at elevated temperatures. The second focus area of this project is to evaluate high temperature proton conducting materials as hydrogen isotope separation membranes. Both computational and experimental approaches will be applied to enhance the knowledge base of hydrogen interactions with metal and metal oxide materials. The common theme between both branches of research is the emphasis on both composition and microstructure influence on the performance of sequestration materials.

Novel Methods of Tritium Sequestration: High Temperature Gettering and Separation Membrane Materials Discovery for Nuclear Energy Systems

International Nuclear Information System (INIS)

2015-01-01

This project is aimed at addressing critical issues related to tritium sequestration in next generation nuclear energy systems. A technical hurdle to the use of high temperature heat from the exhaust produced in the next generation nuclear processes in commercial applications such as nuclear hydrogen production is the trace level of tritium present in the exhaust gas streams. This presents a significant challenge since the removal of tritium from the high temperature gas stream must be accomplished at elevated temperatures in order to subsequently make use of this heat in downstream processing. One aspect of the current project is to extend the techniques and knowledge base for metal hydride materials being developed for the ''hydrogen economy'' based on low temperature absorption/desorption of hydrogen to develop materials with adequate thermal stability and an affinity for hydrogen at elevated temperatures. The second focus area of this project is to evaluate high temperature proton conducting materials as hydrogen isotope separation membranes. Both computational and experimental approaches will be applied to enhance the knowledge base of hydrogen interactions with metal and metal oxide materials. The common theme between both branches of research is the emphasis on both composition and microstructure influence on the performance of sequestration materials.

Application of Ni(II-assisted peptide bond hydrolysis to non-enzymatic affinity tag removal.

Directory of Open Access Journals (Sweden)

Edyta Kopera

Full Text Available In this study, we demonstrate a non-enzymatic method for hydrolytic peptide bond cleavage, applied to the removal of an affinity tag from a recombinant fusion protein, SPI2-SRHWAP-His(6. This method is based on a highly specific Ni(II reaction with (S/TXHZ peptide sequences. It can be applied for the protein attached to an affinity column or to the unbound protein in solution. We studied the effect of pH, temperature and Ni(II concentration on the efficacy of cleavage and developed an analytical protocol, which provides active protein with a 90% yield and ∼100% purity. The method works well in the presence of non-ionic detergents, DTT and GuHCl, therefore providing a viable alternative for currently used techniques.

Effect of the temperature in the sensitivity of CaSO4: Dy

International Nuclear Information System (INIS)

Gonzalez, P.R.; Alcantara, B.C.; Azorin, J.; Furetta, C.

2003-01-01

Sensitivity of a Tl material is one of the most important properties for dosimetry. This property is optimized by means of various processes such as the incorporation of different dopants into the matrix material, the application of certain doses of ionizing radiation and carefully controlled thermal treatments among others. Results obtained of studying the sensitivity of diverse preparations of CaSO 4 : Dy submitted different thermal treatments are presented. The material, which presented the highest sensitivity, was that one it was heated to 800 C during one hour. When the temperature was raised over this value it was observed that sensitivity drops again. In other hand, as the heating rate in lowing down, during the readout, the height of the glow curve decreases proportionally. The temperature of the maximum of the main peak also decreases. (Author)

Role of H2O2 on the kinetics of low-affinity high-capacity Na+-dependent alanine transport in SHR proximal tubular epithelial cells

International Nuclear Information System (INIS)

Pinto, Vanda; Pinho, Maria Joao; Jose, Pedro A.; Soares-da-Silva, Patricio

2010-01-01

Research highlights: → H 2 O 2 in excess is required for the presence of a low-affinity high-capacity component for the Na + -dependent [ 14 C]-L-alanine uptake in SHR PTE cells only. → It is suggested that Na + binding in renal ASCT2 may be regulated by ROS in SHR PTE cells. -- Abstract: The presence of high and low sodium affinity states for the Na + -dependent [ 14 C]-L-alanine uptake in immortalized renal proximal tubular epithelial (PTE) cells was previously reported (Am. J. Physiol. 293 (2007) R538-R547). This study evaluated the role of H 2 O 2 on the Na + -dependent [ 14 C]-L-alanine uptake of ASCT2 in immortalized renal PTE cells from Wistar Kyoto rat (WKY) and spontaneously hypertensive rat (SHR). Na + dependence of [ 14 C]-L-alanine uptake was investigated replacing NaCl with an equimolar concentration of choline chloride in vehicle- and apocynin-treated cells. Na + removal from the uptake solution abolished transport activity in both WKY and SHR PTE cells. Decreases in H 2 O 2 levels in the extracellular medium significantly reduced Na + -K m and V max values of the low-affinity high-capacity component in SHR PTE cells, with no effect on the high-affinity low-capacity state of the Na + -dependent [ 14 C]-L-alanine uptake. After removal of apocynin from the culture medium, H 2 O 2 levels returned to basal values within 1 to 3 h in both WKY and SHR PTE cells and these were found stable for the next 24 h. Under these experimental conditions, the Na + -K m and V max of the high-affinity low-capacity state were unaffected and the low-affinity high-capacity component remained significantly decreased 1 day but not 4 days after apocynin removal. In conclusion, H 2 O 2 in excess is required for the presence of a low-affinity high-capacity component for the Na + -dependent [ 14 C]-L-alanine uptake in SHR PTE cells only. It is suggested that Na + binding in renal ASCT2 may be regulated by ROS in SHR PTE cells.

Polymer-Particle Pressure-Sensitive Paint with High Photostability

Directory of Open Access Journals (Sweden)

Yu Matsuda

2016-04-01

Full Text Available We propose a novel fast-responding and paintable pressure-sensitive paint (PSP based on polymer particles, i.e. polymer-particle (pp-PSP. As a fast-responding PSP, polymer-ceramic (PC-PSP is widely studied. Since PC-PSP generally consists of titanium (IV oxide (TiO2 particles, a large reduction in the luminescent intensity will occur due to the photocatalytic action of TiO2. We propose the usage of polymer particles instead of TiO2 particles to prevent the reduction in the luminescent intensity. Here, we fabricate pp-PSP based on the polystyrene particle with a diameter of 1 μm, and investigate the pressure- and temperature-sensitives, the response time, and the photostability. The performances of pp-PSP are compared with those of PC-PSP, indicating the high photostability with the other characteristics comparable to PC-PSP.

A highly sensitive CMOS digital Hall sensor for low magnetic field applications.

Science.gov (United States)

Xu, Yue; Pan, Hong-Bin; He, Shu-Zhuan; Li, Li

2012-01-01

Integrated CMOS Hall sensors have been widely used to measure magnetic fields. However, they are difficult to work with in a low magnetic field environment due to their low sensitivity and large offset. This paper describes a highly sensitive digital Hall sensor fabricated in 0.18 μm high voltage CMOS technology for low field applications. The sensor consists of a switched cross-shaped Hall plate and a novel signal conditioner. It effectively eliminates offset and low frequency 1/f noise by applying a dynamic quadrature offset cancellation technique. The measured results show the optimal Hall plate achieves a high current related sensitivity of about 310 V/AT. The whole sensor has a remarkable ability to measure a minimum ± 2 mT magnetic field and output a digital Hall signal in a wide temperature range from -40 °C to 120 °C.

Novel high-affinity and selective biaromatic 4-substituted ¿-hydroxybutyric acid (GHB) analogues as GHB ligands

DEFF Research Database (Denmark)

Høg, Signe; Wellendorph, Petrine; Nielsen, Birgitte

2008-01-01

Gamma-hydroxybutyrate (GHB) is a metabolite of gamma-aminobutyric acid (GABA) and has been proposed to function as a neurotransmitter or neuromodulator. GHB is used in the treatment of narcolepsy and is a drug of abuse. GHB binds to both GABA(B) receptors and specific high-affinity GHB sites...

Temperature sensitivity of differential absorption lidar measurements of water vapor in the 720-nm region

Science.gov (United States)

Browell, Edward V.; Ismail, Syed; Grossmann, Benoist E.

1991-01-01

Recently measured properties of water vapor (H2O) absorption lines have been used in calculations to evalute the temperature sensitivity of differential absorption lidar (Dial) H2O measurements. This paper estimates the temperature sensitivity of H2O lines in the 717-733-nm region for both H2O mixing ratio and number density measurements, and discusses the influence of the H2O line ground state energies E-double-prime, the H2O absorption linewidths, the linewidth temperature dependence parameter, and the atmospheric temperature and pressure variations with altitude and location on the temperature sensitivity calculations. Line parameters and temperature sensitivity calculations for 67 H2O lines in the 720-nm band are given which can be directly used in field experiments. Water vapor lines with E-double-prime values in the 100-300/cm range were found to be optimum for Dial measurements of H2O number densities, while E-double-prime values in the 250-500/cm range were found to be optimum for H2O mixing ratio measurements.

Structure-guided development of a high-affinity human Programmed Cell Death-1: Implications for tumor immunotherapy

Energy Technology Data Exchange (ETDEWEB)

Lázár-Molnár, Eszter; Scandiuzzi, Lisa; Basu, Indranil; Quinn, Thomas; Sylvestre, Eliezer; Palmieri, Edith; Ramagopal, Udupi A.; Nathenson, Stanley G.; Guha, Chandan; Almo, Steven C.

