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Sample records for alpha selectively sensitizes

  1. DIFFERENCES IN SENSITIVITY BUT NOT SELECTIVITY OF XENOESTROGEN BINDING TO ALLIGATOR VERSUS HUMAN ESTROGEN RECEPTOR ALPHA

    PubMed Central

    Rider, Cynthia V.; Hartig, Phillip C.; Cardon, Mary C.; Lambright, Christy R.; Bobseine, Kathy L.; Guillette, Louis J.; Gray, L. Earl; Wilson, Vickie S.

    2010-01-01

    Reproductive abnormalities in alligators exposed to contaminants in Lake Apopka, Florida, USA represent a clear example of endocrine disruption in wildlife. Several of these contaminants that are not able to bind to mammalian estrogen receptors (such as atrazine and cyanazine) have previously been reported to bind to the alligator estrogen receptor from oviductal tissue. Binding of known Lake Apopka contaminants to full length estrogen receptors alpha from human (hERα) and alligator (aERα) was assessed in a side-by-side comparison within the same assay system. Baculovirus-expressed recombinant hERα and aERα were used in a competitive binding assay. Atrazine and cyanazine were not able to bind to either receptor. p,p′-Dicofol was able to bind to aERα with a concentration inhibiting 50% of binding (IC50) of 4 μM, while only partially displacing 17β-estradiol (E2) from hERα and yielding a projected IC50 of 45 μM. Chemicals that only partially displaced E2 from either receptor, including some dichlorodiphenyltrichloroethane (DDT) metabolites and trans-nonachlor, appeared to have higher affinity for aERα than hERα. p,p′-Dicofol-mediated transcriptional activation through aERα and hERα was assessed to further explore the preferential binding of p,p′-dicofol to aERα over hERα. p,p′-Dicofol was able to stimulate transcriptional activation in a similar manner with both receptors. However, the in vitro results obtained with p,p′-dicofol were not reflected in an in vivo mammalian model, where Kelthane™ (mixed o,p′-and p,p′-dicofol isomers) did not elicit estrogenic effects. In conclusion, although there was no evidence of exclusively species-specific estrogen receptor binders, some xenoestrogens, especially p,p′-dicofol, had a higher affinity for aERα than for hERα. PMID:20821664

  2. In Vitro Selection of Single-Stranded DNA Molecular Recognition Elements against S. aureus Alpha Toxin and Sensitive Detection in Human Serum

    PubMed Central

    Hong, Ka L.; Battistella, Luisa; Salva, Alysia D.; Williams, Ryan M.; Sooter, Letha J.

    2015-01-01

    Alpha toxin is one of the major virulence factors secreted by Staphylococcus aureus, a bacterium that is responsible for a wide variety of infections in both community and hospital settings. Due to the prevalence of S. aureus related infections and the emergence of methicillin-resistant S. aureus, rapid and accurate diagnosis of S. aureus infections is crucial in benefiting patient health outcomes. In this study, a rigorous Systematic Evolution of Ligands by Exponential Enrichment (SELEX) variant previously developed by our laboratory was utilized to select a single-stranded DNA molecular recognition element (MRE) targeting alpha toxin with high affinity and specificity. At the end of the 12-round selection, the selected MRE had an equilibrium dissociation constant (Kd) of 93.7 ± 7.0 nM. Additionally, a modified sandwich enzyme-linked immunosorbent assay (ELISA) was developed by using the selected ssDNA MRE as the toxin-capturing element and a sensitive detection of 200 nM alpha toxin in undiluted human serum samples was achieved. PMID:25633102

  3. Alpha-particle sensitive test SRAMs

    NASA Technical Reports Server (NTRS)

    Buehler, M. G.; Blaes, B. R.

    1990-01-01

    A bench-level test is being developed to evaluate memory-cell upsets in a test SRAM designed with a cell offset voltage. This offset voltage controls the critical charge needed to upset the cell. The effect is demonstrated using a specially designed 2-micron n-well CMOS 4-kb test SRAM and a Po-208 5.1-MeV 0.61-LET alpha-particle source. This test SRAM has been made sensitive to alpha particles through the use of a cell offset voltage, and this has allowed a bench-level characterization in a laboratory setting. The experimental data are linked to a alpha-particle interaction physics and to SPICE circuit simulations through the alpha-particle collection depth. The collection depth is determined by two methods and found to be about 7 micron. In addition, alpha particles that struck outside the bloated drain were able to flip the SRAM cells. This lateral charge collection was observed to be more than 6 micron.

  4. Sensitivity to alpha-amylcinnamic aldehyde and alpha-amylcinnamic alcohol.

    PubMed

    Guin, J D; Haffley, P

    1983-01-01

    Sensitivity to alpha-amylcinnamic aldehyde (alpha-AcAld) is apparently uncommon, but, like allergy to alpha-amylcinnamic alcohol (alpha-AcAlc), it often accompanies allergy to the perfume in Mycolog cream. Although alpha-AcAlc is a known ingredient, alpha-AcAld is not. However, gas-liquid chromatographic analysis shows alpha-AcAld to be present. Of fourteen persons sensitive to either chemical, ten reacted to both. Of these, one man and three women were markedly sensitive, and all three women had chronic recalcitrant vulvar eczema. That condition might have been the cause as well as the result of sensitization, but reexposure to a suspected product reproduced the eruption in both persons tested. Its use with other potent sensitizers, e.g., ethylenediamine, to treat irritations and chronic eczemas in an area of high absorption may partly explain development of allergy to a relatively weak sensitizer.

  5. Crystallographic variant selection in {alpha}-{beta} brass

    SciTech Connect

    Stanford, N.; Bate, P.S. . E-mail: pete.bate@man.ac.uk

    2005-02-01

    The transformation texture of {alpha}/{beta} brass with a diffusional Widmanstaetten {alpha} growth morphology has been investigated. Electron micrographs and electron backscattered diffraction was used to determine that the orientation relationship between the {beta} phase and the {alpha} associated with nucleation at {beta} grain boundaries was 44.3 deg <1 1 6>. Crystallographic variant selection was observed across those prior {beta}/{beta} grain boundaries, but this has little effect on the transformation texture due to the crystal symmetry. The effect of the crystallographic variant selection on texture is further weakened by nucleation of diffusional transformed {alpha} in the grain interior.

  6. Coefficient alpha and interculture test selection.

    PubMed

    Thurber, Steven; Kishi, Yasuhiro

    2014-04-01

    The internal consistency reliability of a measure can be a focal point in an evaluation of the potential adequacy of an instrument for adaptation to another cultural setting. Cronbach's alpha (α) coefficient is often used as the statistical index for such a determination. However, alpha presumes a tau-equivalent test and may constitute an inaccurate population estimate for multidimensional tests. These notions are expanded and examined with a Japanese version of a questionnaire on nursing attitudes toward suicidal patients, originally constructed in Sweden using the English language. The English measure was reported to have acceptable internal consistency (α) albeit the dimensionality of the questionnaire was not addressed. The Japanese scale was found to lack tau-equivalence. An alternative to alpha, "composite reliability," was computed and found to be below acceptable standards in magnitude and precision. Implications for research application of the Japanese instrument are discussed. PMID:22523134

  7. Entrainment of Human Alpha Oscillations Selectively Enhances Visual Conjunction Search.

    PubMed

    Müller, Notger G; Vellage, Anne-Katrin; Heinze, Hans-Jochen; Zaehle, Tino

    2015-01-01

    The functional role of the alpha-rhythm which dominates the human electroencephalogram (EEG) is unclear. It has been related to visual processing, attentional selection and object coherence, respectively. Here we tested the interaction of alpha oscillations of the human brain with visual search tasks that differed in their attentional demands (pre-attentive vs. attentive) and also in the necessity to establish object coherence (conjunction vs. single feature). Between pre- and post-assessment elderly subjects received 20 min/d of repetitive transcranial alternating current stimulation (tACS) over the occipital cortex adjusted to their individual alpha frequency over five consecutive days. Compared to sham the entrained alpha oscillations led to a selective, set size independent improvement in the conjunction search task performance but not in the easy or in the hard feature search task. These findings suggest that cortical alpha oscillations play a specific role in establishing object coherence through suppression of distracting objects. PMID:26606255

  8. Entrainment of Human Alpha Oscillations Selectively Enhances Visual Conjunction Search

    PubMed Central

    Müller, Notger G.; Vellage, Anne-Katrin; Heinze, Hans-Jochen; Zaehle, Tino

    2015-01-01

    The functional role of the alpha-rhythm which dominates the human electroencephalogram (EEG) is unclear. It has been related to visual processing, attentional selection and object coherence, respectively. Here we tested the interaction of alpha oscillations of the human brain with visual search tasks that differed in their attentional demands (pre-attentive vs. attentive) and also in the necessity to establish object coherence (conjunction vs. single feature). Between pre- and post-assessment elderly subjects received 20 min/d of repetitive transcranial alternating current stimulation (tACS) over the occipital cortex adjusted to their individual alpha frequency over five consecutive days. Compared to sham the entrained alpha oscillations led to a selective, set size independent improvement in the conjunction search task performance but not in the easy or in the hard feature search task. These findings suggest that cortical alpha oscillations play a specific role in establishing object coherence through suppression of distracting objects. PMID:26606255

  9. Object-based attentional selection modulates anticipatory alpha oscillations.

    PubMed

    Knakker, Balázs; Weiss, Béla; Vidnyánszky, Zoltán

    2014-01-01

    Visual cortical alpha oscillations are involved in attentional gating of incoming visual information. It has been shown that spatial and feature-based attentional selection result in increased alpha oscillations over the cortical regions representing sensory input originating from the unattended visual field and task-irrelevant visual features, respectively. However, whether attentional gating in the case of object based selection is also associated with alpha oscillations has not been investigated before. Here we measured anticipatory electroencephalography (EEG) alpha oscillations while participants were cued to attend to foveal face or word stimuli, the processing of which is known to have right and left hemispheric lateralization, respectively. The results revealed that in the case of simultaneously displayed, overlapping face and word stimuli, attending to the words led to increased power of parieto-occipital alpha oscillations over the right hemisphere as compared to when faces were attended. This object category-specific modulation of the hemispheric lateralization of anticipatory alpha oscillations was maintained during sustained attentional selection of sequentially presented face and word stimuli. These results imply that in the case of object-based attentional selection-similarly to spatial and feature-based attention-gating of visual information processing might involve visual cortical alpha oscillations.

  10. Acute ozone exposure increases plasma prostaglandin F2 alpha in ozone-sensitive human subjects

    SciTech Connect

    Schelegle, E.S.; Adams, W.C.; Giri, S.N.; Siefkin, A.D.

    1989-07-01

    Twenty O/sub 3/-sensitive and /sup 2/O O/sub 3/-nonsensitive subjects participated in a study to investigate the effects of disparate O/sub 3/ sensitivity on plasma prostaglandin F2 alpha responses consequent to exposure to ambient O3 concentrations. Subjects were selected from a pool of 75 normal healthy college-aged males who had been previously exposed to 0.35 ppm O3 for 1 h at an exercising VE of 60 L/min. The selection criterion used was the observed decrement in FEV1 after the O/sub 3/ exposure: O/sub 3/-sensitive, FEV1 decrement greater than 24%; O/sub 3/-nonsensitive, FEV1 decrement less than 11%. Each subject was exposed to filtered air and to 0.20 and 0.35 ppm O/sub 3/ for 80 min while exercising at a VE of 50 L/min. These experimental protocols were divided into two 40-min sessions separated by a period of 4 to 10 min. PGF2 alpha, FVC, FEV1, and FEF25-75 were evaluated before, during, and after each protocol. SGaw and Vtg were measured before and after each protocol. Plasma PGF2 alpha was significantly increased in the O/sub 3/-sensitive group during and after the 0.35-ppm O/sub 3/ exposure.

  11. Michrochannel plate for position sensitive alpha particle detection

    SciTech Connect

    Paul Hurley and James Tinsley

    2007-08-31

    This paper will describe the use of a microchannel plate (MCP) as the associated particle detector on a sealed tube neutron generator. The generator produces neutrons and associated alpha particles for use as a probe to locate and identify hidden explosives in associated particle imaging (API). The MCP measures the position in two dimensions and precise timing of the incident alpha particle, information which is then used to calculate the emission time and direction of the corresponding neutron. The MCP replaces the position-sensitive photomultipler tube (PSPMT) which, until recently, had been the only detector available for measuring position and timing for alpha particles in neutron generator applications. Where the PSPMT uses charge division for generating position information, a process that requires a first order correction to each pulse, the MCP uses delay-line timing, which requires no correction. The result is a device with an order of magnitude improvement in both position resolution and timing compared to the PSPMT. Hardware and software development and the measurements made to characterize the MCP for API applications are described.

  12. Arrangement of Kv1 alpha subunits dictates sensitivity to tetraethylammonium.

    PubMed

    Al-Sabi, Ahmed; Shamotienko, Oleg; Dhochartaigh, Sorcha Ni; Muniyappa, Nagesh; Le Berre, Marie; Shaban, Hamdy; Wang, Jiafu; Sack, Jon T; Dolly, J Oliver

    2010-09-01

    Shaker-related Kv1 channels contain four channel-forming alpha subunits. Subfamily member Kv1.1 often occurs oligomerized with Kv1.2 alpha subunits in synaptic membranes, and so information was sought on the influence of their positions within tetramers on the channels' properties. Kv1.1 and 1.2 alpha genes were tandem linked in various arrangements, followed by expression as single-chain proteins in mammalian cells. As some concatenations reported previously seemed not to reliably position Kv1 subunits in their assemblies, the identity of expressed channels was methodically evaluated. Surface protein, isolated by biotinylation of intact transiently transfected HEK-293 cells, gave Kv1.1/1.2 reactivity on immunoblots with electrophoretic mobilities corresponding to full-length concatenated tetramers. There was no evidence of protein degradation, indicating that concatemers were delivered intact to the plasmalemma. Constructs with like genes adjacent (Kv1.1-1.1-1.2-1.2 or Kv1.2-1.2-1.1-1.1) yielded delayed-rectifying, voltage-dependent K(+) currents with activation parameters and inactivation kinetics slightly different from the diagonally positioned genes (Kv1.1-1.2-1.1-1.2 or 1.2-1.1-1.2-1.1). Pore-blocking petidergic toxins, alpha dendrotoxin, agitoxin-1, tityustoxin-Kalpha, and kaliotoxin, were unable to distinguish between the adjacent and diagonal concatamers. Unprecedentedly, external application of the pore-blocker tetraethylammonium (TEA) differentially inhibited the adjacent versus diagonal subunit arrangements, with diagonal constructs having enhanced susceptibility. Concatenation did not directly alter the sensitivities of homomeric Kv1.1 or 1.2 channels to TEA or the toxins. TEA inhibition of currents generated by channels made up from dimers (Kv1.1-1.2 and/or Kv1.2-1.1) was similar to the adjacently arranged constructs. These collective findings indicate that assembly of alpha subunits can be directed by this optimized concatenation, and that subunit

  13. Bimatoprost and prostaglandin F(2 alpha) selectively stimulate intracellular calcium signaling in different cat iris sphincter cells.

    PubMed

    Spada, Clayton S; Krauss, Achim H-P; Woodward, David F; Chen, June; Protzman, Charles E; Nieves, Amelia L; Wheeler, Larry A; Scott, David F; Sachs, George

    2005-01-01

    Bimatoprost is a synthetic analog of prostaglandin F(2 alpha) ethanolamide (prostamide F(2 alpha)), and shares a pharmacological profile consistent with that of the prostamides. Like prostaglandin F(2 alpha) carboxylic acid, bimatoprost potently lowers intraocular pressure in dogs, primates and humans. In order to distinguish its mechanism of action from prostaglandin F(2 alpha), fluorescence confocal microscopy was used to examine the effects of bimatoprost, prostaglandin F(2 alpha) and 17-phenyl prostaglandin F(2 alpha) on calcium signaling in resident cells of digested cat iris sphincter, a tissue which exhibits contractile responses to both agonists. Constant superfusion conditions obviated effective conversion of bimatoprost. Serial challenge with 100 nM bimatoprost and prostaglandin F(2 alpha) consistently evoked responses in different cells within the same tissue preparation, whereas prostaglandin F(2 alpha) and 17-phenyl prostaglandin F(2 alpha) elicited signaling responses in the same cells. Bimatoprost-sensitive cells were consistently re-stimulated with bimatoprost only, and prostaglandin F(2 alpha) sensitive cells could only be re-stimulated with prostaglandin F(2 alpha). The selective stimulation of different cells in the same cat iris sphincter preparation by bimatoprost and prostaglandin F(2 alpha), along with the complete absence of observed instances in which the same cells respond to both agonists, strongly suggests the involvement of distinct receptors for prostaglandin F(2 alpha) and bimatoprost. Further, prostaglandin F(2 alpha) but not bimatoprost potently stimulated calcium signaling in isolated human embryonic kidney cells stably transfected with the feline- and human-prostaglandin F(2 alpha) FP-receptor and in human dermal fibroblast cells, and only prostaglandin F(2 alpha) competed with radioligand binding in HEK-feFP cells. These studies provide further evidence for the existence of a bimatoprost-sensitive receptor that is distinct from

  14. Method for characterizing the upset response of CMOS circuits using alpha-particle sensitive test circuits

    NASA Technical Reports Server (NTRS)

    Buehler, Martin G. (Inventor); Blaes, Brent R. (Inventor); Nixon, Robert H. (Inventor); Soli, George A. (Inventor)

    1995-01-01

    A method for predicting the SEU susceptibility of a standard-cell D-latch using an alpha-particle sensitive SRAM, SPICE critical charge simulation results, and alpha-particle interaction physics. A technique utilizing test structures to quickly and inexpensively characterize the SEU sensitivity of standard cell latches intended for use in a space environment. This bench-level approach utilizes alpha particles to induce upsets in a low LET sensitive 4-k bit test SRAM. This SRAM consists of cells that employ an offset voltage to adjust their upset sensitivity and an enlarged sensitive drain junction to enhance the cell's upset rate.

  15. Sensitivity to the seizure-inducing effects of nicotine is associated with strain-specific variants of the alpha 5 and alpha 7 nicotinic receptor subunit genes.

    PubMed

    Stitzel, J A; Blanchette, J M; Collins, A C

    1998-03-01

    Restriction fragment length polymorphisms (rflps) have been identified for the nicotinic ACh receptor subunit genes alpha 5 and alpha 7 between two mouse strains (C3H/2ibg and DBA/2ibg) that differ in sensitivity to the convulsant effects of nicotine. In the study reported here, F2 animals derived from these two parental stains were tested for their sensitivity to the convulsant effects of nicotine as measured by seizure frequency and overall sensitivity score. Subsequently, the animals were genotyped for the alpha 5 and alpha 7 rflps. In addition, levels of alpha-bungarotoxin (alpha-BTX) binding were measured in four brain regions (colliculi, hippocampus, hypothalamus and striatum) to determine whether there is a correlation among alpha-BTX binding levels, sensitivity to nicotine and nicotinic ACh receptor subunit genotype. A significant relationship was observed between alpha 5 and alpha 7 genotype and sensitivity to nicotine. In addition, the alpha 7 rflp significantly correlated with levels of alpha-BTX binding in hippocampus, colliculi and striatum. The alpha 5 rflp did not correlate with alpha-BTX binding levels in any brain region. Levels of alpha-BTX binding did not correlate with nicotine-induced seizure sensitivity or overall nicotine sensitivity score in any of the four brain regions examined.

  16. Sensitivity of alpha band ERD to individual differences in cognition.

    PubMed

    Neubauer, Aljoscha C; Fink, Andreas; Grabner, Roland H

    2006-01-01

    According to the neural efficiency hypothesis, brighter individuals might be characterized by lower and topographically more differentiated brain activation than less intelligent individuals, presumably reflecting a more specialized recruitment of task-related areas. The findings of several studies analyzing the event-related desynchronization (ERD) in the (upper) alpha frequency band have corroborated and elaborated the original neural efficiency hypothesis. In this chapter, we review classical and recent findings and argue in favor of a more differentiated picture of this phenomenon, emphasizing the role of participants' sex, task complexity, and material specificity, as well as the importance to select an adequate external criterion (intelligence measure). Also, recent ERD findings related to emotional intelligence and creativity as well as recent studies focusing on practice, learning ability, and expertise are presented, which point to the need of a broader neurophysiological ability concept. The reviewed findings point at the high suitability of the ERD method to uncover consistent and stable individual differences in people's brain activation patterns when engaged in performing cognitively demanding tasks. PMID:17071230

  17. Bipolar cells of the chick retina containing alpha-bungarotoxin-sensitive nicotinic acetylcholine receptors.

    PubMed

    Hamassaki-Britto, D E; Brzozowska-Prechtl, A; Karten, H J; Lindstrom, J M

    1994-01-01

    Two cDNA clones for nicotinic acetylcholine receptor (nAChR) subunits sensitive to alpha-bungarotoxin (alpha-Bgt) have been isolated, the so-called alpha-Bgt binding proteins alpha 1 (or alpha 7 nAChR subunit) and alpha 2 (or alpha 8 nAChR subunit). Immunohistochemical experiments have shown that both alpha 7 and alpha 8 subunits, as well as subunits insensitive to alpha-Bgt (beta 2 and alpha 3), are present in amacrine and ganglion cells of the chick retina. However, only the alpha 8 subunit was observed in presumptive bipolar cells. The present study investigated in detail the pattern of distribution of the bipolar cells containing the alpha 8 nAChR subunit and its relation to the pattern of distribution of bipolar cells immunoreactive to protein kinase C (PKC). Presumptive alpha 8- and PKC-like immunoreactive (alpha 8-LI and PKC-LI) bipolar cells were observed sending their dendrites to the outer plexiform layers and their axons to the inner plexiform layer. Whereas alpha 8-LI bipolar cells corresponded to 40-53% of the whole population of bipolar cells, PKC-LI bipolar cells represented only 6-8% of the same population. The soma sizes of the alpha 8-LI bipolar cells were slightly smaller (mean +/- S.D.; 4.9 +/- 0.8 microns) than the soma sizes of the PKC-LI bipolar cells (5.4 +/- 0.9 microns). Double-labeling experiments indicated that probably all PKC-LI bipolar cells also contain alpha 8-LI. This indicates that two distinct groups of cholinoceptive bipolar cells exist in the chick retina, one that contains PKC-LI, and another one that does not.

  18. Unusual diversity in alpha-amanitin sensitivity of RNA polymerases in trichomonads.

    PubMed

    Vanácová, S; Tachezy, J; Ullu, E; Tschudi, C

    2001-07-01

    Previous studies in the parasitic protist Trichomonas vaginalis have revealed that protein coding genes are transcribed by an alpha-amanitin-resistant RNA polymerase (RNAP) II. To investigate whether this unusual property is a general characteristic of trichomonads, we addressed the physiology of RNA synthesis in lysolecithin-permeabilized cells. Unlike in T. vaginalis, RNAP II in Tritrichomonas foetus was highly sensitive to the inhibitor alpha-amanitin. On the other hand, RNAP III, identified by its sensitivity to the specific inhibitor tagetitoxin, was found to be resistant to alpha-amanitin in Tritrichomonas foetus, but showed a typical intermediate sensitivity in T. vaginalis. Extension of this study to an additional seven trichomonad species confirmed this genera specific pattern of alpha-amanitin sensitivity and highlighted an unusual diversity in RNAPs among trichomonads, a closely related group of unicellular eukaryotes. PMID:11420110

  19. Sensitivities of five alpha continuous air monitors for detection of airborne sup 239 Pu

    SciTech Connect

    McIsaac, C.V.; Amaro, C.R.

    1992-07-01

    Results of measurements of the sensitivities of five alpha continuous air monitors (CAMs) for detection of airborne {sup 239}Pu are presented. Four commercially available alpha CAMs (Kurz model 8311, Merlin Gerin Edgar, RADeCO model 452, and Victoreen model 758) and a prototype alpha CAM currently in use at Argonne National Laboratory- West (ANL-W) were tested sampling natural ambient air and laboratory-generated atmospheres laden with either blank dust or dust containing nCi/g concentrations of {sup 239}Pu. Cumulative alpha spectra were stored at 30 or 60 minute intervals during each sampling and were subsequently analyzed using three different commonly used alpha spectrum analysis algorithms. The effect of airborne dust concentration and sample filter porosity on detector resolution and sensitivity for airborne {sup 239}Pu are described.

  20. Sensitivities of five alpha continuous air monitors for detection of airborne {sup 239}Pu

    SciTech Connect

    McIsaac, C.V.; Amaro, C.R.

    1992-07-01

    Results of measurements of the sensitivities of five alpha continuous air monitors (CAMs) for detection of airborne {sup 239}Pu are presented. Four commercially available alpha CAMs (Kurz model 8311, Merlin Gerin Edgar, RADeCO model 452, and Victoreen model 758) and a prototype alpha CAM currently in use at Argonne National Laboratory- West (ANL-W) were tested sampling natural ambient air and laboratory-generated atmospheres laden with either blank dust or dust containing nCi/g concentrations of {sup 239}Pu. Cumulative alpha spectra were stored at 30 or 60 minute intervals during each sampling and were subsequently analyzed using three different commonly used alpha spectrum analysis algorithms. The effect of airborne dust concentration and sample filter porosity on detector resolution and sensitivity for airborne {sup 239}Pu are described.

  1. 5-Amino-pyrazoles as potent and selective p38[alpha] inhibitors

    SciTech Connect

    Das, Jagabandhu; Moquin, Robert V.; Dyckman, Alaric J.; Li, Tianle; Pitt, Sidney; Zhang, Rosemary; Shen, Ding Ren; McIntyre, Kim W.; Gillooly, Kathleen; Doweyko, Arthur M.; Newitt, John A.; Sack, John S.; Zhang, Hongjian; Kiefer, Susan E.; Kish, Kevin; McKinnon, Murray; Barrish, Joel C.; Dodd, John H.; Schieven, Gary L.; Leftheris, Katerina

    2012-02-07

    The synthesis and structure-activity relationships (SAR) of p38{alpha} MAP kinase inhibitors based on a 5-amino-pyrazole scaffold are described. These studies led to the identification of compound 2j as a potent and selective inhibitor of p38{alpha} MAP kinase with excellent cellular potency toward the inhibition of TNF{alpha} production. Compound 2j was highly efficacious in vivo in inhibiting TNF{alpha} production in an acute murine model of TNF{alpha} production. X-ray co-crystallography of a 5-amino-pyrazole analog 2f bound to unphosphorylated p38{alpha} is also disclosed.

  2. Karyopherin alpha2: a control step of glucose-sensitive gene expression in hepatic cells.

    PubMed Central

    Guillemain, Ghislaine; Muñoz-Alonso, Maria J; Cassany, Aurélia; Loizeau, Martine; Faussat, Anne-Marie; Burnol, Anne-Françoise; Leturque, Armelle

    2002-01-01

    Glucose is required for an efficient expression of the glucose transporter GLUT2 and other genes. We have shown previously that the intracytoplasmic loop of GLUT2 can divert a signal, resulting in the stimulation of glucose-sensitive gene transcription. In the present study, by interaction with the GLUT2 loop, we have cloned the rat karyopherin alpha2, a receptor involved in nuclear import. The specificity of the binding was restricted to GLUT2, and not GLUT1 or GLUT4, and to karyopherin alpha2, not alpha1. When rendered irreversible by a cross-linking agent, this transitory interaction was detected in vivo in hepatocytes. A role for karyopherin alpha2 in the transcription of two glucose-sensitive genes was investigated by transfection of native and inactive green fluorescent protein-karyopherin alpha2 in GLUT2-expressing hepatoma cells. The amount of inactive karyopherin alpha2 receptor reduced, in a dose-dependent manner, the GLUT2 and liver pyruvate kinase mRNA levels by competition with endogenous active receptor. In contrast, the overexpression of karyopherin alpha2 did not significantly stimulate GLUT2 and liver pyruvate kinase mRNA accumulation in green fluorescent protein-sorted cells. The present study suggests that, in concert with glucose metabolism, karyopherin alpha2 transmits a signal to the nucleus to regulate glucose-sensitive gene expression. The transitory tethering of karyopherin alpha2 to GLUT2 at the plasma membrane might indicate that the receptor can load the cargo to be imported locally. PMID:11988093

  3. Distribution of alpha-amylase activity in selected broiler tissues.

    PubMed

    Rodeheaver, D P; Wyatt, R D

    1986-02-01

    In an examination of broiler alpha-amylase, significant variation in the serum enzyme activity level was noted, adult levels were lower than those of young chicks. Analysis of alpha-amylase activity in various body fluids and tissues of 11-day and 7-week-old broilers indicated that the liver cannot be considered a source of alpha-amylase, although there was activity in both liver tissue and bile of 10 units/g wet weight and 35 units/100 ml, respectively. Fluid from the oral cavity had low levels of alpha-amylase activity, less than 100 units/100 ml, which decreased with age, indicating that the salivary glands may synthesize some alpha-amylase but are not a primary source. Sonication of the pancreatic homogenates was found to significantly increase the apparent activity of alpha-amylase 35-fold over unsonicated homogenates. The pancreas was the major source of alpha-amylase with activities ranging from 89 X 10(2) to 445 X 10(2) units/g wet weight. The level of activity increased with age of the bird. The electrophoretic zymograms of serum, liver, and pancreatic homogenates indicate a similar pancreatic origin for the alpha-amylase found in each tissue or fluid.

  4. Accumbal noradrenaline that contributes to the alpha-adrenoceptor-mediated release of dopamine from reserpine-sensitive storage vesicles in the nucleus accumbens is derived from alpha-methyl-para-tyrosine-sensitive pools.

    PubMed

    Verheij, M M M; Cools, A R

    2009-04-01

    Alpha-adrenoceptors in the nucleus accumbens are known to inhibit accumbal dopamine release from reserpine-sensitive pools. The aim of this study was to test our previously reported hypothesis that accumbal noradrenaline that controls the dopamine release from these storage vesicles is derived from alpha-methyl-para-tyrosine-sensitive pools. The sensitivity of accumbal alpha-adrenoceptors to noradrenergic agents depends on the amount of noradrenaline that is available in the synapse. In case the synaptic noradrenaline levels decrease, the conformation of alpha-adrenoceptors changes into a state that makes these receptors more sensitive to its agonists. The effects of alpha-methyl-para-tyrosine, respectively reserpine, on the alpha-adrenoceptor-agonist-induced changes of accumbal dopamine release were investigated. Alpha-methyl-para-tyrosine, but not reserpine, made accumbal postsynaptic alpha-adrenoceptors more sensitive to phenylephrine. These results indicate that noradrenaline that inhibits the release of dopamine from reserpine-sensitive storage vesicles, via stimulation of accumbal postsynaptic alpha-adrenoceptors, is derived from alpha-methyl-para-tyrosine-sensitive pools. The clinical impact of these data is discussed.

  5. Human alpha 2-adrenergic receptor subtype distribution: widespread and subtype-selective expression of alpha 2C10, alpha 2C4, and alpha 2C2 mRNA in multiple tissues.

    PubMed

    Eason, M G; Liggett, S B

    1993-07-01

    At present, molecular cloning and pharmacological studies have delineated three human alpha 2-adrenergic receptor (alpha 2AR) subtypes, alpha 2C10, alpha 2C4, and alpha 2C2. Assignment of the alpha 2AR subtypes to specific functions has been limited by an unclear definition of tissue alpha 2AR expression outside of the central nervous system. It has been suggested that alpha 2C4 expression is confined to the brain, that alpha 2C2 expression is only in the liver and kidney, and that there is nearly ubiquitous expression of alpha 2C10. However, this is based on studies of a limited number of rat tissues or on studies using non-species-specific approaches. Therefore, to define alpha 2C10, alpha 2C4, and alpha 2C2 tissue expression, we used reverse transcription of total RNA isolated from 20 human tissues, followed by amplification of alpha 2AR cDNA using the polymerase chain reaction. This technique provided two advantages: high sensitivity and, with the use of subtype-specific oligonucleotide primers and probes, differentiation between the alpha 2AR subtypes. The tissues studied were aorta, vena cava, heart (epicardium and endocardium), lung, skeletal muscle, liver, pancreas (head and tail), fat (perinephric and subcutaneous), kidney (cortex and medulla), prostate, stomach, ileum, jejunum, colon, adrenal gland, and spleen. We found that the majority of these tissues expressed alpha 2C10, with the exceptions being the head of the pancreas, subcutaneous fat, colon, and spleen. In marked distinction to other studies, however, we found a prolific expression of the alpha 2C4 and alpha 2C2 subtypes. Expression of alpha 2C4 was found in all tissues with the exception of liver, fat, stomach, and colon, and a virtually ubiquitous expression of alpha 2C2 was found, with the exception of epicardium. Of all tissues studied, only colon and subcutaneous fat expressed a single alpha 2AR subtype, which was alpha 2C2. Thus, the alpha 2AR subtypes do not have a confined expression but

  6. Human alpha 2-adrenergic receptor subtype distribution: widespread and subtype-selective expression of alpha 2C10, alpha 2C4, and alpha 2C2 mRNA in multiple tissues.

    PubMed

    Eason, M G; Liggett, S B

    1993-07-01

    At present, molecular cloning and pharmacological studies have delineated three human alpha 2-adrenergic receptor (alpha 2AR) subtypes, alpha 2C10, alpha 2C4, and alpha 2C2. Assignment of the alpha 2AR subtypes to specific functions has been limited by an unclear definition of tissue alpha 2AR expression outside of the central nervous system. It has been suggested that alpha 2C4 expression is confined to the brain, that alpha 2C2 expression is only in the liver and kidney, and that there is nearly ubiquitous expression of alpha 2C10. However, this is based on studies of a limited number of rat tissues or on studies using non-species-specific approaches. Therefore, to define alpha 2C10, alpha 2C4, and alpha 2C2 tissue expression, we used reverse transcription of total RNA isolated from 20 human tissues, followed by amplification of alpha 2AR cDNA using the polymerase chain reaction. This technique provided two advantages: high sensitivity and, with the use of subtype-specific oligonucleotide primers and probes, differentiation between the alpha 2AR subtypes. The tissues studied were aorta, vena cava, heart (epicardium and endocardium), lung, skeletal muscle, liver, pancreas (head and tail), fat (perinephric and subcutaneous), kidney (cortex and medulla), prostate, stomach, ileum, jejunum, colon, adrenal gland, and spleen. We found that the majority of these tissues expressed alpha 2C10, with the exceptions being the head of the pancreas, subcutaneous fat, colon, and spleen. In marked distinction to other studies, however, we found a prolific expression of the alpha 2C4 and alpha 2C2 subtypes. Expression of alpha 2C4 was found in all tissues with the exception of liver, fat, stomach, and colon, and a virtually ubiquitous expression of alpha 2C2 was found, with the exception of epicardium. Of all tissues studied, only colon and subcutaneous fat expressed a single alpha 2AR subtype, which was alpha 2C2. Thus, the alpha 2AR subtypes do not have a confined expression but

  7. Sensitive radioimmunoassay for detection of antibodies to recombinant human interferon-alpha A

    SciTech Connect

    Palleroni, A.V.; Trown, P.W.

    1986-12-01

    A radioimmunoassay (RIA) for the detection of antibodies to recombinant human leukocyte interferon A (rHuIFN-alpha A) in human serum has been developed and validated against the standard antiviral neutralization bioassay (ANB). The assay measures the binding of /sup 125/I-labeled rHuIFN-alpha A to immunoglobulins in serum. Aliquots of patients' sera are incubated with /sup 125/I-rHuIFN-alpha A and the complexes formed between antibodies in the sera and the /sup 125/I-rHuIFN-alpha A are precipitated with goat anti-human IgG serum. The radioactivity in the immune precipitate is a measure of the quantity of antibody (if present) in the serum. The sensitivity of this RIA is 5 ng of IgG/ml of serum.

  8. Selective flow path alpha particle detector and method of use

    DOEpatents

    Orr, Christopher Henry; Luff, Craig Janson; Dockray, Thomas; Macarthur, Duncan Whittemore

    2002-01-01

    A method and apparatus for monitoring alpha contamination are provided in which ions generated in the air surrounding the item, by the passage of alpha particles, are moved to a distant detector location. The parts of the item from which ions are withdrawn can be controlled by restricting the air flow over different portions of the apparatus. In this way, detection of internal and external surfaces separately, for instance, can be provided. The apparatus and method are particularly suited for use in undertaking alpha contamination measurements during the commissioning operations.

  9. Transmembrane tumor necrosis factor-alpha sensitizes adipocytes to insulin.

    PubMed

    Zhou, Wenjing; Yang, Peng; Liu, Li; Zheng, Shan; Zeng, Qingling; Liang, Huifang; Zhu, Yazhen; Zhang, Zunyue; Wang, Jing; Yin, Bingjiao; Gong, Feili; Wu, Yiping; Li, Zhuoya

    2015-05-01

    Transmembrane TNF-α (tmTNF-α) acts both as a ligand, delivering 'forward signaling' via TNFR, and as a receptor, transducing 'reverse signaling'. The contradiction of available data regarding the effect of tmTNF-α on insulin resistance may be due to imbalance in both signals. Here, we demonstrated that high glucose-induced impairment of insulin-stimulated glucose uptake by 3T3-L1 adipocytes was concomitant with decreased tmTNF-α expression and increased soluble TNF-α (sTNF-α) secretion. However, when TACE was inhibited, preventing the conversion of tmTNF-α to sTNF-α, this insulin resistance was partially reversed, indicating a salutary role of tmTNF-α. Treatment of 3T3-L1 adipocytes with exogenous tmTNF-α promoted insulin-induced phosphorylation of IRS-1 and Akt, facilitated GLUT4 expression and membrane translocation, and increased glucose uptake while addition of sTNF-α resulted in the opposite effect. Furthermore, tmTNF-α downregulated the production of IL-6 and MCP-1 via NF-κB inactivation, as silencing of A20, an inhibitor for NF-κB, by siRNA, abolished this effect of tmTNF-α. However, tmTNF-α upregulated adiponectin expression through the PPAR-γ pathway, as inhibition of PPAR-γ by GW9662 abrogated both tmTNF-α-induced adiponectin transcription and glucose uptake. Our data suggest that tmTNF-α functions as an insulin sensitizer via forward signaling.

  10. Point mutations in the post-M2 region of human alpha-ENaC regulate cation selectivity.

    PubMed

    Ji, H L; Parker, S; Langloh, A L; Fuller, C M; Benos, D J

    2001-07-01

    We tested the hypothesis that an arginine-rich region immediately following the second transmembrane domain may constitute part of the inner mouth of the epithelial Na+ channel (ENaC) pore and, hence, influence conduction and/or selectivity properties of the channel by expressing double point mutants in Xenopus oocytes. Double point mutations of arginines in this post-M2 region of the human alpha-ENaC (alpha-hENaC) led to a decrease and increase in the macroscopic conductance of alphaR586E,R587Ebetagamma- and alphaR589E,R591Ebetagamma-hENaC, respectively, but had no effect on the single-channel conductance of either double point mutant. However, the apparent equilibrium dissociation constant for Na+ was decreased for both alphaR586E,R587Ebetagamma- and alphaR589E,R591Ebetagamma-hENaC, and the maximum amiloride-sensitive Na+ current was decreased for alphaR586E,R587Ebetagamma-hENaC and increased for alphaR589E,R591Ebetagamma-hENaC. The relative permeabilities of Li+ and K+ vs. Na+ were increased 11.25- to 27.57-fold for alphaR586E,R587Ebetagamma-hENaC compared with wild type. The relative ion permeability of these double mutants and wild-type ENaC was inversely related to the crystal diameter of the permeant ions. Thus the region of positive charge is important for the ion permeation properties of the channel and may form part of the pore itself. PMID:11401828

  11. The HPV-16 E7 oncogene sensitizes malignant cells to IFN-alpha-induced apoptosis

    SciTech Connect

    Wang, Yisong

    2005-10-01

    Interferons (IFNs) exert antitumor effects in several human malignancies, but their mechanism of action is unclear. There is a great variability in sensitivity to IFN treatment depending on both tumor type and the individual patient. The reason for this variable sensitivity is not known. The fact that several IFN-induced anticellular effects are exerted through modulation of proto-oncogenes and tumor suppressor genes may indicate that the malignant genotype may be decisive in the cell's sensitivity to IFN. To determine if a deregulated oncogene could alter the cellular response to IFN, a mouse lymphoma cell line (J3D) was stably transfected with the viral human papillomavirus-16 (HPV-16) E7 oncogene. The E7-transfected cells and their respective mock-transfected sister clones were treated with IFN-{alpha} and examined for possible IFN-induced anticellular effects. We found that the E7-transfected clones were greatly sensitized to IFN-{alpha}-induced apoptosis compared with their mock-transfected counterparts. Induction of apoptosis in the transfected cells correlated with the ability of IFN to activate parts of the proapoptotic machinery specifically in these cells, including activation of caspases and the proapoptotic protein Bak. In summary, our data suggest that transfection of malignant cells with the E7 oncogene can sensitize them to IFN-{alpha}-induced apoptosis. This demonstrates that an oncogenic event may alter the cellular sensitivity to IFN and might also have implications for treatment of HPV related diseases with IFN.

  12. Selective down-regulation of the alpha6-integrin subunit in melanocytes by UVB light.

    PubMed

    Krengel, Sven; Stark, Imke; Geuchen, Christian; Knoppe, Bettina; Scheel, Gabriele; Schlenke, Peter; Gebert, Andreas; Wünsch, Lutz; Brinckmann, Jürgen; Tronnier, Michael

    2005-06-01

    In vivo, melanocytes bind to laminin (LM) molecules of the basement membrane (BM) via the integrins alpha3beta1 and alpha6beta1, and they adhere to neighbouring keratinocytes via E-cadherin. Only few studies have addressed the impact of ultraviolet (UV) light on the interaction of melanocytes with their microenvironment. In this report, we examined the influence of UVB irradiation on the expression of the most important melanocyte-adhesion molecules (E-, N-cadherin, alpha2-, alpha3-, alpha5-, alpha6-, alphaV-, beta1-, beta3-integrins and ICAM-1) in vitro by flow cytometry. We were able to demonstrate that the alpha6-integrin subunit is selectively and reversibly down-regulated by UVB in a dwzm 150ose-dependent manner. In comparison, keratinocytes lacked UVB-inducible alterations in the expression of alpha6-integrin. In the presence of LM-1, the UVB-induced down-regulation of alpha6-integrin in melanocytes was significantly reduced. Moreover, LM-1 increased the resistance of melanocytes to UVB-induced cell death, as measured by annexinV-binding analysis. This effect was reversed by preincubation with an alpha6-integrin-blocking antibody. By immunofluorescence, we could demonstrate that UVB leads to a dose-dependent internalization of alpha6-integrin, providing an obvious explanation for the down-regulation on the outer cell surface observed by flow cytometry. We suggest that adhesion to LM-1 through alpha6-integrin represents a protective mechanism for melanocytes to withstand UVB damage. Through alpha6-integrin internalization, sunburns might alter the interaction between melanocytes and the BM, resulting in apoptosis induced by loss of anchorage (anoikis). Repeated sunburns may then lead to the selection of a population of melanocytes which are capable of anchorage-independent survival, culminating in solar nevogenesis and melanoma development.

  13. Molecular cloning of the alpha-1 subunit of an omega-conotoxin-sensitive calcium channel.

    PubMed Central

    Dubel, S J; Starr, T V; Hell, J; Ahlijanian, M K; Enyeart, J J; Catterall, W A; Snutch, T P

    1992-01-01

    Of the four major types of Ca channel described in vertebrate cells (designated T, L, N, and P), N-type Ca channels are unique in that they are found specifically in neurons, have been correlated with control of neurotransmitter release, and are blocked by omega-conotoxin, a neuropeptide toxin isolated from the marine snail Conus geographus. A set of overlapping cDNA clones were isolated and found to encode a Ca channel alpha-1 subunit, designated rbB-I. Polyclonal antiserum generated against a peptide from the rbB-I sequence selectively immunoprecipitates high-affinity 125I-labeled omega-conotoxin-binding sites from labeled rat forebrain membranes. PCR analysis shows that, like N-type Ca channels, expression of rbB-I is limited to the nervous system and neuronally derived cell lines. This brain Ca channel may mediate the omega-conotoxin-sensitive Ca influx required for neurotransmitter release at many synapses. Images PMID:1317580

  14. Molecularly imprinted solid phase extraction for the selective HPLC determination of alpha-tocopherol in bay leaves.

    PubMed

    Puoci, F; Cirillo, G; Curcio, M; Iemma, F; Spizzirri, U G; Picci, N

    2007-06-19

    A new sorbent for molecularly imprinted solid phase extraction (MISPE) was synthesized to extract and purify alpha-tocopherol (alpha-TP) from vegetable sources. Molecularly imprinted polymers (MIP) were synthesized using methacrylic acid (MAA) as functional monomer and ethylene glycol dimethacrylate (EGDMA) as crosslinking agent using a photo-polymerization procedure. A thermo-polymerization was also performed but no imprinting effect in the resulting materials was raised. The proposed MISPE protocol could overcome the drawback of traditional detection methods, which require pre-treatments of the samples. The possibility to obtain the selective recognition of alpha-TP from natural samples in aqueous mixtures represents one of the main advantages of our materials. Our procedure involves the direct HPLC injection of eluate without any treatment and above all the use of no toxic and biocompatible organic solvents. After the evaluation of the selectivity of the alpha-TP imprinted polymers, the performance of these materials as solid phase extraction (SPE) sorbents was investigated. Our MISPE-HPLC procedure has a high sensitivity, LOD and LOQ were 3.49x10(-7) and 1.16x10(-6) mol L(-1), respectively, as well as good precision (intraday precision below 3.3% and interday precisions below 6.5%) and recovery (60%). Thus, it can be successfully used for the purification of alpha-TP from bay leaves.

  15. Selective presynaptic insectotoxin (alpha-latroinsectotoxin) isolated from black widow spider venom.

    PubMed

    Magazanik, L G; Fedorova, I M; Kovalevskaya, G I; Pashkov, V N; Bulgakov, O V; Grishin, E V

    1992-01-01

    A homogenous protein of 120,000 mol. wt isolated from black widow spider (Lactrodectus mactans tredecimguttatus) venom and referred to as alpha-latroinsectotoxin was highly potent (4 nM) in the induction of an increase of the frequency of miniature excitatory postsynaptic potentials in blowfly (Calliphora vicina) larvae neuromuscular preparations. In the frog nerve ending, however, even 50 nM alpha-latroinsectotoxin failed to affect transmitter release. Pretreatment of insect preparations with alpha-latrotoxin or frog preparations with alpha-latroinsectotoxin did not prevent the specific effect of consequent applications of alpha-latroinsectotoxin (insect) and alpha-latrotoxin (frog), respectively. The binding of labelled [125I]alpha-latroinsectotoxin to insect and [125I]alpha-latrotoxin to bovine membrane preparations was saturable and highly specific. The presynaptic effect, but not the binding of alpha-latroinsectotoxin, was dependent on the presence of divalent cations in the external medium. Mg2+ could readily substitute for Ca2+ and increase of transmitter release induced by alpha-latroinsectotoxin also occurred in Ca(2+)-free solutions. Pretreatment of preparations with 300 micrograms/ml concanavalin A completely abolished both the presynaptic effect of alpha-latroinsectotoxin and its binding to insect membrane preparations. Thus, the phenomenology of alpha-latroinsectotoxin action on insects resembles in general that described for the action of alpha-latrotoxin on vertebrates. The selectivity of alpha-latrotoxin and alpha-latroinsectotoxin seems to be due to differences in the structure of neurotoxin receptors in nerve endings of vertebrates and insects, although the mode of presynaptic action has a great deal in common. PMID:1594101

  16. Structural differences determine the relative selectivity of nicotinic compounds for native alpha 4 beta 2*-, alpha 6 beta 2*-, alpha 3 beta 4*- and alpha 7-nicotine acetylcholine receptors.

    PubMed

    Grady, Sharon R; Drenan, Ryan M; Breining, Scott R; Yohannes, Daniel; Wageman, Charles R; Fedorov, Nikolai B; McKinney, Sheri; Whiteaker, Paul; Bencherif, Merouane; Lester, Henry A; Marks, Michael J

    2010-06-01

    Mammalian brain expresses multiple nicotinic acetylcholine receptor (nAChR) subtypes that differ in subunit composition, sites of expression and pharmacological and functional properties. Among known subtypes of receptors, alpha 4 beta 2* and alpha 6 beta 2*-nAChR have the highest affinity for nicotine (where * indicates possibility of other subunits). The alpha 4 beta 2*-nAChRs are widely distributed, while alpha 6 beta 2*-nAChR are restricted to a few regions. Both subtypes modulate release of dopamine from the dopaminergic neurons of the mesoaccumbens pathway thought to be essential for reward and addiction. alpha 4 beta 2*-nAChR also modulate GABA release in these areas. Identification of selective compounds would facilitate study of nAChR subtypes. An improved understanding of the role of nAChR subtypes may help in developing more effective smoking cessation aids with fewer side effects than current therapeutics. We have screened a series of nicotinic compounds that vary in the distance between the pyridine and the cationic center, in steric bulk, and in flexibility of the molecule. These compounds were screened using membrane binding and synaptosomal function assays, or recordings from GH4C1 cells expressing h alpha 7, to determine affinity, potency and efficacy at four subtypes of nAChRs found in brain, alpha 4 beta 2*, alpha 6 beta 2*, alpha 7 and alpha 3 beta 4*. In addition, physiological assays in gain-of-function mutant mice were used to assess in vivo activity at alpha 4 beta 2* and alpha 6 beta 2*-nAChRs. This approach has identified several compounds with agonist or partial agonist activity that display improved selectivity for alpha 6 beta 2*-nAChR.

  17. Positions under positive selection--key for selectivity and potency of scorpion alpha-toxins.

    PubMed

    Weinberger, Hagar; Moran, Yehu; Gordon, Dalia; Turkov, Michael; Kahn, Roy; Gurevitz, Michael

    2010-05-01

    Alpha-neurotoxins target voltage-gated sodium channels (Na(v)s) and constitute an important component in the venom of Buthidae scorpions. These toxins are short polypeptides highly conserved in sequence and three-dimensional structure, and yet they differ greatly in activity and preference for insect and various mammalian Na(v)s. Despite extensive studies of the structure-function relationship of these toxins, only little is known about their evolution and phylogeny. Using a broad data set based on published sequences and rigorous cloning, we reconstructed a reliable phylogenetic tree of scorpion alpha-toxins and estimated the evolutionary forces involved in the diversification of their genes using maximum likelihood-based methods. Although the toxins are largely conserved, four positions were found to evolve under positive selection, of which two (10 and 18; numbered according to LqhalphaIT and Lqh2 from the Israeli yellow scorpion Leiurus quinquestriatus hebraeus) have been previously shown to affect toxin activity. The putative role of the other two positions (39 and 41) was analyzed by mutagenesis of Lqh2 and LqhalphaIT. Whereas substitution P41K in Lqh2 did not alter its activity, substitution K41P in LqhalphaIT significantly decreased the activity at insect and mammalian Na(v)s. Surprisingly, not only that substitution A39L in both toxins increased their activity by 10-fold but also LqhalphaIT(A39L) was active at the mammalian brain channel rNa(v)1.2a, which otherwise is hardly affected by LqhalphaIT, and Lqh2(A39L) was active at the insect channel, DmNa(v)1, which is almost insensitive to Lqh2. Thus, position 39 is involved not only in activity but also in toxin selectivity. Overall, this study describes evolutionary forces involved in the diversification of scorpion alpha-toxins, highlights the key role of positions under positive selection for selectivity and potency, and raises new questions as to the toxin-channel face of interaction.