2017-03-01

Programmed Cell Death-1 (PD-1) is an inhibitory immune receptor, which plays critical roles in T cell co-inhibition and exhaustion upon binding to its ligands PD-L1 and PD-L2. We report the crystal structure of the human PD-1 ectodomain and the mapping of the PD-1 binding interface. Mutagenesis studies confirmed the crystallographic interface, and resulted in mutant PD-1 receptors with altered affinity and ligand-specificity. In particular, a high-affinity mutant PD-1 (HA PD-1) exhibited 45 and 30-fold increase in binding to PD-L1 and PD-L2, respectively, due to slower dissociation rates. This mutant (A132L) was used to engineer a soluble chimeric Ig fusion protein for cell-based and in vivo studies. HA PD-1 Ig showed enhanced binding to human dendritic cells, and increased T cell proliferation and cytokine production in a mixed lymphocyte reaction (MLR) assay. Moreover, in an experimental model of murine Lewis lung carcinoma, HA PD-1 Ig treatment synergized with radiation therapy to decrease local and metastatic tumor burden, as well as in the establishment of immunological memory responses. Our studies highlight the value of structural considerations in guiding the design of a high-affinity chimeric PD-1 Ig fusion protein with robust immune modulatory properties, and underscore the power of combination therapies to selectively manipulate the PD-1 pathway for tumor immunotherapy.

Structure-guided development of a high-affinity human Programmed Cell Death-1: Implications for tumor immunotherapy

Directory of Open Access Journals (Sweden)

Eszter Lázár-Molnár

2017-03-01

Full Text Available Programmed Cell Death-1 (PD-1 is an inhibitory immune receptor, which plays critical roles in T cell co-inhibition and exhaustion upon binding to its ligands PD-L1 and PD-L2. We report the crystal structure of the human PD-1 ectodomain and the mapping of the PD-1 binding interface. Mutagenesis studies confirmed the crystallographic interface, and resulted in mutant PD-1 receptors with altered affinity and ligand-specificity. In particular, a high-affinity mutant PD-1 (HA PD-1 exhibited 45 and 30-fold increase in binding to PD-L1 and PD-L2, respectively, due to slower dissociation rates. This mutant (A132L was used to engineer a soluble chimeric Ig fusion protein for cell-based and in vivo studies. HA PD-1 Ig showed enhanced binding to human dendritic cells, and increased T cell proliferation and cytokine production in a mixed lymphocyte reaction (MLR assay. Moreover, in an experimental model of murine Lewis lung carcinoma, HA PD-1 Ig treatment synergized with radiation therapy to decrease local and metastatic tumor burden, as well as in the establishment of immunological memory responses. Our studies highlight the value of structural considerations in guiding the design of a high-affinity chimeric PD-1 Ig fusion protein with robust immune modulatory properties, and underscore the power of combination therapies to selectively manipulate the PD-1 pathway for tumor immunotherapy.

Responses to cholinergic agonists of rats selectively bred for differential sensitivity to ethanol.

Science.gov (United States)

de Fiebre, C M; Romm, E; Collins, J T; Draski, L J; Deitrich, R A; Collins, A C

1991-03-01

Alcoholics are almost invariably heavy users of tobacco. Both alcoholism and smoking appear to be influenced by genetic factors but it is not known whether the same or different genes regulate the abuse of ethanol and nicotine. Recent studies have demonstrated that the long-sleep (LS) and short-sleep (SS) mouse lines, which were selectively bred for differences in ethanol-induced anesthesia ("sleep-time"), also differ in several effects of nicotine and the muscarinic agonist, oxotremorine. In order to determine whether or not these differences are due to chance, the relative sensitivities of rat lines which were selectively bred for differences in ethanol-induced sleep-time were determined. The high alcohol sensitivity (HAS) rat line was more sensitive to the locomotor and body temperature depressant effects of nicotine than was the low alcohol sensitivity (LAS) rat line. The control line (CAS) was intermediate in sensitivity. The rat lines did not differ in sensitivity to oxotremorine's hypothermia-producing effects. The numbers and affinities of two classes of brain nicotinic receptors were measured in eight brain regions. No differences among the rat lines were detected. These results suggest that ethanol elicits some of its depressant actions via an effect on brain nicotinic systems, but the differences in sensitivity to ethanol and nicotine are probably not due to differences in the number of brain nicotinic receptors. Perhaps this interaction explains the high correlation between alcoholism and smoking in humans.

High temperature combustion facility: present capabilities and future prospects

International Nuclear Information System (INIS)

Boccio, J.L.; Ginsberg, T.; Ciccarelli, G.

1995-01-01

The high-temperature combustion facility constructed and operated by the Department of Advanced Technology of Brookhaven National Laboratory to support and promote research in the area of hydrogen combustion phenomena in mixtures prototypical to light-water reactor containment atmospheres under potential severe accident conditions is reported. The facility can accommodate combustion research activities encompassing the fields of detonation physics, flame acceleration, and low-speed deflagration in a wide range of combustible gas mixtures at initial temperatures up to 700 K and post-combustion pressures up to 100 atmospheres. Some preliminary test results are presented that provide further evidence that the effect of temperature is to increase the sensitivity of hydrogen-air-steam mixtures to undergo detonation [ru

Preconceptual design of hyfire. A fusion driven high temperature electrolysis plant

International Nuclear Information System (INIS)

Varljen, T.C.; Chi, J.W.H.; Karbowski, J.S.

1983-01-01

Brookhaven National Laboratory has been engaged in a scoping study to investigate the potential merits of coupling a fusion reactor with a high temperature blanket to a high temperature electrolysis (HTE) process to produce hydrogen and oxygen. Westinghouse is assisting this study in the areas of systems design integration, plasma engineering, balance of plant design and electrolyzer technology. The aim of the work done in the past year has been to focus on a reference design point for the plant, which has been designated HYFIRE. In prior work, the STARFIRE commercial tokamak fusion reactor was directly used as the fusion driver. This report describes a new design obtained by scaling the basic STARFIRE design to permit the achievement of a blanket power of 6000 MWt. The high temperature blanket design employs a thermally insulated refractory oxide region which provides high temperature (>1000 deg. C) steam at moderate pressures to high temperature electrolysis units. The electrolysis process selected is based on the high temperature, solid electrolyte fuel cell technology developed by Westinghouse. An initial process design and plant layout has been completed; component cost and plant economics studies are now underway to develop estimates of hydrogen production costs and to determine the sensitivity of this cost to changes in major design parameters. (author)

Highly sensitive room temperature organic vapor sensor based on polybenzoxazine-derived carbon aerogel thin film composite

Energy Technology Data Exchange (ETDEWEB)

Thubsuang, Uthen [Materials Science and Engineering, School of Engineering and Resources, Walailak University, Nakhon Si Thammarat 80160 (Thailand); Sukanan, Darunee [The Petroleum and Petrochemical College and the Center of Excellence on Petrochemical and Materials Technology, Chulalongkorn University, Bangkok 10330 (Thailand); Sahasithiwat, Somboon [National Metal and Materials Technology Center, Thailand Science Park (TSP), Khlong Luang, Pathum Thani 12120 (Thailand); Wongkasemjit, Sujitra [The Petroleum and Petrochemical College and the Center of Excellence on Petrochemical and Materials Technology, Chulalongkorn University, Bangkok 10330 (Thailand); Chaisuwan, Thanyalak, E-mail: thanyalak.c@chula.ac.th [The Petroleum and Petrochemical College and the Center of Excellence on Petrochemical and Materials Technology, Chulalongkorn University, Bangkok 10330 (Thailand)

2015-10-15

Graphical abstract: - Highlights: • Activated carbon aerogel with high surface area can be prepared from polybenzoxazine. • Activated carbon aerogel enhances the adsorption capacity of gas sensor. • Organic vapors with very low concentration can be detected by the as-prepared sensor. • The as-prepared sensor shows impressive short exposure and recovery time. • The response to different organic vapors can be tailored by changing polymer matrix. - Abstract: Gas sensing composites were fabricated using polybenzoxazine-based activated carbon aerogel as a conductive filler. The activated carbon aerogel is a nano-porous material, which has high pore volume of 0.57 cm{sup 3}/g and surface area of 917 m{sup 2}/g. The activated carbon aerogel/polybutadiene composite displayed good response of 11.2 and 6.7 to toluene and n-hexane, respectively, compared to those of graphite/polybutadiene composite. The activated carbon aerogel/polybutadiene composite also showed high sensitivity of 3.09 × 10{sup 2} ppm{sup −1} to toluene. However, the sensitivity of activated carbon aerogel/polybutadiene composite drastically decreased to 1.99 ppm{sup −1} and zero when exposed to acetone and water, respectively. Contrarily, when polyvinyl alcohol was used as a matrix, the sensitivity was about 4.19 ppm{sup −1} to water. While the composite was found to be not sensitive to toluene. The activated carbon aerogel/polybutadiene composite also showed good recovery as the electrical resistance came back to the original value within minutes when exposed to nitrogen gas.

Increase of ozone concentrations, its temperature sensitivity and the precursor factor in South China

Directory of Open Access Journals (Sweden)

Y. C. Lee

2014-08-01

Full Text Available Concerns have been raised about the possible connections between the local and regional photochemical problem and global warming. The current study assesses the trend of ozone in Hong Kong and the Pearl River Delta (PRD in South China and investigates the interannual changes of sensitivity of ozone to air temperature, as well as the trends in regional precursors. Results reveal, at the three monitoring sites from the mid-1990s to 2010, an increase in the mean ozone concentrations from 1.0 to 1.6 µg m−3 per year. The increase occurred in all seasons, with the highest rate in autumn. This is consistent with trends and temperature anomalies in the region. The increase in the sensitivity of ozone to temperature is clearly evident from the correlation between ozone (OMI [Ozone Monitoring Instrument] column amount and surface air temperature (from the Atmospheric Infrared Sounder displayed in the correlation maps for the PRD during the prominently high ozone period of July–September. It is observed to have increased from 2005 to 2010, the latter being the hottest year on record globally. To verify this temporal change in sensitivity, the ground-level trends of correlation coefficients/regression slopes are analysed. As expected, results reveal a statistically significant upward trend over a 14-year period (1997–2010. While the correlation revealed in the correlation maps is in agreement with the corresponding OMI ozone maps when juxtaposed, temperature sensitivity of surface ozone also shows an association with ozone concentration, with R=0.5. These characteristics of ozone sensitivity are believed to have adverse implications for the region. As shown by ground measurements and/or satellite analyses, the decrease in nitrogen oxides (NO2 and NOx in Hong Kong is not statistically significant while NO2 of the PRD has only very slightly changed. However, carbon dioxide has remarkably declined in the whole region. While these observations concerning

Characterization of the temperature-sensitive mutations un-7 and png-1 in Neurospora crassa.