  18. Selective non-steroidal inhibitors of 5 alpha-reductase type 1.

    PubMed

    Occhiato, Ernesto G; Guarna, Antonio; Danza, Giovanna; Serio, Mario

    2004-01-01

    The enzyme 5 alpha-reductase (5 alpha R) catalyses the reduction of testosterone (T) into the more potent androgen dihydrotestosterone (DHT). The abnormal production of DHT is associated to pathologies of the main target organs of this hormone: the prostate and the skin. Benign prostatic hyperplasia (BPH), prostate cancer, acne, androgenetic alopecia in men, and hirsutism in women appear related to the DHT production. Two isozymes of 5 alpha-reductase have been cloned, expressed and characterized (5 alpha R-1 and 5 alpha R-2). They share a poor homology, have different chromosomal localization, enzyme kinetic parameters, and tissue expression patterns. Since 5 alpha R-1 and 5 alpha R-2 are differently distributed in the androgen target organs, a different involvement of the two isozymes in the pathogenesis of prostate and skin disorders can be hypothesized. High interest has been paid to the synthesis of inhibitors of 5 alpha-reductase for the treatment of DHT related pathologies, and the selective inhibition of any single isozyme represents a great challenge for medical and pharmaceutical research in order to have more specific drugs. At present, no 5 alpha R-1 inhibitor is marketed for the treatment of 5 alpha R-1 related pathologies but pharmaceutical research is very active in this field. This paper will review the major classes of 5 alpha R inhibitors focusing in particular on non-steroidal inhibitors and on structural features that enhance the selectivity versus the type 1 isozyme. Biological tests to assess the inhibitory activity towards the two 5 alpha R isozymes will be also discussed. PMID:15026079

  19. Tumour necrosis factor-alpha induces translocation of protein kinase C in tumour necrosis factor-sensitive cell lines.

    PubMed Central

    Matsubara, N; Fuchimoto, S; Orita, K

    1991-01-01

    In this study we investigated whether the anti-proliferative effect of tumour necrosis factor-alpha (TNF-alpha) was associated with the activation of protein kinase C (PKC), using PANC-1 cells (TNF-alpha sensitive) and LoVo cells (TNF-alpha resistant). In combination with 12-0-tetradecanoylphorbol-13-acetate (TPA), a potent activator of PKC, TNF-alpha caused marked inhibition of the growth of LoVo cells. Inhibition of PANC-1 cell growth by TNF-alpha was blocked by pretreatment with TPA for 24 hr, along with down-regulation of PKC activity. Intracellular translocation of PKC from cytosol to membrane was induced by TNF-alpha treatment in PANC-1 cells but not in LoVo cells. PMID:1916896

  20. Sensitization of rhabdo-, lenti-, and spumaviruses to human serum by galactosyl(alpha1-3)galactosylation.

    PubMed

    Takeuchi, Y; Liong, S H; Bieniasz, P D; Jäger, U; Porter, C D; Friedman, T; McClure, M O; Weiss, R A

    1997-08-01

    Vesicular stomatitis virus, human immunodeficiency virus type 2, and human foamy virus, which were produced by cell lines expressing galactosyl(alpha1-3)galactosyl (alphaGal) sugars, were found to be less stable in human serum than those from alphaGal-negative cells, indicating that galactosyl(alpha1-3)galactosylation sensitizes these viruses as well as mammalian type C oncoviruses (Rother et al., J. Exp. Med. 182:1345-1355, 1995; Takeuchi et al., Nature (London) 379:85-88, 1996) to complement killing via natural anti-alphaGal antibodies. Thus, virus killing mediated by anti-alphaGal antibodies may play a role as a barrier to animal-to-human infection of various enveloped viruses. Virus vectors for human in vivo gene therapy based on the viruses mentioned above should be produced from alphaGal-negative cells. PMID:9223512

  1. Increased sensitivity to nicotine-induced seizures in mice expressing the L250T alpha 7 nicotinic acetylcholine receptor mutation.

    PubMed

    Broide, Ron S; Salas, Ramiro; Ji, Daoyun; Paylor, Richard; Patrick, James W; Dani, John A; De Biasi, Mariella

    2002-03-01

    High doses of nicotine, the addictive component of tobacco, induce clonic-tonic seizures in animals. Pharmacological and biochemical data have suggested that alpha 7-containing neuronal nicotinic receptors (nAChRs) contribute to these seizures. To study potential alpha 7 contributions, we examined alpha 7 subunits with a Leu250-to-Thr substitution in the channel domain, which creates a gain-of-function mutation. Previous studies have shown that mice homozygous for the alpha 7 L250T mutation (T/T) die shortly after birth, but animals heterozygous for the mutation (+/T) are viable and grow to adulthood. Hippocampal neurons from the +/T mice exhibited altered alpha 7-type currents with increased amplitudes and slower desensitization kinetics, confirming a partial gain of function for the alpha 7 nAChR. We found that +/T mice were more sensitive to the convulsant effects of nicotine compared with their wild-type (+/+) littermates. Furthermore, although their behavior was normal in basal conditions, +/T mice showed a unique nicotine-induced phenotype, consisting of head-bobbing and paw-tapping movements. Increased sensitivity to nicotine-induced seizures occurred despite a 60% decline in brain alpha 7 nAChR protein levels. There were no changes in the levels of alpha 4, alpha 5, alpha 6, alpha 7, beta 2, and beta 4 mRNA, or in [(125)I]epibatidine and [(3)H]nicotine binding between +/T and +/+ mice. Recent data from our laboratory show that alpha 7-null mice maintain normal sensitivity to nicotine-induced seizures. Hence, these present findings suggest that alterations in the properties rather than absence of alpha 7 nAChRs might affect the mechanisms underlying the convulsive properties of nicotine.

  2. Mechanisms of reduced luteal sensitivity to prostaglandin F2alpha during maternal recognition of pregnancy in ewes.

    PubMed

    Costine, Beth A; Inskeep, E Keith; Blemings, Kenneth P; Flores, Jorge A; Wilson, Matthew E

    2007-02-01

    During maternal recognition of pregnancy, the conceptus stimulates endometrial secretion of PGF2alpha and PGE2. However, PGF2alpha is less effective in causing luteal regression in pregnant than in non-pregnant ewes. Experiments were conducted to elucidate mechanisms for reduced luteal sensitivity to PGF2alpha during maternal recognition of pregnancy. Corpora lutea (CL) were collected from pregnant and non-pregnant ewes 0, 4, or 12h following treatment with PGF2alpha on day 12 after estrus. Luteal PTGHS2 mRNA did not differ due to PGF2alpha or pregnancy status. Luteal PTGES mRNA was reduced in both pregnant and non-pregnant ewes after PGF2alpha treatment; while, luteal PTGFS mRNA was reduced 4h after PGF2alpha in pregnant, but not non-pregnant ewes. The result was a greater ratio of PTGES to PTGFS mRNA in pregnant ewes. Luteal mRNA for HPGD did not differ between pregnant and non-pregnant ewes on day 12. Luteal END1 mRNA was reduced in pregnant as compared to non-pregnant ewes prior to PGF2alpha challenge. Luteal END1 mRNA was increased after PGF2alpha in pregnant and non-pregnant ewes; however, ECE1 mRNA was reduced 4h after PGF2alpha in pregnant, but not non-pregnant ewes. The in vitro conversion of PGF2alpha to PGFM was greater in CL of pregnant than non-pregnant ewes at day 14. Luteal conversion of PGF2alpha to PGFM appears to be regulated post-transcriptionally. During maternal recognition of pregnancy, mechanisms of reduced luteal sensitivity to PGF2alpha may include a shift in prostaglandin production to the luteotropin PGE2, a reduction of ECE1 mRNA, and increased catabolism of PGF2alpha. PMID:16524686

  3. Mutagenicity of tocopheryl quinones: evolutionary advantage of selective accumulation of dietary alpha-tocopherol.

    PubMed

    Cornwell, David G; Williams, Marshall V; Wani, Altaf A; Wani, Gulzar; Shen, Elaine; Jones, Kenneth H

    2002-01-01

    We have shown that phenolic antioxidant tocopherols are oxidized to nonarylating alpha-tocopheryl quinone (alpha-TQ) and arylating gamma- and delta-TQ electrophiles. The arylating quinones stimulate apoptosis and are highly cytotoxic in mammalian cells. Some xenobiotic phenolic antioxidants are mutagens, and it has been suggested that their arylating quinone metabolites are the active agents in mutagenesis related to carcinogenesis. We found that neither alpha- nor gamma-TQ was directly genotoxic in supercoiled-to-nicked circular DNA conversions, but these agents interacted with the cytomegalovirus reporter-driven plasmid and enhanced luciferase transfection, with gamma-TQ > alpha-TQ. The Ames test, using gamma-TQ and a number of Salmonella strains, showed no evidence of bacterial mutagenesis. gamma-TQ was highly cytotoxic and alpha-TQ slightly cytotoxic in eukaryocyte AS52 cells. A guanosine phosphoribosyltransferase gene assay showed that gamma-TQ was highly mutagenic and alpha-TQ slightly mutagenic in AS52 cells. A review of the literature identified associations where a decrease in dietary gamma-tocopherol (gamma-T) diminishes and an increase in dietary gamma-T and its quinone enhances carcinogenicity. Humans and other omnivores selectively accumulate alpha-tocopherol, even though gamma-T is their principal dietary tocopherol. We suggest that this selectivity confers an evolutionary advantage by limiting tissue gamma-T, a putative precursor of the mutagen gamma-TQ.

  4. Evaluation of the pharmacological selectivity profile of alpha 1 adrenoceptor antagonists at prostatic alpha 1 adrenoceptors: binding, functional and in vivo studies.

    PubMed Central

    Kenny, B. A.; Miller, A. M.; Williamson, I. J.; O'Connell, J.; Chalmers, D. H.; Naylor, A. M.

    1996-01-01

    1. The profile of a range of alpha 1 adrenoceptor antagonists was determined in vitro against cloned human alpha 1A, alpha 1B and alpha 1D adrenoceptors and against noradrenaline-mediated contractions of rat aorta and human prostate. The in vivo profile of compounds was determined in an anaesthetized dog model which allowed the simultaneous assessment of antagonist potency against phenylephrine-mediated increases in blood pressure and prostatic pressure. 2. The quinazoline antagonists, prazosin, doxazosin and alfuzosin displayed high affinity but were non selective for the three cloned human alpha 1 adrenoceptors. Indoramin and SNAP 1069 showed selectivity for alpha 1A and alpha 1B adrenoceptors relative to the alpha 1D subtype. Rec 15/2739, WB 4101, SL 89,0591, (+)- and (-)- tamsulosin showed selectivity for alpha 1A and alpha 1D adrenoceptors relative to the alpha 1B subtype. RS 17053 showed high affinity and selectivity for alpha 1A adrenoceptors (pKi 8.6) relative to alpha 1B (pKi = 7.3) and alpha 1D (pKi = 7.1) subtypes. 3. (+)-Tamsulosin, (-)-tamsulosin, SL 89,0591, Rec 15/2739, SNAP 1069 and RS 17053 appeared to act as competitive antagonists of noradrenaline-mediated contractions of rat aorta yielding pA2 affinity estimates which were similar to binding affinities at cloned human alpha 1D adrenoceptors. The following rank order was obtained: prazosin = (-)-tamsulosin > doxazosin > SL 89,0591 = (+)-tamsulosin > Rec 15/2739 > RS 17053 = SNAP 1069. 4. (-)-Tamsulosin was a very potent, insurmountable antagonist of noradrenaline-mediated contractions of human prostate, yielding an approximate pA2 estimate of 9.8 at 1 nM. The corresponding (+)-enantiomer was 30 fold weaker. SL 89,0591, SNAP 1069 and Rec 15/2739 yielded pA2 estimates which compared well with their alpha 1A binding affinities. The affinity estimate for prazosin on human prostate was lower than the corresponding binding affinity determined at alpha 1A adrenoceptors and RS 17053 was a very weak

  5. Selection of novel analogs of thalidomide with enhanced tumor necrosis factor alpha inhibitory activity.

    PubMed Central

    Corral, L. G.; Muller, G. W.; Moreira, A. L.; Chen, Y.; Wu, M.; Stirling, D.; Kaplan, G.

    1996-01-01

    BACKGROUND: Tumor necrosis factor alpha (TNF alpha) is thought to mediate both protective and detrimental manifestations of the inflammatory response. Recently, thalidomide (alpha-N-phthalimidoglutarimide) was shown to partially inhibit monocyte TNF alpha production (by 50-70%) both in vivo and in vitro. More efficient inhibition of TNF alpha may, however, be necessary to rescue the host from more acute and extensive toxicities of TNF alpha-mediated inflammation. MATERIALS AND METHODS: Three structural analogues of thalidomide were selected for study based on increased activity against TNF alpha production. The parent drug and the analogs were tested in vitro in human peripheral blood mononuclear cell cultures for their effects on lipopolysaccharide (LPS) induced cytokine protein and mRNA production using ELISAs and Northern blot hybridization. The in vitro effects of the drugs were then confirmed in vivo in a mouse model of LPS induced lethality. RESULTS: The new compounds (two esters and one amide) showed increased inhibition of TNF alpha production by LPS-stimulated human monocytes, relative to the parent drug thalidomide. The analogs and the parent drug enhanced the production of interleukin 10 (IL-10), but had little effect on IL-6 and IL-1 beta protein and mRNA production. When tested in vivo, the amide analog protected 80% of LPS-treated mice against death from endotoxin induced shock. CONCLUSIONS: Analogs of thalidomide designed to better inhibit TNF alpha production in vitro have correspondingly greater efficacy in vivo. These finding may have therapeutic implication for the treatment of human diseases characterized by acute and extensive TNF alpha production such as tuberculous meningitis or toxic shock. Images FIG. 3 FIG. 4 PMID:8827720

  6. Independent Causal Contributions of Alpha- and Beta-Band Oscillations during Movement Selection

    PubMed Central

    Stolk, Arjen; Marshall, Tom R.; Esterer, Sophie; Sharp, Poppy; Dijkerman, H. Chris; de Lange, Floris P.; Toni, Ivan

    2016-01-01

    To select a movement, specific neuronal populations controlling particular features of that movement need to be activated, whereas other populations are downregulated. The selective (dis)inhibition of cortical sensorimotor populations is governed by rhythmic neural activity in the alpha (8–12 Hz) and beta (15–25 Hz) frequency range. However, it is unclear whether and how these rhythms contribute independently to motor behavior. Building on a recent dissociation of the sensorimotor alpha- and beta-band rhythms, we test the hypothesis that the beta-band rhythm governs the disinhibition of task-relevant neuronal populations, whereas the alpha-band rhythm suppresses neurons that may interfere with task performance. Cortical alpha- and beta-band rhythms were manipulated with transcranial alternating current stimulation (tACS) while human participants selected how to grasp an object. Stimulation was applied at either 10 or 20 Hz and was imposed on the sensorimotor cortex contralaterally or ipsilaterally to the grasping hand. In line with task-induced changes in endogenous spectral power, the effect of the tACS intervention depended on the frequency and site of stimulation. Whereas tACS stimulation generally increased movement selection times, 10 Hz stimulation led to relatively faster selection times when applied to the hemisphere ipsilateral to the grasping hand, compared with other stimulation conditions. These effects occurred selectively when multiple movements were considered. These observations functionally differentiate the causal contribution of alpha- and beta-band oscillations to movement selection. The findings suggest that sensorimotor beta-band rhythms disinhibit task-relevant populations, whereas alpha-band rhythms inhibit neuronal populations that could interfere with movement selection. SIGNIFICANCE STATEMENT This study shows dissociable effects of 10 Hz and 20 Hz tACS on the duration of movement selection. These observations have two elements of

  7. Spectroscopic studies on the riboflavin-sensitized conformational changes of calf lens alpha-crystallin.

    PubMed

    Andley, U P

    1988-04-01

    The change in conformation of calf lens alpha-crystallin by oxidation in the presence of the photosensitizer riboflavin and light has been investigated. Near-UV circular dichroism (CD) spectrum, absorption spectrum, tryptophan fluorescence yield and fluorescence lifetime of the SH-specific fluorescent probe, N-iodoacetyl-N'-(5-sulfo-1-naphtyl) ethylenediamine (1,5-IAEDANS), were significantly altered by irradiation in the presence of RF. In the initial stages of photolysis (1-2 hr), a slight degradation of the protein to lower molecular weight peptides was observed. Upon increased photolysis, intersubunit cross-linking to dimers and other high molecular weight species was observed. To determine the effects of cross-linking on the accessibility of the cysteine residues of the protein, lifetime quenching studies on the IAEDANS-labeled alpha-crystallin were performed. A decrease in the quenching constant (kappa q) in the photolysed sample indicates that the labeled SH groups are less susceptible to collisional quenching, which requires contact between the quencher and the excited state of the fluorophore, due to steric inhibition in the cross-linked protein. Cross-linking and the rate of loss of tryptophan fluorescence of alpha-crystallin diminished under anaerobic conditions and increased when D2O was used in the medium for irradiation. Use of inhibitors and quenchers of active species of oxygen suggests that photo-oxidation probably occurs via the action of singlet oxygen as well as substrate-sensitizer complexation.

  8. Coordinated balancing of muscle oxidative metabolism through PGC-1{alpha} increases metabolic flexibility and preserves insulin sensitivity

    SciTech Connect

    Summermatter, Serge; Santos, Gesa

    2011-04-29

    Highlights: {yields} PGC-1{alpha} enhances muscle oxidative capacity. {yields} PGC-1{alpha} promotes concomitantly positive and negative regulators of lipid oxidation. {yields} Regulator abundance enhances metabolic flexibility and balances oxidative metabolism. {yields} Balanced oxidation prevents detrimental acylcarnitine and ROS generation. {yields} Absence of detrimental metabolites preserves insulin sensitivity -- Abstract: The peroxisome proliferator-activated receptor {gamma} coactivator 1{alpha} (PGC-1{alpha}) enhances oxidative metabolism in skeletal muscle. Excessive lipid oxidation and electron transport chain activity can, however, lead to the accumulation of harmful metabolites and impair glucose homeostasis. Here, we investigated the effect of over-expression of PGC-1{alpha} on metabolic control and generation of insulin desensitizing agents in extensor digitorum longus (EDL), a muscle that exhibits low levels of PGC-1{alpha} in the untrained state and minimally relies on oxidative metabolism. We demonstrate that PGC-1{alpha} induces a strictly balanced substrate oxidation in EDL by concomitantly promoting the transcription of activators and inhibitors of lipid oxidation. Moreover, we show that PGC-1{alpha} enhances the potential to uncouple oxidative phosphorylation. Thereby, PGC-1{alpha} boosts elevated, yet tightly regulated oxidative metabolism devoid of side products that are detrimental for glucose homeostasis. Accordingly, PI3K activity, an early phase marker for insulin resistance, is preserved in EDL muscle. Our findings suggest that PGC-1{alpha} coordinately coactivates the simultaneous transcription of gene clusters implicated in the positive and negative regulation of oxidative metabolism and thereby increases metabolic flexibility. Thus, in mice fed a normal chow diet, over-expression of PGC-1{alpha} does not alter insulin sensitivity and the metabolic adaptations elicited by PGC-1{alpha} mimic the beneficial effects of endurance training

  9. Elucidation of the molecular logic by which misfolded alpha 1-antitrypsin is preferentially selected for degradation.

    PubMed

    Wu, Ying; Swulius, Matthew T; Moremen, Kelley W; Sifers, Richard N

    2003-07-01

    The exocytic pathway provides a physical route through which newly synthesized secretory and membrane proteins are deployed to the eukaryote cell surface. For newly synthesized alpha1-antitrypsin (AAT), the modification of its asparagine-linked oligosaccharides by a slow-acting mannosidase partitions the misfolded monomer into the proteasomal degradation pathway. Herein, we asked whether, and how, modification by endoplasmic reticulum mannosidase I (ERManI) contributes to the preferential selection of the misfolded AAT monomer for proteasomal degradation. Transiently expressed mutant and WT AAT variants underwent rapid destabilization in response to an artificially elevated ERManI concentration in the murine hepatoma cell line, Hepa1a. Based on the mannosidase- and lactacystin-sensitive properties of intracellular turnover, a stochastic model is proposed in which the delayed onset of the glycan modification, relative to the duration of nonnative protein structure, coordinates the preferential degradation of the misfolded monomer and spares the native molecule from destruction. Newly synthesized endogenous transferrin underwent degradation in response to an elevated concentration of ERManI, whereas the nonglycosylated secretory glycoprotein albumin was not affected. Taken together, these findings indicate that efficient conformational maturation might function as the initial quality control standard for a broad population of glycoproteins.

  10. Examination by radioligand binding of the alpha1 adrenoceptors in the mesenteric arterial vasculature during the development of salt-sensitive hypertension.

    PubMed

    Caveney, S W; Taylor, D A; Fleming, W W

    1997-09-01

    Previous experiments have suggested that the vascular smooth muscle of Dahl salt-sensitive (DS) rats may possess a difference in the alpha1-adrenoceptor population or its transduction processes compared to Dahl salt-resistant (DR) rats. The purpose of the current research is to study the role of alpha1-adrenoceptors in the specific supersensitivity to norepinephrine (NE) seen prior to and early in the development of hypertension in the DS rat. Experiments in isolated perfused superior mesenteric arterial vasculature from DS rats chronically fed a high (7%) salt diet for 5 days or 3 weeks, in the absence or presence of an elevation in systolic blood pressure, respectively, demonstrated a specific supersensitivity to NE relative to DR rats. The enhanced responsiveness was specific to NE after 5 days of high salt since no differences in sensitivity of these preparations was observed to either KCl or 5-HT. A small but significant elevation in sensitivity to KCl following 3 weeks of treatment suggests that multiple factors may contribute to tissue responsiveness at this time. Radioligand binding experiments were performed using [125I]-HEAT to study the alpha1-adrenoceptor population and its subtypes. Saturation experiments using membranes prepared from the superior mesenteric arterial vasculature or mesenteric arterial branches showed no significant differences in overall alpha1-adrenoceptor population between DS and DR rats fed a high-salt diet for 5 days or 3 weeks. Competition experiments using membranes prepared from the superior mesenteric arterial branches in the presence of the alpha1A-subtype selective antagonist 5-methylurapidil showed two binding sites (high and low affinity) in these resistance vessels but no significant differences in nature or ratio of these sites between the DS and DR groups. These results suggest that changes in the alpha1-adrenoceptor population are not responsible for the specific supersensitivity to NE, which may be an early event in

  11. A SUCCESSFUL BROADBAND SURVEY FOR GIANT Ly{alpha} NEBULAE. I. SURVEY DESIGN AND CANDIDATE SELECTION

    SciTech Connect

    Prescott, Moire K. M.; Dey, Arjun; Jannuzi, Buell T.

    2012-04-01

    Giant Ly{alpha} nebulae (or Ly{alpha} 'blobs') are likely sites of ongoing massive galaxy formation, but the rarity of these powerful sources has made it difficult to form a coherent picture of their properties, ionization mechanisms, and space density. Systematic narrowband Ly{alpha} nebula surveys are ongoing, but the small redshift range covered and the observational expense limit the comoving volume that can be probed by even the largest of these surveys and pose a significant problem when searching for such rare sources. We have developed a systematic search technique designed to find large Ly{alpha} nebulae at 2 {approx}< z {approx}< 3 within deep broadband imaging and have carried out a survey of the 9.4 deg{sup 2} NOAO Deep Wide-Field Survey Booetes field. With a total survey comoving volume of Almost-Equal-To 10{sup 8} h{sup -3}{sub 70} Mpc{sup 3}, this is the largest volume survey for Ly{alpha} nebulae ever undertaken. In this first paper in the series, we present the details of the survey design and a systematically selected sample of 79 candidates, which includes one previously discovered Ly{alpha} nebula.

  12. The therapeutic effect of TNFR1-selective antagonistic mutant TNF-alpha in murine hepatitis models.

    PubMed

    Shibata, Hiroko; Yoshioka, Yasuo; Ohkawa, Akiko; Abe, Yasuhiro; Nomura, Tetsuya; Mukai, Yohei; Nakagawa, Shinsaku; Taniai, Madoka; Ohta, Tsunetaka; Mayumi, Tadanori; Kamada, Haruhiko; Tsunoda, Shin-ichi; Tsutsumi, Yasuo

    2008-11-01

    Tumor necrosis factor-alpha (TNF-alpha) is critically involved in a wide variety of inflammatory pathologies, such as hepatitis, via the TNF receptor-1 (TNFR1). To develop TNFR1-targeted anti-inflammatory drugs, we have already succeeded in creating a TNFR1-selective antagonistic mutant TNF-alpha (R1antTNF) and shown that R1antTNF efficiently inhibits TNF-alpha/TNFR1-mediated biological activity in vitro. In this study, we examined the therapeutic effect of R1antTNF in acute hepatitis using two independent experimental models, induced by carbon tetrachloride (CCl(4)) or concanavalin A (ConA). In a CCl(4)-induced model, treatment with R1antTNF significantly inhibited elevation in the serum level of ALT (alanine aminotransferase), a marker for liver damage. In a ConA-induced T-cell-mediated hepatitis model, R1antTNF also inhibited the production of serum immune activated markers such as IL-2 and IL-6. These R1antTNF-mediated therapeutic effects were as good as or better than those obtained using conventional anti-TNF-alpha antibody therapy. Our results suggest that R1antTNF may be a clinically useful TNF-alpha antagonist in hepatitis.

  13. Interferon-. alpha. selectively activates the. beta. isoform of protein kinase C through phosphatidylcholine hydrolysis

    SciTech Connect

    Pfeffer, L.M.; Saltiel, A.R. ); Strulovici, B. )

    1990-09-01

    The early events that occur after interferon binds to discrete cell surface receptors remain largely unknown. Human leukocyte interferon (interferon-{alpha}) rapidly increases the binding of ({sup 3}H)phorbol dibutyrate to intact HeLa cells a measure of protein kinase C activation, and induces the selective translocation of the {beta} isoform of protein kinase C from the cytosol to the particulate fraction of HeLa cells. The subcellular distribution of the {alpha} and {epsilon} isoforms is unaffected by interferon-{alpha} treatment. Activation of protein kinase C by phorbol esters mimics the inhibitory action of interferon-{alpha} on HeLa cell proliferation and down-regulation of protein kinase C blocks the induction of antiviral activity by interferon-{alpha} in HeLa cells. Increased phosphatidylcholine hydrolysis and phosphorylcholine production is accompanied by diacylglycerol production in response to interferon. However, inositol phospholipid turnover and free intracellular calcium concentration are unaffected. These results suggest that the transient increase in diacylglycerol, resulting from phosphatidylcholine hydrolysis, may selectively activate the {beta} isoform of protein kinase C. Moreover, the activation of protein kinase C is a necessary element in interferon action on cells.

  14. Simultaneous quantification of GABAergic 3alpha,5alpha/3alpha,5beta neuroactive steroids in human and rat serum.

    PubMed

    Porcu, Patrizia; O'Buckley, Todd K; Alward, Sarah E; Marx, Christine E; Shampine, Lawrence J; Girdler, Susan S; Morrow, A Leslie

    2009-01-01

    The 3alpha,5alpha- and 3alpha,5beta-reduced derivatives of progesterone, deoxycorticosterone, dehydroepiandrosterone and testosterone enhance GABAergic neurotransmission and produce inhibitory neurobehavioral and anti-inflammatory effects. Despite substantial information on the progesterone derivative (3alpha,5alpha)-3-hydroxypregnan-20-one (3alpha,5alpha-THP, allopregnanolone), the physiological significance of the other endogenous GABAergic neuroactive steroids has remained elusive. Here, we describe the validation of a method using gas chromatography-mass spectrometry to simultaneously identify serum levels of the eight 3alpha,5alpha- and 3alpha,5beta-reduced derivatives of progesterone, deoxycorticosterone, dehydroepiandrosterone and testosterone. The method shows specificity, sensitivity and enhanced throughput compared to other methods already available for neuroactive steroid quantification. Administration of pregnenolone to rats and progesterone to women produced selective effects on the 3alpha,5alpha- and 3alpha,5beta-reduced neuroactive steroids, indicating differential regulation of their biosynthetic pathways. Pregnenolone administration increased serum levels of 3alpha,5alpha-THP (+1488%, p<0.001), (3alpha,5alpha)-3,21-dihydroxypregnan-20-one (3alpha,5alpha-THDOC, +205%, p<0.01), (3alpha,5alpha)-3-hydroxyandrostan-17-one (3alpha,5alpha-A, +216%, p<0.001), (3alpha,5alpha,17beta)-androstane-3,17-diol (3alpha,5alpha-A-diol, +190%, p<0.01). (3alpha,5beta)-3-hydroxypregnan-20-one (3alpha,5beta-THP) and (3alpha,5beta)-3-hydroxyandrostan-17-one (3alpha,5beta-A) were not altered, while (3alpha,5beta)-3,21-dihydroxypregnan-20-one (3alpha,5beta-THDOC) and (3alpha,5beta,17beta)-androstane-3,17-diol (3alpha,5beta-A-diol) were increased from undetectable levels to 271+/-100 and 2.4+/-0.9 pg+/-SEM, respectively (5/8 rats). Progesterone administration increased serum levels of 3alpha,5alpha-THP (+1806%, p<0.0001), 3alpha,5beta-THP (+575%, p<0.001), 3alpha,5alpha

  15. Alpha-blockade therapy for benign prostatic hyperplasia: from a nonselective to a more selective alpha1A-adrenergic antagonist.

    PubMed

    Beduschi, M C; Beduschi, R; Oesterling, J E

    1998-06-01

    Benign prostatic hyperplasia (BPH) is very common in older men, causing symptoms that can markedly impair quality of life. Surgical treatment, typically transurethral resection of the prostate (TURP), is highly effective but can be costly and is associated with the risk for significant morbidity. Medical treatments for BPH are targeted toward reducing bladder outlet obstruction either by androgen blockade to reduce prostatic volume or alpha-adrenergic blockade to relax the smooth muscle tone of the prostate. In recent years, understanding of the sympathetic innervation of the prostate has improved. This has been paralleled by the development of alpha-adrenergic blocking agents, from nonselective alpha-antagonists, to selective alpha1-antagonists, to the more selective alpha1A-antagonists. It is anticipated that more specific agents will optimize the therapeutic effectiveness of alpha-adrenergic blockade in the prostate while reducing the side effects associated with alpha-adrenergic blockade in other areas of the body, such as the vascular system. This article reviews the evolution of alpha-blockade therapy in management of BPH, focusing on tamsulosin, an agent targeted toward the alpha1A-adrenoceptor that predominates in the prostate. Clinical trials in Europe and the United States have provided evidence that tamsulosin is effective at doses of 0.4 and 0.8 mg/day. At both doses, tamsulosin is associated with significant improvements in the American Urological Association symptom score and the mean and peak urinary flow rates as compared with placebo. This once-daily alpha1A-adrenergic antagonist is well-tolerated, with a minimal potential for the side effects associated with alphas-blocker therapy.

  16. Antidiabetic Property of Symplocos cochinchinensis Is Mediated by Inhibition of Alpha Glucosidase and Enhanced Insulin Sensitivity

    PubMed Central

    Antu, Kalathookunnel Antony; Riya, Mariam Philip; Mishra, Arvind; Anilkumar, Karunakaran S.; Chandrakanth, Chandrasekharan K.; Tamrakar, Akhilesh K.; Srivastava, Arvind K.; Raghu, K. Gopalan

    2014-01-01

    The study is designed to find out the biochemical basis of antidiabetic property of Symplocos cochinchinensis (SC), the main ingredient of ‘Nisakathakadi’ an Ayurvedic decoction for diabetes. Since diabetes is a multifactorial disease, ethanolic extract of the bark (SCE) and its fractions (hexane, dichloromethane, ethyl acetate and 90% ethanol) were evaluated by in vitro methods against multiple targets relevant to diabetes such as the alpha glucosidase inhibition, glucose uptake, adipogenic potential, oxidative stress, pancreatic beta cell proliferation, inhibition of protein glycation, protein tyrosine phosphatase-1B (PTP-1B) and dipeptidyl peptidase-IV (DPP-IV). Among the extracts, SCE exhibited comparatively better activity like alpha glucosidase inhibition (IC50 value-82.07±2.10 µg/mL), insulin dependent glucose uptake (3 fold increase) in L6 myotubes, pancreatic beta cell regeneration in RIN-m5F (3.5 fold increase) and reduced triglyceride accumulation (22% decrease) in 3T3L1 cells, protection from hyperglycemia induced generation of reactive oxygen species in HepG2 cells (59.57% decrease) with moderate antiglycation and PTP-1B inhibition. Chemical characterization by HPLC revealed the superiority of SCE over other extracts due to presence and quantity of bioactives (beta-sitosterol, phloretin 2′glucoside, oleanolic acid) in addition to minerals like magnesium, calcium, potassium, sodium, zinc and manganese. So SCE has been subjected to oral sucrose tolerance test to evaluate its antihyperglycemic property in mild diabetic and diabetic animal models. SCE showed significant antihyperglycemic activity in in vivo diabetic models. We conclude that SC mediates the antidiabetic activity mainly via alpha glucosidase inhibition, improved insulin sensitivity, with moderate antiglycation and antioxidant activity. PMID:25184241

  17. Selectivity of bevantolol hydrochloride towards alpha- and beta-adrenoceptor subtypes in rat cerebral cortex.

    PubMed

    Takita, M; Kigoshi, S; Muramatsu, I

    1992-02-01

    Selectivity of bevantolol hydrochloride (NC-1400) towards alpha- and beta-adrenoceptor subtypes of rat cerebral cortex was examined in binding experiments and compared with propranolol. Bevantolol biphasically displaced the 3H-dihydroalprenolol binding. The affinity of bevantolol to beta 1-adrenoceptor was equal to that of propranolol. Bevantolol displaced 3H-prazosin binding monophasically but not 3H-p-aminoclonidine binding. These results suggest that bevantolol is a beta 1-adrenoceptor antagonist with a relatively high affinity to alpha 1-adrenoceptor subtypes.

  18. Discovery of an Oxybenzylglycine Based Peroxisome Proliferator Activated Receptor Alpha Selective

    SciTech Connect

    Li, J.; Kennedy, L; Shi, Y; Tao, S; Ye, X; Chen, S; Wang, Y; Hernandez, A; Wang, W; et al.

    2010-01-01

    An 1,3-oxybenzylglycine based compound 2 (BMS-687453) was discovered to be a potent and selective peroxisome proliferator activated receptor (PPAR) {alpha} agonist, with an EC{sub 50} of 10 nM for human PPAR{alpha} and {approx}410-fold selectivity vs human PPAR{gamma} in PPAR-GAL4 transactivation assays. Similar potencies and selectivity were also observed in the full length receptor co-transfection assays. Compound 2 has negligible cross-reactivity against a panel of human nuclear hormone receptors including PPAR{delta}. Compound 2 demonstrated an excellent pharmacological and safety profile in preclinical studies and thus was chosen as a development candidate for the treatment of atherosclerosis and dyslipidemia. The X-ray cocrystal structures of the early lead compound 12 and compound 2 in complex with PPAR{alpha} ligand binding domain (LBD) were determined. The role of the crystal structure of compound 12 with PPAR{alpha} in the development of the SAR that ultimately resulted in the discovery of compound 2 is discussed.

  19. Assessing the SEU resistance of CMOS latches using alpha-particle sensitive test circuits

    NASA Technical Reports Server (NTRS)

    Buehler, M.; Blaes, B.; Nixon, R.

    1990-01-01

    The importance of Cosmic Rays on the performance of integrated circuits (IC's) in a space environment is evident in the upset rate of the Tracking and Data Relay Satellite (TDRS) launched in Apr. 1983. This satellite experiences a single-event-upset (SEU) per day which must be corrected from the ground. Such experience caused a redesign of the Galileo spacecraft with SEU resistant IC's. The solution to the SEU problem continues to be important as the complexity of spacecraft grows, the feature size of IC's decreases, and as space systems are designed with circuits fabricated at non-radiation hardened foundries. This paper describes an approach for verifying the susceptibility of CMOS latches to heavy-ion induced state changes. The approach utilizes alpha particles to induce the upsets in test circuits. These test circuits are standard cells that have offset voltages which sensitize the circuits to upsets. These results are then used to calculate the upsetability at operating voltages. In this study results are presented for the alpha particle upset of a six-transistor static random access memory (SRAM) cell. Then a methodology is described for the analysis of a standard-cell inverter latch.

  20. Fast infrared spectroscopy of protein dynamics: advancing sensitivity and selectivity.

    PubMed

    Koziol, Klemens L; Johnson, Philip J M; Stucki-Buchli, Brigitte; Waldauer, Steven A; Hamm, Peter

    2015-10-01

    2D-IR spectroscopy has matured to a powerful technique to study the structure and dynamics of peptides, but its extension to larger proteins is still in its infancy, the major limitations being sensitivity and selectivity. Site-selective information requires measuring single vibrational probes at sub-millimolar concentrations where most proteins are still stable, which is a severe challenge for conventional (FT)IR spectroscopy. Besides its ultrafast time-resolution, a so far largely underappreciated potential of 2D-IR spectroscopy lies in its sensitivity gain. The present paper sets the goals and outlines strategies how to use that sensitivity gain together with properly designed vibrational labels to make IR spectroscopy a versatile tool to study a wide class of proteins.

  1. Identification of a novel cyclosporin-sensitive element in the human tumor necrosis factor alpha gene promoter

    PubMed Central

    1993-01-01

    Tumor necrosis factor alpha (TNF-alpha), a cytokine with pleiotropic biological effects, is produced by a variety of cell types in response to induction by diverse stimuli. In this paper, TNF-alpha mRNA is shown to be highly induced in a murine T cell clone by stimulation with T cell receptor (TCR) ligands or by calcium ionophores alone. Induction is rapid, does not require de novo protein synthesis, and is completely blocked by the immunosuppressant cyclosporin A (CsA). We have identified a human TNF-alpha promoter element, kappa 3, which plays a key role in the calcium-mediated inducibility and CsA sensitivity of the gene. In electrophoretic mobility shift assays, an oligonucleotide containing kappa 3 forms two DNA protein complexes with proteins that are present in extracts from unstimulated T cells. These complexes appear in nuclear extracts only after T cell stimulation. Induction of the inducible nuclear complexes is rapid, independent of protein synthesis, and blocked by CsA, and thus, exactly parallels the induction of TNF-alpha mRNA by TCR ligands or by calcium ionophore. Our studies indicate that the kappa 3 binding factor resembles the preexisting component of nuclear factor of activated T cells. Thus, the TNF-alpha gene is an immediate early gene in activated T cells and provides a new model system in which to study CsA-sensitive gene induction in activated T cells. PMID:8376940

  2. Imaging metals in biology: balancing sensitivity, selectivity and spatial resolution.

    PubMed

    Hare, Dominic J; New, Elizabeth J; de Jonge, Martin D; McColl, Gawain

    2015-10-01

    Metal biochemistry drives a diverse range of cellular processes associated with development, health and disease. Determining metal distribution, concentration and flux defines our understanding of these fundamental processes. A comprehensive analysis of biological systems requires a balance of analytical techniques that inform on metal quantity (sensitivity), chemical state (selectivity) and location (spatial resolution) with a high degree of certainty. A number of approaches are available for imaging metals from whole tissues down to subcellular organelles, as well as mapping metal turnover, protein association and redox state within these structures. Technological advances in micro- and nano-scale imaging are striving to achieve multi-dimensional and in vivo measures of metals while maintaining the native biochemical environment and physiological state. This Tutorial Review discusses state-of-the-art imaging technology as a guide to obtaining novel insight into the biology of metals, with sensitivity, selectivity and spatial resolution in focus. PMID:26505053

  3. Human fat cell alpha-2 adrenoceptors. I. Functional exploration and pharmacological definition with selected alpha-2 agonists and antagonists

    SciTech Connect

    Galitzky, J.; Mauriege, P.; Berlan, M.; Lafontan, M.

    1989-05-01

    This study was undertaken to investigate more fully the pharmacological characteristics of the human fat cell alpha-2 adrenoceptor. Biological assays were performed on intact isolated fat cells while radioligand binding studies were carried out with (/sup 3/H)yohimbine in membranes. These pharmacological studies brought: (1) a critical definition of the limits of the experimental conditions required for the exploration of alpha-2 adrenergic responsiveness on human fat cells and membranes; (2) an improvement in the pharmacological definition of the human fat cell postsynaptic alpha-2 adrenoceptor. Among alpha-2 agonists, UK-14,304 was the most potent and the relative order of potency was: UK-14,304 greater than p-aminoclonidine greater than clonidine = B-HT 920 greater than rilmenidine. For alpha-2 antagonists, the potency order was: yohimbine greater than idazoxan greater than SK F-86,466 much greater than benextramine; (3) a description of the impact of benextramine (irreversible alpha-1/alpha-2 antagonist) on human fat cell alpha-2 adrenergic receptors and on human fat cell function; the drug inactivates the alpha-2 adrenergic receptors with a minor impact on beta adrenergic receptors and without noticeable alterations of fat cell function as assessed by preservation of beta adrenergic and Al-adenosine receptor-mediated lipolytic responses; and (4) a definition of the relationship existing between alpha-2 adrenergic receptor occupancy, inhibition of adenylate cyclase activity and antilipolysis with full and partial agonists. The existence of a receptor reserve must be taken into account when evaluating alpha-2 adrenergic receptor distribution and regulation of human fat cells.

  4. The role of Pre-H2 domains of alpha- and delta-epithelial Na+ channels in ion permeation, conductance, and amiloride sensitivity.

    PubMed

    Ji, Hong-Long; Bishop, LaToya R; Anderson, Susan J; Fuller, Catherine M; Benos, Dale J

    2004-02-27

    Epithelial Na(+) channels (ENaC) regulate salt and water re-absorption across the apical membrane of absorptive epithelia such as the kidney, colon, and lung. Structure-function studies have suggested that the second transmembrane domain (M2) and the adjacent pre- and post-M2 regions are involved in channel pore formation, cation selectivity, and amiloride sensitivity. Because Na(+) selectivity, unitary Na(+) conductance (gamma(Na)), and amiloride sensitivity of delta-ENaC are strikingly different from those of alpha-ENaC, the hypothesis that the pre-H2 domain may contribute to these characterizations has been examined by swapping the pre-H2, H2, and both (pre-H2+H2) domains of delta- and alpha-ENaCs. Whole-cell and single channel results showed that the permeation ratio of Li(+) and Na(+) (P(Li)/P(Na)) for the swap alpha chimeras co-expressed with betagamma-ENaC in Xenopus oocytes decreased significantly. In contrast, the ratio of P(Li)/P(Na) for the swap delta constructs was not significantly altered. Single channel studies confirmed that swapping of the H2 and the pre-H2+H2 domains increased the gamma(Na) of alpha-ENaC but decreased the gamma(Na) of delta-ENaC. A significant increment in the apparent inhibitory dissociation constant for amiloride (K(i)(amil)) was observed in the alpha chimeras by swapping the pre-H2, H2, and pre-H2+H2 domains. In contrast, a striking decline of K(i)(amil) was obtained in the chimeric delta constructs with substitution of the H2 and pre-H2+H2 domains. Our results demonstrate that the pre-H2 domain, combined with the H2 domain, contributes to the P(Li)/P(Na) ratio, single channel Na(+) conductance, and amiloride sensitivity of alpha- and delta-ENaCs. PMID:14660613

  5. Selective extinction drives taxonomic and functional alpha and beta diversities in island bird assemblages.

    PubMed

    Si, Xingfeng; Baselga, Andrés; Leprieur, Fabien; Song, Xiao; Ding, Ping

    2016-03-01

    Taxonomic diversity considers all species being equally different from each other and thus disregards species' different ecological functions. Exploring taxonomic and functional aspects of biodiversity simultaneously can better understand the processes of community assembly. We analysed taxonomic and functional alpha and beta diversities of breeding bird assemblages on land-bridge islands in the Thousand Island Lake, China. Given the high dispersal ability of most birds at this spatial scale (several kilometres), we predicted (i) selective extinction driving alpha and beta diversities after the creation of land-bridge islands of varying area and (ii) low taxonomic and functional beta diversities that were not correlated to spatial distance. Breeding birds were surveyed on 37 islands annually from 2007 to 2014. We decomposed beta diversity of breeding birds into spatial turnover and nestedness-resultant components, and related taxonomic and functional diversities to island area and isolation using power regression models (for alpha diversity) and multiple regression models on distance matrices (for beta diversity). We then ran simulations to assess the strength of the correlations between taxonomic and functional diversities. Results revealed that both taxonomic and functional alpha diversities increased with island area. The taxonomic nestedness-resultant and turnover components increased and decreased with difference in area, respectively, but functional counterparts did not. Isolation played a minor role in explaining alpha- and beta-diversity patterns. By partitioning beta diversity, we found low levels of overall taxonomic and functional beta diversities. The functional nestedness-resultant component dominated overall functional beta diversity, whereas taxonomic turnover was the dominant component for taxonomic beta diversity. The simulation showed that functional alpha and beta diversities were significantly correlated with taxonomic diversities, and the

  6. Motion words selectively modulate direction discrimination sensitivity for threshold motion

    PubMed Central

    Pavan, Andrea; Skujevskis, Māris; Baggio, Giosuè

    2013-01-01

    Can speech selectively modulate the sensitivity of a sensory system so that, in the presence of a suitable linguistic context, the discrimination of certain perceptual features becomes more or less likely? In this study, participants heard upward or downward motion words followed by a single visual field of random dots moving upwards or downwards. The time interval between the onsets of the auditory and the visual stimuli was varied parametrically. Motion direction could be either discriminable (suprathreshold motion) or non-discriminable (threshold motion). Participants had to judge whether the dots were moving upward or downward. Results show a double dissociation between discrimination sensitivity (d′) and reaction times depending on whether vertical motion was above or at threshold. With suprathreshold motion, responses were faster for congruent directions of words and dots, but sensitivity was equal across conditions. With threshold motion, sensitivity was higher for congruent directions of words and dots, but responses were equally fast across conditions. The observed differences in sensitivity and response times were largest when the dots appeared 450 ms after word onset, that is, consistently with electrophysiology, at the time the up/down semantics of the word had become available. These data suggest that word meanings can alter the balance between signal and noise within the visual system and affect the perception of low-level sensory features. PMID:23596407

  7. Selective electric and magnetic sensitivity of aperture probes.

    PubMed

    Singh, Dilip K; Ahn, Jae Sung; Koo, Sukmo; Kang, Taehee; Kim, Joonyeon; Lee, Sukho; Park, Namkyoo; Kim, Dai-Sik

    2015-08-10

    We report the effect of geometrical factors governing the polarization profiles of near-field scanning optical microscope (NSOM) probes. The most important physical parameter controlling the selective electric or magnetic field sensitivity is found to be the width of the metal rim surrounding aperture. Probes with metal rim width w < λ/2 selectively senses the optical electric field, while those with w > λ/2 selectively senses the optical magnetic field. Intensity variation of optical Hertz standing wave formed upon reflection at oblique incidence shows a phase difference of π/2 between electric and magnetic probes: an analogue of the classical Wiener's experiment. Our work paves way towards electromagnetic engineering of nanostructures. PMID:26367935

  8. Estimating the variation, autocorrelation, and environmental sensitivity of phenotypic selection.

    PubMed

    Chevin, Luis-Miguel; Visser, Marcel E; Tufto, Jarle

    2015-09-01

    Despite considerable interest in temporal and spatial variation of phenotypic selection, very few methods allow quantifying this variation while correctly accounting for the error variance of each individual estimate. Furthermore, the available methods do not estimate the autocorrelation of phenotypic selection, which is a major determinant of eco-evolutionary dynamics in changing environments. We introduce a new method for measuring variable phenotypic selection using random regression. We rely on model selection to assess the support for stabilizing selection, and for a moving optimum that may include a trend plus (possibly autocorrelated) fluctuations. The environmental sensitivity of selection also can be estimated by including an environmental covariate. After testing our method on extensive simulations, we apply it to breeding time in a great tit population in the Netherlands. Our analysis finds support for an optimum that is well predicted by spring temperature, and occurs about 33 days before a peak in food biomass, consistent with what is known from the biology of this species. We also detect autocorrelated fluctuations in the optimum, beyond those caused by temperature and the food peak. Because our approach directly estimates parameters that appear in theoretical models, it should be particularly useful for predicting eco-evolutionary responses to environmental change. PMID:26227394

  9. Long-term reduction of intimal hyperplasia by the selective alpha-1 adrenergic antagonist doxazosin.

    PubMed

    Vashisht, R; Sian, M; Franks, P J; O'Malley, M K

    1992-12-01

    Studies have shown that alpha 1-adrenergic blockade reduces intimal hyperplasia in the rabbit aorta. In this study a selective alpha 1-adrenergic antagonist, doxazosin, has been used to examine whether this effect is persistent and dose dependent. Forty-eight New Zealand White rabbits underwent endothelial denudation of the abdominal aorta using a Fogarty balloon catheter. Test rabbits were given low-dose (2 mg) or high-dose (8 mg) doxazosin daily and all animals killed at either 1 or 12 weeks after the procedure. The aortas were harvested after fixation in situ with 4 per cent glutaraldehyde and neointimal hyperplasia was measured, using an x-y digitizer, as the percentage reduction in luminal cross-sectional area. At 1 week after surgery, rabbits receiving the low dose had a median area reduction of 7.7 per cent and those receiving the high dose a reduction of 8.2 per cent; both had significantly less intimal hyperplasia than control rabbits, which had a median area reduction of 14.8 per cent (P < 0.01). However, at 12 weeks, when compared with the 32.6 per cent reduction in the control group, only those rabbits receiving high-dose doxazosin had significantly less intimal hyperplasia, with a reduction of 5.5 per cent (P < 0.001). It is concluded that selective alpha 1-adrenergic blockade significantly reduces neointimal hyperplasia, that this effect is dose dependent, and that it persists for at least 3 months.

  10. A highly sensitive biosensor for tumor maker alpha fetoprotein based on poly(ethylene glycol) doped conducting polymer PEDOT.

    PubMed

    Cui, Min; Song, Zhiling; Wu, Yumin; Guo, Bing; Fan, Xiaojian; Luo, Xiliang

    2016-05-15

    Biocompatible polymers, such as poly(ethylene glycol) (PEG), are of great significance in the development of bio-interfaces and biosensors, as they possess excellent biocompatibility and are easy for modification. A novel highly biocompatible polymer composite was synthesized herein through electrochemical polymerization of the conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) and a PEG derivative, 4-arm PEG terminated with thiol groups. The electrodeposited conducting polymer composite of PEG doped PEDOT (PEDOT/PEG) exhibited flake-like nanostructure, large surface area and outstanding stability. In order to further immobilize antibodies, gold nanoparticles (AuNPs) were introduced to the PEDOT/PEG composite surface through their unique interaction with the thiol groups. AuNPs modified PEDOT/PEG provided a desirable support for the immobilization of various biomolecules such as antibodies for alpha fetoprotein (AFP), a vital tumor biomarker for liver cancer. The fabricated AFP biosensor demonstrated favorable selectivity, high sensitivity, and ultralow detection limit. Furthermore, owing to the presence of PEG polymers that are highly hydrophilic, such AuNPs/PEDOT/PEG based AFP biosensor also exhibited good anti-fouling ability, and it was capable of assaying target AFP in 10% (V/V) human serum samples, indicating highly feasible potential for clinical diagnosis.

  11. Overexpression of the alpha-2,6-sialyltransferase in MDCK cells increases influenza virus sensitivity to neuraminidase inhibitors.