Science.gov (United States)

Dieterle, Michael G; Wiest, Aric E; Plamann, Mike; McCluskey, Kevin

2010-05-18

The model filamentous fungus Neurospora crassa has been studied for over fifty years and many temperature-sensitive mutants have been generated. While most of these have been mapped genetically, many remain anonymous. The mutation in the N. crassa temperature-sensitive lethal mutant un-7 was identified by a complementation based approach as being in the open reading frame designated NCU00651 on linkage group I. Other mutations in this gene have been identified that lead to a temperature-sensitive morphological phenotype called png-1. The mutations underlying un-7 result in a serine to phenylalanine change at position 273 and an isoleucine to valine change at position 390, while the mutation in png-1 was found to result in a serine to leucine change at position 279 although there were other conservative changes in this allele. The overall morphology of the strain carrying the un-7 mutation is compared to strains carrying the png-1 mutation and these mutations are evaluated in the context of other temperature-sensitive mutants in Neurospora.

Synthesis of hapten and preparation of specific polyclonal antibody with high affinity for lenalidomide, the potent drug for treatment of multiple myeloma

Directory of Open Access Journals (Sweden)

Darwish Ibrahim A

2012-10-01

Full Text Available Abstract Background For therapeutic monitoring and pharmacokinetic studies of lenalidomide (LND, the potent drug for treatment of multiple myeloma (MM, a specific antibody was required for the development of a sensitive immunoassay system for the accurate determination of LND in plasma. Results In this study, a hapten of LND (N-glutaryl-LND was synthesized by introducing the glutaryl moiety, as a spacer, into the primary aromatic amine site of the LND molecular structure. The structure of the hapten (G-LND was confirmed by mass, 1H-NMR, and 13C spectrometric techniques. G-LND was coupled to each of bovine serum albumin (BSA and keyhole limpet hemocyanin (KLH proteins by ethyl-3-(3-dimethylaminopropyl carbodiimide as a coupling reagent. LND-KLH conjugate was used as an immunogen. Four female 2-3 months old New Zealand white rabbits were immunized with an emulsion of LND-KLH with Freund`s adjuvant. The immune response of the rabbits was monitored by direct enzyme-linked immunosorbent assay (ELISA using LND-BSA immobilized onto microwell plates as a solid phase. The rabbit that showed the highest antibody titer and affinity to LND was scarified and its sera were collected. The IgG fraction was isolated and purified by affinity chromatography on protein A column. The specificity of the purified antibody for LND was evaluated by indirect competitive ELISA using dexamethasone as a competitor as it is used with LND in a combination therapy. Conclusions The high affinity of the antibody (IC50 = 10 ng/mL will be useful in the development of an immunoassay system for the determination of plasma LND concentrations. Current research is going to optimize the assay conditions and validate the procedures for the routine application in clinical laboratories.

Synthesis of hapten and preparation of specific polyclonal antibody with high affinity for lenalidomide, the potent drug for treatment of multiple myeloma.

Science.gov (United States)

Darwish, Ibrahim A; Alzoman, Nourh Z; Abuhejail, Reem M; El-Samani, Tilal E

2012-10-26

For therapeutic monitoring and pharmacokinetic studies of lenalidomide (LND), the potent drug for treatment of multiple myeloma (MM), a specific antibody was required for the development of a sensitive immunoassay system for the accurate determination of LND in plasma. In this study, a hapten of LND (N-glutaryl-LND) was synthesized by introducing the glutaryl moiety, as a spacer, into the primary aromatic amine site of the LND molecular structure. The structure of the hapten (G-LND) was confirmed by mass, 1H-NMR, and 13C spectrometric techniques. G-LND was coupled to each of bovine serum albumin (BSA) and keyhole limpet hemocyanin (KLH) proteins by ethyl-3-(3-dimethylaminopropyl) carbodiimide as a coupling reagent. LND-KLH conjugate was used as an immunogen. Four female 2-3 months old New Zealand white rabbits were immunized with an emulsion of LND-KLH with Freund`s adjuvant. The immune response of the rabbits was monitored by direct enzyme-linked immunosorbent assay (ELISA) using LND-BSA immobilized onto microwell plates as a solid phase. The rabbit that showed the highest antibody titer and affinity to LND was scarified and its sera were collected. The IgG fraction was isolated and purified by affinity chromatography on protein A column. The specificity of the purified antibody for LND was evaluated by indirect competitive ELISA using dexamethasone as a competitor as it is used with LND in a combination therapy. The high affinity of the antibody (IC50 = 10 ng/mL) will be useful in the development of an immunoassay system for the determination of plasma LND concentrations. Current research is going to optimize the assay conditions and validate the procedures for the routine application in clinical laboratories.

Hydration, phase separation and nonlinear rheology of temperature-sensitive water-soluble polymers

Energy Technology Data Exchange (ETDEWEB)

Tanaka, Fumihiko; Koga, Tsuyoshi [Department of Polymer Chemistry, Graduate School of Engineering, Kyoto University, Kyoto 615-8510 (Japan); Kaneda, Isamu [Department of Food Science, Rakuno Gakuen University, Ebetsu, Hokkaido 069-8501 (Japan); Winnik, Francoise M, E-mail: ftanaka@phys.polym.kyoto-u.ac.jp [Department of Chemistry and Faculty of Pharmacy, University of Montreal, Montreal, H3C 3J7 (Canada)

2011-07-20

The collapse of a poly(N-isopropylacrylamide) (PNIPAM) chain upon heating and the phase diagrams of aqueous PNIPAM solutions with a very flat lower critical solution temperature (LCST) phase separation line are theoretically studied on the basis of cooperative dehydration (simultaneous dissociation of bound water molecules in a group of correlated sequence), and compared with the experimental observation of temperature-induced coil-globule transition by light scattering methods. The transition becomes sharper with the cooperativity parameter {sigma} of hydration. The reentrant coil-globule-coil transition and cononsolvency in a mixed solvent of water and methanol are also studied from the viewpoint of competitive hydrogen bonds between polymer-water and polymer-methanol. The downward shift of the cloud-point curves (LCST cononsolvency) with the mol fraction of methanol due to the competition is calculated and compared with the experimental data. Aqueous solutions of hydrophobically modified PNIPAM carrying short alkyl chains at both chain ends (telechelic PNIPAM) are theoretically and experimentally studied. The LCST of these solutions is found to shift downward along the sol-gel transition curve as a result of end-chain association (association-induced phase separation), and separate from the coil-globule transition line. Associated structures in the solution, such as flower micelles, mesoglobules, and higher fractal assembly, are studied by ultra small-angle neutron scattering with theoretical modeling of the scattering function. Dynamic-mechanical modulus, nonlinear stationary viscosity, and stress build-up in start-up shear flows of the associated networks are studied on the basis of the affine and non-affine transient network theory. The molecular conditions for thickening, strain hardening, and stress overshoot are found in terms of the nonlinear amplitude A of the chain tension and the tension-dissociation coupling constant g.

Mg/Ca- Δ CO3porewater2- -temperature calibration for Globobulimina spp.: A sensitive paleothermometer for deep-sea temperature reconstruction

Science.gov (United States)

Weldeab, Syee; Arce, Adam; Kasten, Sabine

2016-03-01

Existing benthic foraminiferal Mg/Ca-temperature calibrations are surrounded by substantial uncertainties mainly due to low temperature sensitivity of Mg/Ca in most benthic foraminifers and the effect of carbonate ion concentration on benthic foraminiferal Mg/Ca. Here we present Mg/Ca analysis of Rose Bengal stained and exceptionally well-preserved tests of the infaunal benthic foraminifer Globobulimina spp. from 39 eastern equatorial Atlantic core top samples. Mg/Ca in Globobulimina spp. varies between 2.5 mmol/mol and 9.1 mmol/mol corresponding to bottom water temperatures (BWT) between 1.8 °C and 19.1 °C and Δ CO3pore water2- between 33.7 ± 4 and - 34.3 ± 4 μmol /kg in sediment depths between 1 and 10 cm. Mg/Ca and BWT are linearly correlated with a best fit of Mg/Ca [mmol/mol] = (0.36 ± 0.02) * BWT [°C] + 2.22 ± 0.19 (r2 = 0.92, p-value: 11 *10-20, and n = 39). Using total alkalinity and pH data of pore water samples from 64 Atlantic multi-corer sites, we obtained Δ CO3pore water2- data from the depth habitat range of Globobulimina spp. (≥1 cm ≤ 10 cm below sediment surface). We show that Δ CO3pore waterSUP>2- is significantly lower than and linearly co-varies with the ΔCO2-3 of the overlying bottom water: Δ CO3pore water2- = (0.67 ± 0.05) * Δ CO3bottom water2- - (39.84 ± 1.98); r2 = 0.75, p-value: 6 *10-20, n = 64. We found a Mg/Ca sensitivity of 0.009 ± 0.0044 mmol /mol per μmol/kg Δ CO3pore water2- and Mg/Ca temperature sensitivity of 0.32 ± 0.06 mmol /mol / °C after a correction for the Δ CO3pore water2- effect. This study provides a robust Mg/Ca-temperature calibration, highlights that Δ CO3pore water2- is spatially and most likely temporally variable, and contradicts the notion that infaunal foraminiferal Mg/Ca is relatively immune from ΔCO2-3 changes in the overlying bottom water. Furthermore, comparison of down core Mg/Ca data of Cibicides pachyderma and Globobulimina spp. demonstrates that the high temperature sensitivity of

Mapping of barley alpha-amylases and outer subsite mutants reveals dynamic high-affinity subsites and barriers in the long substrate binding cleft

DEFF Research Database (Denmark)

Kandra, L.; Abou Hachem, Maher; Gyemant, G.