    PubMed

    Matrosovich, Mikhail; Matrosovich, Tatyana; Carr, Jackie; Roberts, Noel A; Klenk, Hans-Dieter

    2003-08-01

    No reliable cell culture assay is currently available for monitoring human influenza virus sensitivity to neuraminidase inhibitors (NAI). This can be explained by the observation that because of a low concentration of sialyl-alpha2,6-galactose (Sia[alpha2,6]Gal)-containing virus receptors in conventional cell lines, replication of human virus isolates shows little dependency on viral neuraminidase. To test whether overexpression of Sia(alpha2,6)Gal moieties in cultured cells could make them suitable for testing human influenza virus sensitivity to NAI, we stably transfected MDCK cells with cDNA of human 2,6-sialyltransferase (SIAT1). Transfected cells expressed twofold-higher amounts of 6-linked sialic acids and twofold-lower amounts of 3-linked sialic acids than parent MDCK cells as judged by staining with Sambucus nigra agglutinin and Maackia amurensis agglutinin, respectively. After transfection, binding of a clinical human influenza virus isolate was increased, whereas binding of its egg-adapted variant which preferentially bound 3-linked receptors was decreased. The sensitivity of human influenza A and B viruses to the neuraminidase inhibitor oseltamivir carboxylate was substantially improved in the SIAT1-transfected cell line and was consistent with their sensitivity in neuraminidase enzyme assay and with the hemagglutinin (HA) receptor-binding phenotype. MDCK cells stably transfected with SIAT1 may therefore be a suitable system for testing influenza virus sensitivity to NAI. PMID:12857911

  12. Selective dimerisation of alpha-olefins using tungsten-based initiators.

    PubMed

    Hanton, Martin J; Daubney, Louisa; Lebl, Tomas; Polas, Stacey; Smith, David M; Willemse, Alex

    2010-08-14

    The selective dimerisation of the alpha-olefins 1-pentene through to 1-nonene is reported using an in situ-generated catalyst derived from tungsten hexachloride, aniline, triethylamine and alkylaluminium halide. The influence of reagent identity and reaction stoichiometry, along with activator, solvent and alpha-olefin substrate choice are probed. The catalyst is found to be highly selective towards dimerisation, minimising the formation of undesired heavier oligomers. Notably, the selectivity within the dimer fraction is found to favour the formation of products with methyl branches. The selectivity towards individual olefin isomers has been determined and the system is found to also produce trace levels of dienes and alkanes. A kinetic study of the system reveals a second order dependence on substrate. Comparison of the products observed, with those expected for metallacyclic and Cossee-type mechanisms, suggests that the latter is in operation, something confirmed by the results of a C(2)H(4)/C(2)D(4) co-dimerisation experiment which showed full isotopic scrambling in the products. Thus a mechanistic proposal is made to account for the observed behaviour of the system, including the diene and alkane formation.

  13. Phage selection restores antibiotic sensitivity in MDR Pseudomonas aeruginosa.

    PubMed

    Chan, Benjamin K; Sistrom, Mark; Wertz, John E; Kortright, Kaitlyn E; Narayan, Deepak; Turner, Paul E

    2016-01-01

    Increasing prevalence and severity of multi-drug-resistant (MDR) bacterial infections has necessitated novel antibacterial strategies. Ideally, new approaches would target bacterial pathogens while exerting selection for reduced pathogenesis when these bacteria inevitably evolve resistance to therapeutic intervention. As an example of such a management strategy, we isolated a lytic bacteriophage, OMKO1, (family Myoviridae) of Pseudomonas aeruginosa that utilizes the outer membrane porin M (OprM) of the multidrug efflux systems MexAB and MexXY as a receptor-binding site. Results show that phage selection produces an evolutionary trade-off in MDR P. aeruginosa, whereby the evolution of bacterial resistance to phage attack changes the efflux pump mechanism, causing increased sensitivity to drugs from several antibiotic classes. Although modern phage therapy is still in its infancy, we conclude that phages, such as OMKO1, represent a new approach to phage therapy where bacteriophages exert selection for MDR bacteria to become increasingly sensitive to traditional antibiotics. This approach, using phages as targeted antibacterials, could extend the lifetime of our current antibiotics and potentially reduce the incidence of antibiotic resistant infections. PMID:27225966

  14. Phage selection restores antibiotic sensitivity in MDR Pseudomonas aeruginosa

    PubMed Central

    Chan, Benjamin K.; Sistrom, Mark; Wertz, John E.; Kortright, Kaitlyn E.; Narayan, Deepak; Turner, Paul E.

    2016-01-01

    Increasing prevalence and severity of multi-drug-resistant (MDR) bacterial infections has necessitated novel antibacterial strategies. Ideally, new approaches would target bacterial pathogens while exerting selection for reduced pathogenesis when these bacteria inevitably evolve resistance to therapeutic intervention. As an example of such a management strategy, we isolated a lytic bacteriophage, OMKO1, (family Myoviridae) of Pseudomonas aeruginosa that utilizes the outer membrane porin M (OprM) of the multidrug efflux systems MexAB and MexXY as a receptor-binding site. Results show that phage selection produces an evolutionary trade-off in MDR P. aeruginosa, whereby the evolution of bacterial resistance to phage attack changes the efflux pump mechanism, causing increased sensitivity to drugs from several antibiotic classes. Although modern phage therapy is still in its infancy, we conclude that phages, such as OMKO1, represent a new approach to phage therapy where bacteriophages exert selection for MDR bacteria to become increasingly sensitive to traditional antibiotics. This approach, using phages as targeted antibacterials, could extend the lifetime of our current antibiotics and potentially reduce the incidence of antibiotic resistant infections. PMID:27225966

  15. Selection of a 2-azabicyclo[2.2.2]octane-based alpha4beta1 integrin antagonist as an inhaled anti-asthmatic agent.

    PubMed

    Lawson, Edward C; Santulli, Rosemary J; Dyatkin, Alexey B; Ballentine, Scott A; Abraham, William M; Rudman, Sandra; Page, Clive P; de Garavilla, Lawrence; Damiano, Bruce P; Kinney, William A; Maryanoff, Bruce E

    2006-06-15

    The alpha4beta1 integrin, expressed on eosinophils and neutrophils, induces inflammation in the lung by facilitating cellular infiltration and activation. From a number of potent alpha4beta1 antagonists that we evaluated for safety and efficacy, 1 was selected as a lead candidate for anti-asthma therapy by the inhalation route. We devised an optimized stereoselective synthesis to facilitate the preparation of a sufficiently large quantity of 1 for assessment in vivo. Administration of 1 to allergen-sensitive sheep by inhalation blocked the late-phase response of asthma and abolished airway hyper-responsiveness at 24h following the antigen challenge. Additionally, the recruitment of inflammatory cells into the lungs was inhibited. Administration of 1 to ovalbumin-sensitized guinea pigs intraperitoneally blocked airway resistance and inhibited the recruitment of inflammatory cells.

  16. Bench-level characterization of a CMOS standard-cell D-latch using alpha-particle sensitive test circuits

    NASA Technical Reports Server (NTRS)

    Blaes, B. R.; Soli, G. A.; Buehler, M. G.

    1991-01-01

    A methodology is described for predicting the SEU susceptibility of a standard-cell D-latch using an alpha-particle sensitive SRAM, SPICE critical charge simulation results, and alpha-particle interaction physics. Measurements were made on a 1.6-micron n-well CMOS 4-kb test SRAM irradiated with an Am-241 alpha-particle source. A collection depth of 6.09 micron was determined using these results and TRIM computer code. Using this collection depth and SPICE derived critical charge results on the latch design, an LET threshold of 34 MeV sq cm/mg was predicted. Heavy ion tests were then performed on the latch and an LET threshold of 41 MeV sq cm/mg was determined.

  17. Choline kinase-alpha by regulating cell aggressiveness and drug sensitivity is a potential druggable target for ovarian cancer

    PubMed Central

    Granata, A; Nicoletti, R; Tinaglia, V; De Cecco, L; Pisanu, M E; Ricci, A; Podo, F; Canevari, S; Iorio, E; Bagnoli, M; Mezzanzanica, D

    2014-01-01

    Background: Aberrant choline metabolism has been proposed as a novel cancer hallmark. We recently showed that epithelial ovarian cancer (EOC) possesses an altered MRS-choline profile, characterised by increased phosphocholine (PCho) content to which mainly contribute over-expression and activation of choline kinase-alpha (ChoK-alpha). Methods: To assess its biological relevance, ChoK-alpha expression was downmodulated by transient RNA interference in EOC in vitro models. Gene expression profiling by microarray analysis and functional analysis was performed to identify the pathway/functions perturbed in ChoK-alpha-silenced cells, then validated by in vitro experiments. Results: In silenced cells, compared with control, we observed: (I) a significant reduction of both CHKA transcript and ChoK-alpha protein expression; (II) a dramatic, proportional drop in PCho content ranging from 60 to 71%, as revealed by 1H-magnetic spectroscopy analysis; (III) a 35–36% of cell growth inhibition, with no evidences of apoptosis or modification of the main cellular survival signalling pathways; (IV) 476 differentially expressed genes, including genes related to lipid metabolism. Ingenuity pathway analysis identified cellular functions related to cell death and cellular proliferation and movement as the most perturbed. Accordingly, CHKA-silenced cells displayed a significant delay in wound repair, a reduced migration and invasion capability were also observed. Furthermore, although CHKA silencing did not directly induce cell death, a significant increase of sensitivity to platinum, paclitaxel and doxorubicin was observed even in a drug-resistant context. Conclusion: We showed for the first time in EOC that CHKA downregulation significantly decreased the aggressive EOC cell behaviour also affecting cells' sensitivity to drug treatment. These observations open the way to further analysis for ChoK-alpha validation as a new EOC therapeutic target to be used alone or in combination with

  18. Induction of sensitivity to the cytotoxic action of tumor necrosis factor alpha by adenovirus E1A is independent of transformation and transcriptional activation.

    PubMed

    Ames, R S; Holskin, B; Mitcho, M; Shalloway, D; Chen, M J

    1990-09-01

    We have previously shown that expression of the adenovirus E1A 12S or 13S products in NIH 3T3 fibroblasts induces susceptibility to the cytotoxic actions of tumor necrosis factor alpha (TNF alpha). A large number of studies have mapped the multiple biological functions of the 12S and 13S products to three highly conserved regions (CR) within the E1A sequence. Here we used plasmids coding for E1A deletion and point mutants in these regions to generate target cell lines for TNF alpha cytotoxicity assays to determine which regions and functions are necessary for the induction of TNF alpha sensitivity. Expression of CR1 was required for the induction of TNF alpha sensitivity. This finding did not reflect a requirement for transforming or transcriptional repression activity, since some mutants that were defective in both of these properties were able to induce TNF alpha sensitivity. CR2 transformation-defective point mutants, but not a CR2/3 region deletion mutant, were also able to induce sensitivity. In addition, NIH 3T3 cells expressing the retroviral transcription activators tat from human immunodeficiency virus type 1 and tax from human T-lymphotropic virus type I were not sensitive to TNF alpha. However, the possibility that E1A-mediated transcriptional activation can augment the induction of TNF alpha sensitivity is not excluded. Comparison of data from previous biological studies with the TNF alpha cytotoxicity assays presented here suggested that the mechanism by which E1A induces sensitivity to TNF alpha in NIH 3T3 cells is independent of many of the known E1A biological functions, including transformation in cooperation with ras, immortalization, induction of DNA synthesis in quiescent cells, and transcriptional repression. A novel E1A-mediated effect may be involved, although our data do not exclude the possibility that sensitization to TNF alpha is mediated through E1A binding to cellular proteins. PMID:2143540

  19. Induction of sensitivity to the cytotoxic action of tumor necrosis factor alpha by adenovirus E1A is independent of transformation and transcriptional activation.

    PubMed Central

    Ames, R S; Holskin, B; Mitcho, M; Shalloway, D; Chen, M J

    1990-01-01

    We have previously shown that expression of the adenovirus E1A 12S or 13S products in NIH 3T3 fibroblasts induces susceptibility to the cytotoxic actions of tumor necrosis factor alpha (TNF alpha). A large number of studies have mapped the multiple biological functions of the 12S and 13S products to three highly conserved regions (CR) within the E1A sequence. Here we used plasmids coding for E1A deletion and point mutants in these regions to generate target cell lines for TNF alpha cytotoxicity assays to determine which regions and functions are necessary for the induction of TNF alpha sensitivity. Expression of CR1 was required for the induction of TNF alpha sensitivity. This finding did not reflect a requirement for transforming or transcriptional repression activity, since some mutants that were defective in both of these properties were able to induce TNF alpha sensitivity. CR2 transformation-defective point mutants, but not a CR2/3 region deletion mutant, were also able to induce sensitivity. In addition, NIH 3T3 cells expressing the retroviral transcription activators tat from human immunodeficiency virus type 1 and tax from human T-lymphotropic virus type I were not sensitive to TNF alpha. However, the possibility that E1A-mediated transcriptional activation can augment the induction of TNF alpha sensitivity is not excluded. Comparison of data from previous biological studies with the TNF alpha cytotoxicity assays presented here suggested that the mechanism by which E1A induces sensitivity to TNF alpha in NIH 3T3 cells is independent of many of the known E1A biological functions, including transformation in cooperation with ras, immortalization, induction of DNA synthesis in quiescent cells, and transcriptional repression. A novel E1A-mediated effect may be involved, although our data do not exclude the possibility that sensitization to TNF alpha is mediated through E1A binding to cellular proteins. Images PMID:2143540

  20. Electrodeposition of selected alpha-emitting nuclides from ammonium acetate electrolyte

    NASA Astrophysics Data System (ADS)

    Lee, Shan C.; Choi, Jae G.; Hodge, Vernon F.

    1994-10-01

    The experimentally optimal conditions of the electrodeposition of selected alpha particle-emitting radionuclides, including Po-208, Ra-226, Th-228, U-238, Pu-239, Am-241 and Cm-(243, 244) with ammonium acetate electrolyte have been determined. This simple method could be used for the determination of the most important actinides in radiological waste and could be applicable to waste treatment. In addition, this method could be used for radium determination instead of the traditional radon emanation technique, which requires approximately 30 days.

  1. Relative sensitivity of some selected aquatic organisms to phenol

    SciTech Connect

    Tisler, T.; Zagorc-Koncan, J.

    1995-05-01

    Although the possibility of adverse effects of materials on organisms was recognized earlier, not until the 1940s or 1950s short-term acute tests with fishes were being studied. The fishes have become the most popular test organism because the effects of toxic substances in streams have been most evident on the fishes. Many other organisms bearing the important role in the environment live in the waters and began to be used in the toxicological research. Nowadays such great variety of test methods and organisms make the selection of the suitable toxicity test difficult. One or two species of test organisms only are often applied in the determination of the toxic substances or wastewaters. The toxicity test with daphnids is widely used due to its sensitivity to the great part of toxic substances. The purpose of our research was to determine the toxicity of phenol to some aquatic organisms from the group of bacteria, algae, crustacea and fishes, and to determine the most sensitive kind. Test organisms have been selected from three basic groups in the food chain (bacteria - decomposers, algae - producers and crustacea, fish - consumers). Phenol, an organic degradable substance that is a frequent contaminant in wastewaters, has been chosen for the test substance. 29 refs., 3 tabs.

  2. Increased sensitivity of estrogen receptor alpha overexpressing antral follicles to methoxychlor and its metabolites.

    PubMed

    Paulose, Tessie; Hernández-Ochoa, Isabel; Basavarajappa, Mallikarjuna S; Peretz, Jackye; Flaws, Jodi A

    2011-04-01

    Methoxychlor (MXC), an organochlorine pesticide, and its metabolites, mono-hydroxy MXC (MOH) and bis-hydroxy MXC (HPTE) are known ovarian toxicants and can cause inhibition of antral follicle growth. Since these chemicals bind to estrogen receptor alpha (ESR1), we hypothesized that ovaries overexpressing ESR1 (ESR1 OE) would be more susceptible to toxicity induced by MXC and its metabolites because the chemicals can bind to more ESR1 in the antral follicles. We cultured antral follicles from controls and ESR1 OE mouse ovaries with either the vehicle dimethylsulfoxide (DMSO), MXC, MOH, or HPTE. The data show that at 96 h, the cultured antral follicles from ESR1 OE antral follicles are more susceptible to toxicity induced by MXC, MOH, and HPTE because low doses of these chemicals cause follicle growth inhibition in ESR1 OE mice but not in control mice. On comparing gene expression levels of nuclear receptors in the cultured antral follicles of ESR1 OE and control follicles, we found differential messenger RNA (mRNA) expression of Esr1, estrogen receptor beta (Esr2), androgen receptor (Ar), progesterone receptor (Pr), and aryl hydrocarbon receptor (Ahr) between the genotypes. We also analyzed mRNA levels of Cyp3a41a, the enzyme metabolizing MOH and HPTE, in the cultured follicles and found that Cyp3a41a was significantly lower in DMSO-treated ESR1 OE follicles compared with controls. In ESR1 OE livers, we found that Cyp3a41a levels were significantly lower compared with control livers. Collectively, these data suggest that MXC and its metabolites cause differential gene expression in ESR1 OE mice compared with controls. The results also suggest that the increased sensitivity of ESR1 OE mouse ovaries to toxicity induced by MXC and its metabolites is due to low clearance of the metabolites by the liver and ovary.

  3. Plasma catecholamine modulation of alpha 2 adrenoreceptor agonist affinity and sensitivity in normotensive and hypertensive human platelets.

    PubMed Central

    Hollister, A S; Onrot, J; Lonce, S; Nadeau, J H; Robertson, D

    1986-01-01

    We measured alpha 2-adrenoreceptor density as well as affinity for and sensitivity to agonist on intact platelets of normotensive and hypertensive subjects before and after physiological increases in plasma catecholamines. In normotensives, posture-induced rises in plasma catecholamines correlated with reduced alpha 2-adrenoreceptor agonist affinity and fewer high affinity state receptors. Platelet aggregation and inhibition of adenylate cyclase by L-epinephrine also was reduced. Hypertensive subjects had similar rises in plasma catecholamines with upright posture, but showed no change in receptor affinity or sensitivity. No change in platelet alpha 2-adrenoreceptor number occurred in these studies. In vitro incubation with L-epinephrine revealed that platelets from hypertensives had slower desensitization than those from normotensives. Binding studies at different temperatures and with varying sodium concentrations found no thermodynamic or sodium-dependent differences between normotensive and hypertensive groups. These studies demonstrate that platelets from hypertensive subjects exhibit a defect in the ability of physiological concentrations of agonist to desensitize the alpha 2-adrenoreceptor. PMID:3009543

  4. Alpha-2-Macroglobulin Is Acutely Sensitive to Freezing and Lyophilization: Implications for Structural and Functional Studies.

    PubMed

    Wyatt, Amy R; Kumita, Janet R; Farrawell, Natalie E; Dobson, Christopher M; Wilson, Mark R

    2015-01-01

    Alpha-2-macroglobulin is an abundant secreted protein that is of particular interest because of its diverse ligand binding profile and multifunctional nature, which includes roles as a protease inhibitor and as a molecular chaperone. The activities of alpha-2-macroglobulin are typically dependent on whether its conformation is native or transformed (i.e. adopts a more compact conformation after interactions with proteases or small nucleophiles), and are also influenced by dissociation of the native alpha-2-macroglobulin tetramer into stable dimers. Alpha-2-macroglobulin is predominately present as the native tetramer in vivo; once purified from human blood plasma, however, alpha-2-macroglobulin can undergo a number of conformational changes during storage, including transformation, aggregation or dissociation. We demonstrate that, particularly in the presence of sodium chloride or amine containing compounds, freezing and/or lyophilization of alpha-2-macroglobulin induces conformational changes with functional consequences. These conformational changes in alpha-2-macroglobulin are not always detected by standard native polyacrylamide gel electrophoresis, but can be measured using bisANS fluorescence assays. Increased surface hydrophobicity of alpha-2-macroglobulin, as assessed by bisANS fluorescence measurements, is accompanied by (i) reduced trypsin binding activity, (ii) increased chaperone activity, and (iii) increased binding to the surfaces of SH-SY5Y neurons, in part, via lipoprotein receptors. We show that sucrose (but not glycine) effectively protects native alpha-2-macroglobulin from denaturation during freezing and/or lyophilization, thereby providing a reproducible method for the handling and long-term storage of this protein. PMID:26103636

  5. Sensitivity of alpha-particle-driven Alfvén eigenmodes to q-profile variation in ITER scenarios

    NASA Astrophysics Data System (ADS)

    Rodrigues, P.; Figueiredo, A. C. A.; Borba, D.; Coelho, R.; Fazendeiro, L.; Ferreira, J.; Loureiro, N. F.; Nabais, F.; Pinches, S. D.; Polevoi, A. R.; Sharapov, S. E.

    2016-11-01

    A perturbative hybrid ideal-MHD/drift-kinetic approach to assess the stability of alpha-particle-driven Alfvén eigenmodes in burning plasmas is used to show that certain foreseen ITER scenarios, namely the {{I}\\text{p}}=15 MA baseline scenario with very low and broad core magnetic shear, are sensitive to small changes in the background magnetic equilibrium. Slight variations (of the order of 1% ) of the safety-factor value on axis are seen to cause large changes in the growth rate, toroidal mode number, and radial location of the most unstable eigenmodes found. The observed sensitivity is shown to proceed from the very low magnetic shear values attained throughout the plasma core, raising issues about reliable predictions of alpha-particle transport in burning plasmas.

  6. Pupil Dilation and EEG Alpha Frequency Band Power Reveal Load on Executive Functions for Link-Selection Processes during Text Reading

    PubMed Central

    Scharinger, Christian; Kammerer, Yvonne; Gerjets, Peter

    2015-01-01

    Executive working memory functions play a central role in reading comprehension. In the present research we were interested in additional load imposed on executive functions by link-selection processes during computer-based reading. For obtaining process measures, we used a methodology of concurrent electroencephalographic (EEG) and eye-tracking data recording that allowed us to compare epochs of pure text reading with epochs of hyperlink-like selection processes in an online reading situation. Furthermore, this methodology allowed us to directly compare the two physiological load-measures EEG alpha frequency band power and pupil dilation. We observed increased load on executive functions during hyperlink-like selection processes on both measures in terms of decreased alpha frequency band power and increased pupil dilation. Surprisingly however, the two measures did not correlate. Two additional experiments were conducted that excluded potential perceptual, motor, or structural confounds. In sum, EEG alpha frequency band power and pupil dilation both turned out to be sensitive measures for increased load during hyperlink-like selection processes in online text reading. PMID:26076026

  7. Pupil Dilation and EEG Alpha Frequency Band Power Reveal Load on Executive Functions for Link-Selection Processes during Text Reading.

    PubMed

    Scharinger, Christian; Kammerer, Yvonne; Gerjets, Peter

    2015-01-01

    Executive working memory functions play a central role in reading comprehension. In the present research we were interested in additional load imposed on executive functions by link-selection processes during computer-based reading. For obtaining process measures, we used a methodology of concurrent electroencephalographic (EEG) and eye-tracking data recording that allowed us to compare epochs of pure text reading with epochs of hyperlink-like selection processes in an online reading situation. Furthermore, this methodology allowed us to directly compare the two physiological load-measures EEG alpha frequency band power and pupil dilation. We observed increased load on executive functions during hyperlink-like selection processes on both measures in terms of decreased alpha frequency band power and increased pupil dilation. Surprisingly however, the two measures did not correlate. Two additional experiments were conducted that excluded potential perceptual, motor, or structural confounds. In sum, EEG alpha frequency band power and pupil dilation both turned out to be sensitive measures for increased load during hyperlink-like selection processes in online text reading.

  8. Selective solid-phase extraction of alpha-tocopherol by functionalized ionic liquid-modified mesoporous SBA-15 adsorbent.

    PubMed

    Li, Min; Pham, Patrisha J; Pittman, Charles U; Li, Tingyu

    2008-10-01

    Ordered mesoporous adsorbents were prepared by physically grafting functionalized ionic liquids onto SBA-15 (a mesoporous siliceous substrate) using incipient wetness immersion method. These adsorbents were successfully applied to the selective extraction and separation of alpha-tocopherol (an isomer of vitamin E) from a model mixture of soybean oil deodorizer distillate. Various parameters affecting adsorption process such as adsorption time, the structures and loadings of ionic liquids, the adsorption isotherm, and the reusability of adsorbent were investigated using liquid-solid extraction. As high as 211 mg/g adsorbent of the adsorption capacity for alpha-tocopherol was obtained through the adsorption isotherm tests using [emim][Gly]/SBA-15 (functionalized ionic liquid 1-ethyl-3-methylimidazolium glycine which was physically coated on SBA-15) as the adsorbent, in which the functionalized ionic liquids contained the amino acid glycine as the anion. The adsorbent [emim][Gly]/SBA-15 also exhibited a very high adsorption selectivity for alpha-tocopherol. The extraction selectivity or the ratio of distribution coefficients between alpha-tocopherol and the major interference component glyceryl triundecanoate (K(d(alpha-tocopherol))/K(d(triglyceride))) was 10.5. The concentration of alpha-tocopherol was significantly increased from 15.6% in original feedstock solution that contained fatty acid methyl ester, triglyceride and alpha-tocopherol to 73.0% after stripping by diethyl ether. Five adsorbent recycle tests showed good reusability of the functionalized ionic liquid-modified mesoporous adsorbent. PMID:18845881

  9. Selective Alpha-Particle Mediated Depletion of Tumor Vasculature with Vascular Normalization

    PubMed Central

    Seshan, Surya V.; Kappel, Barry J.; Chattopadhyay, Debjit; May, Chad; McDevitt, Michael R.; Nolan, Daniel; Mittal, Vivek; Benezra, Robert; Scheinberg, David A.

    2007-01-01

    Background Abnormal regulation of angiogenesis in tumors results in the formation of vessels that are necessary for tumor growth, but compromised in structure and function. Abnormal tumor vasculature impairs oxygen and drug delivery and results in radiotherapy and chemotherapy resistance, respectively. Alpha particles are extraordinarily potent, short-ranged radiations with geometry uniquely suitable for selectively killing neovasculature. Methodology and Principal Findings Actinium-225 (225Ac)-E4G10, an alpha-emitting antibody construct reactive with the unengaged form of vascular endothelial cadherin, is capable of potent, selective killing of tumor neovascular endothelium and late endothelial progenitors in bone-marrow and blood. No specific normal-tissue uptake of E4G10 was seen by imaging or post-mortem biodistribution studies in mice. In a mouse-model of prostatic carcinoma, 225Ac-E4G10 treatment resulted in inhibition of tumor growth, lower serum prostate specific antigen level and markedly prolonged survival, which was further enhanced by subsequent administration of paclitaxel. Immunohistochemistry revealed lower vessel density and enhanced tumor cell apoptosis in 225Ac-E4G10 treated tumors. Additionally, the residual tumor vasculature appeared normalized as evident by enhanced pericyte coverage following 225Ac-E4G10 therapy. However, no toxicity was observed in vascularized normal organs following 225Ac-E4G10 therapy. Conclusions The data suggest that alpha-particle immunotherapy to neovasculature, alone or in combination with sequential chemotherapy, is an effective approach to cancer therapy. PMID:17342201

  10. Sodium selective erythrocyte glycocalyx and salt sensitivity in man.

    PubMed

    Oberleithner, Hans

    2015-06-01

    Negatively charged surfaces of erythrocytes (RBC) reflect properties of the endothelial glycocalyx. Plasma electrolytes counteract these charges and thus control the repulsive forces between RBC and endothelium. Although Na(+) is supposed to exert a rather high affinity to the RBC surface, a direct comparison between Na(+) and K(+) in counteracting the RBC surface has been never made. Therefore, we measured Na(+)/K(+) selectivity of the RBC surface in 20 healthy volunteers applying the previously published salt blood test (SBT). It turned out that the Na(+)/K(+) selectivity ratio of the RBC glycocalyx is on average 6.1 ± 0.39 (ranging from 3 to 9 in different individuals). Considering standard plasma Na(+) and K(+) concentrations, binding probability of Na(+)/K(+) at the RBC surface is about 180:1. The SBT reveals that plasma K(+) counteracts only about 7% of the negative charges in the RBC glycocalyx. As an in vivo proof of principle, a volunteer's blood was continuously tested over 6 months while applying a glycocalyx protective polyphenol-rich natural compound (hawthorn extract). It turned out that RBC Na(+) sensitivity (the inverse of Na(+) buffer capacity) decreased significantly by about 25% while Na(+)/K(+) selectivity of the RBC glycocalyx declined only slightly by about 8 %. Taken together, (i) plasma Na(+) selectively buffers the negative charges of the RBC glycocalyx, (ii) the contribution of K(+) in counteracting these negative surface charges is small, and (iii) natural polyphenols applied in vivo increase RBC surface negativity. In conclusion, low plasma Na(+) is supposed to favor frictionless RBC-slipping through blood vessels.

  11. Elongation factor-1 alpha is a selective regulator of growth factor withdrawal and ER stress-induced apoptosis.

    PubMed

    Talapatra, S; Wagner, J D O; Thompson, C B

    2002-08-01

    To identify genes that contribute to apoptotic resistance, IL-3 dependent hematopoietic cells were transfected with a cDNA expression library and subjected to growth factor withdrawal. Transfected cells were enriched for survivors over two successive rounds of IL-3 withdrawal and reconstitution, resulting in the identification of a full-length elongation factor 1 alpha (EF-1alpha) cDNA. Ectopic EF-1alpha expression conferred protection from growth factor withdrawal and agents that induce endoplasmic reticulum stress, but not from nuclear damage or death receptor signaling. Overexpression of EF-1alpha did not lead to growth factor independent cell proliferation or global alterations in protein levels or rates of synthesis. These findings suggest that overexpression of EF-1alpha results in selective resistance to apoptosis induced by growth factor withdrawal and ER stress. PMID:12107828

  12. Sodium-selective salt sensitivity: its occurrence in blacks.

    PubMed

    Schmidlin, Olga; Forman, Alex; Sebastian, Anthony; Morris, R Curtis

    2007-12-01

    We tested the hypothesis that the Na(+) component of dietary NaCl can have a pressor effect apart from its capacity to complement the extracellular osmotic activity of Cl(-) and, thus, expand plasma volume. We studied 35 mostly normotensive blacks who ingested a low-NaCl diet, 30 mmol/d, for 3 weeks, in the first and third of which Na(+) was loaded orally with either NaHCO(3) or NaCl, in random order (250 mmol/d). In subjects adjudged to be salt sensitive (n=18; Delta mean arterial pressure: >or=5 mm Hg with NaCl load), but not in salt-resistant subjects (n=17), loading with NaHCO(3) was also pressor. The pressor effect of NaHCO(3) was half that of NaCl: mean arterial pressure (millimeters of mercury) increased significantly from 90 on low NaCl to 95 with NaHCO(3) and to 101 with NaCl. The pressor effect of NaCl strongly predicted that of NaHCO(3.) As judged by hematocrit decrease, plasma volume expansion with NaCl was the same in salt-resistant and salt-sensitive subjects and twice that with NaHCO(3), irrespective of the pressor effect. In salt-sensitive subjects, mean arterial pressure varied directly with plasma Na(+) concentration attained with all Na(+) loading. In salt-sensitive but not salt-resistant subjects, NaHCO(3) and NaCl induced decreases in renal blood flow and increases in renal vascular resistance; changes in renal blood flow were not different with the 2 salts. Responses of renal blood flow and renal vascular resistance to NaHCO(3) were strongly predicted by those to NaCl. In establishing the fact of "sodium-selective" salt sensitivity, the current observations demonstrate that the Na(+) component of NaCl can have pressor and renal vasoconstrictive properties apart from its capacity to complement Cl(-) in plasma volume expansion.

  13. Stratification of alpha ganglion cells and ON/OFF directionally selective ganglion cells in the rabbit retina

    PubMed Central

    ZHANG, JIAN; LI, WEI; HOSHI, HIDEO; MILLS, STEPHEN L.; MASSEY, STEPHEN C.

    2007-01-01

    The correlation between cholinergic sensitivity and the level of stratification for ganglion cells was examined in the rabbit retina. As examples, we have used ON or OFF α ganglion cells and ON/OFF directionally selective (DS) ganglion cells. Nicotine, a cholinergic agonist, depolarized ON/OFF DS ganglion cells and greatly enhanced their firing rates but it had modest excitatory effects on ON or OFF α ganglion cells. As previously reported, we conclude that DS ganglion cells are the most sensitive to cholinergic drugs. Confocal imaging showed that ON/OFF DS ganglion cells ramify precisely at the level of the cholinergic amacrine cell dendrites, and co-fasciculate with the cholinergic matrix of starburst amacrine cells. However, neither ON or OFF α ganglion cells have more than a chance association with the cholinergic matrix. Z-axis reconstruction showed that OFF α ganglion cells stratify just below the cholinergic band in sublamina a while ON α ganglion cells stratify just below cholinergic b. The latter is at the same level as the terminals of calbindin bipolar cells. Thus, the calbindin bipolar cell appears to be a prime candidate to provide the bipolar cell input to ON α ganglion cells in the rabbit retina. We conclude that the precise level of stratification is correlated with the strength of cholinergic input. Alpha ganglion cells receive a weak cholinergic input and they are narrowly stratified just below the cholinergic bands. PMID:16212709

  14. Chemically selective polymer substrate based direct isotope dilution alpha spectrometry of Pu.

    PubMed

    Paul, Sumana; Pandey, Ashok K; Shah, R V; Aggarwal, S K

    2015-06-01

    Quantification of actinides in the complex environmental, biological, process and waste streams samples requires multiple steps like selective preconcentration and matrix elimination, solid source preparations generally by evaporation or electrodeposition, and finally alpha spectrometry. To minimize the sample manipulation steps, a membrane based isotope dilution alpha spectrometry method was developed for the determination of plutonium concentrations in the complex aqueous solutions. The advantages of this method are that it is Pu(IV) selective at 3M HNO3, high preconcentration factor can be achieved, and obviates the need of solid source preparation. For this, a thin phosphate-sulfate bifunctional polymer layer was anchored on the surface of microporous poly(ethersulfone) membrane by UV induced surface grafting. The thickness of the bifunctional layer on one surface of the poly(ethersulfone) membrane was optimized. The thickness, physical and chemical structures of the bifunctional layer were studied by secondary ionization mass spectrometry (SIMS), scanning electron microscopy (SEM) and SEM-EDS (energy-dispersive spectroscopy). The optimized membrane was used for preconcentration of Pu(IV) from aqueous solutions having 3-4M HNO3, followed by direct quantification of the preconcentrated Pu(IV) by isotope dilution alpha spectrometry using (238)Pu spike. The chemical recovery efficiency of Pu(IV) was found to be 86±3% below Pu(IV) loading capacity (1.08 μg in 2×1 cm(2)) of the membrane sample. The experiments with single representative actinides indicated that Am(III) did not sorb to significant extent (7%) but U(VI) sorbed with 78±3% efficiency from the solutions having 3M HNO3 concentration. However, Pu(IV) chemical recovery in the membrane remained unaffected from the solution containing 1:1000 wt. proportion of Pu(IV) to U(VI). Pu concentrations in the (U, Pu)C samples and in the irradiated fuel dissolver solutions were determined. The results thus obtained

  15. Chemically selective polymer substrate based direct isotope dilution alpha spectrometry of Pu.

    PubMed

    Paul, Sumana; Pandey, Ashok K; Shah, R V; Aggarwal, S K

    2015-06-01

    Quantification of actinides in the complex environmental, biological, process and waste streams samples requires multiple steps like selective preconcentration and matrix elimination, solid source preparations generally by evaporation or electrodeposition, and finally alpha spectrometry. To minimize the sample manipulation steps, a membrane based isotope dilution alpha spectrometry method was developed for the determination of plutonium concentrations in the complex aqueous solutions. The advantages of this method are that it is Pu(IV) selective at 3M HNO3, high preconcentration factor can be achieved, and obviates the need of solid source preparation. For this, a thin phosphate-sulfate bifunctional polymer layer was anchored on the surface of microporous poly(ethersulfone) membrane by UV induced surface grafting. The thickness of the bifunctional layer on one surface of the poly(ethersulfone) membrane was optimized. The thickness, physical and chemical structures of the bifunctional layer were studied by secondary ionization mass spectrometry (SIMS), scanning electron microscopy (SEM) and SEM-EDS (energy-dispersive spectroscopy). The optimized membrane was used for preconcentration of Pu(IV) from aqueous solutions having 3-4M HNO3, followed by direct quantification of the preconcentrated Pu(IV) by isotope dilution alpha spectrometry using (238)Pu spike. The chemical recovery efficiency of Pu(IV) was found to be 86±3% below Pu(IV) loading capacity (1.08 μg in 2×1 cm(2)) of the membrane sample. The experiments with single representative actinides indicated that Am(III) did not sorb to significant extent (7%) but U(VI) sorbed with 78±3% efficiency from the solutions having 3M HNO3 concentration. However, Pu(IV) chemical recovery in the membrane remained unaffected from the solution containing 1:1000 wt. proportion of Pu(IV) to U(VI). Pu concentrations in the (U, Pu)C samples and in the irradiated fuel dissolver solutions were determined. The results thus obtained

  16. The sensitivity of H alpha profiles to rapid electron beam fluctuations

    NASA Technical Reports Server (NTRS)

    Canfield, Richard C.; Gayley, Kenneth G.

    1986-01-01

    In order to understand the temporal relationship between H alpha and hard X-ray emission predicted by the nonthermal electron thick target model of impulsive-phase energy transport the time-dependent theoretical H alpha profiles were computed for the dynamic model atmospheres of Fisher, Canfield, and McClymont, which simulate the effects of an impulsively initiated power-law beam of electrons. On the basis of the physical analysis it was expected that a very rapid H alpha response to an instantaneous increase in the flux of a nonthermal deka-keV electron beam, as compared to the timescale associated with the propagation of these electrons over characteristic flare coronal loop spatial scales. It was concluded that observational efforts to test the thick target nonthermal electron model through detection of impulsive H alpha brightenings associated with impulsive hard X-ray or microwave bursts should initially focus attention on the H alpha line center. Additional simultaneous blue-wing measurements will have substantial diagnostic potential.

  17. MEC-12, an alpha-tubulin required for touch sensitivity in C. elegans.

    PubMed

    Fukushige, T; Siddiqui, Z K; Chou, M; Culotti, J G; Gogonea, C B; Siddiqui, S S; Hamelin, M

    1999-02-01

    mec-12 is one of a dozen genes required for touch receptor neuron function in Caenorhabditis elegans. Some mec-12 mutants (mechanosensory-defective) lack the large-diameter microtubules that are characteristic of these neurons (15 protofilaments, as opposed to 11). Mutants of mec-7, a alpha-tubulin encoding gene, have a similar phenotype. We have identified the nature of mec-12 by germline transformation rescue and characterization of a point mutation. Sequence analysis of the mec-12 encoded product (MEC-12) indicates that it corresponds to a novel C. elegans alpha-tubulin. MEC-12 is the only identified C. elegans alpha-tubulin that contains a lysine at position 40, a known site of post-translational acetylation. Some mec-12 mutations eliminate microtubule acetylation as assayed immunocyto-chemically; phenotypic rescue using a MEC-12 variant lacking the lysine-40 showed that acetylation is not required for MEC-12 activity. Although functionally needed only in the touch neurons, mec-12 is expressed in several other neuron types. These results support the notion that tubulin isotype diversity contributes to the formation of distinct classes of microtubules; 15-protofilament microtubule assembly requires MEC-12 alpha-tubulin and MEC-7 beta-tubulin, which are both highly expressed in the touch receptor neurons. MEC-12 is the first reported alpha-tubulin isotype that appears to be required in a single class of neuronal microtubules.

  18. Identification of a selective thieno[2,3-c]pyridine inhibitor of COT kinase and TNF-alpha production.

    PubMed

    Cusack, Kevin; Allen, Hamish; Bischoff, Agnieszka; Clabbers, Anca; Dixon, Richard; Fix-Stenzel, Shannon; Friedman, Michael; Gaumont, Yvette; George, Dawn; Gordon, Thomas; Grongsaard, Pintipa; Janssen, Bernd; Jia, Yong; Moskey, Maria; Quinn, Christopher; Salmeron, Andres; Thomas, Christine; Wallace, Grier; Wishart, Neil; Yu, Zhengtian

    2009-03-15

    COT (Tpl2 in mice) is a serine/threonine MAP3 kinase that regulates production of TNF-alpha and other pro-inflammatory cytokines such as IL-1beta via the ERK/MAP kinase pathway. As TNF-alpha and IL-1beta are clinically validated targets for therapeutic intervention in rheumatoid arthritis (RA), blocking COT provides a potential avenue for amelioration of disease. Herein we describe identification of a cellular active selective small molecule inhibitor of COT kinase.

  19. (-)-Discretamine, a selective alpha 1D-adrenoceptor antagonist, isolated from Fissistigma glaucescens.

    PubMed

    Ko, F N; Guh, J H; Yu, S M; Hou, Y S; Wu, Y C; Teng, C M

    1994-08-01

    1. The selectivity of (-)-discretamine for alpha 1-adrenoceptor subtypes was investigated by use of functional and binding studies in rat vas deferens, spleen and aorta, and in cultured DDT1MF-2 and A10 cells. 2. In prostatic portions of rat vas deferens, the competitive antagonists (-)-discretamine, 5-methylurapidil (5-MU) and prazosin inhibited contractions to noradrenaline (NA) with pA2 values of 6.21, 8.71 and 9.27, respectively. The irreversible antagonist, chloroethylclonidine (CEC, 100 microM) failed to affect contractions to NA while nifedipine (1 microM) blocked them almost completely. 3. In rat spleen, the competitive antagonists (-)-discretamine, 5-MU and prazosin inhibited contractions to phenylephrine with pA2 values of 6.44, 7.19 and 9.45, respectively. CEC (100 microM) significantly reduced the maximum contraction to phenylephrine while nifedipine (1 microM) did not affect it. 4. In rat aorta, the competitive antagonists (-)-discretamine, 5-MU and prazosin inhibited contractions to NA with pA2 values of 7.60, 8.00 and 9.40, respectively. CEC also antagonized the contractions to NA in a competitive manner with a pA2 value of 6.10. 5. The specific binding of [3H]-prazosin to DDT1MF-2 and A10 cells was concentration-dependent and saturated at 3-5 nM with KD values of 0.24 +/- 0.02 and 0.20 +/- 0.02 nM, respectively. (-)-Discretamine,5-MU, CEC and prazosin inhibited specific [3H]-prazosin binding to DDTIMF-2 and AlO cells in a concentration-dependent manner with ICso values of 390.8 +/- 20.6, 43.6 +/- 3.9, 200.0 +/- 30.0 and 0.8 +/- 0.1 nM, respectively in DDTIMF-2 cells, and 25.0 +/- 3.2, 8.6 +/- 1.4, 1000.0 +/- 30.8 and 0.52 +/- 0.03 nM, respectively in AlO cells.6. Pretreatment of Al0 cells with CEC (10 MicroM) for 30 min and then washed out thoroughly, reduced specific [3H]-prazosin binding by 30%. The CEC-insensitive [3H]-prazosin binding was inhibited by(-)-discretamine with an IC50 value of 7.0 +/- 0.3 nM.7. 5-MU (100 nM), CEC (1 MicroM) and

  20. Validation of highly specific and sensitive radioimmunoassays for lutropin, follitropin, and free alpha subunit in unextracted urine.

    PubMed

    Landy, H; Schneyer, A L; Whitcomb, R W; Crowley, W F

    1990-02-01

    Measurement of the urinary excretion of lutropin (LH) and follitropin (FSH) and their common free alpha subunit (FAS) assists in monitoring the maturation of the hypothalamic-pituitary-gonadal axis and in understanding the physiology of the pituitary glycoprotein hormones. Here we describe sensitive, specific polyclonal radioimmunoassays for LH and FSH and a monoclonal radioimmunoassay for FAS for use with urine--assays unperturbed by alterations in urinary pH or osmolarity within the broad physiological range encountered in urine. Concordance between LH, FSH, and FAS concentrations in extracted and unextracted urine samples was high. Linearity and parallelism with the standard curves was observed with addition of 25 to 200 microL of unextracted urine. No effect on glycoprotein concentration was seen after as many as 10 freeze-thaw cycles. The need for extraction was further obviated by the high sensitivity of each assay, reflected by minimum detectable doses well below the concentrations encountered in patients' samples. Thus we have measured gonadotropins in unextracted urine as precisely as in extracted urine. We also have demonstrated an equally versatile assay for urinary alpha subunit, using a monoclonal antibody of high specificity for this monomer in its free, uncombined form. These radioimmunoassays complement assays of gonadotropins and free alpha subunit in serum and will allow longitudinal investigations otherwise limited by the constraints of the patient's blood volume.

  1. Selective disturbance of pain sensitivity after social isolation.

    PubMed

    Tuboly, Gabor; Benedek, György; Horvath, Gyöngyi

    2009-01-01

    The effects of social isolation or NMDA-receptor antagonists on pain sensitivity have repeatedly been described. However, the mechanisms underlying the alterations of pain perception in these models still remain a matter of debate. Thus, we aimed to determine the long-lasting effects of subchronic ketamine treatment and social isolation on the C- and Adelta-fiber-mediated nociception. Wistar rats after weaning (21-23 days old) were either housed individually or grouped for 21 days. The animals were treated daily for 14 days with either ketamine (30 mg/kg) or saline. On the 21st day, tail-flick latency was determined at 48 degrees C (C-fiber activation) and 52 degrees C (affects mainly Adelta-fibers), and rats were rehoused. Tail-flick test was repeated 2 and 4 weeks later. On the 5th week, carrageenan-induced heat hyperalgesia was determined on paw-withdrawal test before and after morphine treatment (1, 2 or 3 mg/kg). Regarding tail-flick latencies at 48 degrees C, juvenile isolation, but not ketamine resulted in a significantly enhanced pain threshold (p<0.001) throughout the investigation period, while the changes at 52 degrees C were not significant. In addition, both isolation and ketamine treatments enhanced the antihyperalgesic effect of 2 mg/kg morphine. In summary, juvenile isolation exerts a long-lasting effect on acute heat pain sensitivity, disturbing primarily the C-fiber-linked pain pathways, suggesting a selective disruption in the parallel sensory pathways. Since both social isolation and NMDA treatment are well-known animal models of schizophrenia, our results showed that juvenile isolation but not ketamine administration can simulate hypoalgesia associated with this disease. PMID:18761027

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

    SciTech Connect

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

    1991-04-01

    A highly reproducible, sensitive, and specific sandwich enzyme-linked immunosorbent assay (ELISA) for recombinant human IL-1 {alpha} (rhIL-1 alpha) has been developed. Results from this ELISA have demonstrated that the concentration of rhIL-1 {alpha} 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 {alpha} 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 {alpha} in NHS. Immunoblot analysis indicated that the molecular weight of the binding protein was 150-160K, and the IL-1 {alpha} binding activity was removed and recovered from NHS by Protein-G affinity chromatography; indicating that the binding protein was IL-1 {alpha}-specific IgG. The binding of 125I-labeled IL-1 {alpha} to the serum binding proteins could be inhibited by unlabeled IL-1 alpha (IC50 = 7.4 {times} 10(-11) M) but not by unlabeled IL-1 {beta}. Kinetic analysis with 125I-labeled IL-1 alpha revealed that the average binding affinity of these IL-1 {alpha}-specific IgGs was 4.7 {times} 10(10) M-1. These results suggest that these autoantibodies may interfere with the detection of IL-1 {alpha} in human serum by various assay systems and also could be a regulator of circulating IL-1 {alpha}.

  3. Ocular avirulence of a herpes simplex virus type 1 strain is associated with heightened sensitivity to alpha/beta interferon.

    PubMed Central

    Su, Y H; Oakes, J E; Lausch, R N

    1990-01-01

    BALB/c mice infected on the scarified cornea with herpes simplex virus type 1 strain 35 [HSV-1(35)] rarely developed ocular disease even at challenge doses as high as 10(7) PFU per eye. In contrast, HSV-1(RE) consistently induced stromal keratitis at an inoculum of 2 x 10(4) PFU. The goal of this study was to determine the reason for the difference in virulence between the two HSV strains. Both HSV-1 strains replicated to similar titers in excised corneal "buttons." However, after in vivo infection of the cornea, the growth of strain 35 was evident only during the first 24 h postinfection, whereas the replication of strain RE persisted for at least 4 days. In vitro tests revealed that HSV-1(35) was greater than 10 times more sensitive to alpha/beta interferon (IFN-alpha/beta) than HSV-1(RE). Both strains induced comparable serum levels of IFN after intraperitoneal inoculation. The kinetics of HSV-1(35) clearance from the eye was markedly altered by treatment with rabbit anti-IFN-alpha/beta. Virus titers exceeding 10(4) PFU per eye could be demonstrated 4 to 5 days postinfection in mice given a single inoculation of antiserum 1 h after infection. Furthermore, anti-IFN treatment in 3-week-old mice infected with HSV-1(35) led to the development of clinically apparent corneal disease which subsequently progressed to stromal keratitis in the majority of recipients. These results indicate that the striking difference in the capacity of HSV-1(35) and HSV-1(RE) to induce corneal disease was related to the inherently greater sensitivity of strain 35 to IFN-alpha/beta produced by the host in response to infection. PMID:2157880

  4. cAMP sensitivity conferred to the epithelial Na+ channel by alpha-subunit cloned from guinea-pig colon.

    PubMed

    Schnizler, M; Mastroberardino, L; Reifarth, F; Weber, W M; Verrey, F; Clauss, W

    2000-03-01

    The rate of Na+ (re)absorption across tight epithelia such as in distal kidney nephron and colon is to a large extent controlled at the level of the epithelial Na+ channel (ENaC). In kidney, antidiuretic hormone (ADH, vasopressin) stimulates the expression/activity of this channel by a cAMP/protein-kinase-A- (PKA-) mediated pathway. However, a clear upregulation of ENaC function by cAMP could not be reproduced with cloned channel subunits in the Xenopus oocyte expression system, suggesting the hypothesis that an additional factor is missing. In contrast, we show here that membrane-permeant cAMP can activate ENaC expressed in Xenopus oocytes (3.8-fold) upon replacement of the rat alpha-subunit by a new alpha-subunit cloned from guinea-pig colon (gpalpha). This alpha-subunit is 76% identical with its rat orthologue originating from ADH-insensitive rat colon. The biophysical fingerprints of the hybrid ENaC formed by this guinea-pig alpha-subunit together with rat beta- and gamma-subunits are indistinguishable from those of rat ENaC (rENaC). Injection of the PKA inhibitor PKI-(6-22)-amide into the oocyte had no effect on the basal activity of rat ENaC but inhibited the activity of gpalpha-containing hybrid ENaC and greatly decreased its stimulation by cAMP. This suggests that, unlike for rat ENaC, tonic PKA activity is required for basal function of gpalpha-containing ENaC and that PKA mediates its cAMP-induced activation. This regulatory behaviour is not common to all ENaCs containing an alpha-subunit cloned from an ADH-responsive tissue since xENaC, which was cloned from the ADH-sensitive Xenopus laevis A6 epithelia, is, when expressed in oocytes, resistant to cAMP, similar to rat ENaC. This study demonstrates that the PKA sensitivity of ENaC can depend on the nature of the ENaC alpha-subunit and raises the possibility that cAMP can stimulate ENaCs by different mechanisms. PMID:10764218

  5. Natural ingredients based cosmetics. Content of selected fragrance sensitizers.

    PubMed

    Rastogi, S C; Johansen, J D; Menné, T

    1996-06-01

    In the present study, we have investigated 42 cosmetic products based on natural ingredients for content of 11 fragrance substances: geraniol, hydroxycitronellal, eugenol, isoeugenol, cinnamic aldehyde, cinnamic alcohol, alpha-amylcinnamic aldehyde, citral, coumarin, dihydrocoumarin and alpha-hexylcinnamic aldehyde. The study revealed that the 91% (20/22) of the natural ingredients based perfumes contained 0.027%-7.706% of 1 to 7 of the target fragrances. Between 1 and 5 of the chemically defined synthetic constituents of fragrance mix were found in 82% (18/22) of the perfumes. 35% (7/20) of the other cosmetic products (shampoos, creams, tonics, etc) were found to contain 0.0003-0.0820% of 1 to 3 of the target fragrances. Relatively high concentrations of hydroxycitronellal, coumarin, cinnamic alcohol and alpha-amyl cinnamic aldehyde were found in some of the investigated products. The detection of hydroxycitronellal and alpha-hexylcinnamic aldehyde in some of the products demonstrates that artificial fragrances, i.e., compounds not yet regarded as natural substances, may be present in products claimed to be based on natural ingredients.

  6. Administration of nicotine to adolescent rats evokes regionally selective upregulation of CNS alpha 7 nicotinic acetylcholine receptors.

    PubMed

    Slotkin, Theodore A; Cousins, Mandy M; Seidler, Frederic J

    2004-12-24

    Alpha 7 Nicotinic acetylcholine receptors (nAChRs) play a role in axonogenesis, synaptogenesis and synaptic plasticity, and are therefore targets for developmental neurotoxicants. We administered nicotine to adolescent rats and evaluated the effects on alpha 7 nAChRs in the striatum, brainstem and cerebellum. During the period of nicotine administration (30-47.5 days of age), nicotine elicited alpha 7 nAChR upregulation with a regional hierarchy of striatum>brainstem>cerebellum. Values returned to normal or became slightly subnormal almost immediately after the cessation of treatment (50 days of age) with no further changes through 75 days of age. The temporal and regional patterns of the effects on alpha 7 nAChRs were distinct from those reported earlier for the alpha 4 beta 2 subtype, and neither adult nor fetal/neonatal administration upregulates the alpha 7 subtype in the striatum. Targeting of the striatum is thus unique to nicotine exposure during adolescence and parallels earlier work showing regionally selective effects of this treatment on synaptic signaling. We obtained preliminary evidence for nicotine-induced oxidative stress as a potential contributory mechanism. The present findings reinforce the concept of biologically distinct effects of nicotine in the adolescent brain and provide evidence for a mechanistic involvement of alpha 7 nAChRs in its unique effects during this developmental period.

  7. A possible structural determinant of selectivity of boldine and derivatives for the alpha 1A-adrenoceptor subtype.

    PubMed Central

    Madrero, Y.; Elorriaga, M.; Martinez, S.; Noguera, M. A.; Cassels, B. K.; D'Ocon, P.; Ivorra, M. D.

    1996-01-01

    1. The selectivity of action of boldine and the related aporphine alkaloids, predicentrine (9-O-methylboldine) and glaucine (2,9-O-dimethylboldine) and alpha 1-adrenoceptor subtypes was studied by examining [3H]-prazosin competition binding in rat cerebral cortex. WB 4101 and benoxathian were used as selective alpha 1A-adrenoceptor antagonists. 2. In the competition experiments [3H]-prazosin (0.2 nM) binding was inhibited by WB 4101 and benoxathian. The inhibition curves displayed shallow slopes which could be subdivided into high and low affinity components (pKi = 9.92 and 8.29 for WB 4101, 9.35 and 7.94 for benoxathian). The two antagonists recognized approximately 37% of the sites with high affinity from among the total [3H]-prazosin specific binding sites. 3. Boldine, predicentrine and glaucine also competed for [3H]-prazosin (0.2 nM) binding with shallow and biphasic curves recognizing 30-40% of the sites with high affinity. Drug affinities (pKi) at the high and low affinity sites were, 8.31 and 6.50, respectively, for boldine, 8.13 and 6.39 for predicentrine, and 7.12 and 5.92 for glaucine. The relative order of selectivity for alpha 1A-adrenoceptors was boldine (70 fold alpha 1A-selective) = predicentrine (60 fold, alpha 1A-selective) > glaucine (15 fold, alpha 1A-selective). 4. Pretreatment of rat cerebral cortex membranes with chloroethylclonidine (CEC, 10 microM) for 30 min at 37 degrees C followed by thorough washing out reduced specific [3H]-prazosin binding by approximately 70%. The CEC-insensitive [3H]-prazosin binding was inhibited by boldine monophasically (Hill slope = 0.93) with a single pKi value (7.76). 5. These results suggest that whereas the aporphine structure shared by these alkaloids is responsible for their selectively of action for the alpha 1A-adrenoceptor subtype in rat cerebral cortex, defined functional groups, namely the 2-hydroxy function, induces a significant increase in alpha 1A-subtype selectivity and affinity. PMID:8982502

  8. Differential gene expression in the bovine corpus luteum during transition from early phase to midphase and its potential role in acquisition of luteolytic sensitivity to prostaglandin F2 alpha.

    PubMed

    Goravanahally, Madhusudan P; Salem, Mohamed; Yao, Jianbo; Inskeep, E Keith; Flores, Jorge A

    2009-05-01

    Prostaglandin F2 alpha (PGF(2alpha)) brings about regression of the bovine corpus luteum (CL). This luteolytic property of PGF(2alpha) is used in beef and dairy cattle to synchronize estrus. A limitation of this protocol is insensitivity of the early CL to luteolytic actions of PGF(2alpha). The mechanisms underlying this differential luteal sensitivity are poorly understood. The developing CL has a maximum number of PGF(2alpha) receptors; therefore, differences in signaling events may be responsible for luteal insensitivity. Hence, differential gene expression at two developmental stages of CL, Day 4 (D-4) and D-10 after estrus, might account for differences in signal transduction pathways associated with luteal sensitivity. This possibility was examined in these studies. Microarray analysis (n = 3 cows per stage) identified 167 genes that were differentially expressed (P < 0.05). These were categorized into genes involved in protein biosynthesis and modification (18.5%), transcription regulation and DNA biosynthesis (18.5%), miscellaneous (17.0%), cell signaling (12.0%), steroidogenesis and metabolism (10.2%), extracellular matrix and cytoskeletal proteins (9.5%), unknown functions (6.0%), protein degradation (5.3%), and antioxidant property (3.0%). Real-time PCR confirmed the differential expression of nine selected genes, including tyrosine 3-monooxygenase/tryptophan 5-monooxygense activation protein zeta polypeptide (YWHAZ) and regulator of G protein signaling 2 24-kDa (RGS2), observed in microarray. Furthermore, the in vivo effect of exogenous PGF(2alpha) (n = 3 cows per stage) on selected genes that were found to be differentially expressed during this developmental transition was examined. PGF(2alpha) increased the expression of a guanine nucleotide-binding protein (G protein) beta polypeptide 1 (GNB1) in D-4 CL and calcium/calmodulin-dependent kinase kinase 2 beta (CAMKK2) in D-10 CL. Therefore, GNB1, CAMKK2, YWHAZ, and RGS2 are candidate genes that may

  9. Culture, Personality, Health, and Family Dynamics: Cultural Competence in the Selection of Culturally Sensitive Treatments

    ERIC Educational Resources Information Center

    Sperry, Len

    2010-01-01

    Cultural sensitivity and cultural competence in the selection of culturally sensitive treatments is a requisite for effective counseling practice in working with diverse clients and their families, particularly when clients present with health issues or medical problems. Described here is a strategy for selecting culturally sensitive treatments…

  10. Acidity-controlled selective oxidation of alpha-pinene, isolated from Indonesian pine's turpentine oils (pinus merkusii)

    NASA Astrophysics Data System (ADS)

    Masruri; Farid Rahman, Mohamad; Nurkam Ramadhan, Bagus

    2016-02-01

    Alpha-pinene was isolated in high purity from turpentine oil harvested from Pinus merkusii plantation. The recent investigation on selective oxidation of alpha-pinene using potassium permanganate was undertaken under acidic conditions. The result taught the selective oxidation of alpha-pinene in acidic using potassium permanganate lead to the formation of 2-(3-acetyl-2,2-dimethylcyclobutyl)acetaldehyde or pinon aldehyde. The study method applied reaction in various different buffer conditions i.e. pH 3, 4, 5, and 6, respectively, and each reaction product was monitored using TLC every hour. Product determination was undertaken on spectrometry basis such as infrared, ultra violet-visible, gas chromatography- and liquid chromatography-mass spectrometry.

  11. The selective augmentation by recombinant human tumour necrosis factor-alpha of neutrophil responses to pathogenic Escherichia coli.

    PubMed Central

    Steadman, R; Petersen, M M; Williams, D; Matthews, N; Williams, J D

    1990-01-01

    Endotoxin release may amplify the neutrophil (PMN) responses to bacterial infection through the release of monocyte-derived tumour necrosis factor (TNF). The present study was designed to assess the effect of recombinant human TNF-alpha (rhTNF-alpha) on the in vitro response of human PMN to two defined strains of pathogenic Escherichia coli. In the absence of rhTNF-alpha, a P-fimbriate strain caused significant release of the PMN secondary granule marker vitamin B12-binding protein (B12 BP), and a low level of release of leukotriene B4 (LTB4). Type 1-fimbriate strain 504, however, stimulated the release of the primary granule marker myeloperoxidase (MPO) and PMN chemiluminescence (CL), in addition to B12 BP and LTB4 release. Following rhTNF-alpha (10(-9) M) pretreatment, the release of LTB4 by PMN stimulated with the P-fimbriate strain was synergistically augmented, while B12 BP and MPO release were additively increased. In contrast, rhTNF-alpha did not significantly affect any of the responses by the type 1-fimbriate strain. These results suggest selectivity in the priming of PMN by rhTNF-alpha and confirm the independence of PMN responses to phagocytic stimuli. PMID:2162324

  12. Fitness measures in selection analyses: sensitivity to the overall number of offspring produced in a lifetime.

    PubMed

    Dugdale, H L; Nouvellet, P; Pope, L C; Burke, T; Macdonald, D W

    2010-02-01

    Age at first (alpha) and last (omega) breeding are important life-history traits; however, the direction and strength of selection detected on traits may vary depending on the fitness measure used. We provide the first estimates of lifetime breeding success (LBS) and lambda(ind) (the population growth rate of an individual) of European badgers Meles meles, by genotyping 915 individuals, sampled over 18 years, for 22 microsatellites. Males are slightly larger than females, and the opportunity for selection was slightly greater for males, as predicted. lambda(ind) and LBS both performed well in predicting the number of grand-offspring, and both detected selection for a late omega, until the age of eight. Differential selection (S'(alpha)) for an early alpha, however, was only detected using LBS, not with lambda(ind). In declining populations (lambda(ind) < 1) selection favours reproduction later in life, whereas early reproduction is selected in increasing populations (lambda(ind) > 1). As 41% of badgers were assigned only one offspring (lambda(ind) < 1), whereas 40% were assigned more than two (lambda(ind) > 1), this cancelled out S'(alpha) measured by lambda(ind). PMID:20002246

  13. Selective LXR{alpha} inhibitory effects observed in plant extracts of MEH184 (Parthenocissua tricuspidata) and MEH185 (Euscaphis japonica)

    SciTech Connect

    Kim, Kang Ho; Choi, Seung Hyun; Lee, Thomas S.; Oh, Won Keun; Kim, Dong Sun; Kim, Jae Bum . E-mail: jaebkim@snu.ac.kr

    2006-10-20

    Liver X receptors (LXRs) are nuclear hormone receptors that behave as lipid sensors of cellular cholesterol and fatty acid. Although LXR activation can alleviate hypercholesterolemia by inducing cholesterol efflux, it also results in undesirable effects of fatty acid synthesis, resulting in hepatic steatosis and hyperlipidemia. Therefore, it is critical to identify LXR{alpha} inhibitory agents that would repress fatty acid synthesis and hepatic lipid accumulation. In current study, screening of plant extracts used for traditional oriental medicine resulted in the identification of two candidates demonstrating selective LXR{alpha} inhibitory activity. These were whole leaf methanol extracts of Parthenocissua tricuspidata (MEH184) and Euscaphis japonica (MEH185). Both MEH184 and MEH185 decreased transcriptional activity of LXR{alpha} and the expression of LXR{alpha} target genes, such as FAS and ADD1/SREBP1c. Additionally, MEH184 and MEH184 significantly reduced lipogenesis and adipocyte differentiation. Together, the data imply that MEH184 and MEH185 possess selective antagonistic properties on LXR{alpha} to downregulate lipogenesis.

  14. Expression of tfx and sensitivity to the rhizobial peptide antibiotic trifolitoxin in a taxonomically distinct group of alpha-proteobacteria including the animal pathogen Brucella abortus.

    PubMed Central

    Triplett, E W; Breil, B T; Splitter, G A

    1994-01-01

    Three phylogenetically distinct groups within the alpha-proteobacteria which differ in trifolitoxin sensitivity are described. Trifolitoxin sensitivity was found in strains of Agrobacterium, Brucella, Mycoplana, Ochrobactrum, Phyllobacterium, Rhodobacter, Rhodopseudomonas, Rhodospirillum, and Rhizobium. Strains of Agrobacterium, Brucella, Phyllobacterium, Rhizobium, and Rhodospirillum were capable of producing trifolitoxin upon conjugal transfer of tfxABCDEFG. PMID:7527627

  15. Anthranilate synthase from Ruta graveolens. Duplicated AS alpha genes encode tryptophan-sensitive and tryptophan-insensitive isoenzymes specific to amino acid and alkaloid biosynthesis.

    PubMed Central

    Bohlmann, J; Lins, T; Martin, W; Eilert, U

    1996-01-01

    Anthranilate synthase (AS, EC 4.1.3.27) catalyzes the conversion of chorismate into anthranilate, the biosynthetic precursor of both tryptophan and numerous secondary metabolites, including inducible plant defense compounds. The higher plant Ruta graveolens produces tryptophan and elicitor-inducible, anthranilate-derived alkaloids by means of two differentially expressed nuclear genes for chloroplast-localized AS alpha subunits, AS alpha 1 and AS alpha 2. Mechanisms that partition chorismate between tryptophan and inducible alkaloids thus do not entail chloroplast/cytosol separation of AS isoenzymes and yet might involve differential feedback regulation of pathway-specific AS alpha subunits. The two AS alpha isoenzymes of R. graveolens were expressed as glutathione S-transferase fusion proteins in Escherichia coli deletion mutants defective in AS activity and were purified to homogeneity. Differential sensitivity of the transformed E. coli strains toward 5-methyltryptophan, a false-feedback inhibitor of AS, was demonstrated. Characterization of affinity-purified AS alpha isoenzymes revealed that the noninducible AS alpha 2 of R. graveolens is strongly feedback inhibited by 10 microns tryptophan. In contrast, the elicitor-inducible AS alpha 1 isoenzyme is only slightly affected even by tryptophan concentrations 10-fold higher than those observed in planta. These results are consistent with the hypothesis that chorismate flux into biosynthesis of tryptophan and defense-related alkaloid biosynthesis in R. graveolens is regulated at the site of AS alpha isoenzymes at both genetic and enzymatic levels. PMID:8787026

  16. Delving Deeper into the Solar Dynamo Mechanism: Alpha Effect, Parity Selection and Large Scale Flows.

    NASA Astrophysics Data System (ADS)

    Nandy, D.

    2003-05-01

    Visible manifestations of the 22 year solar magnetic cycle have been the subject of study spanning centuries starting with the telescopic observations of sunspots by Johann Fabricius, Christoph Scheiner and Galileo Galilei in the early 1600s. Coupled with these observations of magnetic features on the solar surface, the advent of the field of helioseismology in recent years has made it possible to map large scale flows in the solar interior - believed to play a crucial role in sustaining the solar cycle. However, a complete understanding of the hydromagnetic dynamo mechanism that powers this solar cycle remains elusive. Here we report studies of the solar dynamo addressing some of the important unresolved questions regarding the nature and location of the alpha effect, solar magnetic parity selection and the role of large scale flows and their variation, with a goal to understand the exact means by which the Sun generates its magnetic cycle. This study was supported by NASA through SR&T grant NAG5-6110.

  17. Cyclosporin A sensitivity of the NF-kappa B site of the IL2R alpha promoter in untransformed murine T cells.

    PubMed Central

    McCaffrey, P G; Kim, P K; Valge-Archer, V E; Sen, R; Rao, A

    1994-01-01

    We have investigated the characteristics of IL2R alpha gene induction in untransformed murine T cells. Induction of IL2R alpha mRNA by TCR/CD3 ligands in a murine T cell clone and in short-term splenic T cell cultures was inhibited by protein synthesis inhibitors and by CsA. This result was contrary to previous observations in JURKAT T leukemia cells and human peripheral blood T cells, suggesting a difference in the mechanisms of IL2R alpha gene induction in these different cell types. The CsA sensitivity of IL2R alpha mRNA induction represented a direct effect on the TCR/CD3 response, and was not due to CsA-sensitive release of the lymphokines IL2 or tumour necrosis factor alpha (TNF alpha) and consequent lymphokine-mediated induction of IL2R alpha mRNA. The NF-kappa B site of the IL2R alpha promoter was essential for gene induction through the TCR/CD3 complex, and the induction of reporter plasmids containing multimers of this site was significantly inhibited by CsA. Northern blotting analysis indicated that while the p65 subunit of NF-kappa B was constitutively expressed and not appreciably induced upon T cell activation, mRNA for the p105 precursor of p50 NF-kappa B was induced in response to TCR/CD3 stimulation and this induction was sensitive to CsA. Electrophoretic mobility shift assays and antiserum against the p50 subunit of NF-kappa B indicated that p50 was a component of the inducible nuclear complex that bound to the IL2R alpha kappa B site. Appearance of the kB-binding proteins was insensitive to CsA at early times after activation (approximately 15 min), but was partially sensitive to CsA at later times. Based on these results, we propose that the NF-kappa B site of the IL2R alpha promoter mediates at least part of the CsA sensitivity of IL2R alpha gene induction in untransformed T cells, possibly because de novo synthesis of p105 NF-kappa B is required for sustained IL2R alpha expression. Images PMID:8029023

  18. Selecting step sizes in sensitivity analysis by finite differences

    NASA Technical Reports Server (NTRS)

    Iott, J.; Haftka, R. T.; Adelman, H. M.

    1985-01-01

    This paper deals with methods for obtaining near-optimum step sizes for finite difference approximations to first derivatives with particular application to sensitivity analysis. A technique denoted the finite difference (FD) algorithm, previously described in the literature and applicable to one derivative at a time, is extended to the calculation of several simultaneously. Both the original and extended FD algorithms are applied to sensitivity analysis for a data-fitting problem in which derivatives of the coefficients of an interpolation polynomial are calculated with respect to uncertainties in the data. The methods are also applied to sensitivity analysis of the structural response of a finite-element-modeled swept wing. In a previous study, this sensitivity analysis of the swept wing required a time-consuming trial-and-error effort to obtain a suitable step size, but it proved to be a routine application for the extended FD algorithm herein.

  19. Sensitivities of Pityrosporum sp. to selected commercial shampoos.

    PubMed

    Butterfield, W; Roberts, M M; Dave, V K

    1987-02-01

    Sensitivity of 25 strains of Pityrosporum sp. to four commercial shampoos was tested using a gel diffusion method and determination of minimum inhibitory concentrations (MICs). All the shampoos when undiluted gave inhibition zones in the gel diffusion test with 13 of the 15 strains tested. Two strains were resistant to 'Polytar'. 'Polytar' was fungistatic, 'Selsun' 'Cetavlon P.C.' and 'Genisol' were fungicidal. MIC results showed the yeast to be most sensitive to 'Cetavlon' and 'Selsun'.

  20. Alpha 1-proteinase inhibitor is more sensitive to inactivation by cigarette smoke than is leukocyte elastase

    SciTech Connect

    Janoff, A.; Dearing, R.

    1982-10-01

    Aqueous solutions of gas phase cigarette smoke were incubated with pure human leukocyte elastase or with crude human leukocyte granule extract, and the effects on enzyme activity were determined using a synthetic amide substrate. Simultaneously, the same smoke solutions were incubated with 10% human serum under identical conditions, and the effects on serum inhibition of purified or crude leukocyte elastase were similarly measured. In addition, aqueous solutions of unfractionated cigarette smoke were incubated with leukocyte elastase or serum, and the abilities of the smoke-treated enzyme to digest elastin and of the smoke-treated serum to inhibit elastin digestion were determined. Both experimental protocols showed that serum elastase-inhibiting capacity (primarily caused by alpha 1-proteinase inhibitor) is more susceptible to inactivation by aqueous solutions of cigarette smoke than is leukocyte elastase, suggesting that elastase inhibition (rather than elastase activity) may be predominantly suppressed by cigarette smoke inhalation in vivo.

  1. Alexa Fluor 546-ArIB[V11L;V16A] is a potent ligand for selectively labeling alpha 7 nicotinic acetylcholine receptors.

    PubMed

    Hone, Arik J; Whiteaker, Paul; Mohn, Jesse L; Jacob, Michele H; McIntosh, J Michael

    2010-08-01

    The alpha7* (*denotes the possible presence of additional subunits) nicotinic acetylcholine receptor (nAChR) subtype is widely expressed in the vertebrate nervous system and implicated in neuropsychiatric disorders that compromise thought and cognition. In this report, we demonstrate that the recently developed fluorescent ligand Cy3-ArIB[V11L;V16A] labels alpha7 nAChRs in cultured hippocampal neurons. However, photobleaching of this ligand during long image acquisition times prompted us to develop a new derivative. In photostability studies, this new ligand, Alexa Fluor 546-ArIB[V11L;V16A], was significantly more resistant to bleaching than the Cy3 derivative. The classic alpha7 ligand alpha-bungarotoxin binds to alpha1* and alpha9* nAChRs. In contrast, Alexa Fluor 546-ArIB[V11L;V16A] potently (IC(50) 1.8 nM) and selectively blocked alpha7 nAChRs but not alpha1* or alpha9* nAChRs expressed in Xenopus oocytes. Selectivity was further confirmed by competition binding studies of native nAChRs in rat brain membranes. The fluorescence properties of Alexa Fluor 546-ArIB[V11L;V16A] were assessed using human embryonic kidney-293 cells stably transfected with nAChRs; labeling was observed on cells expressing alpha7 but not cells expressing alpha3beta2, alpha3beta4, or alpha4beta2 nAChRs. Further imaging studies demonstrate that Alexa Fluor 546-ArIB[V11L;V16A] labels hippocampal neurons from wild-type mice but not from nAChR alpha7 subunit-null mice. Thus, Alexa Fluor 546-ArIB[V11L;V16A] represents a potent and selective ligand for imaging alpha7 nAChRs.

  2. Increased sensitization of acoustic startle response in spasmodic mice with a mutation of the glycine receptor alpha1-subunit gene.

    PubMed

    Plappert, C F; Pilz, P K; Becker, K; Becker, C M; Schnitzler, H U

    2001-06-01

    The spontaneous mutant mouse spasmodic (spd) carries a missense mutation affecting the glycine receptor alpha1-subunit gene. This results in a decreased binding affinity to glycine. Spd mutants show exaggerated acoustic startle responses (ASR). The present study sought to elucidate whether this increased ASR is due to a changed auditory processing or to stronger motor output resulting from a disinhibited motor system or, alternatively, to changes in modulatory influences on the startle pathway, namely in the mechanisms underlying habituation and sensitization. We found that in homozygous spd/spd mutants the startle threshold was lower, and the recorded slope of input/output (i/o) function, which reflects the relation between sensory input and motor output, was steeper. During repetitive presentation of high sound pressure level (SPL) startle stimuli (25 dB above startle threshold), ASR amplitudes did not decrease in spd/spd mutants as they do in the wildtype. In contrast, ASR amplitudes decreased when low SPL startle stimuli were presented. Footshocks presented after high SPL startle stimuli did not cause a further increase in ASR amplitudes of spd/spd mutants as in the wildtype. In heterozygous spd/+ mutants all these parameters were between those of spd/spd mutants and wildtype mice but closer to those of the wildtype. The steeper slope of i/o function in spd/spd mutants may be caused by both an increased sensory input and an increased motor output. The altered course of ASR amplitudes during repetitive stimulation and the deficit in additional footshock sensitization, however, can only be explained by an increased sensitization level in the spd/spd mutants. In accordance with the "dual process theory" strong sensitization evoked by high SPL startle stimuli supposedly counteracts habituation, leading to a constant high ASR amplitude. Furthermore, additional footshock sensitization is prevented. The increased sensitization level may be due to a change in auditory

  3. Selection of the appropriate radionuclide source for the efficiency calibration in methods of determining gross alpha activity in water.

    PubMed

    Corbacho, J A; Zapata-García, D; Montaña, M; Fons, J; Camacho, A; Guillén, J; Serrano, I; Baeza, A; Llauradó, M; Vallés, I

    2016-01-01

    Measuring the gross alpha activity in water samples is a rapid, straightforward way of determining whether the water might contain a radionuclide concentration whose consumption would imply a total indicative dose (TID) greater than some reference limit - currently set at 0.1 mSv/y in Europe. There are several methods used for such measurements. Two of them are desiccation with the salts being deposited on a planchet, and coprecipitation. The main advantage of these two methods is their ease of implementation and low cost of preparing the source to measure. However, there is considerable variability in the selection of the most suitable radioactive reference standard against which to calculate the water's gross alpha activity. The goal of this paper is to propose the most appropriate reference radionuclides to use as standards in determining gross alpha activities with these two methods, taking into account the natural radioactive characteristics of a wide range of waters collected at different points in Spain. Thus, the results will be consistent with each other and representative of the sum of alpha activities of all the alpha-emitters contained in a sample. PMID:26397746

  4. Estrogen alters the diurnal rhythm of alpha 1-adrenergic receptor densities in selected brain regions

    SciTech Connect

    Weiland, N.G.; Wise, P.M.

    1987-11-01

    Norepinephrine regulates the proestrous and estradiol-induced LH surge by binding to alpha 1-adrenergic receptors. The density of alpha 1-receptors may be regulated by estradiol, photoperiod, and noradrenergic neuronal activity. We wished to determine whether alpha 1-receptors exhibit a diurnal rhythm in ovariectomized and/or estradiol-treated female rats, whether estradiol regulates alpha 1-receptors in those areas of brain involved with LH secretion and/or sexual behavior, and whether the concentrations of alpha-receptors vary inversely relative to previously reported norepinephrine turnover patterns. Young female rats, maintained on a 14:10 light-dark cycle were ovariectomized. One week later, half of them were outfitted sc with Silastic capsules containing estradiol. Groups of animals were decapitated 2 days later at 0300, 1000, 1300, 1500, 1800, and 2300 h. Brains were removed, frozen, and sectioned at 20 micron. Sections were incubated with (/sup 3/H)prazosin in Tris-HCl buffer, washed, dried, and exposed to LKB Ultrofilm. The densities of alpha 1-receptors were quantitated using a computerized image analysis system. In ovariectomized rats, the density of alpha 1-receptors exhibited a diurnal rhythm in the suprachiasmatic nucleus (SCN), medial preoptic nucleus (MPN), and pineal gland. In SCN and MPN, receptor concentrations were lowest during the middle of the day and rose to peak levels at 1800 h. In the pineal gland, the density of alpha 1-receptors was lowest at middark phase, rose to peak levels before lights on, and remained elevated during the day. Estradiol suppressed the density of alpha 1 binding sites in the SCN, MPN, median eminence, ventromedial nucleus, and the pineal gland but had no effect on the lateral septum. Estrogen treatment altered the rhythm of receptor densities in MPN, median eminence, and the pineal gland.

  5. (-)-Discretamine, a selective alpha 1D-adrenoceptor antagonist, isolated from Fissistigma glaucescens.

    PubMed Central

    Ko, F. N.; Guh, J. H.; Yu, S. M.; Hou, Y. S.; Wu, Y. C.; Teng, C. M.

    1994-01-01

    1. The selectivity of (-)-discretamine for alpha 1-adrenoceptor subtypes was investigated by use of functional and binding studies in rat vas deferens, spleen and aorta, and in cultured DDT1MF-2 and A10 cells. 2. In prostatic portions of rat vas deferens, the competitive antagonists (-)-discretamine, 5-methylurapidil (5-MU) and prazosin inhibited contractions to noradrenaline (NA) with pA2 values of 6.21, 8.71 and 9.27, respectively. The irreversible antagonist, chloroethylclonidine (CEC, 100 microM) failed to affect contractions to NA while nifedipine (1 microM) blocked them almost completely. 3. In rat spleen, the competitive antagonists (-)-discretamine, 5-MU and prazosin inhibited contractions to phenylephrine with pA2 values of 6.44, 7.19 and 9.45, respectively. CEC (100 microM) significantly reduced the maximum contraction to phenylephrine while nifedipine (1 microM) did not affect it. 4. In rat aorta, the competitive antagonists (-)-discretamine, 5-MU and prazosin inhibited contractions to NA with pA2 values of 7.60, 8.00 and 9.40, respectively. CEC also antagonized the contractions to NA in a competitive manner with a pA2 value of 6.10. 5. The specific binding of [3H]-prazosin to DDT1MF-2 and A10 cells was concentration-dependent and saturated at 3-5 nM with KD values of 0.24 +/- 0.02 and 0.20 +/- 0.02 nM, respectively.(ABSTRACT TRUNCATED AT 250 WORDS) PMID:7952879

  6. Selectively plated infrared-sensitive lead sulfide layers

    NASA Astrophysics Data System (ADS)

    Subramanian, V.; Murali, K. R.; Rangarajan, N.; Lakshmanan, A. S.

    1994-10-01

    Lead chalcogenides are well-known photoconductors in the infrared region. Lead sulfide detectors occupy an important position among infrared detectors owing to their high sensitivity in the near infrared region (1 - 3.2 microns), room temperature operation, low noise and ease of preparation. In this paper, we report for the first time the preparation of infrared sensitive photoconductive lead sulfide films by the swab plating technique. Adherent, non-porous `p' type lead sulfide films of thickness 3 - 4 microns were obtained. These films, after heat- treatment at 550 degree(s)C, showed a dark resistance of 25 - 30 kilo-ohms with a photosensitivity of 0.28. The films have been characterized by XRD, optical and photoconductivity measurements.

  7. Selective detector of cosmic particles based on diamond sensitive elements

    NASA Astrophysics Data System (ADS)

    Altukhov, A. A.; Zaharchenko, K. V.; Kolyubin, V. A.; Lvov, S. A.; Nedosekin, P. G.; Tyurin, E. M.; Ibragimov, R. F.; Kadilin, V. V.; Nikolaev, I. V.

    2016-02-01

    The article describes the device for selective registration of electrons, protons and heavy ions fluxes from the solar and galactic cosmic rays in the twelve energy ranges, built on a base of diamond detector. The use of the diamond detectors allowed for the creation a device for registration of cosmic particles fluxes at the external spacecraft surface with the resource not less than 20 years. Selective detector is aimed for continuous monitoring of radiation situation on board the spacecrafts, in order to predict the residual life of their work and prompt measures to actively protect the spacecraft when the flow of cosmic particles is sharply increased.

  8. On velocity-space sensitivity of fast-ion D-alpha spectroscopy

    NASA Astrophysics Data System (ADS)

    Salewski, M.; Geiger, B.; Moseev, D.; Heidbrink, W. W.; Jacobsen, A. S.; Korsholm, S. B.; Leipold, F.; Madsen, J.; Nielsen, S. K.; Rasmussen, J.; Stejner, M.; Weiland, M.; the ASDEX Upgrade Team

    2014-10-01

    The velocity-space observation regions and sensitivities in fast-ion Dα (FIDA) spectroscopy measurements are often described by so-called weight functions. Here we derive expressions for FIDA weight functions accounting for the Doppler shift, Stark splitting, and the charge-exchange reaction and electron transition probabilities. Our approach yields an efficient way to calculate correctly scaled FIDA weight functions and implies simple analytic expressions for their boundaries that separate the triangular observable regions in (v‖, v⊥)-space from the unobservable regions. These boundaries are determined by the Doppler shift and Stark splitting and could until now only be found by numeric simulation.

  9. Synapsin Is Selectively Required for Anesthesia-Sensitive Memory

    ERIC Educational Resources Information Center

    Knapek, Stephan; Gerber, Bertram; Tanimoto, Hiromu

    2010-01-01

    Odor-shock memory in "Drosophila melanogaster" consists of heterogeneous components each with different dynamics. We report that a null mutant for the evolutionarily conserved synaptic protein Synapsin entails a memory deficit selectively in early memory, leaving later memory as well as sensory motor function unaffected. Notably, a consolidated…

  10. Estrogen receptor alpha overexpressing mouse antral follicles are sensitive to atresia induced by methoxychlor and its metabolites.

    PubMed

    Paulose, Tessie; Hannon, Patrick R; Peretz, Jackye; Craig, Zelieann R; Flaws, Jodi A

    2012-06-01

    Methoxychlor (MXC) and its metabolites bind to estrogen receptors (ESRs) and increase ovarian atresia. To test whether ESR alpha (ESR1) overexpressing (ESR1 OE) antral follicles are more sensitive to atresia compared to controls, we cultured antral follicles with vehicle, MXC (1-100 μg/ml) or metabolites (0.1-10 μg/ml). Results indicate that MXC and its metabolites significantly increase atresia in ESR1 OE antral follicles at lower doses compared to controls. Activity of pro-apoptotic factor caspase-3/7 was significantly higher in ESR1 OE treated antral follicles compared to controls. ESR1 OE mice dosed with MXC 64 mg/kg/day had an increased percentage of atretic antral follicles compared to controls. Furthermore, pro-caspase-3 levels were found to be significantly lower in ESR1 OE ovaries than controls dosed with MXC 64 mg/kg/day. These data suggest that ESR1 OE ovaries are more sensitive to atresia induced by MXC and its metabolites in vitro and in vivo compared to controls.

  11. Pyridoxal 5'-phosphate is a selective inhibitor in vivo of DNA polymerase alpha and epsilon.

    PubMed

    Mizushina, Yoshiyuki; Xu, Xianai; Matsubara, Kiminori; Murakami, Chikako; Kuriyama, Isoko; Oshige, Masahiko; Takemura, Masaharu; Kato, Norihisa; Yoshida, Hiromi; Sakaguchi, Kengo

    2003-12-26

    Vitamin B(6) compounds such as pyridoxal 5(')-phosphate (PLP), pyridoxal (PL), pyridoxine (PN), and pyridoxamine (PM), which reportedly have anti-angiogenic and anti-cancer effects, were thought to be inhibitors of some types of eukaryotic DNA polymerases. PL moderately inhibited only the activities of calf DNA polymerase alpha (pol alpha), while PN and PM had no inhibitory effects on any of the polymerases tested. On the other hand, PLP, a phosphated form of PL, was potentially a strong inhibitor of pol alpha and epsilon from phylogenetic-wide organisms including mammals, fish, insects, plants, and protists. PLP did not suppress the activities of prokaryotic DNA polymerases such as Escherichia coli DNA polymerase I and Taq DNA polymerase, or DNA-metabolic enzymes such as deoxyribonuclease I. For pol alpha and epsilon, PLP acted non-competitively with the DNA template-primer and competitively with the nucleotide substrate. Since PL was converted to PLP in vivo after being incorporated into human cancer cells, the anti-angiogenic and anti-cancer effects caused by PL must have been caused by the inhibition of pol alpha and epsilon activities after conversion to PLP.

  12. The carboxyl terminus of the alpha-subunit of the amiloride-sensitive epithelial sodium channel binds to F-actin.

    PubMed

    Mazzochi, Christopher; Bubien, James K; Smith, Peter R; Benos, Dale J

    2006-03-10

    The activity of the amiloride-sensitive epithelial sodium channel (ENaC) is modulated by F-actin. However, it is unknown if there is a direct interaction between alpha-ENaC and actin. We have investigated the hypothesis that the actin cytoskeleton directly binds to the carboxyl terminus of alpha-ENaC using a combination of confocal microscopy, co-immunoprecipitation, and protein binding studies. Confocal microscopy of Madin-Darby canine kidney cell monolayers stably transfected with wild type, rat isoforms of alpha-, beta-, and gamma-ENaC revealed co-localization of alpha-ENaC with the cortical F-actin cytoskeleton both at the apical membrane and within the subapical cytoplasm. F-actin was found to co-immunoprecipitate with alpha-ENaC from whole cell lysates of this cell line. Gel overlay assays demonstrated that F-actin specifically binds to the carboxyl terminus of alpha-ENaC. A direct interaction between F-actin and the COOH terminus of alpha-ENaC was further corroborated by F-actin co-sedimentation studies. This is the first study to report a direct and specific biochemical interaction between F-actin and ENaC. PMID:16356937

  13. Sensitive And Selective Chemical Sensor With Nanostructured Surfaces.

    DOEpatents

    Pipino, Andrew C. R.

    2003-02-04

    A chemical sensor is provided which includes an optical resonator including a nanostructured surface comprising a plurality of nanoparticles bound to one or more surfaces of the resonator. The nanoparticles provide optical absorption and the sensor further comprises a detector for detecting the optical absorption of the nanoparticles or their environment. In particular, a selective chemical interaction is provided which modifies the optical absorption of the nanoparticles or their environment, and an analyte is detected based on the modified optical absorption. A light pulse is generated which enters the resonator to interrogate the modified optical absorption and the exiting light pulse is detected by the detector.

  14. Sensitivity of Alpha and Beta Oscillations to Sensorimotor Characteristics of Action: An EEG Study of Action Production and Gesture Observation

    PubMed Central

    Quandt, Lorna C.; Marshall, Peter J.; Shipley, Thomas F.; Beilock, Sian L.; Goldin-Meadow, Susan

    2012-01-01

    The sensorimotor experiences we gain when performing an action have been found to influence how our own motor systems are activated when we observe others performing that same action. Here we asked whether this phenomenon applies to the observation of gesture. Would the sensorimotor experiences we gain when performing an action on an object influence activation in our own motor systems when we observe others performing a gesture for that object? Participants were given sensorimotor experience with objects that varied in weight, and then observed video clips of an actor producing gestures for those objects. Electroencephalography (EEG) was recorded while participants first observed either an iconic gesture (pantomiming lifting an object) or a deictic gesture (pointing to an object) for an object, and then grasped and lifted the object indicated by the gesture. We analyzed EEG during gesture observation to determine whether oscillatory activity was affected by the observer’s sensorimotor experiences with the object represented in the gesture. Seeing a gesture for an object previously experienced as light was associated with a suppression of power in alpha and beta frequency bands, particularly at posterior electrodes. A similar pattern was found when participants lifted the light object, but over more diffuse electrodes. Moreover, alpha and beta bands at right parieto-occipital electrodes were sensitive to the type of gesture observed (iconic vs. deictic). These results demonstrate that sensorimotor experience with an object affects how a gesture for that object is processed, as measured by the gesture-observer’s EEG, and suggest that different types of gestures recruit the observer’s own motor system in different ways. PMID:22910276

  15. Enhanced behavioral sensitivity to the competitive GABA agonist, gaboxadol, in transgenic mice over-expressing hippocampal extrasynaptic alpha6beta GABA(A) receptors.

    PubMed

    Saarelainen, Kati S; Ranna, Martin; Rabe, Holger; Sinkkonen, Saku T; Möykkynen, Tommi; Uusi-Oukari, Mikko; Linden, Anni-Maija; Lüddens, Hartmut; Korpi, Esa R

    2008-04-01

    The behavioral and functional significance of the extrasynaptic inhibitory GABA(A) receptors in the brain is still poorly known. We used a transgenic mouse line expressing the GABA(A) receptor alpha6 subunit gene in the forebrain under the Thy-1.2 promoter (Thy1alpha6) mice ectopically expressing alpha6 subunits especially in the hippocampus to study how extrasynaptically enriched alphabeta(gamma2)-type receptors alter animal behavior and receptor responses. In these mice extrasynaptic alpha6beta receptors make up about 10% of the hippocampal GABA(A) receptors resulting in imbalance between synaptic and extrasynaptic inhibition. The synthetic GABA-site competitive agonist gaboxadol (4,5,6,7-tetrahydroisoxazolo[5,4-c]pyridin-3-ol; 3 mg/kg) induced remarkable anxiolytic-like response in the light : dark exploration and elevated plus-maze tests in Thy1alpha6 mice, while being almost inactive in wild-type mice. The transgenic mice also lost quicker and for longer time their righting reflex after 25 mg/kg gaboxadol than wild-type mice. In hippocampal sections of Thy1alpha6 mice, the alpha6beta receptors could be visualized autoradiographically by interactions between gaboxadol and GABA via [(35)S]TBPS binding to the GABA(A) receptor ionophore. Gaboxadol inhibition of the binding could be partially prevented by GABA. Electrophysiology of recombinant GABA(A) receptors revealed that GABA was a partial agonist at alpha6beta3 and alpha6beta3delta receptors, but a full agonist at alpha6beta3gamma2 receptors when compared with gaboxadol. The results suggest strong behavioral effects via selective pharmacological activation of enriched extrasynaptic alphabeta GABA(A) receptors, and the mouse model represents an example of the functional consequences of altered balance between extrasynaptic and synaptic inhibition.

  16. Serum cholesterol selectively regulates glucocorticoid sensitivity through activation of JNK.

    PubMed

    Yang, Nan; Caratti, Giorgio; Ince, Louise M; Poolman, Toryn M; Trebble, Peter J; Holt, Cathy M; Ray, David W; Matthews, Laura C

    2014-11-01

    Glucocorticoids (Gc) are potent anti-inflammatory agents with wide clinical application. We have previously shown that increased serum concentration significantly attenuates regulation of a simple Gc-responsive reporter. We now find that glucocorticoid receptor (GR) regulation of some endogenous transactivated but not transrepressed genes is impaired, suggesting template specificity. Serum did not directly affect GR expression, activity or trafficking, implicating GR crosstalk with other signalling pathways. Indeed, a JNK inhibitor completely abolished the serum effect. We identified the Gc modulating serum component as cholesterol. Cholesterol loading mimicked the serum effect, which was readily reversed by JNK inhibition. Chelation of serum cholesterol with methyl-β-cyclodextrin or inhibition of cellular cholesterol synthesis with simvastatin potentiated the Gc response. To explore the effect in vivo we used ApoE(-/-) mice, a model of hypercholesterolaemia. Consistent with our in vitro studies, we find no impact of elevated cholesterol on the expression of GR, or on the hypothalamic-pituitary-adrenal axis, measured by dexamethasone suppression test. Instead we find selective Gc resistance on some hepatic target genes in ApoE(-/-) mice. Therefore, we have discovered an unexpected role for cholesterol as a selective modulator of Gc action in vivo. Taken together these findings reveal a new environmental constraint on Gc action with relevance to both inflammation and cancer.

  17. Proposed experimental method to determine alpha sensitivity of splitting between ground and 7.6 eV isomeric states in 229Th.

    PubMed

    Berengut, J C; Dzuba, V A; Flambaum, V V; Porsev, S G

    2009-05-29

    The 7.6 eV electromagnetic transition between the nearly degenerate ground state and first excited state in the 229Th nucleus may be very sensitive to potential changes in the fine-structure constant, alpha=e2/variant Planck's over 2pic. However, the sensitivity is not known, and nuclear calculations are currently unable to determine it. We propose measurements of the differences of atomic transition frequencies between thorium atoms (or ions) with the nucleus in the ground state and in the first excited (isomeric) state. This will enable extraction of the change in nuclear charge radius and electric-quadrupole moment between the isomers, and hence the alpha dependence of the isomeric transition frequency with reasonable accuracy.

  18. Sensitivity studies on sensor selection for crack growth investigation

    NASA Astrophysics Data System (ADS)

    Soni, Sunilkumar; Chattopadhyay, Aditi

    2010-10-01

    This work focuses on an unsupervised, data driven, modified k-nearest neighborhood technique to detect and monitor fatigue crack growth in lug joint samples using a surface mounted piezoelectric sensor network. A lug joint is an important structural hotspot in which damage initiates and progresses under fatigue loading. Early detection of fatigue cracks in a lug joint can help in taking preventive measures, thus avoiding any possible structural failure. The lug joint samples used in this study are prepared from an Al 6061 T6 plate with 0.25 inch thickness and are instrumented with a surface mounted piezoelectric actuator/sensor network. Experiments are conducted on lug samples with a single notch and multiple notches that are symmetrically placed. For early initiation of cracks, samples are notched at the shoulders. Under the influence of fatigue loading, the crack growth rate is different even when the notches are symmetrically placed. It is found that although cracks propagate from both the notches, the sample fails from one of the shoulders once the critical crack length is reached. For the given sensor architecture, which is symmetric, the objective of this study is to detect, isolate and monitor fatigue crack growth in each zone. The methodology presented helps in identifying sensors that are most sensitive to the presence of single and multiple cracks. Thus, the computational expense for damage localization studies can be reduced by not making use of redundant sensors.

  19. A Selective Na(+) Aptamer Dissected by Sensitized Tb(3+) Luminescence.

    PubMed

    Zhou, Wenhu; Ding, Jinsong; Liu, Juewen

    2016-08-17

    A previous study of two RNA-cleaving DNAzymes, NaA43 and Ce13d, revealed the possibility of a common Na(+) aptamer motif. Because Na(+) binding to DNA is a fundamental biochemical problem, the interaction between Ce13d and Na(+) was studied in detail by using sensitized Tb(3+) luminescence spectroscopy. Na(+) displaces Tb(3+) from the DNAzyme, and thus quenches the emission from Tb(3+) . The overall requirement for Na(+) binding includes the hairpin and the highly conserved 16-nucleotide loop in the enzyme strand, along with a few unpaired nucleotides in the substrate. Mutation studies indicate good correlation between Na(+) binding and cleavage activity, thus suggesting a critical role of Na(+) binding for the enzyme activity. Ce13d displayed a Kd of ∼20 mm with Na(+) (other monovalent cations: 40-60 mm). The Kd values for other metal ions are mainly due to non-specific competition. With a single nucleotide mutation, the specific Na(+) binding was lost. Another mutant improved Kd to 8 mm with Na(+) . This study has demonstrated a Na(+) aptamer with important biological implications and analytical applications. It has also defined the structural requirements for Na(+) binding and produced an improved mutant. PMID:27238890

  20. Ockham's razor and selective androgen receptor modulators (SARMs): are we overlooking the role of 5alpha-reductase?

    PubMed

    Gao, Wenqing; Dalton, James T

    2007-02-01

    Selective Androgen Receptor Modulators (SARMs) are a novel class of AR ligands that possess tissue-selective pharmacological activities. SARMs of various chemical structures have been discovered and characterized, and lead compounds with much improved specificity for AR, in vivo pharmacokinetic profiles, and higher degree of tissue selectivity have entered clinical development, and are expected to dramatically expand the clinical applications of androgens. With the rapid progress in SARM discovery and increasing demand for mechanism-based drug design, more and more research efforts have been devoted to the mechanisms of action of the observed tissue selectivity of SARMs. There is increasing enthusiasm in adapting the molecular mechanisms of action from SERM research to the SARM field; however, is the SARM story really so complicated? The tissue-specific expression of 5alpha-reductase might provide a simple explanation for this puzzle.

  1. Design of beta-domain swapping, alpha/beta-protein, environmentally sensitive coiled coil and peptide functionalized titania materials

    NASA Astrophysics Data System (ADS)

    Nagarkar, Radhika P.

    2009-12-01

    The objective of this dissertation is to apply rational peptide design to fabricate nanomaterials via self-assembly. This has been demonstrated in structurally diverse systems with an aim of deciphering the underlying principles governing how sequence affects the peptide's ability to adopt a specific secondary structure and ultimate material properties that are realized from the association of these secondary structural elements. Several amyloidogenic proteins have been shown to self-assemble into fibrils using a mechanism known as domain swapping. Here, discreet units of secondary structure are exchanged among discreet proteins during self-assembly to form extended networks with precise three dimensional organization. The possibility of using these mechanisms to design peptides capable of controlled assembly and fibril formation leading to materials with targeted properties is explored. By altering the placement of a beta-turn sequence that varies the size and location of the exchanged strand, twisting, non-twisting and laminated fibrillar nanostructures are obtained. Hydrogels prepared from these strand swapping beta-hairpins have varied rheological properties due to differences in their fibrillar nanostructures. In a second distinct design, alpha/beta-proteins are used to prepare environmentally sensitive hydrogels. Here, multiple distinct motifs for structural integrity and dynamic response within a single self-assembling peptide allow the amyloid-like fibrils formed to controllably alter their nano-topography in response to an external stimulus such as temperature. The development of these self-assembling alpha/beta-protein motifs also necessitated the design of pH sensitive antiparallel coiled coils. Exploring the basic principles responsible for pH dependent conformational changes in coiled coils can lead to new insights in the control of protein structure and function. Lastly, this dissertation discusses the interface between biomolecules and inorganic

  2. Structure of alpha6 beta3 delta GABA(A) receptors and their lack of ethanol sensitivity.

    PubMed

    Baur, Roland; Kaur, Kuldeep H; Sigel, Erwin

    2009-12-01

    Delta (delta) subunit containing GABA(A) receptors are expressed extra-synaptically and mediate tonic inhibition. In cerebellar granule cells, they often form a receptor together with alpha(6) subunits. We were interested to determine the architecture of these receptors. We predefined the subunit arrangement of 24 different GABA(A) receptor pentamers by subunit concatenation. These receptors (composed of alpha(6), beta(3) and delta subunits) were expressed in Xenopus oocytes and their electrophysiological properties analyzed. Currents elicited in response to GABA were determined in presence and absence of 3alpha, 21-dihydroxy-5alpha-pregnan-20-one and to 4,5,6,7-tetrahydroisoxazolo[5,4-c]-pyridin-3-ol. alpha(6)-beta(3)-alpha(6)/delta receptors showed a substantial response to GABA alone. Three receptors, beta(3)-alpha(6)-delta/alpha(6)-beta(3), alpha(6)-beta(3)-alpha(6)/beta(3)-delta and beta(3)-delta-beta(3)/alpha(6)-beta(3), were only uncovered in the combined presence of the neurosteroid 3alpha, 21-dihydroxy-5alpha-pregnan-20-one with GABA. All four receptors were activated by 4,5,6,7-tetrahydroisoxazolo[5,4-c]-pyridin-3-ol. None of the functional receptors was modulated by physiological concentrations (up to 30 mM) of ethanol. GABA concentration response curves indicated that the delta subunit can contribute to the formation of an agonist site. We conclude from the investigated receptors that the delta subunit can assume multiple positions in a receptor pentamer composed of alpha(6), beta(3) and delta subunits.

  3. Correction: A Co(2+)-selective and chirality-sensitive supermolecular metallohydrogel with a nanofiber network skeleton.

    PubMed

    Wang, Xiaojuan; He, Ting; Yang, Lan; Wu, Huiqiong; Zhang, Rui; Zhang, Zhenzhu; Shen, Rujuan; Xiang, Juan; Zhang, Yi; Wei, Chuanwan

    2016-04-21

    Correction for 'A Co(2+)-selective and chirality-sensitive supermolecular metallohydrogel with a nanofiber network skeleton' by Xiaojuan Wang et al., Nanoscale, 2016, DOI: 10.1039/c6nr00822d. PMID:27021200

  4. Highly sensitive and selective bioluminescence based ozone probes and their applications to detect ambient ozone.

    PubMed

    Nam, Younseok; Kim, Beom Seok; Shin, Injae

    2016-01-21

    Highly selective and sensitive bioluminescence based probes, which respond to ozone but not to other ROS, have been developed. These probes were used to determine ozone concentrations in environmental samples.

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

    SciTech Connect

    James, W.M.; Emerick, M.C.; Agnew, W.S. )

    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.