2006-01-01

Subsite affinity maps of long substrate binding clefts in barley alpha-amylases, obtained using a series of maltooligosaccharides of degree of polymerization of 3-12, revealed unfavorable binding energies at the internal subsites -3 and -5 and at subsites -8 and +3/+4 defining these subsites...... as binding barriers. Barley a-amylase I mutants Y105A and T212Y at subsite -6 and +4 resulted in release or anchoring of bound substrate, thus modifying the affinities of other high-affinity subsites (-2 and +2) and barriers. The double mutant Y105A-T212Y displayed a hybrid subsite affinity profile......, converting barriers to binding areas. These findings highlight the dynamic binding energy distribution and the versatility of long maltooligosaccharide derivatives in mapping extended binding clefts in a-amylases....

Sea-ice cover in the Nordic Seas and the sensitivity to Atlantic water temperatures

Science.gov (United States)

Jensen, Mari F.; Nisancioglu, Kerim H.; Spall, Michael A.

2017-04-01

Changes in the sea-ice cover of the Nordic Seas have been proposed to play a key role for the dramatic temperature excursions associated with the Dansgaard-Oeschger events during the last glacial. However, with its proximity to the warm Atlantic water, how a sea-ice cover can persist in the Nordic Seas is not well understood. In this study, we apply an eddy-resolving configuration of the Massachusetts Institute of Technology general circulation model with an idealized topography to study the presence of sea ice in a Nordic Seas-like domain. We assume an infinite amount of warm Atlantic water present in the south by restoring the southern area to constant temperatures. The sea-surface temperatures are restored toward cold, atmospheric temperatures, and as a result, sea ice is present in the interior of the domain. However, the sea-ice cover in the margins of the Nordic Seas, an area with a warm, cyclonic boundary current, is sensitive to the amount of heat entering the domain, i.e., the restoring temperature in the south. When the temperature of the warm, cyclonic boundary current is high, the margins are free of sea ice and heat is released to the atmosphere. We show that with a small reduction in the temperature of the incoming Atlantic water, the Nordic Seas-like domain is fully covered in sea ice. Warm water is still entering the Nordic Seas, however, this happens at depths below a cold, fresh surface layer produced by melted sea ice. Consequently, the heat release to the atmosphere is reduced along with the eddy heat fluxes. Results suggest a threshold value in the amount of heat entering the Nordic Seas before the sea-ice cover disappears in the margins. We study the sensitivity of this threshold to changes in atmospheric temperatures and vertical diffusivity.

High Pressure and Temperature Core Formation as an Alternative to the "Late Veneer" Hypothesis

Science.gov (United States)

Righter, Kevin; Pando, K.; Humayun, M.; Danielson, L.

2011-01-01

The highly siderophile elements (HSE; Re, Au and the Platinum Group Elements - Pd Pt, Rh, Ru, Ir, Os) are commonly utilized to constrain accretion processes in terrestrial differentiated bodies due to their affinity for FeNi metal [1]. These eight elements exhibit highly siderophile behavior, but nonetheless have highly diverse metal-silicate partition coefficients [2]. Therefore the near chondritic relative concentrations of HSEs in the terrestrial and lunar mantles, as well as some other bodies, are attributed to late accretion rather than core formation [1]. Evaluation of competing theories, such as high pressure metal-silicate partitioning or magma ocean hypotheses has been hindered by a lack of relevant partitioning data for this group of eight elements. In particular, systematic studies isolating the effect of one variable (e.g. temperature or melt compositions) are lacking. Here we undertake new experiments on all eight elements, using Fe metal and FeO-bearing silicate melts at fixed pressure, but variable temperatures. These experiments, as well as some additional planned experiments should allow partition coefficients to be more accurately calculated or estimated at the PT conditions and compositions at which core formation is thought to have occurred.

Characterization of glucagon-like peptide-1 receptor beta-arrestin 2 interaction: a high-affinity receptor phenotype

DEFF Research Database (Denmark)

Jorgensen, Rasmus; Martini, Lene; Schwartz, Thue W

2005-01-01

To dissect the interaction between beta-arrestin ((beta)arr) and family B G protein-coupled receptors, we constructed fusion proteins between the glucagon-like peptide 1 receptor and (beta)arr2. The fusion constructs had an increase in apparent affinity selectively for glucagon, suggesting...... that (beta)arr2 interaction locks the receptor in a high-affinity conformation, which can be explored by some, but not all, ligands. The fusion constructs adopted a signaling phenotype governed by the tethered (beta)arr2 with an attenuated G protein-mediated cAMP signal and a higher maximal internalization...... of that which has previously been characterized for family A G protein-coupled receptors, suggesting similarities in the effect of (beta)arr interaction between family A and B receptors also at the molecular level....

Ammonia oxidation at high pressure and intermediate temperatures

DEFF Research Database (Denmark)

Song, Yu; Hashemi, Hamid; Christensen, Jakob Munkholt

2016-01-01

Ammonia oxidation experiments were conducted at high pressure (30 bar and 100 bar) under oxidizing and stoichiometric conditions, respectively, and temperatures ranging from 450 to 925 K. The oxidation of ammonia was slow under stoichiometric conditions in the temperature range investigated. Under...... oxidizing conditions the onset temperature for reaction was 850–875 K at 30 bar, while at 100 bar it was about 800 K, with complete consumption of NH3 at 875 K. The products of reaction were N2 and N2O, while NO and NO2 concentrations were below the detection limit even under oxidizing conditions. The data...... was satisfactory. The main oxidation path for NH3 at high pressure under oxidizing conditions is NH3⟶+OH NH2⟶+HO2,NO2 H2NO⟶+O2 HNO⟶+O2 NO ⟶+NH2 N2. The modeling predictions are most sensitive to the reactions NH2 + NO = NNH + OH and NH2 + HO2 = H2NO + OH, which promote the ammonia consumption by forming OH...

An in vitro-identified high-affinity nucleosome-positioning signal is capable of transiently positioning a nucleosome in vivo

Directory of Open Access Journals (Sweden)

Gracey Lia E

2010-07-01

Full Text Available Abstract Background The physiological function of eukaryotic DNA occurs in the context of nucleosomal arrays that can expose or obscure defined segments of the genome. Certain DNA sequences are capable of strongly positioning a nucleosome in vitro, suggesting the possibility that favorable intrinsic signals might reproducibly structure chromatin segments. As high-throughput sequencing analyses of nucleosome coverage in vitro and in vivo have become possible, a vigorous debate has arisen over the degree to which intrinsic DNA:nucleosome affinities orchestrate the in vivo positions of nucleosomes, thereby controlling physical accessibility of specific sequences in DNA. Results We describe here the in vivo consequences of placing a synthetic high-affinity nucleosome-positioning signal, the 601 sequence, into a DNA plasmid vector in mice. Strikingly, the 601 sequence was sufficient to position nucleosomes during an early phase after introduction of the DNA into the mice (when the plasmid vector transgene was active. This positioning capability was transient, with a loss of strong positioning at a later time point when the transgenes had become silent. Conclusions These results demonstrate an ability of DNA sequences selected solely for nucleosome affinity to organize chromatin in vivo, and the ability of other mechanisms to overcome these interactions in a dynamic nuclear environment.

Photo-sensitive Ge nanocrystal based films controlled by substrate deposition temperature

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Stavarache, Ionel; Maraloiu, Valentin Adrian; Negrila, Catalin; Prepelita, Petronela; Gruia, Ion; Iordache, Gheorghe

2017-10-01

Lowering the temperature of crystallization by deposition of thin films on a heated substrate represents the easiest way to find new means to develop and improve new working devices based on nanocrystals embedded in thin films. The improvements are strongly related with the increasing of operation speed, substantially decreasing the energy consumption and reducing unit fabrication costs of the respective semiconductor devices. This approach avoids major problems, such as those related to diffusion or difficulties in controlling nanocrystallites size, which appear during thermal treatments at high temperatures after deposition. This article reports on a significant progress given by structuring Ge nanocrystals (Ge-NCs) embedded in silicon dioxide (SiO2) thin films by heating the substrate at 400 °C during co-deposition of Ge and SiO2 by magnetron sputtering. As a proof-of-concept, a Si/Ge-NCs:SiO2 photo-sensitive structure was fabricated thereof and characterized. The structure shows superior performance on broad operation bandwidth from visible to near-infrared, as strong rectification properties in dark, significant current rise in the inversion mode when illuminated, high responsivity, high photo-detectivity of 1014 Jones, quick response and significant conversion efficiency with peak value reaching 850% at -1 V and about 1000 nm. This simple preparation approach brings an important contribution to the effort of structuring Ge nanocrystallites in SiO2 thin films at a lower temperature for the purpose of using these materials for devices in optoelectronics, solar cells and electronics on flexible substrates.