  6. IFN-{gamma} sensitizes MIN6N8 insulinoma cells to TNF-{alpha}-induced apoptosis by inhibiting NF-{kappa}B-mediated XIAP upregulation

    SciTech Connect

    Kim, Hun Sik; Kim, Sunshin; Lee, Myung-Shik . E-mail: mslee@smc.samsung.co.kr

    2005-10-28

    Although X-linked inhibitor of apoptosis protein (XIAP) is an important intracellular suppressor of apoptosis in a variety of cell types, its role in cytokine-induced pancreatic {beta}-cell apoptosis remains unclear. Here, we found that: (i) XIAP level was inversely correlated with tumor necrosis factor (TNF)-{alpha}-induced apoptosis in MIN6N8 insulinoma cells; (ii) adenoviral XIAP overexpression abrogated the TNF-{alpha}-induced apoptosis through inhibition of caspase activity; (iii) downregulation of XIAP by antisense oligonucleotide or Smac peptide sensitized MIN6N8 cells to TNF-{alpha}-induced apoptosis; (iv) XIAP expression was induced by TNF-{alpha} through a nuclear factor-{kappa}B (NF-{kappa}B)-dependent pathway, and interferon (IFN)-{gamma} prevented such an induction in a manner independent of NF-{kappa}B, which presents a potential mechanism underlying cytotoxic IFN-{gamma}/TNF-{alpha} synergism. Taken together, our results suggest that XIAP is an important modulator of TNF-{alpha}-induced apoptosis of MIN6N8 cells, and XIAP regulation in pancreatic {beta}-cells might play an important role in pancreatic {beta}-cell apoptosis and in the pathogenesis of type 1 diabetes.

  7. Sensitive amperometric immunosensor for alpha-fetoprotein based on carbon nanotube/gold nanoparticle doped chitosan film.

    PubMed

    Lin, Jiehua; He, Chunyan; Zhang, Lijuan; Zhang, Shusheng

    2009-01-01

    A novel strategy for the fabrication of sensitive immunosensor to detect alpha-fetoprotein (AFP) in human serum has been proposed. The immunosensor was prepared by immobilizing AFP antigen onto the glassy carbon electrode (GC) modified by gold nanoparticles and carbon nanotubes doped chitosan (GNP/CNT/Ch) film. GNP/CNT hybrids were produced by one-step synthesis based on the direct redox reaction. The electrochemical properties of GNP/CNT/Ch films were characterized by impedance spectroscopy and cyclic voltammetry. It was indicated that GNP/CNT nanohybrid acted as an electron promoter and accelerated the electron transfer. Sample AFP, immobilized AFP, and alkaline phosphatase (ALP)-labeled antibody were incubated together for the determination based on a competitive immunoassay format. After the immunoassay reaction, the bound ALP label on the modified GC led to an amperometric response of 1-naphthyl phosphate (1-NP), which was changed with the different antigen concentrations in solution. Under the optimized experimental conditions, the resulting immunosensor could detect AFP in a linear range from 1 to 55 ng ml(-1) with a detection limit of 0.6 ng ml(-1). The proposed immunosensor, by using GNP/CNT/Ch as the immobilization matrix of AFP, offers an excellent amperometric response of ALP-anti-AFP to 1-NP. The immunosensor provided a new alternative to the application of other antigens or other bioactive molecules. PMID:18848914

  8. Highly sensitive, label-free and real-time detection of alpha-fetoprotein using a silicon nanowire biosensor.

    PubMed

    Zhou, Fan; Li, Zengyao; Bao, Zengtao; Feng, Kang; Zhang, Ye; Wang, Tong

    2015-11-01

    Alpha-fetoprotein (AFP) is a tumor-associated fetal protein that can be expressed in large amounts in adult tumor cells, serving as a useful clinical tumor-marker. Silicon nanowire (SiNW) biosensors have emerged as a powerful tool in detecting protein biomarkers, due to their ultrahigh sensitivity, real-time response and label-free detection. We fabricated a SiNW-based field-effect transistor (FET) according to "top-down" methodology. First, anti-AFP antibodies were immobilized onto the surface of the SiNW-FET. A polydimethylsiloxane (PDMS) microchannel was then integrated to the modified SiNW-FET. Various concentrations of AFP were then pumped through the sensing area. We observed a current change that corresponded to binding of AFP onto the surface of our anti-AFP functionalized SiNW-FET biosensor. Concentrations of AFP as low as 0.1 ng/mL were detected. The results implicate our SiNW biosensor as an effective AFP biomarker detector with promising potential in clinical applications. PMID:26205419

  9. Correlation of the ionisation response at selected points of IC sensitive regions with SEE sensitivity parameters under pulsed laser irradiation

    SciTech Connect

    Gordienko, A V; Mavritskii, O B; Egorov, A N; Pechenkin, A A; Savchenkov, D V

    2014-12-31

    The statistics of the ionisation response amplitude measured at selected points and their surroundings within sensitive regions of integrated circuits (ICs) under focused femtosecond laser irradiation is obtained for samples chosen from large batches of two types of ICs. A correlation between these data and the results of full-chip scanning is found for each type. The criteria for express validation of IC single-event effect (SEE) hardness based on ionisation response measurements at selected points are discussed. (laser applications and other topics in quantum electronics)

  10. H-alpha and H-beta Photometry of Selected Open Clusters

    NASA Astrophysics Data System (ADS)

    Joner, Michael D.; Hintz, E. G.

    2012-05-01

    We present new data for the open clusters M35, M67, NGC 188, NGC 752, NGC 869, and NGC 884 secured at the Brigham Young University West Mountain Observatory using standard H-beta filters as well as standard filters for the recently defined BYU H-alpha photometric system. Aside from being well suited for surveys and studies of emmision line objects such as HMXBs and YSOs, the combination of these two reddening independent indices provides a reliable temperature measure for individual stars within a cluster with the H-alpha index reaching stars of slightly cooler temperature than can be found from H-beta measurements. We acknowledge the Brigham Young University College of Physical and Mathematical Sciences for continued support of the research programs at the West Mountain Observatory. Partial supprot for this research was received through NSF grant AST #0618209.

  11. The amiloride-sensitive epithelial sodium channel alpha-subunit is transcriptionally down-regulated in rat parotid cells by the extracellular signal-regulated protein kinase pathway.

    PubMed

    Zentner, M D; Lin, H H; Wen, X; Kim, K J; Ann, D K

    1998-11-13

    Previous studies have shown that an inducible Raf-1 kinase protein, DeltaRaf-1:ER, activates the mitogen-activated protein kinase/extracellular signal-regulated protein kinase (ERK)-signaling pathway, which is required for the transformation of the rat salivary epithelial cell line, Pa-4. Differential display polymerase chain reaction was employed to search for mRNAs repressed by DeltaRaf-1:ER activation. Through this approach, the gene encoding the alpha-subunit of the amiloride-sensitive epithelial sodium channel (alpha-ENaC) was identified as a target of activated Raf-1 kinases. alpha-ENaC down-regulation could also be seen in cells treated with 12-O-tetradecanoyl-1-phorbol-13-acetate (TPA), indicating that the repression of steady-state alpha-ENaC mRNA level was dependent upon the activity of protein kinase C, the target of TPA, as well. Pretreatment of cells with a specific inhibitor of the ERK kinase pathway, PD 98059, markedly abolished the down-regulation of alpha-ENaC expression, consistent with the hypothesis that the ERK kinase-signaling pathway is involved in TPA-mediated repression. Moreover, through the use of transient transfection assays with alpha-ENaC-reporter and activated Raf expression construct(s), we provide the first evidence that activation of the ERK pathway down-regulates alpha-ENaC expression at the transcriptional level. Elucidating the molecular programming that modulates the expression of the alpha-subunit may provide new insights into the modulation of sodium reabsorption across epithelia. PMID:9804854

  12. Oxidative stress disrupts glucocorticoid hormone-dependent transcription of the amiloride-sensitive epithelial sodium channel alpha-subunit in lung epithelial cells through ERK-dependent and thioredoxin-sensitive pathways.

    PubMed

    Wang, H C; Zentner, M D; Deng, H T; Kim, K J; Wu, R; Yang, P C; Ann, D K

    2000-03-24

    The amiloride-sensitive epithelial Na(+) channel (ENaC) plays a critical role in the maintenance of alveolar fluid balance. It is generally accepted that reactive oxygen and nitrogen species can inhibit ENaC activity and aggravate acute lung injury; however, the molecular mechanism for free radical-mediated ENaC inhibition is unclear. Previously, we showed that the expression of the alpha-subunit of ENaC, alpha-ENaC, which is indispensable for ENaC activity, is repressed by Ras activation in salivary epithelial cells. Here, we investigated whether exogenous H(2)O(2) modulates alpha-ENaC gene expression in lung epithelial cells through a similar molecular mechanism. Utilizing transient transfection reporter assays and site-directed mutagenesis analyses, we found that the glucocorticoid response element (GRE), located at -1334 to -1306 base pairs of the alpha-ENaC 5'-flanking region, is the major enhancer for the stimulated alpha-ENaC expression in A549 lung epithelial cells. We further demonstrate that the presence of an intact GRE is necessary and sufficient for oxidants to repress alpha-ENaC expression. Consistent with our hypothesis, exogenous H(2)O(2)-mediated repression of alpha-ENaC GRE activity is partially blocked by either a specific inhibitor for extracellular signal-regulated kinase (ERK) pathway activation, U0126, or dominant negative ERK, suggesting that, in part, activated ERK may mediate the repressive effects of H(2)O(2) on alpha-ENaC expression. In addition, overexpression of thioredoxin restored glucocorticoid receptor action on the alpha-ENaC GRE in the presence of exogenous H(2)O(2). Taken together, we hypothesize that oxidative stress impairs Na(+) transport activity by inhibiting dexamethasone-dependent alpha-ENaC GRE activation via both ERK-dependent and thioredoxin-sensitive pathways. These results suggest a putative mechanism whereby cellular redox potentials modulate the glucocorticoid receptor/dexamethasone effect on alpha-ENaC expression

  13. Selection on Inversion Breakpoints Favors Proximity to Pairing Sensitive Sites in Drosophila melanogaster.

    PubMed

    Corbett-Detig, Russell B

    2016-09-01

    Chromosomal inversions are widespread among taxa, and have been implicated in a number of biological processes including adaptation, sex chromosome evolution, and segregation distortion. Consistent with selection favoring linkage between loci, it is well established that length is a selected trait of inversions. However, the factors that affect the distribution of inversion breakpoints remain poorly understood. "Sensitive sites" have been mapped on all euchromatic chromosome arms in Drosophila melanogaster, and may be a source of natural selection on inversion breakpoint positions. Briefly, sensitive sites are genomic regions wherein proximal structural rearrangements result in large reductions in local recombination rates in heterozygotes. Here, I show that breakpoints of common inversions are significantly more likely to lie within a cytological band containing a sensitive site than are breakpoints of rare inversions. Furthermore, common inversions for which neither breakpoint intersects a sensitive site are significantly longer than rare inversions, but common inversions whose breakpoints intersect a sensitive site show no evidence for increased length. I interpret these results to mean that selection favors inversions whose breakpoints disrupt synteny near to sensitive sites, possibly because these inversions suppress recombination in large genomic regions. To my knowledge this is the first evidence consistent with positive selection acting on inversion breakpoint positions. PMID:27343234

  14. Occurrence of poly(alpha2,8-deaminoneuraminic acid) in mammalian tissues: widespread and developmentally regulated but highly selective expression on glycoproteins.

    PubMed Central

    Ziak, M; Qu, B; Zuo, X; Zuber, C; Kanamori, A; Kitajima, K; Inoue, S; Inoue, Y; Roth, J

    1996-01-01

    In tissues of higher organisms homopolymers of alpha2,8-linked N-acetylneuraminic acid can be found as a posttranslational modification on selected proteins. We report here the discovery of homopolymers of alpha2,8-linked deaminoneuraminic acid [poly(alpha2,8-KDN)] in various tissues derived from all three germ layers in vertebrates including mammals. The monoclonal antibody kdn8kdn in conjunction with a bacterial KDNase permitted the detection of poly(alpha2,8-KDN) by immunohistochemistry and immunoblotting. Further evidence for the existence of poly(alpha2,8-KDN) was obtained by gas/liquid chromatography. The poly(alpha2,8-KDN) glycan was detectable in all tissues studied with the exception of mucus-producing cells present in various organs, the extracellular matrix, and basement membranes. However, in certain organs such as muscle, kidney, lung, and brain its expression was developmentally regulated. Despite its widespread tissue distribution, the poly(alpha2,8-KDN) glycan was detected on a single 150-kDa glycoprotein except for a single >350-kDa glycoprotein in kidney, which makes it most distinctive among polysialic acids. The ubiquitous yet selective expression may be indicative of a general function of the poly(alpha2,8-KDN)-bearing glycoproteins. Images Fig. 1 Fig. 2 Fig. 4 Fig. 5 Fig. 6 PMID:8610115

  15. Estrogen Receptors Alpha (ERα) and Beta (ERβ): Subtype-Selective Ligands and Clinical Potential

    PubMed Central

    Paterni, Ilaria; Granchi, Carlotta; Katzenellenbogen, John A.; Minutolo, Filippo

    2014-01-01

    Estrogen receptors alpha (ERα) and beta (ERβ) are nuclear transcription factors that are involved in the regulation of many complex physiological processes in humans. Modulation of these receptors by prospective therapeutic agents is currently being considered for prevention and treatment of a wide variety of pathological conditions, such as, cancer, metabolic and cardiovascular diseases, neurodegeneration, inflammation, and osteoporosis. This review provides an overview and update of compounds that have been recently reported as modulators of ERs, with a particular focus on their potential clinical applications. PMID:24971815

  16. Thyroid hormone regulation of transmembrane signalling in neonatal rat ventricular myocytes by selective alteration of the expression and coupling of G-protein alpha-subunits.

    PubMed Central

    Bahouth, S W

    1995-01-01

    Thyroid hormone exerts profound effects on the activity of the hormone-sensitive adenylate cyclase system in the heart. Distinct guanine nucleotide-binding regulatory proteins (G-proteins) mediate stimulatory and inhibitory influences on adenylate cyclase activity. To examine whether the effects of thyroid hormone on adenylate cyclase involve specific changes in G-protein subunit expression, the influence of tri-iodothyronine (T3) on the biosynthesis and activity of G-proteins in neonatal rat ventricular myocytes was determined. In myocytes challenged with T3 for 5 days, Gs alpha levels increased by 4 +/- 0.5-fold, whereas Gi2 alpha levels declined by more than 80%. T3 down-regulated Gi2 alpha mRNA by 60% within 3 days, but had no effect on Gs alpha mRNA. The basis for the decline in Gi2 alpha mRNA was the T3-mediated suppression of Gi2 alpha gene transcription by 80 +/- 9% within 4 h. The decline in Gi2 alpha mRNA in response to T3 produced a 2-fold decrease in relative rate of synthesis of Gi2 alpha but not in its half-life (46 +/- 7 h). Gs alpha synthesis was not altered by T3, but the half-life of Gs alpha increased from 50 +/- 6 h in control cells to 72 +/- 8 h in T3-treated cells. In addition, T3 provoked the translocation of Gs alpha from the cytoplasmic to the membranous compartment. Membranous Gs alpha increased from 30 +/- 6% to 61 +/- 7% of total cellular Gs alpha, whereas cytoplasmic Gs alpha declined from 68 +/- 6% to 33 +/- 8% within 1 day of exposure to T3. T3-mediated up-regulation of Gs alpha enhanced the activation of myocardial adenylate cyclase by the stimulatory pathway whereas the down-regulation of Gi2 alpha attenuated the deactivation of myocardial adenylate cyclase by the inhibitory pathway. Images Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 PMID:7741715

  17. Simulating alpha/beta selectivity at the human thyroid hormone receptor: consensus scoring using multidimensional QSAR.

    PubMed

    Vedani, Angelo; Zumstein, Martin; Lill, Markus A; Ernst, Beat

    2007-01-01

    We present a consensus-scoring study on the human thyroid hormone receptor alpha and beta using two receptor-modeling concepts (software Quasar and Raptor) that are based on multidimensional QSAR and allow for the explicit simulation of induced fit. The binding mode of 82 agonists and indirect antagonists, spanning an activity range of seven orders of magnitude in K(i), was identified through flexible docking to the respective X-ray crystal structures (Yeti software) and represented by a 4D data set with up to four conformations per compound. The receptor surrogates for the thyroid alpha receptor converged at a cross-validated r(2) of 0.846/0.919 (64 training compounds; for Quasar and Raptor, respectively) and yielded a predictive r(2) of 0.812/0.814 (18 test compounds); the models for the thyroid beta receptor resulted in a cross-validated r(2) of 0.823/0.909 and a predictive r(2) of 0.665/0.796, respectively. Consensus was achieved as, on average, the calculated activities of the training set differ only by a factor of 2.2 in K(i) and those of the test set by a factor of 2.8 when predicted by Quasar and Raptor, respectively.

  18. High-sensitivity C-reactive protein levels in cerebrospinal fluid and serum in severe head injury: relationship to tumor necrosis factor-alpha and interleukin-6.

    PubMed

    Is, Merih; Coskun, Abdurrahman; Sanus, Galip Zihni; Tanriverdi, Taner; Kafadar, Ali Metin; Hanimoglu, Hakan; Tanriover, Necmettin; Gezen, Ferruh; Uzan, Mustafa

    2007-12-01

    Recent studies have demonstrated the role of high-sensitivity C-reactive protein (hsCRP) in inflammatory diseases; however, it is unclear whether this molecule has a role after severe head injury (SHI). Our aim was to evaluate the levels of hsCRP in both cerebrospinal fluid (CSF) and serum from patients after SHI. The study focused on 11 patients with SHI, and evaluated CSF and serum levels of hsCRP, tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6) in a 10-day period following the head trauma. The values were compared with those from nine control patients, who had normal pressure hydrocephalus. In the CSF and serum of the patients after SHI, HsCRP was found to be significantly higher, at all times, than in the controls; TNF-alpha and IL-6 levels were also higher in these patients. However, hsCRP levels did not correlate with either TNF-alpha or IL-6. TNF-alpha and IL-6 increased during the period immediately following the SHI, and intrathecal levels were always higher than those of the serum. This study demonstrates for the first time that hsCRP reaches high levels in both CSF and serum in patients with SHI, and it may therefore be used as an inflammatory index. This finding suggests a need for further studies in this area, which are larger in scope than the present study.

  19. Na(+)-, ouabain-, Ca(2+)-, and thapsigargin-sensitive ATPase activity expressed in chimeras between the calcium and the sodium pump alpha subunits.

    PubMed Central

    Ishii, T; Lemas, M V; Takeyasu, K

    1994-01-01

    Using the chicken sarcoplasmic/endoplasmic reticulum Ca2+ (SERCA)-ATPase as a parental molecule and replacing various portions with the corresponding portions of the chicken Na+,K(+)-ATPase alpha 1 subunit, Ca2+/thapsigargin- and Na+/ouabain-sensitive domains critical for these P-type ATPase activities were identified. In the chimera, [n/c]CC, the amino-terminal amino acids Met-1 to Asp-162 of the SERCA (isoform 1) (SERCA1) ATPase were replaced with the corresponding portion (Met-1-Asp-200) of the Na+,K(+)-ATPase alpha 1 subunit. In the chimera CC[c/n], the carboxyl-terminal amino acids (Ser-830 to COOH) of the SERCA1 ATPase were replaced with the corresponding segment (Leu-861 to COOH) of the Na+,K(+)-ATPase alpha 1 subunit, and in the chimera CNC, the middle part (Gly-354-Lys-712) of the SERCA1 ATPase was exchanged with the Na+,K(+)-ATPase alpha 1 subunit (Gly-378-Lys-724). None of the chimeric molecules exhibited any detectable ouabain-sensitive Na+,K(+)-ATPase activity, but they did exhibit thapsigargin-sensitive Ca(2+)-ATPase activity. Therefore, the segments Ile-163-Gly-354 and Lys-712-Ser-830 of the SERCA1 ATPase are sufficient for Ca2+ and thapsigargin sensitivity. The SERCA1-ATPase activity of [n/c]CC, but not of CCC, CNC, or CC[c/n], was further stimulated by addition of Na+ in the assay medium containing Ca2+. This additional stimulation of SERCA1-ATPase activity by Na+ was abolished when the amino-terminal region (Met-1-Leu-69) of [n/c]CC was deleted ([delta n/c]CC). In the absence of Na+, the SERCA1-ATPase activity of [n/c]CC was inhibited by ouabain, and, in the presence of Na+, its activity was stimulated by this drug. On the other hand, the ATPase activity of [delta n/c]CC was not affected by ouabain, although [delta n/c]CC can still bind [3H]ouabain. These results suggest that a distinct Na(+)-sensitive domain (Na+ sensor) located within the restricted amino-terminal region (Met-1-Leu-69) of the Na+,K(+)-ATPase alpha 1 subunit regulates ATPase

  20. Synthesis and biological evaluation of fluorine-18 labeled RS-15385-197 analogs: Potent and selective alpha-2 adrenergic receptor radioligands for PET

    SciTech Connect

    Enas, J.D.; VanBrocklin, H.F.; Budinger, T.F.; Clark, R.D.

    1997-12-31

    Aberrations in the {alpha}{sub 2}-adrenergic receptor system have been implicated in a number of disease states including hypertension, drug abuse, depression, and neurodegenerative disorders such as Alzheimer`s Disease. RS-15385-FP (1) and RS-15385-FPh (2) are analogs of the {alpha}{sub 2}-adrenergic receptor antagonist RS- 15385-197 which display a high receptor binding affinity (K{sub i} = 0.2 and 0.5 nM, respectively) as well as a high degree of {alpha}{sub 2}/{alpha}{sub 1} selectivity (7000:1 and 2000:1, respectively). We synthesized [F-18]-2 was synthesized by fluoro-for-nitro exchange on the corresponding nitrophenyl derivative which was produced in two steps from the hydroxypropyl sulfonamide. In vivo distribution studies in rats and PET studies in monkeys demonstrate uptake in {alpha}{sub 2}-adrenergic receptor rich regions of the brain, particularly the locus coeruleus.

  1. Resistance to extinction of fear-relevant stimuli: preparedness or selective sensitization?

    PubMed

    Lovibond, P F; Siddle, D A; Bond, N W

    1993-12-01

    Human conditioning research has revealed an apparent resistance to extinction of aversive conditioning to pictures of fear-relevant stimuli such as snakes and spiders, supporting M. E. P. Seligman's (1971) preparedness theory of fears and phobias. This article examines an alternative account based on activation of preexisting response tendencies under threat (selective sensitization). Two experiments demonstrate that selective sensitization of electrodermal responses is attenuated when a fear-relevant stimulus serves as a negative conditioned stimulus (CS-), but is maintained when it serves as a positive conditioned stimulus (CS+). Previous extinction results may therefore be due to preservation of initial responding to CS+ but not CS-. Selective sensitization offers a model for the nonassociative activation of fears and phobias to prepotent stimuli under conditions of stress or threat. Possible genetic and cognitive mechanisms are discussed.

  2. Recent progress in research on peroxisome proliferator-activated receptor alpha-selective ligands.

    PubMed

    Miyachi, Hiroyuki

    2004-08-01

    The understanding of the functions of the nuclear receptor peroxisome proliferator-activated receptor a (PPARalpha) as a regulator of lipid and lipoprotein homeostasis, and the rapid development of parallel high-throughput screening assays to evaluate the activity toward other PPAR subtypes (PPARdelta and PPARgamma), have provided an opportunity to develop novel PPARalpha-selective, PPARalpha/gamma dual and PPAR pan agonists for the treatment of various metabolic diseases. This review focuses on the molecular pharmacology of PPARalpha, and summarizes recent literature and patent applications disclosing medicinal chemistry strategies to identify new PPARalpha-selective agonists. The species selectivity of some classes of PPARalpha-selective agonists in response to in vitro PPARalpha transactivation activity is also reported. PMID:15334308

  3. Development of Solution Algorithm and Sensitivity Analysis for Random Fuzzy Portfolio Selection Model

    NASA Astrophysics Data System (ADS)

    Hasuike, Takashi; Katagiri, Hideki

    2010-10-01

    This paper focuses on the proposition of a portfolio selection problem considering an investor's subjectivity and the sensitivity analysis for the change of subjectivity. Since this proposed problem is formulated as a random fuzzy programming problem due to both randomness and subjectivity presented by fuzzy numbers, it is not well-defined. Therefore, introducing Sharpe ratio which is one of important performance measures of portfolio models, the main problem is transformed into the standard fuzzy programming problem. Furthermore, using the sensitivity analysis for fuzziness, the analytical optimal portfolio with the sensitivity factor is obtained.

  4. Alpha-helical hydrophobic polypeptides form proton-selective channels in lipid bilayers.

    PubMed Central

    Oliver, A E; Deamer, D W

    1994-01-01

    Proton translocation is important in membrane-mediated processes such as ATP-dependent proton pumps, ATP synthesis, bacteriorhodopsin, and cytochrome oxidase function. The fundamental mechanism, however, is poorly understood. To test the theoretical possibility that bundles of hydrophobic alpha-helices could provide a low energy pathway for ion translocation through the lipid bilayer, polyamino acids were incorporated into extruded liposomes and planar lipid membranes, and proton translocation was measured. Liposomes with incorporated long-chain poly-L-alanine or poly-L-leucine were found to have proton permeability coefficients 5 to 7 times greater than control liposomes, whereas short-chain polyamino acids had relatively little effect. Potassium permeability was not increased markedly by any of the polyamino acids tested. Analytical thin layer chromatography measurements of lipid content and a fluorescamine assay for amino acids showed that there were approximately 135 polyleucine or 65 polyalanine molecules associated with each liposome. Fourier transform infrared spectroscopy indicated that a major fraction of the long-chain hydrophobic peptides existed in an alpha-helical conformation. Single-channel recording in both 0.1 N HCl and 0.1 M KCl was also used to determine whether proton-conducting channels formed in planar lipid membranes (phosphatidylcholine/phosphatidylethanolamine, 1:1). Poly-L-leucine and poly-L-alanine in HCl caused a 10- to 30-fold increase in frequency of conductive events compared to that seen in KCl or by the other polyamino acids in either solution. This finding correlates well with the liposome observations in which these two polyamino acids caused the largest increase in membrane proton permeability but had little effect on potassium permeability. Poly-L-leucine was considerably more conductive than poly-L-alanine due primarily to larger event amplitudes and, to a lesser extent, a higher event frequency. Poly-L-leucine caused two

  5. Alpha-helical hydrophobic polypeptides form proton-selective channels in lipid bilayers

    NASA Technical Reports Server (NTRS)

    Oliver, A. E.; Deamer, D. W.

    1994-01-01

    Proton translocation is important in membrane-mediated processes such as ATP-dependent proton pumps, ATP synthesis, bacteriorhodopsin, and cytochrome oxidase function. The fundamental mechanism, however, is poorly understood. To test the theoretical possibility that bundles of hydrophobic alpha-helices could provide a low energy pathway for ion translocation through the lipid bilayer, polyamino acids were incorporated into extruded liposomes and planar lipid membranes, and proton translocation was measured. Liposomes with incorporated long-chain poly-L-alanine or poly-L-leucine were found to have proton permeability coefficients 5 to 7 times greater than control liposomes, whereas short-chain polyamino acids had relatively little effect. Potassium permeability was not increased markedly by any of the polyamino acids tested. Analytical thin layer chromatography measurements of lipid content and a fluorescamine assay for amino acids showed that there were approximately 135 polyleucine or 65 polyalanine molecules associated with each liposome. Fourier transform infrared spectroscopy indicated that a major fraction of the long-chain hydrophobic peptides existed in an alpha-helical conformation. Single-channel recording in both 0.1 N HCl and 0.1 M KCl was also used to determine whether proton-conducting channels formed in planar lipid membranes (phosphatidylcholine/phosphatidylethanolamine, 1:1). Poly-L-leucine and poly-L-alanine in HCl caused a 10- to 30-fold increase in frequency of conductive events compared to that seen in KCl or by the other polyamino acids in either solution. This finding correlates well with the liposome observations in which these two polyamino acids caused the largest increase in membrane proton permeability but had little effect on potassium permeability. Poly-L-leucine was considerably more conductive than poly-L-alanine due primarily to larger event amplitudes and, to a lesser extent, a higher event frequency. Poly-L-leucine caused two

  6. Phenotypic and transcriptional response to selection for alcohol sensitivity in Drosophila melanogaster

    PubMed Central

    Morozova, Tatiana V; Anholt, Robert RH; Mackay, Trudy FC

    2007-01-01

    Background Alcoholism is a complex disorder determined by interactions between genetic and environmental risk factors. Drosophila represents a powerful model system to dissect the genetic architecture of alcohol sensitivity, as large numbers of flies can readily be reared in defined genetic backgrounds and under controlled environmental conditions. Furthermore, flies exposed to ethanol undergo physiological and behavioral changes that resemble human alcohol intoxication, including loss of postural control, sedation, and development of tolerance. Results We performed artificial selection for alcohol sensitivity for 35 generations and created duplicate selection lines that are either highly sensitive or resistant to ethanol exposure along with unselected control lines. We used whole genome expression analysis to identify 1,678 probe sets with different expression levels between the divergent lines, pooled across replicates, at a false discovery rate of q < 0.001. We assessed to what extent genes with altered transcriptional regulation might be causally associated with ethanol sensitivity by measuring alcohol sensitivity of 37 co-isogenic P-element insertional mutations in 35 candidate genes, and found that 32 of these mutants differed in sensitivity to ethanol exposure from their co-isogenic controls. Furthermore, 23 of these novel genes have human orthologues. Conclusion Combining whole genome expression profiling with selection for genetically divergent lines is an effective approach for identifying candidate genes that affect complex traits, such as alcohol sensitivity. Because of evolutionary conservation of function, it is likely that human orthologues of genes affecting alcohol sensitivity in Drosophila may contribute to alcohol-associated phenotypes in humans. PMID:17973985

  7. Sensitive radioimmunoassay of. cap alpha. human atrial natriuretic polypeptide using monoclonal antibody recognizing human form ring structure

    SciTech Connect

    Naomi, S.; Umeda, T.; Iwaoka, T.; Miura, F.; Ohno, M.; Sasaki, M.; Oishi, S.; Sato, T.; Takatsu, K.

    1988-01-01

    A monoclonal antibody (C351) against ..cap alpha.. human atrial natriuretic polypeptide (..cap alpha..hANP) recognizing human form ring structure was established and applied to a radioimmunoassay of plasma ..cap alpha..hANP. The minimum detectable amount in terms of 10 % radioligand displacement relative to zero dose were 0.28 fmol/tube, corresponding to 0.7 fmol/ml in plasma after extraction using Sep-Pak C18 cartridges. When the mean plasma levels at recumbent position in fasted morning were compared in 10 young (<30 years) and 10 elderly (greater than or equal to50 years) healthy subjects taking normal sodium diet, it was slightly higher in the latter. After i.v. infusion of hypertonic saline at a rate of 0.24 ml/kg/min for 20 min in 6 normal subjects (26 to 35 years), it was increased from 4.1 +/- 0.4 to 5.9 +/- 0.7 fmol/ml (p<0.01). In 6 patients with essential hypertension (34 to 57 years), it was elevated with high salt intake. From these results, the radioimmunoassay of plasma IR-..cap alpha..hANP using MAb C351 seems to be quite suitable to detect rather small changes at low plasma concentrations and to investigate a physiological importance of ..cap alpha..hANP in man.

  8. Reaction rate sensitivity of 44Ti production in massive stars and implications of a thick target yield measurement of 40Ca(alpha,gamma)44Ti

    SciTech Connect

    Hoffman, R D; Sheets, S A; Burke, J T; Scielzo, N D; Rauscher, T; Norman, E B; Tumey, S; Brown, T A; Grant, P G; Hurst, A M; Phair, L; Stoyer, M A; Wooddy, T; Fisker, J L; Bleuel, D

    2010-02-16

    We evaluate two dominant nuclear reaction rates and their uncertainties that affect {sup 44}Ti production in explosive nucleosynthesis. Experimentally we develop thick-target yields for the {sup 40}Ca({alpha},{gamma}){sup 44}Ti reaction at E{sub {alpha}} = 4.13, 4.54, and 5.36 MeV using {gamma}-ray spectroscopy. At the highest beam energy, we also performed an activation measurement which agrees with the thick target result. From the measured yields a stellar reaction rate was developed that is smaller than current statistical-model calculations and recent experimental results, which would suggest lower {sup 44}Ti production in scenarios for the {alpha}-rich freeze out. Special attention has been paid to assessing realistic uncertainties of stellar reaction rates produced from a combination of experimental and theoretical cross sections. With such methods, we also develop a re-evaluation of the {sup 44}Ti({alpha},p){sup 47}V reaction rate. Using these two rates we carry out a sensitivity survey of {sup 44}Ti synthesis in eight expansions representing peak temperature and density conditions drawn from a suite of recent supernova explosion models. Our results suggest that the current uncertainty in these two reaction rates could lead to as large an uncertainty in {sup 44}Ti synthesis as that produced by different treatments of stellar physics.

  9. A highly sensitive and selective immunoassay for the detection of tetrabromobisphenol A in soil and sediment

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Tetrabromobisphenol A (TBBPA) is the most widely used brominated flame retardant. A sensitive and selective enzyme-linked immunosorbent assay (ELISA) for the detection of TBBPA was developed. Six haptens (T1-T6) mimicking different structural elements of TBBPA were synthesized and coupled to keyhole...

  10. Highly Sensitive and Fast Anion-Selective InN Quantum Dot Electrochemical Sensors

    NASA Astrophysics Data System (ADS)

    Hassan Alvi, Naveed ul; Rodriguez, Paul E. D. Soto; Gómez, Victor J.; Kumar, Praveen; Willander, Magnus; Nötzel, Richard

    2013-11-01

    Epitaxial InN quantum dots (QDs) are demonstrated as ion-selective electrode for potentiometric anion concentration measurements. The sensor reveals high sensitivity above 90 mV/decade for the detection of chlorine and hydroxyl ions in sodium chloride (NaCl), calcium chloride (CaCl2), and sodium hydroxide (NaOH) solutions. The response time is less than two seconds after which the signal is very stable and repeatable. The sensitivity for the InN QDs is about two times that for a reference InN thin film and the response time is about five times shorter. In pH buffer solutions the sensor reveals no clear response to cations. A model is presented for the high sensitivity, fast response, and ion selectivity based on the unique electronic properties of the InN surface together with the zero-dimensional nature of the QDs.

  11. Low-sensitivity, frequency-selective amplifier circuits for hybrid and bipolar fabrication.

    NASA Technical Reports Server (NTRS)

    Pi, C.; Dunn, W. R., Jr.

    1972-01-01

    A network is described which is suitable for realizing a low-sensitivity high-Q second-order frequency-selective amplifier for high-frequency operation. Circuits are obtained from this network which are well suited for realizing monolithic integrated circuits and which do not require any process steps more critical than those used for conventional monolithic operational and video amplifiers. A single chip version using compatible thin-film techniques for the frequency determination elements is then feasible. Center frequency and bandwidth can be set independently by trimming two resistors. The frequency selective circuits have a low sensitivity to the process variables, and the sensitivity of the center frequency and bandwidth to changes in temperature is very low.

  12. A novel sensitive and selective ligation-based ATP assay using a molecular beacon.

    PubMed

    Ma, Changbei; Tang, Zhiwen; Wang, Kemin; Yang, Xiaohai; Tan, Weihong

    2013-05-21

    In this paper, a novel, facile fluorometric ATP assay with high sensitivity and excellent selectivity has been reported. This approach utilizes a molecular beacon, T4 DNA ligase and two short oligonucleotides. In the presence of ATP, the T4 DNA ligase catalyzes the ligation reaction and the ligation product restores the fluorescence of the molecular beacon. Owing to the high sensitivity of the molecular beacon and T4 DNA ligase's high substrate dependence, this novel ATP assay demonstrates exceptional selectivity and high sensitivity down to 0.14 nM in homogeneous solution. Cellular ATP concentrations in several cell lines have been determined by measuring the lysate sample containing 8.0 × 10(3) cells.

  13. Sensitivity analysis of standardization procedures in drought indices to varied input data selections

    NASA Astrophysics Data System (ADS)

    Liu, Yi; Ren, Liliang; Hong, Yang; Zhu, Ye; Yang, Xiaoli; Yuan, Fei; Jiang, Shanhu

    2016-07-01

    Reasonable input data selection is of great significance for accurate computation of drought indices. In this study, a comprehensive comparison is conducted on the sensitivity of two commonly used standardization procedures (SP) in drought indices to datasets, namely the probability distribution based SP and the self-calibrating Palmer SP. The standardized Palmer drought index (SPDI) and the self-calibrating Palmer drought severity index (SC-PDSI) are selected as representatives of the two SPs, respectively. Using meteorological observations (1961-2012) in the Yellow River basin, 23 sub-datasets with a length of 30 years are firstly generated with the moving window method. Then we use the whole time series and 23 sub-datasets to compute two indices separately, and compare their spatiotemporal differences, as well as performances in capturing drought areas. Finally, a systematic investigation in term of changing climatic conditions and varied parameters in each SP is conducted. Results show that SPDI is less sensitive to data selection than SC-PDSI. SPDI series derived from different datasets are highly correlated, and consistent in drought area characterization. Sensitivity analysis shows that among the three parameters in the generalized extreme value (GEV) distribution, SPDI is most sensitive to changes in the scale parameter, followed by location and shape parameters. For SC-PDSI, its inconsistent behaviors among different datasets are primarily induced by the self-calibrated duration factors (p and q). In addition, it is found that the introduction of the self-calibrating procedure for duration factors further aggravates the dependence of drought index on input datasets compared with original empirical algorithm that Palmer uses, making SC-PDSI more sensitive to variations in data sample. This study clearly demonstrate the impacts of dataset selection on sensitivity of drought index computation, which has significant implications for proper usage of drought

  14. Sulindac sulfide selectively increases sensitivity of ABCC1 expressing tumor cells to doxorubicin and glutathione depletion

    PubMed Central

    Whitt, Jason D.; Keeton, Adam B.; Gary, Bernard D.; Sklar, Larry A.; Sodani, Kamlesh; Chen, Zhe-Sheng; Piazza, Gary A.

    2016-01-01

    Abstract ATP-binding cassette (ABC) transpo rters ABCC1 (MRP1), ABCB1 (P-gp), and ABCG2 (BCRP) contribute to chemotherapy failure. The primary goals of this study were to characterize the efficacy and mechanism of the non­steroidal anti-inflammatory drug (NSAID), sulindac sulfide, to reverse ABCC1 mediated resistance to chemother­apeutic drugs and to determine if sulindac sulfide can influence sensitivity to chemotherapeutic drugs independently of drug efflux. Cytotoxicity assays were performed to measure resistance of ABC-expressing cell lines to doxoru­bicin and other chemotherapeutic drugs. NSAIDs were tested for the ability to restore sensitivity to resistance selected tumor cell lines, as well as a large panel of standard tumor cell lines. Other experiments characterized the mechanism by which sulindac sulfide inhibits ABCC1 substrate and co-substrate (GSH) transport in isolated membrane vesicles and intact cells. Selective reversal of multi-drug resistance (MDR), decreased efflux of doxor­ubicin, and fluorescent substrates were demonstrated by sulindac sulfide and a related NSAID, indomethacin, in resistance selected and engineered cell lines expressing ABCC1, but not ABCB1 or ABCG2. Sulindac sulfide also inhibited transport of leukotriene C4 into membrane vesicles. Sulindac sulfide enhanced the sensitivity to doxoru­bicin in 24 of 47 tumor cell lines, including all melanoma lines tested (7-7). Sulindac sulfide also decreased intra­cellular GSH in ABCC1 expressing cells, while the glutathione synthesis inhibitor, BSO, selectively increased sensitivity to sulindac sulfide induced cytotoxicity. Sulindac sulfide potently and selectively reverses ABCC1-mediated MDR at clinically achievable concentrations. ABCC1 expressing tumors may be highly sensitive to the direct cytotoxicity of sulindac sulfide, and in combination with chemotherapeutic drugs that induce oxidative stress.

  15. The amino-terminal 200 amino acids of the plasma membrane Na+,K+-ATPase alpha subunit confer ouabain sensitivity on the sarcoplasmic reticulum Ca(2+)-ATPase.

    PubMed Central

    Ishii, T; Takeyasu, K

    1993-01-01

    Cardiac glycosides such as G-strophanthin (ouabain) bind to and inhibit the plasma membrane Na+,K(+)-ATPase but not the sarcoplasmic reticulum (SR) Ca(2+)-ATPase, whereas thapsigargin specifically blocks the SR Ca(2+)-ATPase. The chimera [n/c]CC, in which the amino-terminal amino acids Met1 to Asp162 of the SR Ca(2+)-ATPase (SERCA1) were replaced with the corresponding portion of the Na+,K(+)-ATPase alpha 1 subunit (Met1 to Asp200), retained thapsigargin- and Ca(2+)-sensitive ATPase activity, although the activity was lower than that of the wild-type SR Ca(2+)-ATPase. Moreover, this Ca(2+)-sensitive ATPase activity was inhibited by ouabain. The chimera NCC, in which Met1-Gly354 of the SR Ca(2+)-ATPase were replaced with the corresponding portion of the Na+,K(+)-ATPase, lost the thapsigargin-sensitive Ca(2+)-ATPase activity seen in CCC and [n/c]CC. [3H]Ouabain binding to [n/c]CC and NCC demonstrated that the affinity for this inhibitor seen in the wild-type chicken Na+,K(+)-ATPase was restored in these chimeric molecules. Thus, the ouabain-binding domains are distinct from the thapsigargin sites; ouabain binds to the amino-terminal portion (Met1 to Asp200) of the Na+,K(+)-ATPase alpha 1 subunit, whereas thapsigargin interacts with the regions after Asp162 of the Ca(2+)-ATPase. Moreover, the amino-terminal 200 amino acids of the Na+,K(+)-ATPase alpha 1 subunit are sufficient to exert ouabain-dependent inhibition even after incorporation into the corresponding portion of the Ca(2+)-ATPase, and the segment Ile163 to Gly354 of the SR Ca(2+)-ATPase is critical for thapsigargin- and Ca(2+)-sensitive ATPase activity. Images Fig. 5 PMID:8415625

  16. Improvement of endothelial dysfunction by selective estrogen receptor-alpha stimulation in ovariectomized SHR.

    PubMed

    Widder, Julian; Pelzer, Theo; von Poser-Klein, Christine; Hu, Kai; Jazbutyte, Virginija; Fritzemeier, Karl-Heinrich; Hegele-Hartung, Christa; Neyses, Ludwig; Bauersachs, Johann

    2003-11-01

    Both known estrogen receptors, ERalpha and ERbeta, are expressed in blood vessels. To gain further insight into the role of ERalpha in a functional setting, we investigated the effect of the novel highly selective ERalpha agonist Cpd1471 on vascular reactivity in ovariectomized spontaneously hypertensive rats (SHR). After ovariectomy or sham operation, 12-week-old female SHR received either 17beta-estradiol (E2, 2 microg/kg body wt per day), the selective ERalpha agonist Cpd1471 (30 microg/kg body wt per day), or placebo. Acetylcholine-induced endothelium-dependent vasorelaxation was significantly blunted in aortas from ovariectomized rats (Rmax, 53%+/-3% versus sham, 79%+/-2%; P<0.001). Treatment with E2 or Cpd1471 significantly augmented acetylcholine-induced relaxation in ovariectomized rats (Rmax, 70%+/-2%; resp, 73%+/-2%). Endothelium-independent relaxation induced by sodium nitroprusside was not different among the four groups. The contractile response induced by the nitric oxide (NO) synthase inhibitor Nomega-nitro-l-arginine, an index of basal NO formation, was significantly lower in ovariectomized rats compared with sham-operated animals (53+/-2% versus 77%+/-5%; P<0.01) and was normalized by both E2 (70%+/-2%) and Cpd1471 (70%+/-3%). Aortic endothelial NO synthase (eNOS) expression and phosphorylation of the vasodilator-stimulated phosphoprotein, an index of NO/cGMP-signaling, was reduced in ovariectomized SHR and normalized by E2 and Cpd1471. In SHR after ovariectomy, endothelium-dependent NO-mediated vasorelaxation and eNOS expression are attenuated. The novel selective ERalpha agonist Cpd1471 prevented these pathophysiological changes to a similar extent as E2. Thus, the pharmacological principle of selective ERalpha activation mediates positive vascular effects.

  17. Sensitive ChIP-DSL technology reveals an extensive estrogen receptor alpha-binding program on human gene promoters.

    PubMed

    Kwon, Young-Soo; Garcia-Bassets, Ivan; Hutt, Kasey R; Cheng, Christine S; Jin, Mingjie; Liu, Dongyan; Benner, Chris; Wang, Dong; Ye, Zhen; Bibikova, Marina; Fan, Jian-Bing; Duan, Lingxun; Glass, Christopher K; Rosenfeld, Michael G; Fu, Xiang-Dong

    2007-03-20

    ChIP coupled with microarray provides a powerful tool to determine in vivo binding profiling of transcription factors to deduce regulatory circuitries in mammalian cells. Aiming at improving the specificity and sensitivity of such analysis, we developed a new technology called ChIP-DSL using the DNA selection and ligation (DSL) strategy, permitting robust analysis with much reduced materials compared with standard procedures. We profiled general and sequence-specific DNA binding transcription factors using a full human genome promoter array based on the ChIP-DSL technology, revealing an unprecedented number of the estrogen receptor (ERalpha) target genes in MCF-7 cells. Coupled with gene expression profiling, we found that only a fraction of these direct ERalpha target genes were highly responsive to estrogen and that the expression of those ERalpha-bound, estrogen-inducible genes was associated with breast cancer progression in humans. This study demonstrates the power of the ChIP-DSL technology in revealing regulatory gene expression programs that have been previously invisible in the human genome.

  18. Pertussis toxin-sensitive G-protein mediates the alpha 2-adrenergic receptor inhibition of melatonin release in photoreceptive chick pineal cell cultures

    SciTech Connect

    Pratt, B.L.; Takahashi, J.S.

    1988-07-01

    The avian pineal gland is a photoreceptive organ that has been shown to contain postjunctional alpha 2-adrenoceptors that inhibit melatonin synthesis and/or release upon receptor activation. Physiological response and (32P)ADP ribosylation experiments were performed to investigate whether pertussis toxin-sensitive guanine nucleotide-binding proteins (G-proteins) were involved in the transduction of the alpha 2-adrenergic signal. For physiological response studies, the effects of pertussis toxin on melatonin release in dissociated cell cultures exposed to norepinephrine were assessed. Pertussis toxin blocked alpha 2-adrenergic receptor-mediated inhibition in a dose-dependent manner. Pertussis toxin-induced blockade appeared to be noncompetitive. One and 10 ng/ml doses of pertussis toxin partially blocked and a 100 ng/ml dose completely blocked norepinephrine-induced inhibition. Pertussis toxin-catalyzed (32P)ADP ribosylation of G-proteins in chick pineal cell membranes was assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Membranes were prepared from cells that had been pretreated with 0, 1, 10, or 100 ng/ml pertussis toxin. In the absence of pertussis toxin pretreatment, two major proteins of 40K and 41K mol wt (Mr) were labeled by (32P)NAD. Pertussis toxin pretreatment of pineal cells abolished (32P) radiolabeling of the 40K Mr G-protein in a dose-dependent manner. The norepinephrine-induced inhibition of both cAMP efflux and melatonin release, as assessed by RIA of medium samples collected before membrane preparation, was also blocked in a dose-dependent manner by pertussis toxin. Collectively, these results suggest that a pertussis toxin-sensitive 40K Mr G-protein labeled by (32P)NAD may be functionally associated with alpha 2-adrenergic signal transduction in chick pineal cells.

  19. Topical alpha-selective p38 MAP kinase inhibition reduces acute skin inflammation in guinea pig

    PubMed Central

    Medicherla, Satyanarayana; Ma, Jing Ying; Reddy, Mamtha; Esikova, Irina; Kerr, Irene; Movius, Fabiola; Higgins, Linda S; Protter, Andrew A

    2010-01-01

    Certain skin pathologies, including psoriasis, are thought to be immune-mediated inflammatory diseases. Available literature clearly indicates the involvement of inflammatory cells (neutrophils, T cells, and macrophages), their cytokines, and the p38 mitogen-activated protein kinase (MAPK) signaling pathway in the pathophysiology of psoriasis. Neutrophils play an important role in the formation of acute inflammatory changes in psoriasis. Acute inflammation or acute flares in psoriasis remain poorly addressed in clinical medicine. In this communication, we first establish a simple and reproducible model for studying neutrophil-mediated acute skin inflammation. Using the hairless guinea pig, due to the similarity of skin architecture to that of human, acute inflammation was induced with an intradermal injection of 50 μg/mL lipopolysaccharide (LPS) in 50 μL solution. Myeloperoxidase (MPO) activity was measured by MPO-positive neutrophils and shown to increase for 24-hours post-injection. Simultaneously, the level of phosphorylated p38 MAPK was documented for 48-hours post-LPS injection in the skin. Next, we used this model to examine the therapeutic potential of an α-selective p38 MAPK inhibitor, SCIO-469. A comparison of topical application of SCIO-469 at 5 mg/mL or 15 mg/mL to vehicle revealed that SCIO-469 dose-dependently reduces acute skin inflammation and that this effect is statistically significant at the higher dose. Further examination of tissues that received this dose also revealed statistically significant reduction of MPO activity, phosphorylated p38 MAPK, interleukin-6, and cyclooxygenase-2. These data suggest that the α-selective p38 MAPK inhibitor, SCIO-469, acts as a topical anti-inflammatory agent via the p38 MAPK pathway to reduce neutrophil induced acute inflammation in the skin. These observations suggest that α-selective p38 MAPK inhibition may be an effective therapeutic strategy to manage acute skin inflammation PMID:22096353

  20. Pyrazolo-Pyrimidines: A Novel Heterocyclic Scaffold for Potent and Selective p38alpha Inhibitors

    SciTech Connect

    Das,J.; Moquin, R.; Pitt, S.; Zhang, R.; Shen, D.; McIntyre, K.; Gillooly, K.; Doweyko, A.; Sack, J.; et al

    2008-01-01

    The synthesis and structure-activity relationships (SAR) of p38a MAP kinase inhibitors based on a pyrazolo-pyrimidine scaffold are described. These studies led to the identification of compound 2x as a potent and selective inhibitor of p38a MAP kinase with excellent cellular potency toward the inhibition of TNFa production. Compound 2x was highly efficacious in vivo in inhibiting TNFa production in an acute murine model of TNFa production. X-ray co-crystallography of a pyrazolo-pyrimidine analog 2b bound to unphosphorylated p38a is also disclosed.

  1. Characterization of beta-R1, a gene that is selectively induced by interferon beta (IFN-beta) compared with IFN-alpha.

    PubMed

    Rani, M R; Foster, G R; Leung, S; Leaman, D; Stark, G R; Ransohoff, R M

    1996-09-13

    We report preliminary characterization of a gene designated beta-R1, which is selectively expressed in response to interferon beta (IFN-beta) compared with IFN-alpha. In human astrocytoma cells, beta-R1 was induced to an equivalent extent by 10 IU/mL IFN-beta or 2500 IU/mL IFN-alpha2. To address the mechanism of this differential response, we analyzed induction of the beta-R1 gene in fibrosarcoma cells and derivative mutant cells lacking components required for signaling by type I IFNs. beta-R1 was readily induced by IFN-beta in the parental 2fTGH cell line, but not by recombinant IFN-alpha2, IFN-alpha Con1, or a mixture of IFN-alpha subtypes. IFN-alpha8 induced beta-R1 weakly. beta-R1 was not induced by IFN-beta in mutant cell lines U2A, U3A, U4A, and U6A, which lack, respectively, p48, STAT1, JAK1, and STAT2. U5A cells, which lack the Ifnar 2.2 component of the IFN-alpha and -beta receptor, also failed to express beta-R1. U1A cells are partially responsive to IFN-beta and IFN-alpha8 but lacked beta-R1 expression, indicating that TYK2 protein is essential for induction of this gene. Taken together, these results suggest that the expression of beta-R1 in response to type I IFN requires IFN-stimulated gene factor 3 plus an additional component, which is more efficiently formed on induction by IFN-beta compared with IFN-alpha.

  2. Perceptual load manipulation reveals sensitivity of the face-selective N170 to attention.

    PubMed

    Mohamed, Tarik N; Neumann, Markus F; Schweinberger, Stefan R

    2009-05-27

    It has been controversial whether the face-sensitive N170 is affected by selective attention. We manipulated attention sensu Lavie's perceptual load theory to short (200 ms) presentations of task-irrelevant unfamiliar faces or houses, while participants identified superimposed target letters 'X' versus 'N'. These targets were strings of either six identical (low load) or six different letters (high load). Under low load, we found a prominent face-selective N170 response. Under high load, however, we not only observed a dramatic reduction of the face N170 but also an unexpected enhancement of the house N170, such that face selectivity was almost completely lost. We conclude that the early stages of face processing indexed by the N170 strongly depend on selective attention. PMID:19369907

  3. Perceptual load manipulation reveals sensitivity of the face-selective N170 to attention.

    PubMed

    Mohamed, Tarik N; Neumann, Markus F; Schweinberger, Stefan R

    2009-05-27

    It has been controversial whether the face-sensitive N170 is affected by selective attention. We manipulated attention sensu Lavie's perceptual load theory to short (200 ms) presentations of task-irrelevant unfamiliar faces or houses, while participants identified superimposed target letters 'X' versus 'N'. These targets were strings of either six identical (low load) or six different letters (high load). Under low load, we found a prominent face-selective N170 response. Under high load, however, we not only observed a dramatic reduction of the face N170 but also an unexpected enhancement of the house N170, such that face selectivity was almost completely lost. We conclude that the early stages of face processing indexed by the N170 strongly depend on selective attention.

  4. Benzo[c]quinolizin-3-ones: a novel class of potent and selective nonsteroidal inhibitors of human steroid 5alpha-reductase 1.

    PubMed

    Guarna, A; Machetti, F; Occhiato, E G; Scarpi, D; Comerci, A; Danza, G; Mancina, R; Serio, M; Hardy, K

    2000-10-01

    The synthesis and biological evaluation of a series of novel, selective inhibitors of isoenzyme 1 of human 5alpha-reductase (5alphaR) (EC 1.3.99.5) are reported. The inhibitors are 4aH- (19-29) or 1H-tetrahydrobenzo[c]quinolizin-3-ones (35-47) bearing at positions 1, 4, 5, and 6 a methyl group and at position 8 a hydrogen, methyl group, or chlorine atom. All these compounds were tested toward 5alphaR-1 and 5alphaR-2 expressed in CHO cells (CHO 1827 and CHO 1829, respectively) resulting in selective inhibitors of the type 1 isoenzyme, with inhibitory potencies (IC(50)) ranging from 7.6 to 9100 nM. The inhibitors of the 4aH-series, having a double bond at position 1,2, were generally less active than the corresponding inhibitors of the 1H-series having the double bond at position 4,4a on the A ring. The presence of a methyl group at position 4 (as in compounds 39-40 and 45-47), associated with a substituent at position 8, determined the highest inhibition potency (IC(50) from 7.6 to 20 nM). Compounds 39 and 40, having K(i) values of 5.8+/-1.8 and 2.7+/-0.6 nM, respectively, toward 5alphaR-1 expressed in CHO cells, were also tested toward native 5alphaR-1 in human scalp and 5alphaR-2 in human prostate homogenates, in comparison with finasteride and the known 5alphaR-1-selective inhibitor LY191704, and their mechanism of inhibition was determined. They both inhibited the enzyme through a reversible competitive mechanism and again were selective inhibitors of 5alphaR-1 with IC(50) values of 41 nM. These specific features make these inhibitors suitable candidates for further development as drugs in the treatment of DHT-dependent disorders such as acne and androgenic alopecia in men and hirsutism in women. PMID:11020287

  5. Antireflection treatment of thickness sensitive spectrally selective (TSSS) paints for thermal solar absorbers

    SciTech Connect

    Lundh, M.; Waeckelgaard, E.; Blom, T.

    2010-01-15

    There are several methods to produce solar absorbers, and one cheap alternative is painted absorbers, preferably painted with a spectrally selective paint. The optical properties of Thickness Sensitive Spectrally Selective (TSSS) paints are, however, limited by the thickness of the paint layer. In this study it is shown that the solar absorptance of two commercial TSSS paints can be increased between 0.01 and 0.02 units with an antireflection treatment using a silicon dioxide layer deposited from silica-gel. It was found that the thermal emittance (100 C) did not change significantly after the treatment. (author)

  6. Prothymosin alpha selectively enhances estrogen receptor transcriptional activity by interacting with a repressor of estrogen receptor activity.

    PubMed

    Martini, P G; Delage-Mourroux, R; Kraichely, D M; Katzenellenbogen, B S

    2000-09-01

    We find that prothymosin alpha (PTalpha) selectively enhances transcriptional activation by the estrogen receptor (ER) but not transcriptional activity of other nuclear hormone receptors. This selectivity for ER is explained by PTalpha interaction not with ER, but with a 37-kDa protein denoted REA, for repressor of estrogen receptor activity, a protein that we have previously shown binds to ER, blocking coactivator binding to ER. We isolated PTalpha, known to be a chromatin-remodeling protein associated with cell proliferation, using REA as bait in a yeast two-hybrid screen with a cDNA library from MCF-7 human breast cancer cells. PTalpha increases the magnitude of ERalpha transcriptional activity three- to fourfold. It shows lesser enhancement of ERbeta transcriptional activity and has no influence on the transcriptional activity of other nuclear hormone receptors (progesterone receptor, glucocorticoid receptor, thyroid hormone receptor, or retinoic acid receptor) or on the basal activity of ERs. In contrast, the steroid receptor coactivator SRC-1 increases transcriptional activity of all of these receptors. Cotransfection of PTalpha or SRC-1 with increasing amounts of REA, as well as competitive glutathione S-transferase pulldown and mammalian two-hybrid studies, show that REA competes with PTalpha (or SRC-1) for regulation of ER transcriptional activity and suppresses the ER stimulation by PTalpha or SRC-1, indicating that REA can function as an anticoactivator in cells. Our data support a model in which PTalpha, which does not interact with ER, selectively enhances the transcriptional activity of the ER but not that of other nuclear receptors by recruiting the repressive REA protein away from ER, thereby allowing effective coactivation of ER with SRC-1 or other coregulators. The ability of PTalpha to directly interact in vitro and in vivo with REA, a selective coregulator of the ER, thereby enabling the interaction of ER with coactivators, appears to explain

  7. Mechanical stimulation of skeletal muscle increases prostaglandin F2(alpha) synthesis and cyclooxygenase activity by a pertussis toxin sensitive mechanism

    NASA Technical Reports Server (NTRS)

    Vandenburgh, Herman H.; Shansky, Janet; Solerssi, Rosa; Chromiak, Joseph

    1992-01-01

    Repetitive mechanical stimulation of differentiated skeletal muscle in tissue culture increases the production of prostaglandin F(sub 2(alpha)), an anabolic stimulator of myofiber growth. Within 4 h of initiating mechanical activity, the activity of cyclooxygenase, a regulatory enzyme in prostaglandin synthesis, was increased 82% (P is less than .005), and this increase was maintained for at least 24 h. Kinetic analysis of the stretch-activated cyclooxygenase indicated a two to three-fold decrease in the enzyme's K(sub m) with no change in V(sub max). The stretch-induced increase in enzymatic activity was not inhibited by cycloheximide, was independent of cellular electrical activity (tetrodotoxin-insensitive), but was prevented by the G protein inhibitor pertussis toxin. Pertussis toxin also inhibited the stretch-induced increases in PGF(sub 2(alpha)) production, and cell growth. It is concluded that stretch of skeletal muscle increases the synthesis of the anabolic modulator PGF(sub 2(alpha)) by a G protein-dependent process which involves activation of cyclooxygenase by a posttranslational mechanism.

  8. Trypsin-stabilized fluorescent gold nanocluster for sensitive and selective Hg2+ detection.

    PubMed

    Kawasaki, Hideya; Yoshimura, Kouta; Hamaguchi, Kenji; Arakawa, Ryuichi

    2011-01-01

    We report on trypsin-stabilized fluorescent gold nanoclusters (Au NCs) for the sensitive and selective detection of Hg(2+) ions. The Au NCs have an average size of 1 nm and show a red emission at 645 nm. The photostable properties of the trypsin-stabilized Au NCs were examined, and their photochemical stability was found to be similar to that of CdSe quantum dots. The fluorescence was particularly quenched by Hg(2+), and therefore the Au NCs can be used as fluorescent sensors for sensitive and selective Hg(2+) detection to a detection limit of 50 ± 10 nM and the quantitative detection of Hg(2+) in wide and low concentration range of 50-600 nM.

  9. Anticancer chemosensitivity changes between the original and recurrent tumors after successful chemotherapy selected according to the sensitivity assay.

    PubMed Central

    Nio, Y; Tamura, K; Tsubono, M; Kawabata, K; Masai, Y; Hayashi, H; Ishigami, S; Araya, S; Imamura, M

    1995-01-01

    OBJECTIVE: The authors compare and characterize the changes in chemosensitivity between the original tumors before chemotherapy and recurrent tumors after responses. SUMMARY BACKGROUND DATA: The drug resistance in clinical chemotherapy appears to be different from that in experimental chemotherapy, and the profile and mechanisms of clinical drug resistance in recurrent tumors, especially after successful chemotherapy has scarcely been studied. METHODS: Applied chemotherapies were selected out of four agents, cisplatin (CDDP), adriamycin (ADR), mitomycin-C (MMC) and 5-fluorouracil (5-FU), singly or in combinations by a DNA synthesis inhibition assay, by which the sensitivity of recurrent tumors was assessed. Responses were defined according to the standard criteria, and successful chemotherapy indicates complete response (CR) or partial response (PR) for solid tumors and complete disappearance for malignant effusion. RESULTS: In 37 patients, the effectiveness of four agents were compared between before chemotherapy and after recurrence, and the response lasted between 2 and 26 months (mean +/- SD, 7.7 +/- 5.5). The results suggest that locally recurred tumors may become resistant to the agents previously administered; by contrast, distantly recurred tumors may not necessarily become resistant to the agents administered. The recurrent tumors are suggested to be sensitive to the agents as follows: locally recurrent solid tumors, 5-FU; distantly recurrent solid tumors, 5-FU and CDDP; locally recurrent effusion, CDDP; distantly recurrent effusion, ADR. Twenty-three of 37 recurrent tumors were re-treated with chemotherapies selected according to the sensitivity assay, singly or in combination with a biologic response modifier (BRM)--a streptococcal preparation, OK-432, or interferon-alpha. Responses were seen in 1 of 13 solid recurrent tumors and in 6 of 10 recurrent effusions. Responses were seen only when the patients were treated with a combination of chemotherapy and

  10. Virtually complete E-selective alpha,beta-unsaturated ester synthesis by Hg(OTf)2-catalyzed hydration of sec-ethoxyalkynyl acetate.

    PubMed

    Nishizawa, Mugio; Hirakawa, Hiroko; Nakagawa, Yuki; Yamamoto, Hirofumi; Namba, Kosuke; Imagawa, Hiroshi

    2007-12-20

    The reaction of alkyl-substituted sec-ethoxyalkynyl acetates with water catalyzed by Hg(OTf)2 afforded alpha,beta-unsaturated esters in excellent yield with high catalytic turnover up to 1000 times under very mild reaction conditions with virtually complete E-selectivity, superior even to that of the HWE reaction.

  11. Selectivity and sensitivity of some thin-layer chromatographic detection systems.

    PubMed

    Fodor-Csorba, K; Dutka, F

    1986-09-19

    The selectivity and sensitivity of some thin-layer chromatographic detection systems widely used internationally and developed in our laboratory were studied. Halogenated organophosphorus pesticides were found to interfere with the detection of organochlorine pesticides when using silver nitrate-2-phenoxyethanol. The stability of colours formed by the 4-(4'-nitrobenzyl)pyridine-tetraethylenepentamine system was enhanced by spraying with acetic acid and allowed densitometric evaluation. The most sensitive detection method for thiocarbamates is the reaction with 2,6-dichlorobenzoquinone-N-chloroimine or its dibromo analogue (50 ng). Application of this method for sulphur-containing organophosphorus insecticides results in the same or better sensitivity. Quantitation of these compounds was carried out by densitometry.

  12. The C-terminal 165 amino acids of the plasma membrane Ca(2+)-ATPase confer Ca2+/calmodulin sensitivity on the Na+,K(+)-ATPase alpha-subunit.

    PubMed Central

    Ishii, T; Takeyasu, K

    1995-01-01

    The C-terminal 165 amino acids of the rat brain plasma membrane (PM) Ca(2+)-ATPase II containing the calmodulin binding auto-inhibitory domain was connected to the C-terminus of the ouabain sensitive chicken Na+,K(+)-ATPase alpha 1 subunit. Expression of this chimeric molecule in ouabain resistant mouse L cells was assured by the high-affinity binding of [3H]ouabain. In the presence of Ca2+/calmodulin, this chimeric molecule exhibited ouabain inhibitable Na+,K(+)-ATPase activity; the putative chimeric ATPase activity was absent in the absence of Ca2+/calmodulin and activated by Ca2+/calmodulin in a dose-dependent manner. Furthermore, this chimeric molecule could bind monoclonal IgG 5 specific to the chicken Na+,K(+)-ATPase alpha 1 subunit only in the presence of Ca2+/calmodulin, suggesting that the epitope for IgG 5 in this chimera is masked in the absence of Ca2+/calmodulin and uncovered in their presence. These results propose a direct interaction between the calmodulin binding auto-inhibitory domain of the PM Ca(2+)-ATPase and the specific regions of the Na+,K(+)-ATPase alpha 1 subunit that are structurally homologous to the PM Ca(2+)-ATPase. A comparison of the deduced amino acid sequences revealed several possible regions within the Na+,K(+)-ATPase that might interact with the auto-inhibitory domain of the PM Ca(2+)-ATPase. Images PMID:7828596

  13. Structural insights into selective agonist actions of tamoxifen on human Estrogen Receptor alpha

    PubMed Central

    Chakraborty, Sandipan; Biswas, P. K.

    2014-01-01

    Tamoxifen, an anti-estrogenic ligand in breast tissues and being used as a first-line treatment in ER-positive breast cancers, is found to develop resistance followed by resumption of growth of the tumor in about 30% of cases. Whether tamoxifen starts assisting in proliferation in such cases or there exists any ligand-independent pathways to transcription is not fully understood; also, no ERα mutants have been detected so far which could lead to tamoxifen resistance. Performing in-silico conformational analysis of ERα ligand binding domain, in the absence and presence of selective agonist (Diethylstilbestrol; DES), antagonist (Faslodex; ICI), and SERM (4-hydroxy tamoxifen; 4-OHT) ligands, we elucidated ligand-responsive structural modulations of ERα-LBD dimer in their agonist and antagonist complexes and address the issue of “tamoxifen resistance”. We found DES and ICI to stabilize the dimer in their agonist and antagonist conformations, respectively. The ERα-LBD dimer without the presence of any bound ligand also leads to a stable structure in agonist conformation. However, the binding of 4-OHT to antagonist structure is found to lead to a flexible conformation allowing the protein visiting conformations populated by agonists as are evident from principal component analysis and radius of gyration plots. Further, the relaxed conformations of the 4-OHT bound protein is found to exhibit a diminished size of the co-repressor binding pocket at LBD, thus signaling a partial blockage of the co-repressor binding motif. Thus, the ability of 4-OHT bound ERα-LBD to assume flexible conformations visited by agonists and reduced co-repressor binding surface at LBD provide crucial structural insights into tamoxifen-resistance complementing our existing understanding. PMID:25060147

  14. Perceived egocentric distance sensitivity and invariance across scene-selective cortex.

    PubMed

    Persichetti, Andrew S; Dilks, Daniel D

    2016-04-01

    Behavioral studies in many species and studies in robotics have demonstrated two sources of information critical for visually-guided navigation: sense (left-right) information and egocentric distance (proximal-distal) information. A recent fMRI study found sensitivity to sense information in two scene-selective cortical regions, the retrosplenial complex (RSC) and the occipital place area (OPA), consistent with hypotheses that these regions play a role in human navigation. Surprisingly, however, another scene-selective region, the parahippocampal place area (PPA), was not sensitive to sense information, challenging hypotheses that this region is directly involved in navigation. Here we examined how these regions encode egocentric distance information (e.g., a house seen from close up versus far away), another type of information crucial for navigation. Using fMRI adaptation and a regions-of-interest analysis approach in human adults, we found sensitivity to egocentric distance information in RSC and OPA, while PPA was not sensitive to such information. These findings further support that RSC and OPA are directly involved in navigation, while PPA is not, consistent with the hypothesis that scenes may be processed by distinct systems guiding navigation and recognition.

  15. Nonmaternal Care's Association With Mother's Parenting Sensitivity: A Case of Self-Selection Bias?

    PubMed

    Nomaguchi, Kei M; Demaris, Alfred

    2013-06-01

    Although attachment theory posits that the use of nonmaternal care undermines quality of mothers' parenting, empirical evidence for this link is inconclusive. Using data from the National Institute of Child Health and Human Development Study of Early Child Care and Youth Development (N = 1,233), the authors examined the associations between nonmaternal care characteristics and maternal sensitivity during the first 3 years of children's lives, with special attention to selection effects and moderation by resource levels. Findings from fixed-effects regression models suggested that, on average, there is little relationship between nonmaternal care characteristics and maternal sensitivity, once selection factors are held constant. Some evidence of moderation effects was found, however. Excellent-quality care is related to more sensitivity for mothers with lower family income. Poor-quality care is related to lower sensitivity for single mothers, but not partnered mothers. In sum, nonmaternal care characteristics do not seem to have as much influence on mothers' parenting as attachment theory claims.

  16. Perceived egocentric distance sensitivity and invariance across scene-selective cortex.

    PubMed

    Persichetti, Andrew S; Dilks, Daniel D

    2016-04-01

    Behavioral studies in many species and studies in robotics have demonstrated two sources of information critical for visually-guided navigation: sense (left-right) information and egocentric distance (proximal-distal) information. A recent fMRI study found sensitivity to sense information in two scene-selective cortical regions, the retrosplenial complex (RSC) and the occipital place area (OPA), consistent with hypotheses that these regions play a role in human navigation. Surprisingly, however, another scene-selective region, the parahippocampal place area (PPA), was not sensitive to sense information, challenging hypotheses that this region is directly involved in navigation. Here we examined how these regions encode egocentric distance information (e.g., a house seen from close up versus far away), another type of information crucial for navigation. Using fMRI adaptation and a regions-of-interest analysis approach in human adults, we found sensitivity to egocentric distance information in RSC and OPA, while PPA was not sensitive to such information. These findings further support that RSC and OPA are directly involved in navigation, while PPA is not, consistent with the hypothesis that scenes may be processed by distinct systems guiding navigation and recognition. PMID:26963085

  17. Nonmaternal Care's Association With Mother's Parenting Sensitivity: A Case of Self-Selection Bias?

    PubMed

    Nomaguchi, Kei M; Demaris, Alfred

    2013-06-01

    Although attachment theory posits that the use of nonmaternal care undermines quality of mothers' parenting, empirical evidence for this link is inconclusive. Using data from the National Institute of Child Health and Human Development Study of Early Child Care and Youth Development (N = 1,233), the authors examined the associations between nonmaternal care characteristics and maternal sensitivity during the first 3 years of children's lives, with special attention to selection effects and moderation by resource levels. Findings from fixed-effects regression models suggested that, on average, there is little relationship between nonmaternal care characteristics and maternal sensitivity, once selection factors are held constant. Some evidence of moderation effects was found, however. Excellent-quality care is related to more sensitivity for mothers with lower family income. Poor-quality care is related to lower sensitivity for single mothers, but not partnered mothers. In sum, nonmaternal care characteristics do not seem to have as much influence on mothers' parenting as attachment theory claims. PMID:23772093

  18. Highly sensitive and doubly orientated selective molecularly imprinted electrochemical sensor for Cu(2.).

    PubMed

    Li, Jianping; Zhang, Lianming; Wei, Ge; Zhang, Yun; Zeng, Ying

    2015-07-15

    Studies on molecularly imprinted electrochemical sensors for metal ions determination have been widely reported. However, the sensitivity and selectivity of the sensors needs to be improved urgently. In the current work, a novel molecularly imprinted electrochemical sensor was originally developed for selective determination of ultratrace Cu(2+) by combining the metal-ligand chelate orientated recognition with enzyme amplification effect. The detection relied on a competition reaction between Cu(2+)-glycine (Cu-Gly) and horse radish peroxidase (HRP)-labeled Cu-Gly on the imprinted polymer membrane modified electrode. The sensitivity of this sensor was promoted by enzyme amplification. Selectivity was improved by the double-specificity derived from ligand-to-metal ion and metal-ligand chelate orientated recognition of 3D imprinted cavities. This technique was quantitatively sensitive to Cu(2+) concentrations ranging from 0.5nmol/L to 30nmol/L, with a detection limit of 42.4pmol/L. which was lower than those in most of the reported methods. The allowable amounts of interference ions were higher when it compared to other common molecularly imprinted sensors. Moreover, the results of assaying several real samples have proven its feasibility for practical applications. PMID:25771304

  19. Selecting focal species as surrogates for imperiled species using relative sensitivities derived from occupancy analysis

    USGS Publications Warehouse

    Silvano, Amy; Guyer, Craig; Steury, Todd; Grand, James B.

    2017-01-01

    Most imperiled species are rare or elusive and difficult to detect, which makes gathering data to estimate their response to habitat restoration a challenge. We used a repeatable, systematic method for selecting focal species using relative sensitivities derived from occupancy analysis. Our objective was to select suites of focal species that would be useful as surrogates when predicting effects of restoration of habitat characteristics preferred by imperiled species. We developed 27 habitat profiles that represent general habitat relationships for 118 imperiled species. We identified 23 regularly encountered species that were sensitive to important aspects of those profiles. We validated our approach by examining the correlation between estimated probabilities of occupancy for species of concern and focal species selected using our method. Occupancy rates of focal species were more related to occupancy rates of imperiled species when they were sensitive to more of the parameters appearing in profiles of imperiled species. We suggest that this approach can be an effective means of predicting responses by imperiled species to proposed management actions. However, adequate monitoring will be required to determine the effectiveness of using focal species to guide management actions.

  20. Discovery of alpha,gamma-diketo acids as potent selective and reversible inhibitors of hepatitis C virus NS5b RNA-dependent RNA polymerase.

    PubMed

    Summa, Vincenzo; Petrocchi, Alessia; Pace, Paola; Matassa, Victor G; De Francesco, Raffaele; Altamura, Sergio; Tomei, Licia; Koch, Uwe; Neuner, Philippe

    2004-01-01

    alpha,gamma-Diketo acids (DKA) were discovered from screening as selective and reversible inhibitors of hepatitis C virus NS5b RNA-dependent RNA polymerase. The diketo acid moiety proved essential for activity, while substitution on the gamma position was necessary for selectivity and potency. Optimization led to the identification of a DKA inhibitor of NS5b polymerase with IC(50) = 45 nM, one of the most potent HCV NS5b polymerase inhibitors reported.

  1. THE FIRST SYSTEMATIC SURVEY FOR Ly{alpha} EMITTERS AT z = 7.3 WITH RED-SENSITIVE SUBARU/SUPRIME-CAM

    SciTech Connect

    Shibuya, Takatoshi; Kashikawa, Nobunari; Iye, Masanori; Ota, Kazuaki; Ouchi, Masami; Furusawa, Hisanori; Shimasaku, Kazuhiro; Hattori, Takashi

    2012-06-20

    We have performed deep imaging surveys for Ly{alpha} emitters (LAEs) at redshift {approx}7.3 in two blank fields, the Subaru Deep Field (SDF) and the Subaru/XMM-Newton Deep survey Field (SXDF), using the Subaru/Suprime-Cam equipped with new red-sensitive CCDs and a new narrowband filter, NB1006 ({lambda}{sub c} = 10052 Angstrom-Sign , FWHM {Delta}{lambda} = 214 A). We identified four objects as LAE candidates that exhibit luminosity excess in NB1006. By carrying out deep follow-up spectroscopy for three of them using Subaru/FOCAS and Keck/DEIMOS, a definitively asymmetric emission line is detected for one of them, SXDF-NB1006-2. Assuming this line is Ly{alpha}, this object is an LAE at z = 7.215 which has a luminosity of 1.2{sup +1.5}{sub -0.6} Multiplication-Sign 10{sup 43} erg s{sup -1} and a weighted skewness S{sub {omega}} = 4.90 {+-} 0.86. Another object, SDF-NB1006-2, shows variable photometry and is thus probably a quasar (QSO) or an active galactic nucleus. It shows an asymmetric emission line at 10076 A which may be due to either Ly{alpha} at z = 7.288 or [O II] at z = 1.703. The third object, SDF-NB1006-1, is likely a galaxy with temporal luminosity enhancement associated with a supernova explosion, as the brightness of this object varies between the observed epochs. Its spectrum does not show any emission lines. The inferred decrease in the number density of LAEs toward higher redshift is n{sup z={sup {sup 7.3}{sub Ly{alpha}}}}/n{sub Ly{alpha}}{sup z={sup {sup 5.7}}} = 0.05{sup +0.11}{sub -0.05} from z = 5.7 to 7.3 down to L{sup Ly{alpha}} = 1.0 Multiplication-Sign 10{sup 43} erg s{sup -1}. The present result is consistent with the interpretation in previous studies that the neutral hydrogen fraction is rapidly increasing from z = 5.7 to 7.3.

  2. A simple and rapid chemiluminescence aptasensor for acetamiprid in contaminated samples: Sensitivity, selectivity and mechanism.

    PubMed

    Qi, Yingying; Xiu, Fu-Rong; Zheng, Minfeng; Li, Baoxin

    2016-09-15

    Ultralow concentration and selective detection of pesticide residue is important to evaluate the environmental and biological pollution and the threat to human health which single component pesticide can bring. Herein, we report an amplified chemiluminescence (CL) sensing platform for ultrasensitive and selective acetamiprid (widely used pesticide) detection. It is based on aptamer's high binding affinity to target and the relevance between AuNPs' morphology and its catalytic effect to stimulate the generation of CL in the presence of H2O2 and luminol. Moreover, AuNPs morphological slight change induced by aptamers' conformation during targets binding could lead to the significant change of catalytic properties. Therefore, the proposed sensing platform for pesticide residue exhibited a high sensitivity toward acetamiprid with a detection limit of 62pM, which was about 100-fold lower than that of other aptamer-based sensor for acetamiprid detection. Because of the intrinsic specificity of aptamer's recognization, this sensing platform has high selectivity. So, this sensing platform provides a label-free and cost-effective approach for sensitive and selective detection of single component pesticide residue. More importantly, this CL method was successfully used to determine acetamiprid in real contaminated samples. PMID:27131997

  3. Dihydropyridine-sensitive calcium channels in cardiac and skeletal muscle membranes: studies with antibodies against the. cap alpha. subunits

    SciTech Connect

    Takahashi, M.; Catterall, W.A.

    1987-08-25

    Polyclonal antibodies (PAC-2) against the purified skeletal muscle calcium channel were prepared and shown to be directed against ..cap alpha.. subunits of this protein by immunoblotting and immunoprecipitation. These polypeptides have an apparent molecular weight of 162,000 without reduction of disulfide bonds. Under conditions where the functional properties of the purified skeletal muscle calcium channel are retained, ..beta.. subunits (M/sub r/ 50,000) and lambda subunits (M/sub r/ 33,000) are coprecipitated, demonstrating specific noncovalent association of these three polypeptides in the purified skeletal muscle channel. PAC-2 immunoprecipitated cardiac calcium channels labeled with (/sup 3/H)isopropyl 4-(2,1,3-benzoxadiazol-4-yl)-1,4-dihydro-2,6-dimethyl-5-(methoxycarbonyl)pyridine-3-carboxylate ((/sup 3/H)PN200-110) at a 3-fold higher concentration than skeletal muscle channels. Preincubation with cardiac calcium channels blocked only 49% of the immunoreactivity of PAC-2 toward skeletal muscle channels, indicating that these two proteins have both homologous and distinct epitopes. The immunoreactive component of the cardiac calcium channel was identified by immunoprecipitation and polyacrylamide gel electrophoresis as a polypeptide with an apparent molecular weight of 170,000 before reduction of disulfide bonds and 141,000 after reduction, in close analogy with the properties of the ..cap alpha../sub 2/ subunits of the skeletal muscle channel. The calcium channels were radiolabeled with /sup 32/P and /sup 125/I. It is concluded that these two calcium channels have a homologous, but distinct, ..cap alpha.. subunit as a major polypeptide component.

  4. Implication of potassium trimolybdate nanowires as highly sensitive and selective ammonia sensor at room temperature

    NASA Astrophysics Data System (ADS)

    Joshi, Aditee C.; Gangal, S. A.

    2016-09-01

    Potassium trimolybdate nanowires are demonstrated as unique and highly selective NH3 sensing materials at room temperature. The nanowires were synthesized by using chemical route under normal ambient conditions and subsequently characterized by scanning electron microscopy (SEM) and x-ray diffraction (XRD). Gas sensors based on nanowires were fabricated by isolating and aligning nanowires between microspaced electrodes using dielectrophoresis. Room temperature gas sensing studies for different vapors indicated excellent selectivity for NH3 and capability to detect NH3 at concentrations down to ppb level. The sensors exhibited higher sensitivity for concentration range much below toxic limit of NH3 from 500 ppb up to 25 ppm. Since nanowires are isolated and aligned, the gas sensing reaction is rapid, and the availability of abundant oxide and hydroxyl surface groups on nanowires surface makes the reaction significantly prominent and selective with highly reducing nature of NH3.

  5. Selection tool alpha-fetoprotein for patients waiting for liver transplantation: How to easily manage a fractal algorithm

    PubMed Central

    Lai, Quirino; Levi Sandri, Giovanni Battista; Lerut, Jan

    2015-01-01

    Alpha-fetoprotein (AFP) behavior in patients with hepatocellular carcinoma (HCC) waiting for liver transplant (LT) represents a perfect biological example of a fractal model in which its progressive modification and possible future prediction of its values are very hard to capture. As a consequence, AFP represents a useful but poorly manageable tool to increase the ability to better select HCC patients waiting for LT. Trying to find a “fil-rouge” in the recent literature, no definitive answers can be done to several open questions: (1) the best AFP value to adopt; (2) the best cut-off measurement; and (3) the best way to comfortably capture the effective, time-related, fluctuations of this biological marker. More, structured and prospective, studies using serial determination of AFP values within and without the context of locoregional therapies are needed in order to find the “ideal” (static and dynamic) cut-off values allowing to respond to all the still open questions in this field of transplant oncology. PMID:26244064

  6. A nonionic surfactant-decorated liquid crystal sensor for sensitive and selective detection of proteins.

    PubMed

    Wang, Yi; Hu, Qiongzheng; Tian, Tongtong; Gao, Yan'an; Yu, Li

    2016-09-21

    Proteins are responsible for most biochemical events in human body. It is essential to develop sensitive and selective methods for the detection of proteins. In this study, liquid crystal (LC)-based sensor for highly selective and sensitive detection of lysozyme, concanavalin A (Con A), and bovine serum albumin (BSA) was constructed by utilizing the LC interface decorated with a nonionic surfactant, dodecyl β-d-glucopyranoside. A change of the LC optical images from bright to dark appearance was observed after transferring dodecyl β-d-glucopyranoside onto the aqueous/LC interface due to the formation of stable self-assembled surfactant monolayer, regardless of pH and ion concentrations studied in a wide range. The optical images turned back from dark to bright appearance after addition of lysozyme, Con A and BSA, respectively. Noteworthy is that these proteins can be further distinguished by adding enzyme inhibitors and controlling incubation temperature of the protein solutions based on three different interaction mechanisms between proteins and dodecyl β-d-glucopyranoside, viz. enzymatic hydrolysis, specific saccharide binding, and physical absorption. The LC-based sensor decorated with dodecyl β-d-glucopyranoside shows high sensitivity for protein detection. The limit of detection (LOD) for lysozyme, Con A and BSA reaches around 0.1 μg/mL, 0.01 μg/mL and 0.001 μg/mL, respectively. These results might provide new insights into increasing selectivity and sensitivity of LC-based sensors for the detection of proteins.

  7. Sensitive and selective tumor imaging with novel and highly activatable fluorescence strategies

    NASA Astrophysics Data System (ADS)

    Urano, Yasuteru

    2008-02-01

    Nowadays, several tumor imaging modalities such as MRI, PET and fluorescence imaging techniques have been extensively investigated. One of the central problems associated with these conventional tumor-targeted imaging methods, however, is the fact that the signal contrast between tumor and surrounding tissues relies on the efficient targeting to the tumor and the rapid sequestration or excretion of unbound agent. Among these modalities, only fluorescence imaging technique has a significant feature, in that great signal activation could be achieved which potentially leads to the selective imaging of cancer with higher tumor-to-background ratio. In this symposium, I will present some examples of fluorescence cancer imaging based on highly activatable strategies with using precisely designed novel fluorescence probes. Recently, we developed highly sensitive fluorescence probes for β-galactosidase which is applicable for living cell system. By utilizing these probes, we could establish a novel and highly activatable strategy for sensitive and selective optical imaging of imbedded tumor in the peritoneum. We took a two step procedure in that a lectin is used to localize β-galactosidase to cancer cells as an activating enzyme, and subsequent administration of a highly-sensitive fluorescence probe for the enzyme have afforded remarkable fluorescence activation selectively in tumor mass. Since the tumor-targeted enzyme can catalyze numerous substrate turnovers, a great number of fluorescent molecules could be produced and hence the rapid and sensitive detection of tumor in vivo with high tumor-to-background ratio could be achieved. Moreover, the consequent close-up investigation using fluorescence microscopy revealed that cancer microfoci as small as 200 μm could be successfully visualized.

  8. A nonionic surfactant-decorated liquid crystal sensor for sensitive and selective detection of proteins.

    PubMed

    Wang, Yi; Hu, Qiongzheng; Tian, Tongtong; Gao, Yan'an; Yu, Li

    2016-09-21

    Proteins are responsible for most biochemical events in human body. It is essential to develop sensitive and selective methods for the detection of proteins. In this study, liquid crystal (LC)-based sensor for highly selective and sensitive detection of lysozyme, concanavalin A (Con A), and bovine serum albumin (BSA) was constructed by utilizing the LC interface decorated with a nonionic surfactant, dodecyl β-d-glucopyranoside. A change of the LC optical images from bright to dark appearance was observed after transferring dodecyl β-d-glucopyranoside onto the aqueous/LC interface due to the formation of stable self-assembled surfactant monolayer, regardless of pH and ion concentrations studied in a wide range. The optical images turned back from dark to bright appearance after addition of lysozyme, Con A and BSA, respectively. Noteworthy is that these proteins can be further distinguished by adding enzyme inhibitors and controlling incubation temperature of the protein solutions based on three different interaction mechanisms between proteins and dodecyl β-d-glucopyranoside, viz. enzymatic hydrolysis, specific saccharide binding, and physical absorption. The LC-based sensor decorated with dodecyl β-d-glucopyranoside shows high sensitivity for protein detection. The limit of detection (LOD) for lysozyme, Con A and BSA reaches around 0.1 μg/mL, 0.01 μg/mL and 0.001 μg/mL, respectively. These results might provide new insights into increasing selectivity and sensitivity of LC-based sensors for the detection of proteins. PMID:27590553

  9. Crystal Structures of Trypanosoma brucei Sterol 14[alpha]-Demethylase and Implications for Selective Treatment of Human Infections

    SciTech Connect

    Lepesheva, Galina I.; Park, Hee-Won; Hargrove, Tatiana Y.; Vanhollebeke, Benoit; Wawrzak, Zdzislaw; Harp, Joel M.; Sundaramoorthy, Munirathinam; Nes, W. David; Pays, Etienne; Chaudhuri, Minu; Villalta, Fernando; Waterman, Michael R.

    2010-01-25

    Sterol 14{alpha}-demethylase (14DM, the CYP51 family of cytochrome P450) is an essential enzyme in sterol biosynthesis in eukaryotes. It serves as a major drug target for fungal diseases and can potentially become a target for treatment of human infections with protozoa. Here we present 1.9 {angstrom} resolution crystal structures of 14DM from the protozoan pathogen Trypanosoma brucei, ligand-free and complexed with a strong chemically selected inhibitor N-1-(2,4-dichlorophenyl)-2-(1H-imidazol-1-yl)ethyl-4-(5-phenyl-1,3,4-oxadi-azol-2-yl)benzamide that we previously found to produce potent antiparasitic effects in Trypanosomatidae. This is the first structure of a eukaryotic microsomal 14DM that acts on sterol biosynthesis, and it differs profoundly from that of the water-soluble CYP51 family member from Mycobacterium tuberculosis, both in organization of the active site cavity and in the substrate access channel location. Inhibitor binding does not cause large scale conformational rearrangements, yet induces unanticipated local alterations in the active site, including formation of a hydrogen bond network that connects, via the inhibitor amide group fragment, two remote functionally essential protein segments and alters the heme environment. The inhibitor binding mode provides a possible explanation for both its functionally irreversible effect on the enzyme activity and its selectivity toward the 14DM from human pathogens versus the human 14DM ortholog. The structures shed new light on 14DM functional conservation and open an excellent opportunity for directed design of novel antiparasitic drugs.

  10. Selective increase of alpha 1-adrenoceptors and muscarinic cholinergic receptors in rat cerebral cortex after chronic haloperidol.

    PubMed

    Pazo, J H; Levi de Stein, M; Jerusalinsky, D; Novas, M L; Raskovsky, S; Tumilasci, O R; Medina, J H; De Robertis, E

    1987-06-30

    The effect of chronic administration of haloperidol on alpha 1-, alpha 2-, and beta-adrenoceptors, cholinergic muscarinic, GABAA and benzodiazepine receptors in the cerebral cortex of the rat was investigated. Doses of 0.3 and 2 mg/kg of haloperidol during 7 days increased markedly the density of alpha 1-adrenoceptors without changes in affinity. The alpha 2- and beta-adrenoceptors were not modified after neuroleptic administration. The number of muscarinic receptors were also increased after haloperidol treatment (2 mg/kg/day). However, the GABAA and benzodiazepine binding sites remained unchanged. In the brainstem an increment in the alpha 1-, but not the beta-adrenoceptors was observed. The well known increase in the dopamine receptors in the striatum was confirmed. These observations demonstrate a multireceptor effect of haloperidol in the cerebral cortex.

  11. In vitro protective effect of bacteria-derived bovine alpha interferon I1 against selected bovine viruses.

    PubMed

    Gillespie, J H; Robson, D S; Scott, F W; Schiff, E I

    1985-12-01

    We used bacteria-derived bovine alpha-interferon I1 (Bo IFN-alpha I1) to study its antiviral effect in a bovine turbinate cell line on bovine diarrhea virus, infectious bovine rhinotracheitis virus, parainfluenza 3 virus, and pseudorabies virus. We based our study upon replicate tests for each strain by using a block titration system with various concentrations of Bo IFN-alpha I1 against various concentrations of virus. The data were compiled in two-axis tables (replicate X concentration) and were statistically analyzed by the Spearman-Kärber method. An increase in the concentration of Bo IFN-alpha I1 enhanced its protective effect against every test virus strain. Bo IFN-alpha I1 had a marked in vitro effect on the bovine diarrhea viral strains. It demonstrated less protection against the pseudorabies and parainfluenza 3 viruses. Its effectiveness against the two infectious bovine rhinotracheitis viral strains was lesser and of a low order.

  12. Improved selectivity and sensitivity in the visualization of neurofibrillary tangles, plaques and neuropil threads.

    PubMed

    Cullen, K M; Halliday, G M; Cartwright, H; Kril, J J

    1996-06-01

    Stain sensitivity is a key factor in estimating the frequency of plaques and neurofibrillary tangles in Alzheimer's disease (AD), making it essential that the sensitivity and selectivity of detection methods for identifying these lesions is maximized. Several new, improved techniques have recently been described, although these methods have not been compared quantitatively with those techniques currently recommended for use in standardized diagnostic protocols. In the present study, eight different stains were examined for their selectivity and sensitivity in detecting plaques and tangles in serial tissue sections from AD and control brains. Techniques compared were immunohistochemistry for tau and beta-amyloid, thioflavin S, nickel peroxidase method, and four silver impregnation techniques (Gallyas silver iodide, Campbell-Switzer-Martin, Garvey's modified Bielschowsky and methenamine silver methods). Among these eight staining techniques, the nickel peroxidase proved the most reliable method for the demonstration of the histopathological lesions of AD. This method labels all morphological types of plaques and tangles within a single tissue section, and provides several advantages for the analysis of lesion progression and distribution.

  13. Advancing the sensitivity of selected reaction monitoring-based targeted quantitative proteomics

    SciTech Connect

    Shi, Tujin; Su, Dian; Liu, Tao; Tang, Keqi; Camp, David G.; Qian, Weijun; Smith, Richard D.

    2012-04-01

    Selected reaction monitoring (SRM)—also known as multiple reaction monitoring (MRM)—has emerged as a promising high-throughput targeted protein quantification technology for candidate biomarker verification and systems biology applications. A major bottleneck for current SRM technology, however, is insufficient sensitivity for e.g., detecting low-abundance biomarkers likely present at the pg/mL to low ng/mL range in human blood plasma or serum, or extremely low-abundance signaling proteins in the cells or tissues. Herein we review recent advances in methods and technologies, including front-end immunoaffinity depletion, fractionation, selective enrichment of target proteins/peptides or their posttranslational modifications (PTMs), as well as advances in MS instrumentation, which have significantly enhanced the overall sensitivity of SRM assays and enabled the detection of low-abundance proteins at low to sub- ng/mL level in human blood plasma or serum. General perspectives on the potential of achieving sufficient sensitivity for detection of pg/mL level proteins in plasma are also discussed.

  14. Molecular imprinting ratiometric fluorescence sensor for highly selective and sensitive detection of phycocyanin.

    PubMed

    Wang, Xiaoyan; Yu, Jialuo; Kang, Qi; Shen, Dazhong; Li, Jinhua; Chen, Lingxin

    2016-03-15

    A facile strategy was developed to prepare molecular imprinting ratiometric fluorescence sensor for highly selective and sensitive detection of phycocyanin (PC) based on fluorescence resonance energy transfer (FRET), via a sol-gel polymerization process using nitrobenzoxadiazole (NBD) as fluorescent signal source. The ratio of two fluorescence peak emission intensities of NBD and PC was utilized to determine the concentration of PC, which could effectively reduce the background interference and fluctuation of diverse conditions. As a result, this sensor obtained high sensitivity with a low detection limit of 0.14 nM within 6 min, and excellent recognition specificity for PC over its analogues with a high imprinting factor of 9.1. Furthermore, the sensor attained high recoveries in the range of 93.8-110.2% at three spiking levels of PC, with precisions below 4.7% in seawater and lake water samples. The developed sensor strategy demonstrated simplicity, reliability, rapidity, high selectivity and high sensitivity, proving to be a feasible way to develop high efficient fluorescence sensors and thus potentially applicable for ultratrace analysis of complicated matrices. PMID:26485176

  15. Soluble interleukin-15 receptor alpha (IL-15R alpha)-sushi as a selective and potent agonist of IL-15 action through IL-15R beta/gamma. Hyperagonist IL-15 x IL-15R alpha fusion proteins.

    PubMed

    Mortier, Erwan; Quéméner, Agnès; Vusio, Patricia; Lorenzen, Inken; Boublik, Yvan; Grötzinger, Joachim; Plet, Ariane; Jacques, Yannick

    2006-01-20

    Interleukin-15 (IL-15) is crucial for the generation of multiple lymphocyte subsets (natural killer (NK), NK-T cells, and memory CD8 T cells), and transpresentation of IL-15 by monocytes and dendritic cells has been suggested to be the dominant activating process of these lymphocytes. We have previously shown that a natural soluble form of IL-15R alpha chain corresponding to the entire extracellular domain of IL-15R alpha behaves as a high affinity IL-15 antagonist. In sharp contrast with this finding, we demonstrate in this report that a recombinant, soluble sushi domain of IL-15R alpha, which bears most of the binding affinity for IL-15, behaves as a potent IL-15 agonist by enhancing its binding and biological effects (proliferation and protection from apoptosis) through the IL-15R beta/gamma heterodimer, whereas it does not affect IL-15 binding and function of the tripartite IL-15R alpha/beta/gamma membrane receptor. Our results suggest that, if naturally produced, such soluble sushi domains might be involved in the IL-15 transpresentation mechanism. Fusion proteins (RLI and ILR), in which IL-15 and IL-15R alpha-sushi are attached by a flexible linker, are even more potent than the combination of IL-15 plus sIL-15R alpha-sushi. After binding to IL-15R beta/gamma, RLI is internalized and induces a biological response very similar to the IL-15 high affinity response. Such hyper-IL-15 fusion proteins appear to constitute potent adjuvants for the expansion of lymphocyte subsets.

  16. Long-Observation-Window Band-Selective Homonuclear Decoupling: Increased Sensitivity and Resolution in Solid-State NMR Spectroscopy of Proteins

    PubMed Central

    Struppe, Jochem O.; Yang, Chen; Wang, Yachong; Hernandez, Roy V.; Shamansky, Lisa M.; Mueller, Leonard J.

    2013-01-01

    Sensitivity and resolution are the two fundamental obstacles to extending solid-state nuclear magnetic resonance to even larger protein systems. Here, a novel long-observation-window band-selective homonuclear decoupling (LOW BASHD) scheme is introduced that increases resolution up to a factor of 3 and sensitivity up to 1.8 by decoupling backbone alpha-carbon (Cα) and carbonyl (C′) nuclei in U-13C-labeled proteins during direct 13C acquisition. This approach introduces short (<200 μs) pulse breaks into much longer (~8 ms) sampling windows to efficiently refocus the J-coupling interaction during detection while avoiding the deleterious effects on sensitivity inherent in rapid stroboscopic band-selective homonuclear decoupling techniques. A significant advantage of LOW BASHD detection is that it can be directly incorporated into existing correlation methods, as illustrated here for 2D CACO, NCO, and NCA correlation spectroscopy applied to the β1 immunoglobulin binding domain of protein G and 3D CBCACO correlation spectroscopy applied to the α-subunit of tryptophan synthase. PMID:24095840

  17. Intraluteal prostaglandin biosynthesis and signaling are selectively directed towards PGF2alpha during luteolysis but towards PGE2 during the establishment of pregnancy in sheep.

    PubMed

    Lee, JeHoon; McCracken, John A; Stanley, Jone A; Nithy, Thamizh K; Banu, Sakhila K; Arosh, Joe A

    2012-10-01

    In ruminants, endometrial prostalgandin (PG) F(2alpha) causes functional luteolysis, whereas luteal synthesis of PGF(2alpha) is required for structural luteolysis. PGE(2) is considered to be a luteoprotective mediator. Molecular aspects of luteal PGF(2alpha) and PGE(2) biosynthesis and signaling during the estrous cycle and establishment of pregnancy are largely unknown. The objectives of the present study were 1) to determine the regulation of proteins involved in PGF(2alpha) and PGE(2) biosynthesis, catabolism, transport and signaling in the corpus luteum (CL); 2) to investigate the transport of interferon tau (IFNT), PGF(2alpha), and PGE(2) from the uterus to the ovary through the vascular utero-ovarian plexus (UOP); and 3) to compare the intraluteal production of PGF(2alpha) and PGE(2) on Days 12, 14, and 16 of the estrous cycle and pregnancy in sheep. Our results indicate that luteal PG biosynthesis is selectively directed towards PGF(2alpha) at the time of luteolysis and towards PGE(2) during the establishment of pregnancy. Moreover, the ability of the CL of early pregnancy to resist luteolysis is due to increased intraluteal biosynthesis of PGE(2) and PGE(2) receptor (PTGER) 2 (also known as EP2)- and PTGER4 (also known as EP4)-mediated signaling. We also found that IFNT protein is not transported through the UOP from the uterus to the ovary; in contrast, a large proportion of endometrial PGE(2) is transported from the uterus to the ovary through the UOP. These results indicate that endometrial PGE(2) stimulated by pregnancy is transported locally to the ovary, which increases luteal PGE(2) biosynthesis and hence activates luteal PTGER2 and PTGER4 signaling, thus protecting the CL during the establishment of pregnancy in sheep.

  18. Highly sensitive and selective sugar detection by terahertz nano-antennas

    PubMed Central

    Lee, Dong-Kyu; Kang, Ji-Hun; Lee, Jun-Seok; Kim, Hyo-Seok; Kim, Chulki; Hun Kim, Jae; Lee, Taikjin; Son, Joo-Hiuk; Park, Q-Han; Seo, Minah

    2015-01-01

    Molecular recognition and discrimination of carbohydrates are important because carbohydrates perform essential roles in most living organisms for energy metabolism and cell-to-cell communication. Nevertheless, it is difficult to identify or distinguish various carbohydrate molecules owing to the lack of a significant distinction in the physical or chemical characteristics. Although there has been considerable effort to develop a sensing platform for individual carbohydrates selectively using chemical receptors or an ensemble array, their detection and discrimination limits have been as high in the millimolar concentration range. Here we show a highly sensitive and selective detection method for the discrimination of carbohydrate molecules using nano-slot-antenna array-based sensing chips which operate in the terahertz (THz) frequency range (0.5–2.5 THz). This THz metamaterial sensing tool recognizes various types of carbohydrate molecules over a wide range of molecular concentrations. Strongly localized and enhanced terahertz transmission by nano-antennas can effectively increase the molecular absorption cross sections, thereby enabling the detection of these molecules even at low concentrations. We verified the performance of nano-antenna sensing chip by both THz spectra and images of transmittance. Screening and identification of various carbohydrates can be applied to test even real market beverages with a high sensitivity and selectivity. PMID:26494203

  19. Highly sensitive and selective sugar detection by terahertz nano-antennas.

    PubMed

    Lee, Dong-Kyu; Kang, Ji-Hun; Lee, Jun-Seok; Kim, Hyo-Seok; Kim, Chulki; Kim, Jae Hun; Lee, Taikjin; Son, Joo-Hiuk; Park, Q-Han; Seo, Minah

    2015-10-23

    Molecular recognition and discrimination of carbohydrates are important because carbohydrates perform essential roles in most living organisms for energy metabolism and cell-to-cell communication. Nevertheless, it is difficult to identify or distinguish various carbohydrate molecules owing to the lack of a significant distinction in the physical or chemical characteristics. Although there has been considerable effort to develop a sensing platform for individual carbohydrates selectively using chemical receptors or an ensemble array, their detection and discrimination limits have been as high in the millimolar concentration range. Here we show a highly sensitive and selective detection method for the discrimination of carbohydrate molecules using nano-slot-antenna array-based sensing chips which operate in the terahertz (THz) frequency range (0.5-2.5 THz). This THz metamaterial sensing tool recognizes various types of carbohydrate molecules over a wide range of molecular concentrations. Strongly localized and enhanced terahertz transmission by nano-antennas can effectively increase the molecular absorption cross sections, thereby enabling the detection of these molecules even at low concentrations. We verified the performance of nano-antenna sensing chip by both THz spectra and images of transmittance. Screening and identification of various carbohydrates can be applied to test even real market beverages with a high sensitivity and selectivity.