High and low temperatures have unequal reinforcing properties in Drosophila spatial learning.

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Zars, Melissa; Zars, Troy

2006-07-01

Small insects regulate their body temperature solely through behavior. Thus, sensing environmental temperature and implementing an appropriate behavioral strategy can be critical for survival. The fly Drosophila melanogaster prefers 24 degrees C, avoiding higher and lower temperatures when tested on a temperature gradient. Furthermore, temperatures above 24 degrees C have negative reinforcing properties. In contrast, we found that flies have a preference in operant learning experiments for a low-temperature-associated position rather than the 24 degrees C alternative in the heat-box. Two additional differences between high- and low-temperature reinforcement, i.e., temperatures above and below 24 degrees C, were found. Temperatures equally above and below 24 degrees C did not reinforce equally and only high temperatures supported increased memory performance with reversal conditioning. Finally, low- and high-temperature reinforced memories are similarly sensitive to two genetic mutations. Together these results indicate the qualitative meaning of temperatures below 24 degrees C depends on the dynamics of the temperatures encountered and that the reinforcing effects of these temperatures depend on at least some common genetic components. Conceptualizing these results using the Wolf-Heisenberg model of operant conditioning, we propose the maximum difference in experienced temperatures determines the magnitude of the reinforcement input to a conditioning circuit.

Species composition and forest structure explain the temperature sensitivity patterns of productivity in temperate forests

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F. J. Bohn

2018-03-01

Full Text Available Rising temperatures due to climate change influence the wood production of forests. Observations show that some temperate forests increase their productivity, whereas others reduce their productivity. This study focuses on how species composition and forest structure properties influence the temperature sensitivity of aboveground wood production (AWP. It further investigates which forests will increase their productivity the most with rising temperatures. We described forest structure by leaf area index, forest height and tree height heterogeneity. Species composition was described by a functional diversity index (Rao's Q and a species distribution index (ΩAWP. ΩAWP quantified how well species are distributed over the different forest layers with regard to AWP. We analysed 370 170 forest stands generated with a forest gap model. These forest stands covered a wide range of possible forest types. For each stand, we estimated annual aboveground wood production and performed a climate sensitivity analysis based on 320 different climate time series (of 1-year length. The scenarios differed in mean annual temperature and annual temperature amplitude. Temperature sensitivity of wood production was quantified as the relative change in productivity resulting from a 1 °C rise in mean annual temperature or annual temperature amplitude. Increasing ΩAWP positively influenced both temperature sensitivity indices of forest, whereas forest height showed a bell-shaped relationship with both indices. Further, we found forests in each successional stage that are positively affected by temperature rise. For such forests, large ΩAWP values were important. In the case of young forests, low functional diversity and small tree height heterogeneity were associated with a positive effect of temperature on wood production. During later successional stages, higher species diversity and larger tree height heterogeneity were an advantage. To achieve such a

Species composition and forest structure explain the temperature sensitivity patterns of productivity in temperate forests

Science.gov (United States)

Bohn, Friedrich J.; May, Felix; Huth, Andreas

2018-03-01

Rising temperatures due to climate change influence the wood production of forests. Observations show that some temperate forests increase their productivity, whereas others reduce their productivity. This study focuses on how species composition and forest structure properties influence the temperature sensitivity of aboveground wood production (AWP). It further investigates which forests will increase their productivity the most with rising temperatures. We described forest structure by leaf area index, forest height and tree height heterogeneity. Species composition was described by a functional diversity index (Rao's Q) and a species distribution index (ΩAWP). ΩAWP quantified how well species are distributed over the different forest layers with regard to AWP. We analysed 370 170 forest stands generated with a forest gap model. These forest stands covered a wide range of possible forest types. For each stand, we estimated annual aboveground wood production and performed a climate sensitivity analysis based on 320 different climate time series (of 1-year length). The scenarios differed in mean annual temperature and annual temperature amplitude. Temperature sensitivity of wood production was quantified as the relative change in productivity resulting from a 1 °C rise in mean annual temperature or annual temperature amplitude. Increasing ΩAWP positively influenced both temperature sensitivity indices of forest, whereas forest height showed a bell-shaped relationship with both indices. Further, we found forests in each successional stage that are positively affected by temperature rise. For such forests, large ΩAWP values were important. In the case of young forests, low functional diversity and small tree height heterogeneity were associated with a positive effect of temperature on wood production. During later successional stages, higher species diversity and larger tree height heterogeneity were an advantage. To achieve such a development, one could plant

Evolution of vertebrate transient receptor potential vanilloid 3 channels: opposite temperature sensitivity between mammals and western clawed frogs.

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Shigeru Saito

2011-04-01

Full Text Available Transient Receptor Potential (TRP channels serve as temperature receptors in a wide variety of animals and must have played crucial roles in thermal adaptation. The TRP vanilloid (TRPV subfamily contains several temperature receptors with different temperature sensitivities. The TRPV3 channel is known to be highly expressed in skin, where it is activated by warm temperatures and serves as a sensor to detect ambient temperatures near the body temperature of homeothermic animals such as mammals. Here we performed comprehensive comparative analyses of the TRPV subfamily in order to understand the evolutionary process; we identified novel TRPV genes and also characterized the evolutionary flexibility of TRPV3 during vertebrate evolution. We cloned the TRPV3 channel from the western clawed frog Xenopus tropicalis to understand the functional evolution of the TRPV3 channel. The amino acid sequences of the N- and C-terminal regions of the TRPV3 channel were highly diversified from those of other terrestrial vertebrate TRPV3 channels, although central portions were well conserved. In a heterologous expression system, several mammalian TRPV3 agonists did not activate the TRPV3 channel of the western clawed frog. Moreover, the frog TRPV3 channel did not respond to heat stimuli, instead it was activated by cold temperatures. Temperature thresholds for activation were about 16 °C, slightly below the lower temperature limit for the western clawed frog. Given that the TRPV3 channel is expressed in skin, its likely role is to detect noxious cold temperatures. Thus, the western clawed frog and mammals acquired opposite temperature sensitivity of the TRPV3 channel in order to detect environmental temperatures suitable for their respective species, indicating that temperature receptors can dynamically change properties to adapt to different thermal environments during evolution.

Enhancement of NH3 gas sensitivity at room temperature by carbon nanotube-based sensor coated with Co nanoparticles.

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Nguyen, Lich Quang; Phan, Pho Quoc; Duong, Huyen Ngoc; Nguyen, Chien Duc; Nguyen, Lam Huu

2013-01-30

Multi-walled carbon nanotube (MWCNT) film has been fabricated onto Pt-patterned alumina substrates using the chemical vapor deposition method for NH(3) gas sensing applications. The MWCNT-based sensor is sensitive to NH(3) gas at room temperature. Nanoclusters of Co catalysts have been sputtered on the surface of the MWCNT film to enhance gas sensitivity with respect to unfunctionalized CNT films. The gas sensitivity of Co-functionalized MWCNT-based gas sensors is thus significantly improved. The sensor exhibits good repeatability and high selectivity towards NH(3), compared with alcohol and LPG.

Sensitivity of LUCC on the Surface Temperature of Tibetan Plateau

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Qi, W.; Deng, X.; Wu, F.

2016-12-01

The Tibetan Plateau has an important effect on the ecological security in China, even in Asia, which makes the region become the hot spot in recently research. Under the joint influence of global change and human activities, ecosystem destabilizing and the increasing pressure on resources and environment emerge on the Tibetan Plateau, but the potential spatial sensitivity of land use and land cover changes(LUCC) on surface temperature has not been quantitatively analyzed. This study analyzed the mainly types of LUCC, urbanization, grassland degradation, deforestation on Tibetan Plateau along with Representative Concentration Pathways (RCPs) of the Intergovernmental Panel on Climate Change (IPCC). The LUCC in recent decades was first quantitatively analyzed in this study to give the basic fact with a significant increase in temperatures, reduced precipitation and increased evaporation. This study focused on the future spatio-temporal heterogeneity of the temperature and precipitation. Finally, the influencing factors with LUCC on Tibetan Plateau were simulated with the Weather Research and Forecasting (WRF) model, and the sensitivity of different land use types was spatially analyzed with Singular Value Decomposition (SVD). The results indicate that the large-area alpine grassland plays a more important role in alleviating global warming than other vegetation types do. The changes of the landscape structure resulting from the urban expansion play a significant role in intensifying regional temperature increase. In addition, the effects of LUCC on monthly average temperature change would vary from month to month with obviously spatial heterogeneity.

High-affinity binding of two molecules of cysteine proteinases to low-molecular-weight kininogen.

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Turk, B.; Stoka, V.; Björk, I.; Boudier, C.; Johansson, G.; Dolenc, I.; Colic, A.; Bieth, J. G.; Turk, V.

1995-01-01

Human low-molecular-weight kininogen (LK) was shown by fluorescence titration to bind two molecules of cathepsins L and S and papain with high affinity. By contrast, binding of a second molecule of cathepsin H was much weaker. The 2:1 binding stoichiometry was confirmed by titration monitored by loss of enzyme activity and by sedimentation velocity experiments. The kinetics of binding of cathepsins L and S and papain showed the two proteinase binding sites to have association rate constants kass,1 = 10.7-24.5 x 10(6) M-1 s-1 and kass,2 = 0.83-1.4 x 10(6) M-1 s-1. Comparison of these kinetic constants with previous data for intact LK and its separated domains indicate that the faster-binding site is also the tighter-binding site and is present on domain 3, whereas the slower-binding, lower-affinity site is on domain 2. These results also indicate that there is no appreciable steric hindrance for the binding of proteinases between the two binding sites or from the kininogen light chain. PMID:8528085

Continuous affine processes

DEFF Research Database (Denmark)

Buchardt, Kristian

2016-01-01

Affine processes possess the property that expectations of exponential affine transformations are given by a set of Riccati differential equations, which is the main feature of this popular class of processes. In this paper we generalise these results for expectations of more general transformati...

Affinity in electrophoresis.

Science.gov (United States)

Heegaard, Niels H H

2009-06-01

The journal Electrophoresis has greatly influenced my approaches to biomolecular affinity studies. The methods that I have chosen as my main tools to study interacting biomolecules--native gel and later capillary zone electrophoresis--have been the topic of numerous articles in Electrophoresis. Below, the role of the journal in the development and dissemination of these techniques and applications reviewed. Many exhaustive reviews on affinity electrophoresis and affinity CE have been published in the last few years and are not in any way replaced by the present deliberations that are focused on papers published by the journal.

Detection-Guided Fast Affine Projection Channel Estimator for Speech Applications

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Yan Wu Jennifer

2007-04-01

Full Text Available In various adaptive estimation applications, such as acoustic echo cancellation within teleconferencing systems, the input signal is a highly correlated speech. This, in general, leads to extremely slow convergence of the NLMS adaptive FIR estimator. As a result, for such applications, the affine projection algorithm (APA or the low-complexity version, the fast affine projection (FAP algorithm, is commonly employed instead of the NLMS algorithm. In such applications, the signal propagation channel may have a relatively low-dimensional impulse response structure, that is, the number m of active or significant taps within the (discrete-time modelled channel impulse response is much less than the overall tap length n of the channel impulse response. For such cases, we investigate the inclusion of an active-parameter detection-guided concept within the fast affine projection FIR channel estimator. Simulation results indicate that the proposed detection-guided fast affine projection channel estimator has improved convergence speed and has lead to better steady-state performance than the standard fast affine projection channel estimator, especially in the important case of highly correlated speech input signals.

Peptides in headlock – a novel high-affinity and versatile peptide-binding nanobody for proteomics and microscopy

Science.gov (United States)

Braun, Michael B.; Traenkle, Bjoern; Koch, Philipp A.; Emele, Felix; Weiss, Frederik; Poetz, Oliver; Stehle, Thilo; Rothbauer, Ulrich

2016-01-01

Nanobodies are highly valuable tools for numerous bioanalytical and biotechnical applications. Here, we report the characterization of a nanobody that binds a short peptide epitope with extraordinary affinity. Structural analysis reveals an unusual binding mode where the extended peptide becomes part of a β-sheet structure in the nanobody. This interaction relies on sequence-independent backbone interactions augmented by a small number of specificity-determining side chain contacts. Once bound, the peptide is fastened by two nanobody side chains that clamp it in a headlock fashion. Exploiting this unusual binding mode, we generated a novel nanobody-derived capture and detection system. Matrix-coupled nanobody enables the fast and efficient isolation of epitope-tagged proteins from prokaryotic and eukaryotic expression systems. Additionally, the fluorescently labeled nanobody visualizes subcellular structures in different cellular compartments. The high-affinity-binding and modifiable peptide tag of this system renders it a versatile and robust tool to combine biochemical analysis with microscopic studies. PMID:26791954

Isolation of temperature-sensitive mutants of 16 S rRNA in Escherichia coli

DEFF Research Database (Denmark)

Triman, K; Becker, E; Dammel, C

1989-01-01

Temperature-sensitive mutants have been isolated following hydroxylamine mutagenesis of a plasmid containing Escherichia coli rRNA genes carrying selectable markers for spectinomycin resistance (U1192 in 16 S rRNA) and erythromycin resistance (G2058 in 23 S rRNA). These antibiotic resistance....... The mutations were localized by in vitro restriction fragment replacement followed by in vivo marker rescue and were identified by DNA sequence analysis. We report here seven single-base alterations in 16 S rRNA (A146, U153, A350, A359, A538, A1292 and U1293), five of which produce temperature......-sensitive spectinomycin resistance and two that produce unconditional loss of resistance. In each case, loss of ribosomal function can be accounted for by disruption of base-pairing in the secondary structure of 16 S rRNA. For the temperature-sensitive mutants, there is a lag period of about two generations between...

Blockage of High-Affinity Choline Transporter Increases Visceral Hypersensitivity in Rats with Chronic Stress

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Chen Zhao

2018-01-01

Full Text Available Background. Visceral hypersensitivity is a common feature of irritable bowel syndrome. Cholinergic system involves in the development of visceral hypersensitivity, and high-affinity choline transporter (CHT1 is of crucial importance in choline uptake system. However, involvement of CHT1 in visceral hypersensitivity remains unknown. The research aimed to study the CHT1 expression in dorsal root ganglions (DRGs and the role of CHT1 in visceral hypersensitivity. Methods. Repetitive water avoidance stress (WAS was used to induce visceral hypersensitivity in rats. Colorectal distension (CRD was determined, and the abdominal withdrawal reflex (AWR and threshold intensity data were recorded to measure the visceral sensitivity. After intraperitoneal injection of hemicholinium-3 (HC-3, the specific inhibitor of CHT1, CRD data were also recorded. The CHT1 expression of DRGs was investigated by Western blotting, immunohistochemistry, and quantitative RT-PCR. Acetylcholine levels in the DRGs were detected by the assay kit. Results. Repetitive WAS increased the AWR score of CRD at high distension pressure and decreased the mean threshold of rats. The CHT1 expression and acetylcholine concentration of DRG were significantly increased in WAS rats. After the administration of HC-3, the AWR score in WAS group was significantly increased at higher distension pressure while the threshold intensity was significantly reduced compared to the normal saline group. Acetylcholine concentration was significantly lower than the normal saline rats. Conclusion. Our research firstly reports that CHT1 is overexpressed in noninflammatory visceral hypersensitivity, and blockage of CHT1 can enhance the visceral hypersensitivity. CHT1 may play an inhibitory role in visceral hypersensitivity.

Pyrolyzed feather fibers for adsorbent and high temperature applications

Science.gov (United States)

Senoz, Erman

Chicken feather fibers (CFF) are problematic and costly for the poultry industry in terms of managing maintenance and disposal. Considering their great availability, low cost, and unique protein structure, CFF can be an environmentally friendly and bio-renewable candidate to replace petroleum products. CFF's low degradation and melting temperature render them useless at high temperatures. Pyrolysis methods were developed for CFF by using two temperature steps to convert them into high temperature resistant and adsorbent fibers while retaining their original physical appearance and affine dimensions. An intermolecular crosslinking mechanism in the first step of pyrolysis at 215 ºC for 24 h provided an intact fibrous structure with no subsequent melting. The evidence obtained from the thermal, bulk, and surface analysis techniques was indication of the simultaneous side chain degradation, polypeptide backbone scission, disulfide bond cleavage, and isopeptide crosslinking. The variation in the reaction kinetics of disulfide bond cleavage and isopeptide crosslinking played an important role in the melting transition. Consequently, long-lasting heat treatments below the melting point provided sufficient crosslinks in the protein matrix to keep the fibrous structure intact. Water-insoluble and crosslinked CFF reinforced the triglyceride-fatty acid based composites by providing a 15 fold increase in storage and tensile modulus at room temperature. These thermally stable fibers can be used instead of CFF in composites which may require high temperature compounding and molding processes. The second step of pyrolysis at 400--450 ºC for 1 h resulted in microporous fibers with a micropore volume of ˜0.18 cm3/g STP and with a narrower pore size distribution than commercial activated carbons through thermal degradation. Nearly all accessible pores in the microporous pyrolyzed chicken feather fibers (PCFF) had diameters less than 1 nm and therefore, showed a potential to be

Passive Wireless Temperature Sensors with Enhanced Sensitivity and Range, Phase I

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National Aeronautics and Space Administration — This proposal describes the development of passive surface acoustic wave (SAW) temperature sensors with enhanced sensitivity and detection range for NASA application...

High-Temperature Sensor Based on Fabry-Perot Interferometer in Microfiber Tip

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Zhenshi Chen

2018-01-01

Full Text Available A miniaturized tip Fabry-Perot interferometer (tip-FPI is proposed for high-temperature sensing. It is simply fabricated for the first time by splicing a short length of microfiber (MF to the cleaved end of a standard single mode fiber (SMF with precise control of the relative cross section position. Such a MF acts as a Fabry-Perot (FP cavity and serves as a tip sensor. A change in temperature modifies the length and refractive index of the FP cavity, and then a corresponding change in the reflected interference spectrum can be observed. High temperatures of up to 1000 °C are measured in the experiments, and a high sensitivity of 13.6 pm/°C is achieved. This compact sensor, with tip diameter and length both of tens of microns, is suitable for localized detection, especially in harsh environments.

The metric-affine gravitational theory as the gauge theory of the affine group

International Nuclear Information System (INIS)

Lord, E.A.

1978-01-01

The metric-affine gravitational theory is shown to be the gauge theory of the affine group, or equivalently, the gauge theory of the group GL(4,R) of tetrad deformations in a space-time with a locally Minkowskian metric. The identities of the metric-affine theory, and the relationship between them and those of general relativity and Sciama-Kibble theory, are derived. (Auth.)

Cyclic GMP-AMP Containing Mixed Phosphodiester Linkages Is An Endogenous High Affinity Ligand for STING

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Zhang, Xu; Shi, Heping; Wu, Jiaxi; Zhang, Xuewu; Sun, Lijun; Chen, Chuo; Chen, Zhijian J.