  20. Highly sensitive and selective sugar detection by terahertz nano-antennas.

    PubMed

    Lee, Dong-Kyu; Kang, Ji-Hun; Lee, Jun-Seok; Kim, Hyo-Seok; Kim, Chulki; Kim, Jae Hun; Lee, Taikjin; Son, Joo-Hiuk; Park, Q-Han; Seo, Minah

    2015-01-01

    Molecular recognition and discrimination of carbohydrates are important because carbohydrates perform essential roles in most living organisms for energy metabolism and cell-to-cell communication. Nevertheless, it is difficult to identify or distinguish various carbohydrate molecules owing to the lack of a significant distinction in the physical or chemical characteristics. Although there has been considerable effort to develop a sensing platform for individual carbohydrates selectively using chemical receptors or an ensemble array, their detection and discrimination limits have been as high in the millimolar concentration range. Here we show a highly sensitive and selective detection method for the discrimination of carbohydrate molecules using nano-slot-antenna array-based sensing chips which operate in the terahertz (THz) frequency range (0.5-2.5 THz). This THz metamaterial sensing tool recognizes various types of carbohydrate molecules over a wide range of molecular concentrations. Strongly localized and enhanced terahertz transmission by nano-antennas can effectively increase the molecular absorption cross sections, thereby enabling the detection of these molecules even at low concentrations. We verified the performance of nano-antenna sensing chip by both THz spectra and images of transmittance. Screening and identification of various carbohydrates can be applied to test even real market beverages with a high sensitivity and selectivity. PMID:26494203

  1. A highly selective electrochemical impedance spectroscopy-based aptasensor for sensitive detection of acetamiprid.

    PubMed

    Fan, Lifang; Zhao, Guohua; Shi, Huijie; Liu, Meichuan; Li, Zhengxin

    2013-05-15

    A simple aptasensor for sensitive and selective detection of acetamiprid has been developed based on electrochemical impedance spectroscopy (EIS). To improve sensitivity of the aptasensor, gold nanoparticles (AuNPs) were electrodeposited on the bare gold electrode surface by cycle voltammetry (CV), which was employed as a platform for aptamer immobilization. With the addition of acetamiprid, the formation of acetamiprid-aptamer complex on the AuNPs-deposited electrode surface resulted in an increase of electron transfer resistance (Ret). The change of Ret strongly depends on acetamiprid concentration, which is applied for acetamiprid quantification. A wide linear range was obtained from 5 to 600nM with a low detection limit of 1nM. The control experiments performed by employing the pesticides that may coexist or have similar structure with acetamiprid demonstrate that the aptasensor has only specific recognition to acetamiprid, resulting in high selectivity of the aptasensor. The dissociation constant, Kd of 23.41nM for acetamiprid-aptamer complex has been determined from the differential capacitance (Cd) by assuming a Langmuir isotherm, which indicates strong interaction between acetamiprid and aptamer, further proving high selectivity of the aptasensor. Besides, the applicability of the developed aptasensor has been successfully evaluated by determining acetamiprid in the real samples, wastewater and tomatoes. PMID:23274191

  2. A Catalytic Beacon Sensor for Uranium with Parts-per-Trillion Sensitivity and Millionfold Selectivity

    SciTech Connect

    Liu, Juewen; Brown, Andrea K.; Meng, Xiangli; Cropek, Donald M.; IstokD., Jonathan; Watson, David B; Liu, Yi

    2007-01-01

    Here, we report a catalytic beacon sensor for uranyl (UO{sub 2}{sup 2+}) based on an in vitro-selected UO{sub 2}{sup 2+}-specific DNAzyme. The sensor consists of a DNA enzyme strand with a 3' quencher and a DNA substrate with a ribonucleotide adenosine (rA) in the middle and a fluorophore and a quencher at the 5' and 3' ends, respectively. The presence of UO{sub 2}{sup 2+} causes catalytic cleavage of the DNA substrate strand at the rA position and release of the fluorophore and thus dramatic increase of fluorescence intensity. The sensor has a detection limit of 11 parts per trillion (45 pM), a dynamic range up to 400 nM, and selectivity of >1-million-fold over other metal ions. The most interfering metal ion, Th(IV), interacts with the fluorescein fluorophore, causing slightly enhanced fluorescence intensity, with an apparent dissociation constant of {approx}230 {micro}M. This sensor rivals the most sensitive analytical instruments for uranium detection, and its application in detecting uranium in contaminated soil samples is also demonstrated. This work shows that simple, cost-effective, and portable metal sensors can be obtained with similar sensitivity and selectivity as much more expensive and sophisticated analytical instruments. Such a sensor will play an important role in environmental remediation of radionuclides such as uranium.

  3. Selective and Sensitive Sensing of Flame Retardant Chemicals Through Phage Display Discovered Recognition Peptide.

    PubMed

    Jin, Hyo-Eon; Zueger, Chris; Chung, Woo-Jae; Wong, Winnie; Lee, Byung Yang; Lee, Seung-Wuk

    2015-11-11

    We report a highly selective and sensitive biosensor for the detection of an environmentally toxic molecule, decabrominated diphenyl ether (DBDE), one of the most common congeners of the polybrominated frame retardants (polybrominated diphenyl ether (PBDE)), using newly discovered DBDE peptide receptors integrated with carbon nanotube field-effect transistors (CNT-FET). The specific DBDE peptide receptor was identified using a high-throughput screening process of phage library display. The resulting binding peptide carries an interesting consensus binding pocket with two Trp-His/Asn-Trp repeats, which binds to the DBDE in a multivalent manner. We integrated the novel DBDE binding peptide onto the CNT-FET using polydiacetylene coating materials linked through cysteine-maleimide click chemistry. The resulting biosensor could detect the desired DBDE selectively with a 1 fM detection limit. Our combined approaches of selective receptor discovery, material nanocoating through click chemistry, and integration onto a sensitive CNT-FET electronic sensor for desired target chemicals will pave the way toward the rapid development of portable and easy-to-use biosensors for desired chemicals to protect our health and environment. PMID:26455834

  4. Selective and sensitive colorimetric detection of stringent alarmone ppGpp with Fenton-like reagent.

    PubMed

    Zheng, Lin Ling; Huang, Cheng Zhi

    2014-12-01

    Stringent alarmone, namely, guanosine 3'-diphosphate-5'-diphosphate (ppGpp), is a global regulator that plays a critical role in the survival, growth, metabolism, and many other vital processes of microorganisms. Because of its structural similarity to normal nucleotides, it is also a challenge for the selective and sensitive detection of ppGpp nowadays. Herein, we developed a colorimetric method for the selective detection of ppGpp by inhibiting the redox reaction between Fenton-like reagent (composed of Fe(3+) and H2O2) with 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS). Owing to the strong coordination affinity between ppGpp and Fe(3+), the chromogenic reaction between ABTS and Fenton-like reagent, occurred in aqueous medium at 37 °C and resulted in a bluish-green solution, which was inhibited with the addition of ppGpp. This phenomenon forms the basis for the colorimetric detection of ppGpp, with a detection limit of 0.19 μM and good selectivity for ppGpp over other nucleotides and anions. Furthermore, the results could be visualized by the naked eye, and the sensitivity of the naked-eye observation could even be further improved with the aid of the introduction of a background color.

  5. A catalytic beacon sensor for uranium with parts-per-trillion sensitivity and millionfold selectivity.

    PubMed

    Liu, Juewen; Brown, Andrea K; Meng, Xiangli; Cropek, Donald M; Istok, Jonathan D; Watson, David B; Lu, Yi

    2007-02-13

    Here, we report a catalytic beacon sensor for uranyl (UO2(2+)) based on an in vitro-selected UO2(2+)-specific DNAzyme. The sensor consists of a DNA enzyme strand with a 3' quencher and a DNA substrate with a ribonucleotide adenosine (rA) in the middle and a fluorophore and a quencher at the 5' and 3' ends, respectively. The presence of UO2(2+) causes catalytic cleavage of the DNA substrate strand at the rA position and release of the fluorophore and thus dramatic increase of fluorescence intensity. The sensor has a detection limit of 11 parts per trillion (45 pM), a dynamic range up to 400 nM, and selectivity of >1-million-fold over other metal ions. The most interfering metal ion, Th(IV), interacts with the fluorescein fluorophore, causing slightly enhanced fluorescence intensity, with an apparent dissociation constant of approximately 230 microM. This sensor rivals the most sensitive analytical instruments for uranium detection, and its application in detecting uranium in contaminated soil samples is also demonstrated. This work shows that simple, cost-effective, and portable metal sensors can be obtained with similar sensitivity and selectivity as much more expensive and sophisticated analytical instruments. Such a sensor will play an important role in environmental remediation of radionuclides such as uranium.

  6. Highly sensitive and selective sugar detection by terahertz nano-antennas

    NASA Astrophysics Data System (ADS)

    Lee, Dong-Kyu; Kang, Ji-Hun; Lee, Jun-Seok; Kim, Hyo-Seok; Kim, Chulki; Hun Kim, Jae; Lee, Taikjin; Son, Joo-Hiuk; Park, Q.-Han; Seo, Minah

    2015-10-01

    Molecular recognition and discrimination of carbohydrates are important because carbohydrates perform essential roles in most living organisms for energy metabolism and cell-to-cell communication. Nevertheless, it is difficult to identify or distinguish various carbohydrate molecules owing to the lack of a significant distinction in the physical or chemical characteristics. Although there has been considerable effort to develop a sensing platform for individual carbohydrates selectively using chemical receptors or an ensemble array, their detection and discrimination limits have been as high in the millimolar concentration range. Here we show a highly sensitive and selective detection method for the discrimination of carbohydrate molecules using nano-slot-antenna array-based sensing chips which operate in the terahertz (THz) frequency range (0.5-2.5 THz). This THz metamaterial sensing tool recognizes various types of carbohydrate molecules over a wide range of molecular concentrations. Strongly localized and enhanced terahertz transmission by nano-antennas can effectively increase the molecular absorption cross sections, thereby enabling the detection of these molecules even at low concentrations. We verified the performance of nano-antenna sensing chip by both THz spectra and images of transmittance. Screening and identification of various carbohydrates can be applied to test even real market beverages with a high sensitivity and selectivity.

  7. Highly sensitive and selective optical chemosensor for determination of Cu2+ in aqueous solution.

    PubMed

    Xiang, Yu; Li, Zifan; Chen, Xiaotong; Tong, Aijun

    2008-02-15

    A highly selective and sensitive rhodamine-based colorimetric chemosensor (1) for quantification of divalent copper in aqueous solution has been investigated in this work. It was designed using salicylaldehyde hydrazone and rhodamine 6G as copper-chelating and signal-reporting groups, respectively. In environmentally friendly media (50%(v/v) water/ethanol and 10mM NaAc-HAc neutral buffer (pH 7.0)), the sensor exhibited selective absorbance enhancement to Cu(2+) over other metal ions at 529 nm, with a dynamic working range of 0.05-5.00 microM and a detection limit of 10nM Cu(2+), respectively. To achieve fluorometric determination of Cu(2+), the Cu(2+)-induced absorbance enhancement of 1 was efficiently converted to fluorescence quenching by fluorescence inner filter effects using rhodamine B (RB) as a fluorophore. The selectivity and sensitivity of fluorescence analysis were similar to those of absorptiometric measurement. Both absorptiometric and fluorometric methods were successfully applied to the detection of Cu(2+) in three water samples. PMID:18371763

  8. Selective and Sensitive Sensing of Flame Retardant Chemicals Through Phage Display Discovered Recognition Peptide.

    PubMed

    Jin, Hyo-Eon; Zueger, Chris; Chung, Woo-Jae; Wong, Winnie; Lee, Byung Yang; Lee, Seung-Wuk

    2015-11-11

    We report a highly selective and sensitive biosensor for the detection of an environmentally toxic molecule, decabrominated diphenyl ether (DBDE), one of the most common congeners of the polybrominated frame retardants (polybrominated diphenyl ether (PBDE)), using newly discovered DBDE peptide receptors integrated with carbon nanotube field-effect transistors (CNT-FET). The specific DBDE peptide receptor was identified using a high-throughput screening process of phage library display. The resulting binding peptide carries an interesting consensus binding pocket with two Trp-His/Asn-Trp repeats, which binds to the DBDE in a multivalent manner. We integrated the novel DBDE binding peptide onto the CNT-FET using polydiacetylene coating materials linked through cysteine-maleimide click chemistry. The resulting biosensor could detect the desired DBDE selectively with a 1 fM detection limit. Our combined approaches of selective receptor discovery, material nanocoating through click chemistry, and integration onto a sensitive CNT-FET electronic sensor for desired target chemicals will pave the way toward the rapid development of portable and easy-to-use biosensors for desired chemicals to protect our health and environment.

  9. Highly sensitive and selective gas sensor using hydrophilic and hydrophobic graphenes.

    PubMed

    Some, Surajit; Xu, Yang; Kim, Youngmin; Yoon, Yeoheung; Qin, Hongyi; Kulkarni, Atul; Kim, Taesung; Lee, Hyoyoung

    2013-01-01

    New hydrophilic 2D graphene oxide (GO) nanosheets with various oxygen functional groups were employed to maintain high sensitivity in highly unfavorable environments (extremely high humidity, strong acidic or basic). Novel one-headed polymer optical fiber sensor arrays using hydrophilic GO and hydrophobic reduced graphene oxide (rGO) were carefully designed, leading to the selective sensing of volatile organic gases for the first time. The two physically different surfaces of GO and rGO could provide the sensing ability to distinguish between tetrahydrofuran (THF) and dichloromethane (MC), respectively, which is the most challenging issue in the area of gas sensors. The eco-friendly physical properties of GO allowed for faster sensing and higher sensitivity when compared to previous results for rGO even under extreme environments of over 90% humidity, making it the best choice for an environmentally friendly gas sensor. PMID:23736838

  10. Highly Sensitive and Selective Gas Sensor Using Hydrophilic and Hydrophobic Graphenes

    NASA Astrophysics Data System (ADS)

    Some, Surajit; Xu, Yang; Kim, Youngmin; Yoon, Yeoheung; Qin, Hongyi; Kulkarni, Atul; Kim, Taesung; Lee, Hyoyoung

    2013-06-01

    New hydrophilic 2D graphene oxide (GO) nanosheets with various oxygen functional groups were employed to maintain high sensitivity in highly unfavorable environments (extremely high humidity, strong acidic or basic). Novel one-headed polymer optical fiber sensor arrays using hydrophilic GO and hydrophobic reduced graphene oxide (rGO) were carefully designed, leading to the selective sensing of volatile organic gases for the first time. The two physically different surfaces of GO and rGO could provide the sensing ability to distinguish between tetrahydrofuran (THF) and dichloromethane (MC), respectively, which is the most challenging issue in the area of gas sensors. The eco-friendly physical properties of GO allowed for faster sensing and higher sensitivity when compared to previous results for rGO even under extreme environments of over 90% humidity, making it the best choice for an environmentally friendly gas sensor.

  11. Sensitive hydrogen sensor based on selectively infiltrated photonic crystal fiber with Pt-loaded WO₃ coating.

    PubMed

    Wang, Ying; Wang, D N; Yang, Fan; Li, Zhi; Yang, Minghong

    2014-07-01

    A sensitive hydrogen sensing device based on a selectively infiltrated photonic crystal fiber (PCF) coated with Pt-loaded WO₃ is demonstrated. With Pt-loaded WO₃ coating acting as the catalytic layer, hydrogen undergoes an exothermic reaction with oxygen and releases heat when the device is exposed to gas mixtures of air and hydrogen, which induces local temperature change in the PCF and hence leads to the resonant wavelength shift of the proposed device. The maximum wavelength shift of 98.5 nm is obtained with a 10-mm-long infiltrated PCF for 4% (v/v) H₂ concentration, and a hydrogen sensitivity of 32.3 nm/% (v/v) H₂ is achieved within the range of 1%-4% (v/v) H₂ in air. PMID:24978759

  12. Highly Sensitive and Selective Gas Sensor Using Hydrophilic and Hydrophobic Graphenes

    PubMed Central

    Some, Surajit; Xu, Yang; Kim, Youngmin; Yoon, Yeoheung; Qin, Hongyi; Kulkarni, Atul; Kim, Taesung; Lee, Hyoyoung

    2013-01-01

    New hydrophilic 2D graphene oxide (GO) nanosheets with various oxygen functional groups were employed to maintain high sensitivity in highly unfavorable environments (extremely high humidity, strong acidic or basic). Novel one-headed polymer optical fiber sensor arrays using hydrophilic GO and hydrophobic reduced graphene oxide (rGO) were carefully designed, leading to the selective sensing of volatile organic gases for the first time. The two physically different surfaces of GO and rGO could provide the sensing ability to distinguish between tetrahydrofuran (THF) and dichloromethane (MC), respectively, which is the most challenging issue in the area of gas sensors. The eco-friendly physical properties of GO allowed for faster sensing and higher sensitivity when compared to previous results for rGO even under extreme environments of over 90% humidity, making it the best choice for an environmentally friendly gas sensor. PMID:23736838

  13. Leveraging zinc interstitials and oxygen vacancies for sensitive biomolecule detection through selective surface functionalization

    NASA Astrophysics Data System (ADS)

    Radha Shanmugam, Nandhinee; Muthukumar, Sriram; Chaudhry, Shajee; Prasad, Shalini

    2015-03-01

    In this study, functionally engineered EIS technique was implemented to investigate the influence of surface functionalization on sensitivity of biomolecule detection using nanostructured ZnO platform. Organic molecules with thiol and carboxylic functional groups were chosen to control biomolecule immobilization on zinc and oxygen-terminated 2D planar and 1D nanostructured ZnO surfaces. The amount of functionalization and its influence on charge perturbations at the ZnO-electrolyte interface were studied using fluorescence and EIS measurements. We observed the dependence of charge transfer on both the polarity of platform and concentration of cross-linker molecules. Such selectively modified surfaces were used for detection of cortisol, a major stress indicator. Results demonstrated preferential binding of thiol groups to Zn terminations and thus leveraging ZnO interstitials increases the sensitivity of detection over larger dynamic range with detection limit at 10fg/mL.

  14. TU-C-BRE-08: IMRT QA: Selecting Meaningful Gamma Criteria Based On Error Detection Sensitivity

    SciTech Connect

    Steers, J; Fraass, B

    2014-06-15

    Purpose: To develop a strategy for defining meaningful tolerance limits and studying the sensitivity of IMRT QA gamma criteria by inducing known errors in QA plans. Methods: IMRT QA measurements (ArcCHECK, Sun Nuclear) were compared to QA plan calculations with induced errors. Many (>24) gamma comparisons between data and calculations were performed for each of several kinds of cases and classes of induced error types with varying magnitudes (e.g. MU errors ranging from -10% to +10%), resulting in over 3,000 comparisons. Gamma passing rates for each error class and case were graphed against error magnitude to create error curves in order to represent the range of missed errors in routine IMRT QA using various gamma criteria. Results: This study demonstrates that random, case-specific, and systematic errors can be detected by the error curve analysis. Depending on location of the peak of the error curve (e.g., not centered about zero), 3%/3mm threshold=10% criteria may miss MU errors of up to 10% and random MLC errors of up to 5 mm. Additionally, using larger dose thresholds for specific devices may increase error sensitivity (for the same X%/Ymm criteria) by up to a factor of two. This analysis will allow clinics to select more meaningful gamma criteria based on QA device, treatment techniques, and acceptable error tolerances. Conclusion: We propose a strategy for selecting gamma parameters based on the sensitivity of gamma criteria and individual QA devices to induced calculation errors in QA plans. Our data suggest large errors may be missed using conventional gamma criteria and that using stricter criteria with an increased dose threshold may reduce the range of missed errors. This approach allows quantification of gamma criteria sensitivity and is straightforward to apply to other combinations of devices and treatment techniques.

  15. Inhibitors of tumor progression loci-2 (Tpl2) kinase and tumor necrosis factor alpha (TNF-alpha) production: selectivity and in vivo antiinflammatory activity of novel 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles.

    PubMed

    Green, Neal; Hu, Yonghan; Janz, Kristin; Li, Huan-Qiu; Kaila, Neelu; Guler, Satenig; Thomason, Jennifer; Joseph-McCarthy, Diane; Tam, Steve Y; Hotchandani, Rajeev; Wu, Junjun; Huang, Adrian; Wang, Qin; Leung, Louis; Pelker, Jefferey; Marusic, Suzana; Hsu, Sang; Telliez, Jean-Baptiste; Hall, J Perry; Cuozzo, John W; Lin, Lih-Ling

    2007-09-20

    Tumor progression loci-2 (Tpl2) (Cot/MAP3K8) is a serine/threonine kinase in the MAP3K family directly upstream of MEK. Recent studies using Tpl2 knockout mice have indicated an important role for Tpl2 in the lipopolysaccharide (LPS) induced production of tumor necrosis factor alpha (TNF-alpha) and other proinflammatory cytokines involved in diseases such as rheumatoid arthritis. Initial 4-anilino-6-aminoquinoline-3-carbonitrile leads showed poor selectivity for Tpl2 over epidermal growth factor receptor (EGFR) kinase. Using molecular modeling and crystallographic data of the EGFR kinase domain with and without an EGFR kinase-specific 4-anilinoquinazoline inhibitor (erlotinib, Tarceva), we hypothesized that we could diminish the inhibition of EGFR kinase by substitution at the C-8 position of our 4-anilino-6-aminoquinoline-3-carbonitrile leads. The 8-substituted-4-anilino-6-aminoquinoline-3-carbonitriles were prepared from the appropriate 2-substituted 4-nitroanilines. Modifications to the C-6 and C-8 positions led to the identification of compounds with increased inhibition of TNF-alpha release from LPS-stimulated rat and human blood, and these analogues were also highly selective for Tpl2 kinase over EGFR kinase. Further structure-activity based modifications led to the identification of 8-bromo-4-(3-chloro-4-fluorophenylamino)-6-[(1-methyl-1H-imidazol-4-yl)methylamino]quinoline-3-carbonitrile, which demonstrated in vitro as well as in vivo efficacy in inhibition of LPS-induced TNF-alpha production. PMID:17715908

  16. Intrinsic Selectivity and Structure Sensitivity of Rhodium Catalysts for C(2+) Oxygenate Production.

    PubMed

    Yang, Nuoya; Medford, Andrew J; Liu, Xinyan; Studt, Felix; Bligaard, Thomas; Bent, Stacey F; Nørskov, Jens K

    2016-03-23

    Synthesis gas (CO + H2) conversion is a promising route to converting coal, natural gas, or biomass into synthetic liquid fuels. Rhodium has long been studied as it is the only elemental catalyst that has demonstrated selectivity to ethanol and other C2+ oxygenates. However, the fundamentals of syngas conversion over rhodium are still debated. In this work a microkinetic model is developed for conversion of CO and H2 into methane, ethanol, and acetaldehyde on the Rh (211) and (111) surfaces, chosen to describe steps and close-packed facets on catalyst particles. The model is based on DFT calculations using the BEEF-vdW functional. The mean-field kinetic model includes lateral adsorbate-adsorbate interactions, and the BEEF-vdW error estimation ensemble is used to propagate error from the DFT calculations to the predicted rates. The model shows the Rh(211) surface to be ∼6 orders of magnitude more active than the Rh(111) surface, but highly selective toward methane, while the Rh(111) surface is intrinsically selective toward acetaldehyde. A variety of Rh/SiO2 catalysts are synthesized, tested for catalytic oxygenate production, and characterized using TEM. The experimental results indicate that the Rh(111) surface is intrinsically selective toward acetaldehyde, and a strong inverse correlation between catalytic activity and oxygenate selectivity is observed. Furthermore, iron impurities are shown to play a key role in modulating the selectivity of Rh/SiO2 catalysts toward ethanol. The experimental observations are consistent with the structure-sensitivity predicted from theory. This work provides an improved atomic-scale understanding and new insight into the mechanism, active site, and intrinsic selectivity of syngas conversion over rhodium catalysts and may also guide rational design of alloy catalysts made from more abundant elements. PMID:26958997

  17. Selective and sensitive detection of chromium(VI) in waters using electrospray ionization mass spectrometry.

    PubMed

    Weldy, Effie; Wolff, Chloe; Miao, Zhixin; Chen, Hao

    2013-09-01

    From 2000 through 2011, there were 14 criminal cases of violations of the Clean Water Act involving the discharge of chromium, a toxic heavy metal, into drinking and surface water sources. As chromium(VI), a potential carcinogen present in the environment, represents a significant safety concern, it is currently the subject of an EPA health risk assessment. Therefore, sensitive and selective detection of this species is highly desired. This study reports the analysis of chromium(VI) in water samples by electrospray ionization mass spectrometry (ESI-MS) following its reduction and complexation with ammonium pyrrolidinedithiocarbamate (APDC). The reduction and subsequent complexation produce a characteristic [Cr(III)O]-PDC complex which can be detected as a protonated ion of m/z 507 in the positive ion mode. The detection is selective to chromium(VI) under acidic pH, even in the presence of chromium(III) and other metal ions, providing high specificity. Different water samples were examined, including deionized, tap, and river waters, and sensitive detection was achieved. In the case of deionized water, quantification over the concentration range of 3.7 to 148ppb gave an excellent correlation coefficient of 0.9904 using the enhanced MS mode scan. Using the single-reaction monitoring (SRM) mode (monitoring the characteristic fragmentation of m/z 507 to m/z 360), the limit of detection (LOD) was found to be 0.25ppb. The LOD of chromium(VI) for both tap and river water samples was determined to be 2.0ppb. A preconcentration strategy using simple vacuum evaporation of the aqueous sample was shown to further improve the ESI signal by 15 fold. This method, with high sensitivity and selectivity, should provide a timely solution for the real-world analysis of toxic chromium(VI). PMID:23937937

  18. Selective and sensitive detection of chromium(VI) in waters using electrospray ionization mass spectrometry.

    PubMed

    Weldy, Effie; Wolff, Chloe; Miao, Zhixin; Chen, Hao

    2013-09-01

    From 2000 through 2011, there were 14 criminal cases of violations of the Clean Water Act involving the discharge of chromium, a toxic heavy metal, into drinking and surface water sources. As chromium(VI), a potential carcinogen present in the environment, represents a significant safety concern, it is currently the subject of an EPA health risk assessment. Therefore, sensitive and selective detection of this species is highly desired. This study reports the analysis of chromium(VI) in water samples by electrospray ionization mass spectrometry (ESI-MS) following its reduction and complexation with ammonium pyrrolidinedithiocarbamate (APDC). The reduction and subsequent complexation produce a characteristic [Cr(III)O]-PDC complex which can be detected as a protonated ion of m/z 507 in the positive ion mode. The detection is selective to chromium(VI) under acidic pH, even in the presence of chromium(III) and other metal ions, providing high specificity. Different water samples were examined, including deionized, tap, and river waters, and sensitive detection was achieved. In the case of deionized water, quantification over the concentration range of 3.7 to 148ppb gave an excellent correlation coefficient of 0.9904 using the enhanced MS mode scan. Using the single-reaction monitoring (SRM) mode (monitoring the characteristic fragmentation of m/z 507 to m/z 360), the limit of detection (LOD) was found to be 0.25ppb. The LOD of chromium(VI) for both tap and river water samples was determined to be 2.0ppb. A preconcentration strategy using simple vacuum evaporation of the aqueous sample was shown to further improve the ESI signal by 15 fold. This method, with high sensitivity and selectivity, should provide a timely solution for the real-world analysis of toxic chromium(VI).

  19. Genetics of Microenvironmental Sensitivity of Body Weight in Rainbow Trout (Oncorhynchus mykiss) Selected for Improved Growth

    PubMed Central

    Janhunen, Matti; Kause, Antti; Vehviläinen, Harri; Järvisalo, Otso

    2012-01-01

    Microenvironmental sensitivity of a genotype refers to the ability to buffer against non-specific environmental factors, and it can be quantified by the amount of residual variation in a trait expressed by the genotype’s offspring within a (macro)environment. Due to the high degree of polymorphism in behavioral, growth and life-history traits, both farmed and wild salmonids are highly susceptible to microenvironmental variation, yet the heritable basis of this characteristic remains unknown. We estimated the genetic (co)variance of body weight and its residual variation in 2-year-old rainbow trout (Oncorhynchus mykiss) using a multigenerational data of 45,900 individuals from the Finnish national breeding programme. We also tested whether or not microenvironmental sensitivity has been changed as a correlated genetic response when genetic improvement for growth has been practiced over five generations. The animal model analysis revealed the presence of genetic heterogeneity both in body weight and its residual variation. Heritability of residual variation was remarkably lower (0.02) than that for body weight (0.35). However, genetic coefficient of variation was notable in both body weight (14%) and its residual variation (37%), suggesting a substantial potential for selection responses in both traits. Furthermore, a significant negative genetic correlation (−0.16) was found between body weight and its residual variation, i.e., rapidly growing genotypes are also more tolerant to perturbations in microenvironment. The genetic trends showed that fish growth was successfully increased by selective breeding (an average of 6% per generation), whereas no genetic change occurred in residual variation during the same period. The results imply that genetic improvement for body weight does not cause a concomitant increase in microenvironmental sensitivity. For commercial production, however, there may be high potential to simultaneously improve weight gain and increase its

  20. Cheat sensitive quantum bit commitment via pre- and post-selected quantum states

    NASA Astrophysics Data System (ADS)

    Li, Yan-Bing; Wen, Qiao-Yan; Li, Zi-Chen; Qin, Su-Juan; Yang, Ya-Tao

    2014-01-01

    Cheat sensitive quantum bit commitment is a most important and realizable quantum bit commitment (QBC) protocol. By taking advantage of quantum mechanism, it can achieve higher security than classical bit commitment. In this paper, we propose a QBC schemes based on pre- and post-selected quantum states. The analysis indicates that both of the two participants' cheat strategies will be detected with non-zero probability. And the protocol can be implemented with today's technology as a long-term quantum memory is not needed.

  1. A sensitive and selective fluorescence sensor for the detection of arsenic(III) in organic media.

    PubMed

    Ezeh, Vivian C; Harrop, Todd C

    2012-02-01

    Arsenic contamination is a leading environmental problem. As such, levels of this toxic metalloid must be constantly monitored by reliable and low-cost methodologies. Because the currently accepted upper limit for arsenic in water is 10 ppb, very sensitive and selective detection strategies must be developed. Herein we describe the synthesis and characterization of a fluorescent chemical probe, namely, ArsenoFluor1, which is the first example of a chemosensor for As(3+) detection in organic solvents at 298 K. AF1 exhibits a 25-fold fluorescence increase in the presence of As(3+) at λ(em) = 496 nm in THF, which is selective for As(3+) over other biologically relevant ions (such as Na(+), Mg(2+), Fe(2+), and Zn(2+)) and displays a sub-ppb detection limit.

  2. Sensitivity- and effort-gain analysis: multilead ECG electrode array selection for activation time imaging.

    PubMed

    Hintermüller, Christoph; Seger, Michael; Pfeifer, Bernhard; Fischer, Gerald; Modre, Robert; Tilg, Bernhard

    2006-10-01

    Methods for noninvasive imaging of electric function of the heart might become clinical standard procedure the next years. Thus, the overall procedure has to meet clinical requirements as an easy and fast application. In this paper, we propose a new electrode array which improves the resolution of methods for activation time imaging considering clinical constraints such as easy to apply and compatibility with routine leads. For identifying the body-surface regions where the body surface potential (BSP) is most sensitive to changes in transmembrane potential (TMP), a virtual array method was used to compute local linear dependency (LLD) maps. The virtual array method computes a measure for the LLD in every point on the body surface. The most suitable number and position of the electrodes within the sensitive body surface regions was selected by constructing effort gain (EG) plots. Such a plot depicts the relative attainable rank of the leadfield matrix in relation to the increase in number of electrodes required to build the electrode array. The attainable rank itself was computed by a detector criterion. Such a criterion estimates the maximum number of source space eigenvectors not covered by noise when being mapped to the electrode space by the leadfield matrix and recorded by a detector. From the sensitivity maps, we found that the BSP is most sensitive to changes in TMP on the upper left frontal and dorsal body surface. These sensitive regions are covered best by an electrode array consisting of two L-shaped parts of approximately 30 cm x 30 cm and approximately 20 cm x 20 cm. The EG analysis revealed that the array meeting clinical requirements best and improving the resolution of activation time imaging consists of 125 electrodes with a regular horizontal and vertical spacing of 2-3 cm.

  3. Kinetic interconversion of rat and bovine homologs of the alpha subunit of an amiloride-sensitive Na+ channel by C-terminal truncation of the bovine subunit.

    PubMed

    Fuller, C M; Ismailov, I I; Berdiev, B K; Shlyonsky, V G; Benos, D J

    1996-10-25

    We have recently cloned the alpha subunit of a bovine amiloride-sensitive Na+ channel (alphabENaC). This subunit shares extensive homology with both rat and human alphaENaC subunits but shows marked divergence at the C terminus beginning at amino acid 584 of the 697-residue sequence. When incorporated into planar lipid bilayers, alphabENaC almost exclusively exhibits a main transition to 39 picosiemens (pS) with very rare 13 pS step transitions to one of two subconductance states (26 and 13 pS). In contrast, the alpha subunit of the rat renal homolog of ENaC (alpharENaC) has a main transition step to 13 pS that is almost constituitively open, with a second stepwise transition of 26 to 39 pS. A deletion mutant of alphabENaC, encompassing the entire C-terminal region (R567X), converts the kinetic behavior of alphabENaC to that of alpharENaC, i. e. a transition to 13 pS followed by a second 26 pS transition to 39 pS. Chemical cross-linking of R567X restores the wild-type alphabENaC gating pattern, whereas treatment with the reducing agent dithiothreitol produced only 13 pS transitions. In contrast, an equivalent C-terminal truncation of alpharENaC (R613X) had no effect on the gating pattern of alpharENaC. These results are consistent with the hypothesis that interactions between the C termini of alphabENaC account for the different kinetic behavior of this member of the ENaC family of Na+ channels. PMID:8900133

  4. Role of the nitric oxide synthase pathway in inhibition of growth of interferon-sensitive and interferon-resistant Rickettsia prowazekii strains in L929 cells treated with tumor necrosis factor alpha and gamma interferon.

    PubMed Central

    Turco, J; Winkler, H H

    1993-01-01

    The ability of tumor necrosis factor alpha (TNF-alpha) alone and in combination with gamma interferon (IFN-gamma) to inhibit the growth of interferon-sensitive and -resistant Rickettsia prowazekii strains in mouse L929 cells was examined, and the possible role of the nitric oxide synthase pathway in the suppression of rickettsial growth induced by TNF-alpha, IFN-gamma, or both cytokines was evaluated. TNF-alpha inhibited the growth of strains Madrid E (IFN-gamma sensitive and alpha/beta interferon [IFN-alpha/beta] sensitive) and Breinl (IFN-gamma sensitive and IFN-alpha/beta resistant), but not that of strain 83-2P (IFN-gamma resistant and IFN-alpha/beta resistant), in L929 cells. Inhibition of the growth of the Madrid E strain in L929 cells treated with TNF-alpha and IFN-gamma in combination was greater than that observed with either TNF-alpha or IFN-gamma alone. Similarly, inhibition of the growth of the Breinl strain in L929 cells treated with both cytokines was greater than that observed with TNF-alpha alone; however, it did not differ significantly from the inhibition observed with IFN-gamma alone. Although strain 83-2P was resistant to TNF-alpha or IFN-gamma alone, its growth was inhibited in L929 cells treated with TNF-alpha and IFN-gamma in combination. Nitrite production was measured in mock-infected and infected L929 cell cultures, and the nitric oxide synthase inhibitors NG-methyl-L-arginine (NGMA) and aminoguanidine were used to evaluate the role of the nitric oxide synthase pathway in cytokine-induced inhibition of rickettsial growth. Nitrite production was induced in mock-infected or R. prowazekii-infected L929 cell cultures treated with IFN-gamma plus TNF-alpha, but not in mock-infected cultures that were untreated or treated with IFN-gamma or TNF-alpha alone. Nitrite production was also not induced in untreated, R. prowazekii-infected cultures; however, in some instances, it was induced in infected cultures treated with IFN-gamma or TNF-alpha alone

  5. Anaplasma marginale msp1alpha genotypes evolved under positive selection pressure but are not markers for geographic isolates.

    PubMed

    de la Fuente, José; Van Den Bussche, Ronald A; Prado, Tulio M; Kocan, Katherine M

    2003-04-01

    and protein sequences of 13 isolates from Oklahoma in comparison with those of 7 Latin American and 13 isolates from the United States by MP and ML analyses demonstrated no geographic clustering and provided evidence that this gene is under positive selection pressure. The results indicate that msp1alpha is not a marker for the characterization of A. marginale geographic isolates and suggest that the genetic heterogeneity observed among isolates of A. marginale within Oklahoma could be explained by cattle movement and the maintenance of different genotypes by independent transmission events. PMID:12682152

  6. Highly selective and sensitive nanoprobes for cyanide based on gold nanoclusters with red fluorescence emission

    NASA Astrophysics Data System (ADS)

    Zhang, Guomei; Qiao, Yunyun; Xu, Ting; Zhang, Caihong; Zhang, Yan; Shi, Lihong; Shuang, Shaomin; Dong, Chuan

    2015-07-01

    We report a novel and environmentally friendly fluorescent probe for detecting the cyanide ion (CN-) using l-amino acid oxidase (LAAOx)-protected Au nanoclusters (LAAOx@AuNCs) with red emission. The fluorescence-based sensing behaviour of LAAOx@AuNCs towards anions was investigated in buffered aqueous media. Among the anions studied, CN- was found to effectively quench the fluorescence emission of AuNCs based on CN- induced Au core decomposition. Excellent sensitivity and selectivity toward the detection of CN- in aqueous solution were observed. The CN- detection limit was determined to be approximately 180 nM, which is 15 times lower than the maximum level (2700 nM) of CN- in drinking water permitted by the World Health Organization (WHO). A linear relationship between the fluorescence intensity and CN- concentration was observed in two ranges of CN- concentration, including 3.2 × 10-6 to 3.4 × 10-5 mol L-1 and 3.81 × 10-5 to 1.04 × 10-4 mol L-1. The high sensitivity and selectivity to CN- among the 17 types of anions make the AuNCs good candidates for use in fluorescent nanoprobes of CN-.

  7. Synthesis of Au/Graphene Oxide Composites for Selective and Sensitive Electrochemical Detection of Ascorbic Acid

    PubMed Central

    Song, Jian; Xu, Lin; Xing, Ruiqing; Li, Qingling; Zhou, Chunyang; Liu, Dali; Song, Hongwei

    2014-01-01

    In this work, we present a novel ascorbic acid (AA) sensor applied to the detection of AA in human sera and pharmaceuticals. A series of Au nanoparticles (NPs) and graphene oxide sheets (Au NP/GO) composites were successfully synthesized by reduction of gold (III) using sodium citrate. Then the Au NP/GO composites were used to construct nonenzymatic electrodes in practical AA measurement. The electrode that has the best performance presents attractive analytical features, such as a low working potential of +0.15 V, a high sensitivity of 101.86 μA mM−1 cm−2 to AA, a low detection limit of 100 nM, good reproducibility and excellent selectivity. And more,it was also employed to accurately and practically detect AA in human serum and clinical vitamin C tablet with the existence of some food additive. The enhanced AA electrochemical properties of the Au NP/GO modified electrode in our work can be attributed to the improvement of electroactive surface area of Au NPs and the synergistic effect from the combination of Au NPs and GO sheets. This work shows that the Au NP/GO/GCEs hold the prospect for sensitive and selective determination of AA in practical clinical application. PMID:25515430

  8. Aptamer-based colorimetric sensing of acetamiprid in soil samples: sensitivity, selectivity and mechanism.

    PubMed

    Shi, Huijie; Zhao, Guohua; Liu, Meichuan; Fan, Lifang; Cao, Tongcheng

    2013-09-15

    A simple and selective aptamer-based colorimetric method was developed for highly sensitive detection of acetamiprid, taking advantages of the sensitive target-induced colour changes that arisen from the interparticle plasmon coupling during the aggregation of Au nanoparticles (NPs). The results showed that the established method could be applied to detect acetamiprid in the linear range between 75 nM to 7.5 μM, with a low detection limit of 5 nM. Meanwhile, by employing an "artificial antibody" acetamiprid-binding aptamer (ABA) as recognition element, highly selective and specific colorimetric visualization of acetamiprid was obtained. It indicated that pesticides which may coexist with acetamiprid could not interfere with the detection of acetamiprid even that had similar structure with acetamiprid, such as imidacloprid and chlorpyrifos. Mechanism study suggested that it could be attributed to the specific supramolecular interaction between ABA and acetamiprid, as well as the resulted target-binding event induced conformation changes of ABA from random coil to hairpin structure. The practical application of the colorimetric method was realized for detecting acetamiprid in real soil samples and monitoring its natural degradation process. PMID:23846126

  9. Synthesis of Au/Graphene Oxide Composites for Selective and Sensitive Electrochemical Detection of Ascorbic Acid

    NASA Astrophysics Data System (ADS)

    Song, Jian; Xu, Lin; Xing, Ruiqing; Li, Qingling; Zhou, Chunyang; Liu, Dali; Song, Hongwei

    2014-12-01

    In this work, we present a novel ascorbic acid (AA) sensor applied to the detection of AA in human sera and pharmaceuticals. A series of Au nanoparticles (NPs) and graphene oxide sheets (Au NP/GO) composites were successfully synthesized by reduction of gold (III) using sodium citrate. Then the Au NP/GO composites were used to construct nonenzymatic electrodes in practical AA measurement. The electrode that has the best performance presents attractive analytical features, such as a low working potential of +0.15 V, a high sensitivity of 101.86 μA mM-1 cm-2 to AA, a low detection limit of 100 nM, good reproducibility and excellent selectivity. And more,it was also employed to accurately and practically detect AA in human serum and clinical vitamin C tablet with the existence of some food additive. The enhanced AA electrochemical properties of the Au NP/GO modified electrode in our work can be attributed to the improvement of electroactive surface area of Au NPs and the synergistic effect from the combination of Au NPs and GO sheets. This work shows that the Au NP/GO/GCEs hold the prospect for sensitive and selective determination of AA in practical clinical application.

  10. Synthesis of Au/graphene oxide composites for selective and sensitive electrochemical detection of ascorbic acid.

    PubMed

    Song, Jian; Xu, Lin; Xing, Ruiqing; Li, Qingling; Zhou, Chunyang; Liu, Dali; Song, Hongwei

    2014-01-01

    In this work, we present a novel ascorbic acid (AA) sensor applied to the detection of AA in human sera and pharmaceuticals. A series of Au nanoparticles (NPs) and graphene oxide sheets (Au NP/GO) composites were successfully synthesized by reduction of gold (III) using sodium citrate. Then the Au NP/GO composites were used to construct nonenzymatic electrodes in practical AA measurement. The electrode that has the best performance presents attractive analytical features, such as a low working potential of +0.15 V, a high sensitivity of 101.86 μA mM(-1) cm(-2) to AA, a low detection limit of 100 nM, good reproducibility and excellent selectivity. And more,it was also employed to accurately and practically detect AA in human serum and clinical vitamin C tablet with the existence of some food additive. The enhanced AA electrochemical properties of the Au NP/GO modified electrode in our work can be attributed to the improvement of electroactive surface area of Au NPs and the synergistic effect from the combination of Au NPs and GO sheets. This work shows that the Au NP/GO/GCEs hold the prospect for sensitive and selective determination of AA in practical clinical application. PMID:25515430

  11. Synthesis of Au/graphene oxide composites for selective and sensitive electrochemical detection of ascorbic acid.

    PubMed

    Song, Jian; Xu, Lin; Xing, Ruiqing; Li, Qingling; Zhou, Chunyang; Liu, Dali; Song, Hongwei

    2014-12-17

    In this work, we present a novel ascorbic acid (AA) sensor applied to the detection of AA in human sera and pharmaceuticals. A series of Au nanoparticles (NPs) and graphene oxide sheets (Au NP/GO) composites were successfully synthesized by reduction of gold (III) using sodium citrate. Then the Au NP/GO composites were used to construct nonenzymatic electrodes in practical AA measurement. The electrode that has the best performance presents attractive analytical features, such as a low working potential of +0.15 V, a high sensitivity of 101.86 μA mM(-1) cm(-2) to AA, a low detection limit of 100 nM, good reproducibility and excellent selectivity. And more,it was also employed to accurately and practically detect AA in human serum and clinical vitamin C tablet with the existence of some food additive. The enhanced AA electrochemical properties of the Au NP/GO modified electrode in our work can be attributed to the improvement of electroactive surface area of Au NPs and the synergistic effect from the combination of Au NPs and GO sheets. This work shows that the Au NP/GO/GCEs hold the prospect for sensitive and selective determination of AA in practical clinical application.

  12. MWCNT-polymer composites as highly sensitive and selective room temperature gas sensors

    NASA Astrophysics Data System (ADS)

    Mangu, Raghu; Rajaputra, Suresh; Singh, Vijay P.

    2011-05-01

    Multi-walled carbon nanotubes (MWCNTs)-polymer composite-based hybrid sensors were fabricated and integrated into a resistive sensor design for gas sensing applications. Thin films of MWCNTs were grown onto Si/SiO2 substrates via xylene pyrolysis using the chemical vapor deposition technique. Polymers like PEDOT:PSS and polyaniline (PANI) mixed with various solvents like DMSO, DMF, 2-propanol and ethylene glycol were used to synthesize the composite films. These sensors exhibited excellent response and selectivity at room temperature when exposed to low concentrations (100 ppm) of analyte gases like NH3 and NO2. The effect of various solvents on the sensor response imparting selectivity to CNT-polymer nanocomposites was investigated extensively. Sensitivities as high as 28% were observed for an MWCNT-PEDOT:PSS composite sensor when exposed to 100 ppm of NH3 and - 29.8% sensitivity for an MWCNT-PANI composite sensor to 100 ppm of NO2 when DMSO was used as a solvent. Additionally, the sensors exhibited good reversibility.

  13. Ultra-sensitive and selective Hg{sup 2+} detection based on fluorescent carbon dots

    SciTech Connect

    Liu, Ruihua; Li, Haitao; Kong, Weiqian; Liu, Juan; Liu, Yang; Tong, Cuiyan; Zhang, Xing; Kang, Zhenhui

    2013-07-15

    Graphical abstract: Fluorescent carbon dots were efficiently synthesized by one-step sodium hydroxide-assisted reflux method from PEG and demonstrated to show high selectivity toward Hg2+ ions detection. - Highlights: • FCDs were synthesized by one-step sodium hydroxide-assisted reflux method from PEG. • The FCDs emit blue photoluminescence and have upconversion fluorescent property. • The FCDs show ultra-sensitive detective ability for Hg{sup 2+} ions. - Abstract: Fluorescent carbon dots (FCDs) were efficiently synthesized by one-step sodium hydroxide-assisted reflux method from poly(ethylene glycol) (PEG). The obtained FCDs exhibit excellent water-solubility and high stability. Under the UV irradiation, the FCDs could emit bright blue photoluminescence, and also they were found to show excellent up-conversion fluorescence. It was further demonstrated that such FCDs can serve as effective fluorescent sensing platform for Hg{sup 2+} ions detection with ultra-sensitivity and selectivity. The sensing system achieved a limit of detection as low as 1 fM, which is much lower than all the previous reported sensing systems for Hg{sup 2+} ions detection. This FCDs sensing system has been successfully applied for the analysis of Hg{sup 2+} ions in water samples from river, lake, and tap water, showing good practical feasibility.

  14. A Selective Small Molecule DNA2 Inhibitor for Sensitization of Human Cancer Cells to Chemotherapy.

    PubMed

    Liu, Wenpeng; Zhou, Mian; Li, Zhengke; Li, Hongzhi; Polaczek, Piotr; Dai, Huifang; Wu, Qiong; Liu, Changwei; Karanja, Kenneth K; Popuri, Vencat; Shan, Shu-Ou; Schlacher, Katharina; Zheng, Li; Campbell, Judith L; Shen, Binghui

    2016-04-01

    Cancer cells frequently up-regulate DNA replication and repair proteins such as the multifunctional DNA2 nuclease/helicase, counteracting DNA damage due to replication stress and promoting survival. Therefore, we hypothesized that blocking both DNA replication and repair by inhibiting the bifunctional DNA2 could be a potent strategy to sensitize cancer cells to stresses from radiation or chemotherapeutic agents. We show that homozygous deletion of DNA2 sensitizes cells to ionizing radiation and camptothecin (CPT). Using a virtual high throughput screen, we identify 4-hydroxy-8-nitroquinoline-3-carboxylic acid (C5) as an effective and selective inhibitor of DNA2. Mutagenesis and biochemical analysis define the C5 binding pocket at a DNA-binding motif that is shared by the nuclease and helicase activities, consistent with structural studies that suggest that DNA binding to the helicase domain is necessary for nuclease activity. C5 targets the known functions of DNA2 in vivo: C5 inhibits resection at stalled forks as well as reducing recombination. C5 is an even more potent inhibitor of restart of stalled DNA replication forks and over-resection of nascent DNA in cells defective in replication fork protection, including BRCA2 and BOD1L. C5 sensitizes cells to CPT and synergizes with PARP inhibitors. PMID:27211550

  15. Ultrathin pH-sensitive nanoporous membranes for superfast size-selective separation.

    PubMed

    Zhang, Qiu Gen; Deng, Chao; Liu, Rong Rong; Lin, Zhen; Li, Hong Mei; Zhu, Ai Mei; Liu, Qing Lin

    2015-05-01

    Stimuli-responsive nanoporous membranes have attracted increasing interest in various fields due to their abrupt changes of permeation/separation in response to the external environment. Here we report ultrathin pH-sensitive nanoporous membranes that are easily fabricated by the self-assembly of poly(acrylic acid) (PAA) in a metal hydroxide nanostrand solution. PAA-adsorbed nanostrands (2.5-5.0 nm) and PAA-Cu(II) nanogels (2.0-2.5 nm) grow competitively during self-assembly. The PAA-adsorbed nanostrands are deposited on a porous support to fabricate ultrathin PAA membranes. The membranes display ultrafast water permeation and good rejection as well as significant pH-sensitivity. The 28 nm-thick membrane has a water flux decrease from 3740 to 1350 L m(-1)  h(-1)  bar(-1) (pH 2.0 to 7.0) with a sharp decrease at pH 5.0. This newly developed pH-sensitive nanoporous membranes may find a wide range of applications such as controlled release and size- and charge-selective separation. PMID:25736206

  16. Selective induction of apoptosis in the hamster flank sebaceous gland organ by a topical liposome 5-alpha-reductase inhibitor: a treatment strategy for acne.

    PubMed

    Li, Lingna; Tang, Li; Baranov, Eugene; Yang, Meng; Amoh, Yasuyuki; Katsuoka, Kensei; Hoffman, Robert M

    2010-02-01

    Acne is a very widespread cosmesis problem. Isotretinoin, a synthetic oral retinoid is used to treat acne, which is androgen dependent. Numerous side-effects occur from this treatment. 5-alpha-Reductase plays a critical role in normal and pathological androgen-dependent processes. We have taken the approach to develop a selective, effective, topically-applied 5-alpha-reductase inhibitor to modify unwanted or pathological processes in the pilosebaceous unit such as acne. Toward this goal, we have previously developed a selective liposome hair follicle targeting system. We demonstrate in this report that the 5-alpha-reductase inhibitor N,N-diethyl-4-methyl-3-oxo-4-aza-5alpha-androstane-17beta-carboxamide (4-MA) incorporated into liposomes induces apoptosis and inhibits growth of the dihydrotestosterone (DHT)-dependent hamster flank organ sebaceous gland. We have compared topical application of liposome 4-MA and solvent-formulated 4-MA and observed selective efficacy of topical application of liposome 4-MA by the reduction of size and induction of apoptosis only in the treated hamster flank organ. Apoptosis induced by liposome 4-MA in the treated flank organ sebaceous gland cells was observed both by assays for DNA fragments (transferase deoxytidyl uridine end labeling) and by observation of condensed and fragmented nuclei. When 4-MA was topically applied formulated in ethanol and glycerol without liposomes, the selective efficacy was lost. Liposome 4-MA did not significantly affect prostate weight, testosterone/DHT ratios or bodyweight gain compared to controls indicating safety as well as efficacy of topical application of liposome 4-MA for pathological processes such as acne. PMID:20175850

  17. Investigation of the sensitivity, selectivity, and reversibility of the chemically-sensitive field-effect transistor (CHEMFET) to detect nitrogen dioxide, dimethyl methylphosphonate, and boron trifluoride. Master's thesis

    SciTech Connect

    Hauschild, N.T.