2013-01-01

The presence of microbial or self DNA in the cytoplasm of mammalian cells is a danger signal detected by the DNA sensor cyclic-GMP-AMP (cGAMP) synthase (cGAS), which catalyzes the production of cGAMP that in turn serves as a second messenger to activate innate immune responses. Here we show that endogenous cGAMP in mammalian cells contains two distinct phosphodiester linkages, one between 2′-OH of GMP and 5′-phosphate of AMP, and the other between 3′-OH of AMP and 5′-phosphate of GMP. This molecule, termed 2′3′-cGAMP, is unique in that it binds to the adaptor protein STING with a much greater affinity than cGAMP molecules containing other combinations of phosphodiester linkages. The crystal structure of STING bound to 2′3′-cGAMP revealed the structural basis of this high-affinity binding and a ligand-induced conformational change in STING that may underlie its activation. PMID:23747010

Cyclic GMP-AMP containing mixed phosphodiester linkages is an endogenous high-affinity ligand for STING.

Science.gov (United States)

Zhang, Xu; Shi, Heping; Wu, Jiaxi; Zhang, Xuewu; Sun, Lijun; Chen, Chuo; Chen, Zhijian J

2013-07-25

The presence of microbial or self DNA in the cytoplasm of mammalian cells is a danger signal detected by the DNA sensor cyclic-GMP-AMP (cGAMP) synthase (cGAS), which catalyzes the production of cGAMP that in turn serves as a second messenger to activate innate immune responses. Here we show that endogenous cGAMP in mammalian cells contains two distinct phosphodiester linkages, one between 2'-OH of GMP and 5'-phosphate of AMP, and the other between 3'-OH of AMP and 5'-phosphate of GMP. This molecule, termed 2'3'-cGAMP, is unique in that it binds to the adaptor protein STING with a much greater affinity than cGAMP molecules containing other combinations of phosphodiester linkages. The crystal structure of STING bound to 2'3'-cGAMP revealed the structural basis of this high-affinity binding and a ligand-induced conformational change in STING that may underlie its activation. Copyright © 2013 Elsevier Inc. All rights reserved.

Solution structure of the Grb2 SH2 domain complexed with a high-affinity inhibitor

International Nuclear Information System (INIS)

Ogura, Kenji; Shiga, Takanori; Yokochi, Masashi; Yuzawa, Satoru; Burke, Terrence R.; Inagaki, Fuyuhiko

2008-01-01

The solution structure of the growth factor receptor-bound protein 2 (Grb2) SH2 domain complexed with a high-affinity inhibitor containing a non-phosphorus phosphate mimetic within a macrocyclic platform was determined by nuclear magnetic resonance (NMR) spectroscopy. Unambiguous assignments of the bound inhibitor and intermolecular NOEs between the Grb2 SH2 domain and the inhibitor was accomplished using perdeuterated Grb2 SH2 protein. The well-defined solution structure of the complex was obtained and compared to those by X-ray crystallography. Since the crystal structure of the Grb2 SH2 domain formed a domain-swapped dimer and several inhibitors were bound to a hinge region, there were appreciable differences between the solution and crystal structures. Based on the binding interactions between the inhibitor and the Grb2 SH2 domain in solution, we proposed a design of second-generation inhibitors that could be expected to have higher affinity

The ketamine analogue methoxetamine and 3- and 4-methoxy analogues of phencyclidine are high affinity and selective ligands for the glutamate NMDA receptor.

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Bryan L Roth

Full Text Available In this paper we determined the pharmacological profiles of novel ketamine and phencyclidine analogues currently used as 'designer drugs' and compared them to the parent substances via the resources of the National Institute of Mental Health Psychoactive Drug Screening Program. The ketamine analogues methoxetamine ((RS-2-(ethylamino-2-(3-methoxyphenylcyclohexanone and 3-MeO-PCE (N-ethyl-1-(3-methoxyphenylcyclohexanamine and the 3- and 4-methoxy analogues of phencyclidine, (1-[1-(3-methoxyphenylcyclohexyl]piperidine and 1-[1-(4-methoxyphenylcyclohexyl]piperidine, were all high affinity ligands for the PCP-site on the glutamate NMDA receptor. In addition methoxetamine and PCP and its analogues displayed appreciable affinities for the serotonin transporter, whilst the PCP analogues exhibited high affinities for sigma receptors. Antagonism of the NMDA receptor is thought to be the key pharmacological feature underlying the actions of dissociative anaesthetics. The novel ketamine and PCP analogues had significant affinities for the NMDA receptor in radioligand binding assays, which may explain their psychotomimetic effects in human users. Additional actions on other targets could be important for delineating side-effects.

Temperature sensitivity and enzymatic mechanisms of soil organic matter decomposition along an altitudinal gradient on Mount Kilimanjaro

Science.gov (United States)

Blagodatskaya, Еvgenia; Blagodatsky, Sergey; Khomyakov, Nikita; Myachina, Olga; Kuzyakov, Yakov

2016-02-01

Short-term acceleration of soil organic matter decomposition by increasing temperature conflicts with the thermal adaptation observed in long-term studies. Here we used the altitudinal gradient on Mt. Kilimanjaro to demonstrate the mechanisms of thermal adaptation of extra- and intracellular enzymes that hydrolyze cellulose, chitin and phytate and oxidize monomers (14C-glucose) in warm- and cold-climate soils. We revealed that no response of decomposition rate to temperature occurs because of a cancelling effect consisting in an increase in half-saturation constants (Km), which counteracts the increase in maximal reaction rates (Vmax with temperature). We used the parameters of enzyme kinetics to predict thresholds of substrate concentration (Scrit) below which decomposition rates will be insensitive to global warming. Increasing values of Scrit, and hence stronger canceling effects with increasing altitude on Mt. Kilimanjaro, explained the thermal adaptation of polymer decomposition. The reduction of the temperature sensitivity of Vmax along the altitudinal gradient contributed to thermal adaptation of both polymer and monomer degradation. Extrapolating the altitudinal gradient to the large-scale latitudinal gradient, these results show that the soils of cold climates with stronger and more frequent temperature variation are less sensitive to global warming than soils adapted to high temperatures.

Molecular cloning of a second subunit of the receptor for human granulocyte - macrophage colony-stimulating factor (GM-CSF): Reconstitution of a high-affinity GM-CSF receptor

International Nuclear Information System (INIS)

Hayashida, Kazuhiro; Kitamura, Toshio; Gorman, D.M.; Miyajima, Atsushi; Arai, Kenichi; Yokota, Takashi

1990-01-01

Using the mouse interleukin 3 (IL-3) receptor cDNA as a probe, the authors obtained a monologous cDNA (KH97) from a cDNA library of a human hemopoietic cell line, TF-1. The protein encoded by the KH97 cDNA has 56% amino acid sequence identity with the mouse IL-3 receptor and retains features common to the family of cytokine receptors. Fibroblasts transfected with the KH97 cDNA expressed a protein of 120 kDa but did not bind any human cytokines, including IL-3 and granulocyte - macrophage colony-stimulating factor (GM-CSF). Interestingly, cotransfection of cDNAs for KH97 and the low-affinity human GM-CSF receptor in fibroblasts resulted in formation of a high-affinity receptor for GM-CSF. The dissociation rate of GM-CSF from the reconstituted high-affinity receptor was slower than that from the low-affinity site, whereas the association rate was unchanged. Cross-linking of 125 I-labeled GM-CSF to fibroblasts cotransfected with both cDNAs revealed the same cross-linking patterns as in TF-1 cells - i.e., two major proteins of 80 and 120 kDa which correspond to the low-affinity GM-CSF receptor and the KH97 protein, respectively. These results indicate that the high-affinity GM-CSF receptor is composed of at least two components in a manner analogous to the IL-2 receptor. They therefore propose to designate the low-affinity GM-CSF receptor and the KH97 protein as the α and β subunits of the GM-CSF receptor, respectively

High order depletion sensitivity analysis

International Nuclear Information System (INIS)

Naguib, K.; Adib, M.; Morcos, H.N.

2002-01-01

A high order depletion sensitivity method was applied to calculate the sensitivities of build-up of actinides in the irradiated fuel due to cross-section uncertainties. An iteration method based on Taylor series expansion was applied to construct stationary principle, from which all orders of perturbations were calculated. The irradiated EK-10 and MTR-20 fuels at their maximum burn-up of 25% and 65% respectively were considered for sensitivity analysis. The results of calculation show that, in case of EK-10 fuel (low burn-up), the first order sensitivity was found to be enough to perform an accuracy of 1%. While in case of MTR-20 (high burn-up) the fifth order was found to provide 3% accuracy. A computer code SENS was developed to provide the required calculations

A rhodamine-labeled citalopram analogue as a high-affinity fluorescent probe for the serotonin transporter

DEFF Research Database (Denmark)

Zhang, Peng; Jørgensen, Trine Nygaard; Løland, Claus Juul

2013-01-01

A novel fluorescent ligand was synthesized as a high-affinity, high specificity probe for visualizing the serotonin transporter (SERT). The rhodamine fluorophore was extended from an aniline substitution on the 5-position of the dihydroisobenzofuran ring of citalopram (2, 1-(3-(dimethylamino......)propyl)-1-(4-fluorophenyl)-1,3-dihydroisobenzofuran-5-carbonitrile), using an ethylamino linker. The resulting rhodamine-labeled ligand 8 inhibited [3H]5-HT uptake in COS-7 cells (Ki = 225 nM) with similar potency to the tropane-based JHC 1-064 (1), but with higher specificity towards the SERT relative...

Low-temperature fabrication of TiO2 nanocrystalline film electrodes for dye-sensitized solar cells

Energy Technology Data Exchange (ETDEWEB)

Shan, G.; Lee, K.E.; Charboneau, C.; Demopoulos, G.P.; Gauvin, R. [McGill Univ., Montreal, PQ (Canada). Dept. of Materials Engineering; Savadogo, O. [Ecole Polytechnique de Montreal, PQ (Canada). Dept. de Genie Chimique

2008-07-01

Dye-sensitized solar cells (DSSCs) have the potential to render solar energy widely accessible. The deposition of titania nano-crystalline powders on a substrate is an important step in the manufacture of the DSSC. The deposition forms a mesoporous thin film that is followed by thermal treatment and sensitization. Usually titania films are deposited on glass by screen printing and then annealed at temperatures as high as 530 degrees C to provide a good electrical contact between the semiconductor particles and crystallization of the anatase phase. Several research and development efforts have focused on the deposition of titania film on flexible plastic substrates that will simplify the whole manufacturing process in terms of flexibility, weight, application and cost. Lower temperature processing is needed for the preparation of plastic-based titania film electrodes, but this has proven to be counterproductive when it comes to the cell's conversion efficiency. This paper presented a comprehensive evaluation of the different coating and annealing techniques at low temperature as well as important processing factors for improvement. To date, these techniques include pressing, hydrothermal process, electrodeposition, electrophoretic deposition, microwave or UV irradiation, and lift-off technique.

A temperature monitor circuit with small voltage sensitivity using a topology-reconfigurable ring oscillator

Science.gov (United States)

Kishimoto, Tadashi; Ishihara, Tohru; Onodera, Hidetoshi

2018-04-01

In this paper, we propose a temperature monitor circuit that exhibits a small supply voltage sensitivity adopting a circuit topology of a reconfigurable ring oscillator. The circuit topology of the monitor is crafted such that the oscillation frequency is determined by the amount of subthreshold leakage current, which has an exponential dependence on temperature. Another important characteristic of the monitor is its small supply voltage sensitivity. The measured oscillation frequency of a test chip fabricated in a 65 nm CMOS process varies only 2.6% under a wide range of supply voltages from 0.4 to 1.0 V at room temperature. The temperature estimation error ranges from -0.3 to 0.4 °C over a temperature range of 10 to 100 °C.

Online micro-solid-phase extraction based on boronate affinity monolithic column coupled with high-performance liquid chromatography for the determination of monoamine neurotransmitters in human urine.

Science.gov (United States)

Yang, Xiaoting; Hu, Yufei; Li, Gongke

2014-05-16

Quantification of monoamine neurotransmitters is very important in diagnosing and monitoring of patients with neurological disorders. We developed an online analytical method to selectively determine urinary monoamine neurotransmitters, which coupled the boronate affinity monolithic column micro-solid-phase extraction with high-performance liquid chromatography (HPLC). The boronate affinity monolithic column was prepared by in situ polymerization of vinylphenylboronic acid (VPBA) and N,N'-methylenebisacrylamide (MBAA) in a stainless capillary column. The prepared monolithic column showed good permeability, high extraction selectivity and capacity. The column-to-column reproducibility was satisfactory and the enrichment factors were 17-243 for four monoamine neurotransmitters. Parameters that influence the online extraction efficiency, including pH of sample solution, flow rate of extraction and desorption, extraction volume and desorption volume were investigated. Under the optimized conditions, the developed method exhibited low limit of detection (0.06-0.80μg/L), good linearity (with R(2) between 0.9979 and 0.9993). The recoveries in urine samples were 81.0-105.5% for four monoamine neurotransmitters with intra- and inter-day RSDs of 2.1-8.2% and 3.7-10.6%, respectively. The online analytical method was sensitive, accurate, selective, reliable and applicable to analysis of trace monoamine neurotransmitters in human urine sample. Copyright © 2014 Elsevier B.V. All rights reserved.

Ratiometric highly sensitive luminescent nanothermometers working in the room temperature range. Applications to heat propagation in nanofluids

Science.gov (United States)

Brites, Carlos D. S.; Lima, Patrícia P.; Silva, Nuno J. O.; Millán, Angel; Amaral, Vitor S.; Palacio, Fernando; Carlos, Luís D.

2013-07-01

There is an increasing demand for accurate, non-invasive and self-reference temperature measurements as technology progresses into the nanoscale. This is particularly so in micro- and nanofluidics where the comprehension of heat transfer and thermal conductivity mechanisms can play a crucial role in areas as diverse as energy transfer and cell physiology. Here we present two luminescent ratiometric nanothermometers based on a magnetic core coated with an organosilica shell co-doped with Eu3+ and Tb3+ chelates. The design of the hybrid host and chelate ligands permits the working of the nanothermometers in a nanofluid at 293-320 K with an emission quantum yield of 0.38 +/- 0.04, a maximum relative sensitivity of 1.5% K-1 at 293 K and a spatio-temporal resolution (constrained by the experimental setup) of 64 × 10-6 m/150 × 10-3 s (to move out of 0.4 K - the temperature uncertainty). The heat propagation velocity in the nanofluid, (2.2 +/- 0.1) × 10-3 m s-1, was determined at 294 K using the nanothermometers' Eu3+/Tb3+ steady-state spectra. There is no precedent of such an experimental measurement in a thermographic nanofluid, where the propagation velocity is measured from the same nanoparticles used to measure the temperature.There is an increasing demand for accurate, non-invasive and self-reference temperature measurements as technology progresses into the nanoscale. This is particularly so in micro- and nanofluidics where the comprehension of heat transfer and thermal conductivity mechanisms can play a crucial role in areas as diverse as energy transfer and cell physiology. Here we present two luminescent ratiometric nanothermometers based on a magnetic core coated with an organosilica shell co-doped with Eu3+ and Tb3+ chelates. The design of the hybrid host and chelate ligands permits the working of the nanothermometers in a nanofluid at 293-320 K with an emission quantum yield of 0.38 +/- 0.04, a maximum relative sensitivity of 1.5% K-1 at 293 K and a spatio

A multiplex microplatform for the detection of multiple DNA methylation events using gold-DNA affinity.

Science.gov (United States)

Sina, Abu Ali Ibn; Foster, Matthew Thomas; Korbie, Darren; Carrascosa, Laura G; Shiddiky, Muhammad J A; Gao, Jing; Dey, Shuvashis; Trau, Matt

2017-10-07

We report a new multiplexed strategy for the electrochemical detection of regional DNA methylation across multiple regions. Using the sequence dependent affinity of bisulfite treated DNA towards gold surfaces, the method integrates the high sensitivity of a micro-fabricated multiplex device comprising a microarray of gold electrodes, with the powerful multiplexing capability of multiplex-PCR. The synergy of this combination enables the monitoring of the methylation changes across several genomic regions simultaneously from as low as 500 pg μl -1 of DNA with no sequencing requirement.

Growth and Development Temperature Influences Level of Tolerance to High Light Stress 1

Science.gov (United States)

Steffen, Kenneth L.; Palta, Jiwan P.

1989-01-01

The influence of growth and development temperature on the relative tolerance of photosynthetic tissue to high light stress at chilling temperatures was investigated. Two tuber-bearing potato species, Solanum tuberosum L. cv Red Pontiac and Solanum commersonii were grown for 4 weeks, at either 12 or 24°C with 12 hours of about 375 micromoles per second per square meter of photosynthetically active radiation. Paired leaf discs were cut from directly across the midvein of leaflets of comparable developmental stage and light environment from each species at each growth temperature treatment. One disc of each pair was exposed to 1°C and about 1000 micromoles per second per square meter photosynthetically active radiation for 4 hours, and the other disc was held at 1°C in total darkness for the same duration. Photosynthetic tissue of S. tuberosum, developed at 12°C, was much more tolerant to high light and low temperature stress than tissue developed under 24°C conditions. Following the high light treatment, 24°C-grown S. tuberosum tissue demonstrated light-limited and light-saturated rates that were approximately 50% of their paired dark controls. In contrast, the 12°C-grown tissue from S. tuberosum that was subjected to the light stress showed only a 18 and 6% reduction in light-limited and light-saturated rates of photosynthetic oxygen evolution, respectively. Tissue from 24°C-grown S. commersonii was much less sensitive to the light stress than was tissue from S. tuberosum grown under the same conditions. The results presented here demonstrate that: (a) acclimation of S. tuberosum to lower temperature growth conditions with a constant light environment, results in the increased capacity of photosynthetic tissue to tolerate high light stress at chilling temperature and (b) following growth and development at relatively high temperatures S. commersonii, a frost- and heat-tolerant wild species, has a much greater tolerance to the high light stress at chilling

Novel and high affinity fluorescent ligands for the serotonin transporter based on (s)-citalopram

DEFF Research Database (Denmark)

Kumar, Vivek; Rahbek-Clemmensen, Troels; Billesbølle, Christian B

2014-01-01

Novel rhodamine-labeled ligands, based on (S)-citalopram, were synthesized and evaluated for uptake inhibition at the human serotonin, dopamine, and norepinephrine transporters (hSERT, hDAT, and hNET, respectively) and for binding at SERT, in transiently transfected COS7 cells. Compound 14 demons...... demonstrated high affinity binding and selectivity for SERT (K i = 3 nM). Visualization of SERT, using confocal laser scanning microscopy, validated compound 14 as a novel tool for studying SERT expression and distribution in living cells....

A Generalized Affine Isoperimetric Inequality

OpenAIRE

Chen, Wenxiong; Howard, Ralph; Lutwak, Erwin; Yang, Deane; Zhang, Gaoyong

2004-01-01

A purely analytic proof is given for an inequality that has as a direct consequence the two most important affine isoperimetric inequalities of plane convex geometry: The Blaschke-Santalo inequality and the affine isoperimetric inequality of affine differential geometry.