    1993-09-01

    This study investigated the sensitivity, selectivity, and reversibility of a chemically-sensitive field-effect transistor (CHEMFET) gas microsensor. Various physical operating parameters were tested to determine which produced the most significant sensitivity, selectivity, and reversibility which were computed from response changes generated from electrical conductivity modulations when exposed to challenge gases. The variable operating parameters included: thinfilm material, film thickness, challenge gas specie, challenge gas concentration, and operating temperature. Copper phthalocyanine and lead phthalocyanine were used as thin films to detect the following challenge gases: nitrogen dioxide, dimethyl methylphosphonate, boron trifluoride, methanol, carbon monoxide, vinyl chloride, and trichloroethylene. Tests revealed that copper phthalocyanine was the most sensitive to dimethyl methylphosphonate and boron trifluoride, whereas lead phthalocyanine was the most sensitive to the remaining challenge gases. The CHEMFET was selective to the binary challenge gas combinations. The films were most selective for nitrogen dioxide. The CHEMFET was fully reversibly, and the time duration for full reversibility increased with increasing challenge gas concentrations and increasing time of exposure.

  18. In Vitro Selection of a Single-Stranded DNA Molecular Recognition Element against Clostridium difficile Toxin B and Sensitive Detection in Human Fecal Matter

    PubMed Central

    Maher, Eamonn; Williams, Ryan M.; Sooter, Letha J.

    2015-01-01

    Toxin B is one of the major virulence factors of Clostridium difficile, a bacterium that is responsible for a significant number of diarrhea cases in acute care settings. Due to the prevalence of C. difficile induced diarrhea, rapid and correct diagnosis is crucial in the disease management. In this study, we have employed a stringent in vitro selection method to identify single-stranded DNA molecular recognition elements (MRE) specific for toxin B. At the end of the 12-round selection, one MRE with high affinity (Kd = 47.3 nM) for toxin B was identified. The selected MRE demonstrated low cross binding activities on negative targets: bovine serum albumin, Staphylococcus aureus alpha toxin, Pseudomonas aeruginosa exotoxin A, and cholera toxin of Vibrio cholera. A modified sandwich ELISA assay was developed utilizing the selected ssDNA MRE as the antigen capturing element and achieved a sensitive detection of 50 nM of toxin B in human fecal preparations. PMID:25734010

  19. [Chromatographic determination of bile acids in biological fluids with sensitive and selective detection].

    PubMed

    Goto, J

    1990-11-01

    Separation and determination of bile acids in biological fluids by high-performance liquid chromatography are reviewed. The capacity ratios of bile acids on an ODS column were affected by the number, position and configuration of the hydroxyl group on the steroid nucleus, and the chromatographic behavior was markedly influenced by the pH of a mobile phase according to the conjugated form at C-24. A new pre-column derivatization reagent, 1-anthroyl nitrile, was developed and applied to the analysis of bile acids in biological fluids. Bile acids were derivatized through the 3 alpha-hydroxyl group into the corresponding esters, separated on an ODS column, and monitored by a fluorescence detector with detection limit of 20 fmol. The sensitive method for the determination of bile acids in biological materials by gas chromatography (GC) in combination with negative ion chemical ionization (NICI) mass spectrometry is also described. Of various derivatives for the carboxyl group, the pentafluorobenzyl (PFB) ester provided the highest value of the ratio of the negative to positive ion current. A characteristic carboxylate anion [M-181]- was produced as the most abundant ion by the loss of the PFB group in NICI. PFB esters of bile acids were further derivatized into the dimethylethylsilyl ethers and then separated by GC. The detection limit was 2 fg when the characteristic anion was monitored in the NICI mode. The preparation of 18O-labelled bile acids, as the internal standard for the trace analysis or the tracer for the metabolic study, was developed. Finally, the clean-up procedure for bile acids in biological fluids was investigated. The combined use of solid-phase extraction with a Sep-pak C18 or Bond Elut cartridge and group separation on a lipophilic ion-exchange gel, piperidinohydroxypropyl Sephadex LH-20, was found most effective for this purpose. PMID:2082011

  20. Surface enhanced Raman scattering based sensitive detection of histone demethylase activity using a formaldehyde-selective reactive probe.

    PubMed

    Wang, Yu; Deng, Xianghua; Liu, Jinwen; Tang, Hao; Jiang, Jianhui

    2013-10-01

    A novel surface enhanced Raman scattering (SERS) based assay using a formaldehyde-selective reactive probe for sensitive detection of activity of histone demethylases (HDMs) by direct observation of by-product formaldehyde was reported.

  1. A simple "molecular beacon"-based fluorescent sensing strategy for sensitive and selective detection of mercury (II).

    PubMed

    Xu, Huifeng; Zhu, Xi; Ye, Hongzhi; Yu, Lishuang; Liu, Xianxiang; Chen, Guonan

    2011-11-28

    A novel fluorescent sensor for the detection of Hg(2+) in aqueous media was developed. The method takes advantages of the highly selective thymine-Hg(2+)-thymine coordination and the sensitive "signal-on" structure-switching molecular beacon.

  2. Imaging alpha particle detector

    DOEpatents

    Anderson, D.F.

    1980-10-29

    A method and apparatus for detecting and imaging alpha particles sources is described. A dielectric coated high voltage electrode and a tungsten wire grid constitute a diode configuration discharge generator for electrons dislodged from atoms or molecules located in between these electrodes when struck by alpha particles from a source to be quantitatively or qualitatively analyzed. A thin polyester film window allows the alpha particles to pass into the gas enclosure and the combination of the glass electrode, grid and window is light transparent such that the details of the source which is imaged with high resolution and sensitivity by the sparks produced can be observed visually as well. The source can be viewed directly, electronically counted or integrated over time using photographic methods. A significant increase in sensitivity over other alpha particle detectors is observed, and the device has very low sensitivity to gamma or beta emissions which might otherwise appear as noise on the alpha particle signal.

  3. Imaging alpha particle detector

    DOEpatents

    Anderson, David F.

    1985-01-01

    A method and apparatus for detecting and imaging alpha particles sources is described. A conducting coated high voltage electrode (1) and a tungsten wire grid (2) constitute a diode configuration discharge generator for electrons dislodged from atoms or molecules located in between these electrodes when struck by alpha particles from a source (3) to be quantitatively or qualitatively analyzed. A thin polyester film window (4) allows the alpha particles to pass into the gas enclosure and the combination of the glass electrode, grid and window is light transparent such that the details of the source which is imaged with high resolution and sensitivity by the sparks produced can be observed visually as well. The source can be viewed directly, electronically counted or integrated over time using photographic methods. A significant increase in sensitivity over other alpha particle detectors is observed, and the device has very low sensitivity to gamma or beta emissions which might otherwise appear as noise on the alpha particle signal.

  4. A Highly Sensitive ESIPT-Based Ratiometric Fluorescence Sensor for Selective Detection of Al(3.).

    PubMed

    Sinha, Sanghamitra; Chowdhury, Bijit; Ghosh, Pradyut

    2016-09-19

    An excited-state intramolecular proton transfer (ESIPT)-based highly sensitive ratiometric fluorescence sensor, 1H was developed for selective detection of aluminum (Al(3+)) in acetonitrile as well as in 90% aqueous system. Single-crystal X-ray diffraction analysis reveals almost planar and conjugated structure of 1H. Photophysical properties of the sensor as well as its selectivity toward Al(3+) are explored using UV-visible, steady-state, and time-resolved fluorescence spectroscopic studies. The bright cyan (λem = 445 nm) fluorescence of 1H in acetonitrile turns into deep blue (λem = 412 nm) with ∼2.3-fold enhancement in emission intensity, in the presence of parts per billion level Al(3+) (detection limit = 0.5 nM). Interestingly, the probe 1H exhibits increased selectivity toward Al(3+) in H2O/acetonitrile (9:1 v/v) solvent system with a change in fluorescence color from pale green to deep blue associated with ca. sixfold enhancement in emission intensity. Density functional theoretical (DFT) calculations provide the ground- and excited-state energy optimized structures and properties of the proposed aluminum complex [Al(1) (OH)]2(2+), which is in harmony with the solution-state experimental findings and also supports the occurrence of ESIPT process in 1H. The ESIPT mechanism was also ascertained by comparing the basic photophysical properties of 1H with a similar O-methylated analogue, 1'Me. PMID:27571218

  5. Selective blockade of CaMKII-alpha inhibits NMDA-induced caspase-3-dependent cell death but does not arrest PARP-1 activation or loss of plasma membrane selectivity in rat retinal neurons.

    PubMed

    Goebel, Dennis J

    2009-02-23

    Calcium/calmodulin-dependent protein kinase II-alpha (CaMKII-alpha) has been implicated in a number of receptor mediated events in neurons. Pharmacological blockade of CaMKII-alpha has been shown to prevent phosphorylation of NMDA-R2A and R2B receptor subunits, suggesting that this enzyme may be linked to receptor trafficking of glutamate receptors and serve as a regulatory protein for neuronal cell death. In the retina, inhibition of CaMKII-alpha has been reported to be neuroprotective against NMDA-induced cell death by preventing the activation of the caspase-3 dependent pathway. However, the effects of CaMKII-alpha blockade on the caspase-3 independent, PARP-1 dependent and the non-programmed cell death pathways have not previously been investigated. In the present study, blockade of CaMKII-alpha with the highly specific antagonist myristoylated autocamtide-2-related inhibitory peptide (AIP) was used in a rat in vivo model of retinal toxicity to compare the effects of on NMDA-induced caspase-3-dependent, PARP-1 dependent and the non-programmed (necrosis) cell death pathways. Results confirmed that AIP fully attenuates caspase-3 activation for at least 8 h following NMDA insult and also significantly improves retinal ganglion cell survival. However, this blockade had little effect on reducing the loss of plasma membrane selectivity (LPMS, e.g. necrosis) in cells located in the ganglion cell and inner nuclear layers and did not alter NMDA-induced PARP-1 hyperactivation, or prevent TUNEL labeling following a moderate NMDA-insult. These findings support a specific role for CaMKII-alpha in mediating the caspase-3 dependent cell death pathway and provide evidence that it is not directly linked to the signaling of either the PARP-1 dependent or the non-programmed cell death pathways.

  6. Further evaluation of quantitative structure--activity relationship models for the prediction of the skin sensitization potency of selected fragrance allergens.

    PubMed

    Patlewicz, Grace Y; Basketter, David A; Pease, Camilla K Smith; Wilson, Karen; Wright, Zoe M; Roberts, David W; Bernard, Guillaume; Arnau, Elena Giménez; Lepoittevin, Jean-Pierre

    2004-02-01

    Fragrance substances represent a very diverse group of chemicals; a proportion of them are associated with the ability to cause allergic reactions in the skin. Efforts to find substitute materials are hindered by the need to undertake animal testing for determining both skin sensitization hazard and potency. One strategy to avoid such testing is through an understanding of the relationships between chemical structure and skin sensitization, so-called structure-activity relationships. In recent work, we evaluated 2 groups of fragrance chemicals -- saturated aldehydes and alpha,beta-unsaturated aldehydes. Simple quantitative structure-activity relationship (QSAR) models relating the EC3 values [derived from the local lymph node assay (LLNA)] to physicochemical properties were developed for both sets of aldehydes. In the current study, we evaluated an additional group of carbonyl-containing compounds to test the predictive power of the developed QSARs and to extend their scope. The QSAR models were used to predict EC3 values of 10 newly selected compounds. Local lymph node assay data generated for these compounds demonstrated that the original QSARs were fairly accurate, but still required improvement. Development of these QSAR models has provided us with a better understanding of the potential mechanisms of action for aldehydes, and hence how to avoid or limit allergy. Knowledge generated from this work is being incorporated into new/improved rules for sensitization in the expert toxicity prediction system, deductive estimation of risk from existing knowledge (DEREK).

  7. Tumor Necrosis Factor Alpha Inhibits L-Type Ca2+ Channels in Sensitized Guinea Pig Airway Smooth Muscle through ERK 1/2 Pathway

    PubMed Central

    Reyes-García, Jorge; Flores-Soto, Edgar; Solís-Chagoyán, Héctor; Sommer, Bettina; Díaz-Hernández, Verónica; García-Hernández, Luz María

    2016-01-01

    Tumor necrosis factor alpha (TNF-α) is a potent proinflammatory cytokine that plays a significant role in the pathogenesis of asthma by inducing hyperresponsiveness and airway remodeling. TNF-α diminishes the L-type voltage dependent Ca2+ channel (L-VDCC) current in cardiac myocytes, an observation that seems paradoxical. In guinea pig sensitized tracheas KCl responses were lower than in control tissues. Serum from sensitized animals (Ser-S) induced the same phenomenon. In tracheal myocytes from nonsensitized (NS) and sensitized (S) guinea pigs, an L-VDCC current (ICa) was observed and diminished by Ser-S. The same decrease was detected in NS myocytes incubated with TNF-α, pointing out that this cytokine might be present in Ser-S. We observed that a small-molecule inhibitor of TNF-α (SMI-TNF) and a TNF-α receptor 1 (TNFR1) antagonist (WP9QY) reversed ICa decrease induced by Ser-S in NS myocytes, confirming the former hypothesis. U0126 (a blocker of ERK 1/2 kinase) also reverted the decrease in ICa. Neither cycloheximide (a protein synthesis inhibitor) nor actinomycin D (a transcription inhibitor) showed any effect on the TNF-α-induced ICa reduction. We found that CaV1.2 and CaV1.3 mRNA and proteins were expressed in tracheal myocytes and that sensitization did not modify them. In cardiac myocytes, ERK 1/2 phosphorylates two sites of the L-VDCC, augmenting or decreasing ICa; we postulate that, in guinea pig tracheal smooth muscle, TNF-α diminishes ICa probably by phosphorylating the L-VDCC site that reduces its activity through the ERK1/2 MAP kinase pathway. PMID:27445440

  8. Tumor Necrosis Factor Alpha Inhibits L-Type Ca(2+) Channels in Sensitized Guinea Pig Airway Smooth Muscle through ERK 1/2 Pathway.

    PubMed

    Reyes-García, Jorge; Flores-Soto, Edgar; Solís-Chagoyán, Héctor; Sommer, Bettina; Díaz-Hernández, Verónica; García-Hernández, Luz María; Montaño, Luis M

    2016-01-01

    Tumor necrosis factor alpha (TNF-α) is a potent proinflammatory cytokine that plays a significant role in the pathogenesis of asthma by inducing hyperresponsiveness and airway remodeling. TNF-α diminishes the L-type voltage dependent Ca(2+) channel (L-VDCC) current in cardiac myocytes, an observation that seems paradoxical. In guinea pig sensitized tracheas KCl responses were lower than in control tissues. Serum from sensitized animals (Ser-S) induced the same phenomenon. In tracheal myocytes from nonsensitized (NS) and sensitized (S) guinea pigs, an L-VDCC current (ICa) was observed and diminished by Ser-S. The same decrease was detected in NS myocytes incubated with TNF-α, pointing out that this cytokine might be present in Ser-S. We observed that a small-molecule inhibitor of TNF-α (SMI-TNF) and a TNF-α receptor 1 (TNFR1) antagonist (WP9QY) reversed ICa decrease induced by Ser-S in NS myocytes, confirming the former hypothesis. U0126 (a blocker of ERK 1/2 kinase) also reverted the decrease in ICa. Neither cycloheximide (a protein synthesis inhibitor) nor actinomycin D (a transcription inhibitor) showed any effect on the TNF-α-induced ICa reduction. We found that CaV1.2 and CaV1.3 mRNA and proteins were expressed in tracheal myocytes and that sensitization did not modify them. In cardiac myocytes, ERK 1/2 phosphorylates two sites of the L-VDCC, augmenting or decreasing ICa; we postulate that, in guinea pig tracheal smooth muscle, TNF-α diminishes ICa probably by phosphorylating the L-VDCC site that reduces its activity through the ERK1/2 MAP kinase pathway.

  9. Tumor Necrosis Factor Alpha Inhibits L-Type Ca(2+) Channels in Sensitized Guinea Pig Airway Smooth Muscle through ERK 1/2 Pathway.

    PubMed

    Reyes-García, Jorge; Flores-Soto, Edgar; Solís-Chagoyán, Héctor; Sommer, Bettina; Díaz-Hernández, Verónica; García-Hernández, Luz María; Montaño, Luis M

    2016-01-01

    Tumor necrosis factor alpha (TNF-α) is a potent proinflammatory cytokine that plays a significant role in the pathogenesis of asthma by inducing hyperresponsiveness and airway remodeling. TNF-α diminishes the L-type voltage dependent Ca(2+) channel (L-VDCC) current in cardiac myocytes, an observation that seems paradoxical. In guinea pig sensitized tracheas KCl responses were lower than in control tissues. Serum from sensitized animals (Ser-S) induced the same phenomenon. In tracheal myocytes from nonsensitized (NS) and sensitized (S) guinea pigs, an L-VDCC current (ICa) was observed and diminished by Ser-S. The same decrease was detected in NS myocytes incubated with TNF-α, pointing out that this cytokine might be present in Ser-S. We observed that a small-molecule inhibitor of TNF-α (SMI-TNF) and a TNF-α receptor 1 (TNFR1) antagonist (WP9QY) reversed ICa decrease induced by Ser-S in NS myocytes, confirming the former hypothesis. U0126 (a blocker of ERK 1/2 kinase) also reverted the decrease in ICa. Neither cycloheximide (a protein synthesis inhibitor) nor actinomycin D (a transcription inhibitor) showed any effect on the TNF-α-induced ICa reduction. We found that CaV1.2 and CaV1.3 mRNA and proteins were expressed in tracheal myocytes and that sensitization did not modify them. In cardiac myocytes, ERK 1/2 phosphorylates two sites of the L-VDCC, augmenting or decreasing ICa; we postulate that, in guinea pig tracheal smooth muscle, TNF-α diminishes ICa probably by phosphorylating the L-VDCC site that reduces its activity through the ERK1/2 MAP kinase pathway. PMID:27445440

  10. Potent and highly selective human immunodeficiency virus type 1 (HIV-1) inhibition by a series of alpha-anilinophenylacetamide derivatives targeted at HIV-1 reverse transcriptase.

    PubMed Central

    Pauwels, R; Andries, K; Debyser, Z; Van Daele, P; Schols, D; Stoffels, P; De Vreese, K; Woestenborghs, R; Vandamme, A M; Janssen, C G

    1993-01-01

    In vitro evaluation of a large chemical library of pharmacologically acceptable prototype compounds in a high-capacity, cellular-based screening system has led to the discovery of another family of human immunodeficiency virus type 1 (HIV-1) inhibitors. Through optimization of a lead compound, several alpha-anilinophenylacetamide (alpha-APA) derivatives have been identified that inhibit the replication of several HIV-1 strains (IIIB/LAI, RF, NDK, MN, HE) in a variety of host cell types at concentrations that are 10,000- to 100,000-fold lower than their cytotoxic concentrations. The IC50 of the alpha-APA derivative R 89439 for HIV-1 cytopathicity in MT-4 cells was 13 nM. The median 90% inhibitory concentration (IC90) in a variety of host cells was 50-100 nM. Although these alpha-APA derivatives are active against a tetrahydroimidazo [4,5,1-jk][1,4]benzodiazepin-2(1H)-thione-(TIBO)-resistant HIV-1 strain, they do not inhibit replication of HIV-2 (strains ROD and EHO) or simian immunodeficiency virus (strains Mac251, mndGB1, and agm3). An HIV-1 strain containing the Tyr181-->Cys mutation in the reverse transcriptase region displayed reduced sensitivity. alpha-APA derivative R 89439 inhibited virion and recombinant reverse transcriptase of HIV-1 but did not inhibit that of HIV-2. Reverse transcriptase inhibition depended upon the template/primer used. The relatively uncomplicated synthesis of R 89439, its potent anti-HIV-1 activity, and its favorable pharmacokinetic profile make R 89439 a good candidate for clinical studies. PMID:7680476

  11. Effects of selective inhibition of N-acetylated-alpha-linked-acidic dipeptidase (NAALADase) on mice in learning and memory tasks.

    PubMed

    Lukawski, Krzysztof; Kamiński, Rafał M; Czuczwar, Stanisław J

    2008-01-28

    The purpose of the present study was to examine the effects of 2-(phosphonomethyl)-pentanedioic acid (2-PMPA), a selective inhibitor of N-acetylated-alpha-linked-acidic dipeptidase (NAALADase, glutamate carboxypeptidase II), an enzyme catalyzing the cleavage of glutamate from the neuropeptide N-acetyl-aspartyl-glutamate (NAAG), on memory processes in mice. Long-term memory was evaluated in step-through passive avoidance task while alternation behavior, as a measure involving spatial working memory, was assessed in Y-maze task. Additionally, horizontal activity was evaluated by means of electronically monitored locomotor activity system. The mice were treated with either 2-PMPA (50, 100 and 150 mg/kg i.p.) or N-methyl-d-aspartate (NMDA) receptor antagonist, (5R,10S)-(+)-5-methyl-10,11-dihydro-5H-dibenzo[a,d]cyclo-hepten-5,10-imine hydrogen maleate (MK-801) at doses of: 0.05, 0.1, 0.15 and 0.2 mg/kg i.p., as a comparator. In the passive avoidance task, the drugs were administered once before or immediately after training, and before retention test. 2-PMPA at the doses used did not affect retention of passive avoidance; however, it increased the latency to enter the dark box during the training day. In the Y-maze task, 2-PMPA (150 mg/kg i.p.) impaired spontaneous alternation and reduced locomotion while the lower dose of 100 mg/kg was ineffective. In the locomotor activity test, 2-PMPA (100 and 150 mg/kg i.p.) did not significantly affect horizontal activity. MK-801 (0.2 mg/kg i.p.) injected before training reduced retention in the passive avoidance task. In the Y-maze task, MK-801 (0.1 mg/kg i.p.) impaired alternation behavior and considerably increased locomotion in the Y-maze and locomotor activity test. These results indicate that NAALADase inhibition may impair alternation behavior.

  12. The alpha 1 Na(+)-K+ pump of the Dahl salt-sensitive rat exhibits altered Na+ modulation of K+ transport in red blood cells.

    PubMed

    Canessa, M; Romero, J R; Ruiz-Opazo, N; Herrera, V L

    1993-06-01

    The properties of the alpha 1 Na(+)-K+ pump were compared in Dahl salt-sensitive (DS) and salt-resistant (DR) strains by measuring ouabain-sensitive fluxes (mmol/liter cell x hr = FU, Mean +/- SE) in red blood cells (RBCs) and varying internal (i) and external (o) Na+ and K+ concentrations. Kinetic parameters of several modes of operation, i.e., Na+/K+, K+/K+, Na+/Na+ exchanges, were characterized and analyzed for curve-fitting using the Enzfitter computer program. In unidirectional flux studies (n = 12 rats of each strain) into fresh cells incubated in 140 mM Na(+) + 5 mM K+, ouabain-sensitive K+ influx was substantially lower in the DS than in DR RBCs, while ouabain-sensitive Na+ efflux and Nai were similar in both strains. Thus, the coupling ratio between unidirectional Na+:K+ fluxes was significantly higher in DS than in DR cells at similar RBC Na+ content. In the presence of 140 mM Nao, activation of ouabain-sensitive K+ influx by Ko had a lower Km and Vmax in DS as estimated by the Garay equation (N = 2.70 +/- 0.33, Km 0.74 +/- 0.09 mM; Vmax 2.87 +/- 0.09 FU) than in DR rats (N = 1.23 +/- 0.36, Km 2.31 +/- 0.16 mM; Vmax 5.70 +/- 0.52 FU). However, the two kinetic parameters were similar following Nao removal. The activation of ouabain-sensitive K+ influx by Nai had significantly lower Vmax in DS (9.3 +/- 0.4 FU) than in DR (14.5 +/- 0.6 FU) RBCs but similar Km. These data suggest that the low K+ influx in DS cells is caused by a defect in modulation by Nao and Nai. Na+ efflux showed no differences in Nai activation or trans effects by Nao and Ko, thus accounting for the different Na+:K+ coupling ratio in the Dahl strains. Further evidence for the differences in the coupling of ouabain-sensitive fluxes was found in studies of net Na+ and K+ fluxes, where the net ouabain-sensitive Na+ losses showed similar magnitudes in the two Dahl strains while the net ouabain-sensitive K+ gains were significantly greater in the DR than the DS RBCs. Ouabain-sensitive Na

  13. Spin-Sensitive and Angular Dependent Detection of Resonant Excitations at the K Absorption Pre-Edge of {alpha}-Fe2O3

    SciTech Connect

    Glatzel, Pieter; Mirone, Alessandro; Eeckhout, Sigrid G.; Sikora, Marcin; Giuli, Gabriele

    2007-02-02

    An experimental and theoretical study of the K absorption pre-edge in hematite ({alpha}-Fe2O3) is presented. Resonant inelastic X-ray scattering with a 3p hole in the final states was used to obtain spin-selective absorption spectra. Spectral variations with changing the orientation of the incident X-ray polarization vector with respect to the crystal c-axis in single crystalline hematite are discussed. The experimental results can be successfully modeled using a band-structure approach (WIEN2k with LDA+U). A pre-edge absorption feature is assigned to unoccupied p electronic states due to Fe-Fe interactions, i.e. they are due to non-local transitions.

  14. Receptor-selective retinoids implicate retinoic acid receptor alpha and gamma in the regulation of bmp-2 and bmp-4 in F9 embryonal carcinoma cells.

    PubMed

    Rogers, M B

    1996-01-01

    The effect of retinoids on malignant cells and embryos indicates that retinoids influence the expression of growth factors or alter the response of cells to growth factors. The bone morphogenetic proteins, Bmp-2 and Bmp-4, are candidates for such growth factors because retinoic acid (RA) treatment of F9 embryonal carcinoma cells induced Bmp-2 mRNA, while simultaneously repressing Bmp-4 levels. Also, recombinant Bmp-2 affected the growth and differentiation of these cells. Regulation of each gene was concentration dependent and required continuous RA treatment. The short half-lives of the Bmp-2 (75 +/- 11 min) and Bmp-4 (70 +/- 4 min) mRNAs suggest that their abundance is primarily controlled at the transcriptional level. To determine which RA receptor (RAR) controls bmp-2 and bmp-4 expression, F9 cells were exposed to various receptor-selective retinoids. RAR alpha- and gamma-selective retinoids induced Bmp-2 and repressed Bmp-4 equally as well as all-trans RA. In contrast, a RAR beta-selective retinoid had little effect on Bmp-2 induction but repressed Bmp-4. A RAR alpha-selective antagonist inhibited all-trans RA stimulation of Bmp-2, although not as dramatically as a RAR beta gamma-selective antagonist. No differences were observed between Bmp levels in all-trans RA and 9-cis RA-treated cells, indicating that the RXRs play little part in controlling these genes. The results are consistent with RAR alpha and gamma-controlled Bmp-2 and Bmp-4 regulation.

  15. Conducting Polymer-Based Nanohybrid Transducers: A Potential Route to High Sensitivity and Selectivity Sensors

    PubMed Central

    Park, Seon Joo; Kwon, Oh Seok; Lee, Ji Eun; Jang, Jyongsik; Yoon, Hyeonseok

    2014-01-01

    The development of novel sensing materials provides good opportunities to realize previously unachievable sensor performance. In this review, conducting polymer-based nanohybrids are highlighted as innovative transducers for high-performance chemical and biological sensing devices. Synthetic strategies of the nanohybrids are categorized into four groups: (1) impregnation, followed by reduction; (2) concurrent redox reactions; (3) electrochemical deposition; (4) seeding approach. Nanocale hybridization of conducting polymers with inorganic components can lead to improved sorption, catalytic reaction and/or transport behavior of the material systems. The nanohybrids have thus been used to detect nerve agents, toxic gases, volatile organic compounds, glucose, dopamine, and DNA. Given further advances in nanohybrids synthesis, it is expected that sensor technology will also evolve, especially in terms of sensitivity and selectivity. PMID:24561406

  16. Highly functionalized tetrahydropyridines are cytotoxic and selective inhibitors of human puromycin sensitive aminopeptidase.

    PubMed

    Aeluri, Raghunath; Ganji, Roopa Jones; Marapaka, Anil Kumar; Pillalamarri, Vijaykumar; Alla, Manjula; Addlagatta, Anthony

    2015-12-01

    Efficient one-pot five-component synthetic protocols for highly functionalized tetrahydropyridines (THPs) and their biological evaluation have been illustrated. Synthesis of novel functionalized tetrahydropyridines containing differential substitutions at 2,6-positions has been achieved via a modified MCR. Cytotoxic studies of 23 synthesized compounds have been carried out against three different cell lines, namely A-549, HeLa and HepG2, wherein some compounds have displayed appreciable cytotoxicity. Further, investigation of enzyme inhibition by the synthesized THPs has been carried out against four members of M1 family aminopeptidases. Several compounds have selectively inhibited only one member of this enzyme family i.e., human puromycin sensitive aminopeptidase (hPSA). Among the compounds; 4b, 9b, 9e and 10a demonstrated best inhibition against hPSA.

  17. ZnO-nanowires modified polypyrrole films as highly selective and sensitive chlorine sensors

    NASA Astrophysics Data System (ADS)

    Joshi, Aditee; Aswal, D. K.; Gupta, S. K.; Yakhmi, J. V.; Gangal, S. A.

    2009-03-01

    Room temperature sensing characteristics of the polypyrrole (PPy) films modified with different weight percentages of ZnO nanowires (ZnO-NWs) have been studied for various oxidizing (NO2 and Cl2) as well as reducing (H2S, NH3, CH4, and CO) gases. We demonstrate that ZnO-NW (50 wt %):PPy composite films are highly selective to Cl2 along with high-sensitivity (40 at 10 ppm), fast-response (55 s), and highly reproducible response curves. It has been shown that negatively charged O2- surfaces of ZnO-NW transfer electrons to PPy, making it in a highly reduced form. A strong localization of charge carriers in the reduced PPy makes composite film highly resistive (>1010 Ω cm) as well insensitive to interaction with most of the gases except Cl2. Cl2 being highly oxidizing gas interacts with composite films and causes a sharp reduction in its resistivity.

  18. Highly selective, sensitive and fast-responsive fluorescent sensor for Hg2 +

    NASA Astrophysics Data System (ADS)

    Niu, Qingfen; Wu, Xingxing; Li, Tianduo; Cui, Yuezhi; Zhang, Shanshan; Li, Xiaoyan

    2016-06-01

    A phenylamine-oligothiophene-based fluorescent sensor 2TBEA was reported. This sensor exhibited highly selective, sensitive and rapid detection of Hg2 + ion in THF/H2O (7/3, v/v) solution through fluorescence quenching. The detection was unaffected by the coexistence of other competitive metal cations including Na+, K+, Ag+, Ca2 +, Fe3 +, Al3 +, Co2 +, Cu2 +, Ni2 +, Zn2 +, Pb2 +, Cd2 +, Fe2 + and Cr3 +. A1:1 binding ratio for 2TBEA - Hg2 + was demonstrated by Job's plot and mole-ratio curves. The coordination process was chemically reversible with EDTA. The detection limit was evaluated to be as low as 6.164 × 10- 8 M.

  19. Sensitive and Selective Detection of HIV-1 RRE RNA Using Vertical Silicon Nanowire Electrode Array

    NASA Astrophysics Data System (ADS)

    Lee, Jaehyung; Hong, Min-Ho; Han, Sanghun; Na, Jukwan; Kim, Ilsoo; Kwon, Yong-Joon; Lim, Yong-beom; Choi, Heon-Jin

    2016-07-01

    In this study, HIV-1 Rev response element (RRE) RNA was detected via an Au-coated vertical silicon nanowire electrode array (VSNEA). The VSNEA was fabricated by combining bottom-up and top-down approaches and then immobilized by artificial peptides for the recognition of HIV-1 RRE. Differential pulse voltammetry (DPV) analysis was used to measure the electrochemical response of the peptide-immobilized VSNEA to the concentration and types of HIV-1 RRE RNA. DPV peaks showed linearity to the concentration of RNA with a detection limit down to 1.513 fM. It also showed the clear different peaks to the mutated HIV-1 RRE RNA. The high sensitivity and selectivity of VSNEA for the detection of HIV-1 RRE RNA may be attributed to the high surface-to-volume ratio and total overlap diffusion mode of ions of the one-dimensional nanowire electrodes.

  20. Sensitive and Selective Detection of HIV-1 RRE RNA Using Vertical Silicon Nanowire Electrode Array.

    PubMed

    Lee, Jaehyung; Hong, Min-Ho; Han, Sanghun; Na, Jukwan; Kim, Ilsoo; Kwon, Yong-Joon; Lim, Yong-Beom; Choi, Heon-Jin

    2016-12-01

    In this study, HIV-1 Rev response element (RRE) RNA was detected via an Au-coated vertical silicon nanowire electrode array (VSNEA). The VSNEA was fabricated by combining bottom-up and top-down approaches and then immobilized by artificial peptides for the recognition of HIV-1 RRE. Differential pulse voltammetry (DPV) analysis was used to measure the electrochemical response of the peptide-immobilized VSNEA to the concentration and types of HIV-1 RRE RNA. DPV peaks showed linearity to the concentration of RNA with a detection limit down to 1.513 fM. It also showed the clear different peaks to the mutated HIV-1 RRE RNA. The high sensitivity and selectivity of VSNEA for the detection of HIV-1 RRE RNA may be attributed to the high surface-to-volume ratio and total overlap diffusion mode of ions of the one-dimensional nanowire electrodes.

  1. A new diketopyrrolopyrrole-based probe for sensitive and selective detection of sulfite in aqueous solution

    NASA Astrophysics Data System (ADS)

    Yang, Xiaofeng; Cui, Yu; Li, Yexin; Zheng, Luyi; Xie, Lijun; Ning, Rui; Liu, Zheng; Lu, Junling; Zhang, Gege; Liu, Chunxiang; Zhang, Guangyou

    2015-02-01

    A new probe was synthesized by incorporating an α,β -unsaturated ketone to a diketopyrrolopyrrole fluorophore. The probe had exhibited a selective and sensitive response to the sulfite against other thirteen anions and biothiols (Cys, Hcy and GSH), through the nucleophilic addition of sulfite to the alkene of probe with the detection limit of 0.1 μM in HEPES (10 mM, pH 7.4) THF/H2O (1:1, v/v). Meanwhile, it could be easily observed that the probe for sulfite changed from pink to colorless by the naked eye, and from pink to blue under UV lamp after the sulfite was added for 20 min. The NMR and Mass spectral analysis demonstrated the expected addition of sulfite to the Cdbnd C bonds.

  2. Surface-selective laser sintering of thermolabile polymer particles using water as heating sensitizer

    NASA Astrophysics Data System (ADS)

    Antonov, E. N.; Krotova, L. I.; Minaev, N. V.; Minaeva, S. A.; Mironov, A. V.; Popov, V. K.; Bagratashvili, V. N.

    2015-11-01

    We report the implementation of a novel scheme for surface-selective laser sintering (SSLS) of polymer particles, based on using water as a sensitizer of laser heating and sintering of particles as well as laser radiation at a wavelength of 1.94 μm, corresponding to the strong absorption band of water. A method of sintering powders of poly(lactide-co-glycolide), a hydrophobic bioresorbable polymer, after modifying its surface with an aqueous solution of hyaluronic acid is developed. The sintering thresholds for wetted polymer are by 3 - 4 times lower than those for sintering in air. The presence of water restricts the temperature of the heated polymer, preventing its thermal destruction. Polymer matrices with a developed porous structure are obtained. The proposed SSLS method can be applied to produce bioresorbable polymer matrices for tissue engineering.

  3. Highly sensitive and selective electrochemical dopamine sensing properties of multilayer graphene nanobelts

    NASA Astrophysics Data System (ADS)

    Karthick Kannan, Padmanathan; Moshkalev, Stanislav A.; Sekhar Rout, Chandra

    2016-02-01

    In the present study, we report the electrochemical sensing property of multi-layer graphene nanobelts (GNBs) towards dopamine (DA). GNBs are synthesized from natural graphite and characterized by using techniques like field-emission scanning electron microscopy, atomic force microscopy and Raman spectroscopy. An electrochemical sensor based on GNBs is developed for the detection of DA. From the cyclic voltammetry and amperometry studies, it is found that GNBs possess excellent electrocatalytic activity towards DA molecules. The developed DA sensor showed a sensitivity value of 0.95 μA μM-1 cm-2 with a linear range of 2 μM to 0.2 mM. The interference data exhibited that GNB is highly selective to DA even in the presence of common interfering species like ascorbic acid, uric acid, glucose and lactic acid.

  4. A new hydroxynaphthyl benzothiazole derived fluorescent probe for highly selective and sensitive Cu2 + detection

    NASA Astrophysics Data System (ADS)

    Tang, Lijun; He, Ping; Zhong, Keli; Hou, Shuhua; Bian, Yanjiang

    2016-12-01

    A new reactive probe, 1-(benzo[d]thiazol-2-yl)naphthalen-2-yl-picolinate (BTNP), was designed and synthesized. BTNP acts as a highly selective probe to Cu2 + in DMSO/H2O (7/3, v/v, Tris-HCl 10 mM, pH = 7.4) solution based on Cu2 + catalyzed hydrolysis of the picolinate ester moiety in BTNP, which leads to the formation of an ESIPT active product with dual wavelength emission enhancement. The probe also possesses the advantages of simple synthesis, rapid response and high sensitivity. The pseudo-first-order reaction rate constant was calculated to be 0.205 min- 1. Moreover, application of BTNP to Cu2 + detection in living cells and real water samples was also explored.

  5. A conserved membrane attachment site in alpha-SNAP facilitates N-ethylmaleimide-sensitive factor (NSF)-driven SNARE complex disassembly.

    PubMed

    Winter, Ulrike; Chen, Xiong; Fasshauer, Dirk

    2009-11-13

    The ATPase NSF (N-ethylmaleimide-sensitive factor) and its SNAP (soluble N-ethylmaleimide-sensitive factor attachment protein) cofactor constitute the ubiquitous enzymatic machinery responsible for recycling of the SNARE (SNAP receptor) membrane fusion machinery. The enzyme uses the energy of ATP hydrolysis to dissociate the constituents of the SNARE complex, which is formed during the fusion of a transport vesicle with the acceptor membrane. However, it is still unclear how NSF and the SNAP adaptor work together to take the tight SNARE bundle apart. SNAPs have been reported to attach to membranes independently from SNARE complex binding. We have investigated how efficient the disassembly of soluble and membrane-bound substrates are, comparing the two. We found that SNAPs support disassembly of membrane-bound SNARE complexes much more efficiently. Moreover, we identified a putative, conserved membrane attachment site in an extended loop within the N-terminal domain of alpha-SNAP. Mutation of two highly conserved, exposed phenylalanine residues on the extended loop prevent SNAPs from facilitating disassembly of membrane-bound SNARE complexes. This implies that the disassembly machinery is adapted to attack membrane-bound SNARE complexes, probably in their relaxed cis-configuration.

  6. Gold-silver-graphene hybrid nanosheets-based sensors for sensitive amperometric immunoassay of alpha-fetoprotein using nanogold-enclosed titania nanoparticles as labels.

    PubMed

    Su, Biling; Tang, Dianping; Li, Qunfang; Tang, Juan; Chen, Guonan

    2011-04-29

    A new sandwich-type electrochemical immunosensor with enhanced sensitivity was developed for detection of alpha-fetoprotein (AFP, as a model analyte) in biological fluids by using nanogold-enclosed titania nanoparticle (AuTi)-labeled secondary antibody on a gold-silver-graphene hybrid nanosheet (AuAgGP)-functionalized glassy carbon electrode (GCE). The presence of the AuAgGP nanosheets not only enhanced the immobilized amount of biomolecules, but also improved the electrochemical properties of the immunosensor. With the aid of AuTi nanolabels, the electrochemical signal was greatly amplified in comparison with pure nanogold or titania-based labels. Under optimal conditions, the sensitivity and dynamic range of the immunosensor were evaluated by using the labeled horseradish peroxidase on the AuTi as trace and H(2)O(2) as enzyme substrate, and exhibited a wide dynamic range of 0.001-200 ng mL(-1) with a low detection limit (LOD) of 0.5 pg mL(-1) AFP (at 3σ). Both the intra- and inter-assay coefficients of variation were less than 10%. The current of the immunosensor at 13th day was as much as 90% of the initial current. In addition, the methodology was evaluated for 8 positive serum specimens obtained from hepatocarcinoma patients and 19 negative sera, and validated with the commercially available Roche 2010 Electrochemiluminescent (ECL) Automatic Analyzer. No significant differences at the 95% confidence level were encountered between two methods.

  7. Highly sensitive and selective spectrophotometric and spectrofluorimetric methods for the determination of ropinirole hydrochloride in tablets.

    PubMed

    Aydoğmuş, Zeynep

    2008-06-01

    Three sensitive, selective, accurate spectrophotometric and spectrofluorimetric methods have been developed for the determination of ropinirole hydrochloride in tablets. The first method was based on measuring the absorbance of drug solution in methanol at 250 nm. The Beer's law was obeyed in the concentration range 2.5-24 microg ml(-1). The second method was based on the charge transfer reaction of drug, as n-electron donor with 7,7,8,8-tetracyanoquinodimethane (TCNQ), as pi-acceptor in acetonitrile to give radical anions that are measured at 842 nm. The Beer's law was obeyed in the concentration range 0.6-8 microg ml(-1). The third method was based on derivatization reaction with 4-chloro-7-nitrobenzofurazan (NBD-Cl) in borate buffer of pH 8.5 followed by measuring the fluorescence intensity at 525 nm with excitation at 464 nm in chloroform. Beer's law was obeyed in the concentration range 0.01-1.3 microg ml(-1). The derivatization reaction product of drug with NBD-Cl was characterized by IR, 1H NMR and mass spectroscopy. The developed methods were validated. The following analytical parameters were investigated: the molar absorptivity (epsilon), limit of detection (LOD, microg ml(-1)) and limit of quantitation (LOQ, microg ml(-1)), precision, accuracy, recovery, and Sandell's sensitivity. Selectivity was validated by subjecting stock solution of ropinirole to acidic, basic, oxidative, and thermal degradation. No interference was observed from common excipients present in formulations. The proposed methods were successfully applied for determination of drug in tablets. The results of these proposed methods were compared with each other statistically. PMID:17719838

  8. Highly sensitive and selective spectrophotometric and spectrofluorimetric methods for the determination of ropinirole hydrochloride in tablets

    NASA Astrophysics Data System (ADS)

    Aydoğmuş, Zeynep

    2008-06-01

    Three sensitive, selective, accurate spectrophotometric and spectrofluorimetric methods have been developed for the determination of ropinirole hydrochloride in tablets. The first method was based on measuring the absorbance of drug solution in methanol at 250 nm. The Beer's law was obeyed in the concentration range 2.5-24 μg ml -1. The second method was based on the charge transfer reaction of drug, as n-electron donor with 7,7,8,8-tetracyanoquinodimethane (TCNQ), as π-acceptor in acetonitrile to give radical anions that are measured at 842 nm. The Beer's law was obeyed in the concentration range 0.6-8 μg ml -1. The third method was based on derivatization reaction with 4-chloro-7-nitrobenzofurazan (NBD-Cl) in borate buffer of pH 8.5 followed by measuring the fluorescence intensity at 525 nm with excitation at 464 nm in chloroform. Beer's law was obeyed in the concentration range 0.01-1.3 μg ml -1. The derivatization reaction product of drug with NBD-Cl was characterized by IR, 1H NMR and mass spectroscopy. The developed methods were validated. The following analytical parameters were investigated: the molar absorptivity ( ɛ), limit of detection (LOD, μg ml -1) and limit of quantitation (LOQ, μg ml -1), precision, accuracy, recovery, and Sandell's sensitivity. Selectivity was validated by subjecting stock solution of ropinirole to acidic, basic, oxidative, and thermal degradation. No interference was observed from common excipients present in formulations. The proposed methods were successfully applied for determination of drug in tablets. The results of these proposed methods were compared with each other statistically.

  9. Selective Fragmentation of Radiation-Sensitive Novel Polymeric Resist Materials by Inner-Shell Irradiation.

    PubMed

    Chagas, Gabriela Ramos; Satyanarayana, Vardhineedi Sri Venkata; Kessler, Felipe; Belmonte, Guilherme Kretzmann; Gonsalves, Kenneth E; Weibel, Daniel Eduardo

    2015-08-01

    Two key concepts in extreme ultraviolet lithography (EUVL) are important for it to be a candidate for the mass production of future integrated circuits: the polymer formulation and the photofragmentation process. In this work, both concepts were carefully studied. The design and synthesis of radiation-sensitive organic polymeric materials based on the inclusion of a radiation-sensitive tetrahydrothiophenium functional group are outlined. A 1-(4-methacryloyoxy)naphthalene-1-yl)tetrahydro-1H-thiophenium trifluoromethanesulfonate (MANTMS) monomer containing the tetrahydrothiophenium group undergoes homo- and copolymerizations using free-radical polymerization with a 2,2'-azobis(isobutyronitrile) initiator. The surface photodegradation and oxidation of these novel polymeric materials were investigated using highly monochromatized soft X-rays from synchrotron radiation at the carbon K-edge excitation region. An efficient functionalization was observed when the excitation energy was tuned to C 1s → π*C═C. A high rate of defluorination and a loss of sulfonated groups as a result of an increase in the irradiation time for the MANTMS homopolymer thin films were mainly observed under the π*C═C excitation of the naphthyl functional groups. On the contrary, excitation similar to C 1s → π*C═O or C 1s → σ*C-F did not produce important degradation, showing a highly selective process of bond breaking. Additionally, the presence of methyl methacrylate copolymer in the original MANTMS yielded a much higher degree of stability against inner-shell radiation damage. Our results highlight the importance of choosing the right polymer formulation and excitation energy to produce a sensitive material for EUVL without using the concept of chemical amplification. PMID:26158557

  10. Force-sensitive detents improve user performance for linear selection tasks.

    PubMed

    Berdahl, E; Smith, J O; Weinzierl, S; Niemeyer, G

    2013-01-01

    Haptic technology, providing force cues and creating a programmable interface, can assist users in more accurately using an interface. This paper investigates haptic assistance in combination with auditory feedback instead of visual feedback. A user test is carried out in which participants select fundamental frequencies from a continuous range to play brief musical melodies. Two control conditions are compared with two detent-based haptic assistance conditions. The detents gently guide the users toward locations of equal tempered fundamental frequencies. Results from the user test confirm improved accuracy brought about by the detents. It is further helpful to provide regulation of the strength of haptic assistance in real time, allowing the user to remain always in control. This concept motivated the force-sensitive detent condition, which enables the user to adjust the strength of the haptic assistance in real time by changing the downward force applied to the haptic device. The work implies that users of graphical user interfaces could similarly benefit from force-sensitive detents and more generally real-time regulation of the strength of haptic assistance. PMID:24808304

  11. Novel fluorescent silver nanoparticles: sensitive and selective turn off sensor for cadmium ions

    NASA Astrophysics Data System (ADS)

    Makwana, Bharat A.; Vyas, Disha J.; Bhatt, Keyur D.; Darji, Savan; Jain, Vinod K.

    2016-04-01

    The synthesis of metal nanoparticles by eco-friendly and reliable processes is an important aspect in many fields. In this study, octamethoxy resorcin [4] arene tetrahydrazide (OMRTH)-reduced and stabilized silver nanoparticles were synthesized via a simple one-pot method. Synthesized silver nanoparticles were characterized by UV-visible spectroscopy, transmission electron microscopy (TEM) and particle size analyzer (PSA). Furthermore, the application of OMRTH-AgNps as a simple, cost-effective and sensitive fluorescent sensor for rapid detection of cadmium was explored. Under optimum conditions, the fluorescence intensity of OMRTH-AgNps was inversely proportional to the cadmium concentration. Using OMRTH-AgNps as a selective and sensitive fluorescent probe, cadmium can be detected at a minimum concentration level of 10-8 M in a facile way of fluorescence quenching, i.e., by a "turn off" mechanism. The method has been successfully applied for determination of Cd[II] ions in groundwater and industrial effluent wastewater samples.

  12. Sulfated alpha-L-galactans from the sea urchin ovary: selective 6-desulfation as eggs are spawned.

    PubMed

    Cinelli, Leonardo P; Andrade, Leonardo; Valente, Ana Paula; Mourão, Paulo A S

    2010-06-01

    The sea urchin eggs are surrounded by a jelly coat, which contains sulfated polysaccharides with unique structures. These molecules are responsible for inducing the species-specific acrosome reaction, an obligatory event for the binding of sperm and fusion with the egg. The mechanism of biosynthesis of these sulfated polysaccharides is virtually unknown. The egg jelly of the sea urchin Echinometra lucunter contains a simple 2-sulfated, 3-linked alpha-L-galactan. Here, we pulse labeled the sea urchin ovary in vitro with (35)S-sulfate to follow the biosynthesis of the sulfated alpha-L-galactan. We found that the ovary contains a 2,6-disulfated, 3-linked alpha-L-galactan, which incorporates (35)S-sulfate more avidly than the 2-sulfated isoform. The 2,6-disulfated alpha-L-galactan was purified by anion exchange chromatography, analyzed by electrophoresis and characterized by 1D and 2D nuclear magnetic resonance spectra. We also investigated the location of the sulfated polysaccharides on the oocytes using histochemical procedures. The stain revealed high amounts of sulfated polysaccharide in mature oocytes and accessory cells. The amount of intracellular sulfated polysaccharides decreased as oocytes are spawned. We speculate that 2,6-disulfated galactan is initially synthesized in the ovary and that 6-sulfate ester is removed when the polysaccharide is secreted into the egg jelly. Similar events related to remodeling of sulfated polysaccharides have been reported in other biological systems.

  13. Sensitive and Selective Ratiometric Fluorescence Probes for Detection of Intracellular Endogenous Monoamine Oxidase A.

    PubMed

    Wu, Xiaofeng; Li, Lihong; Shi, Wen; Gong, Qiuyu; Li, Xiaohua; Ma, Huimin

    2016-01-19

    Monoamine oxidase A (MAO-A) is known to widely exist in most cell lines in the body, and its dysfunction (unusually high or low levels of MAO-A) is thought to be responsible for several psychiatric and neurological disorders. Thus, a sensitive and selective method for evaluating the relative MAO-A levels in different live cells is urgently needed to better understand the function of MAO-A, but to our knowledge such a method is still lacking. Herein, we rationally design two new ratiometric fluorescence probes (1 and 2) that can sensitively and selectively detect MAO-A. The probes are constructed by incorporating a recognition group of propylamine into the fluorescent skeleton of 1,8-naphthalimide, and the detection mechanism is based on amine oxidation and β-elimination to release the fluorophore (4-hydroxy-N-butyl-1,8-naphthalimide), which is verified by HPLC analysis. Reaction of the probes with MAO-A produces a remarkable fluorescence change from blue to green, and the ratio of fluorescence intensity at 550 and 454 nm is directly proportional to the concentration of MAO-A in the ranges of 0.5-1.5 and 0.5-2.5 μg/mL with detection limits of 1.1 and 10 ng/mL (k = 3) for probes 1 and 2, respectively. Surprisingly, these probes show strong fluorescence responses to MAO-A but almost none to MAO-B (one of two isoforms of MAO), indicating superior ability to distinguish MAO-A from MAO-B. The high specificity of the probes for MAO-A over MAO-B is further supported by different inhibitor experiments. Moreover, probe 1 displays higher sensitivity than probe 2 and is thus investigated to image the relative MAO-A levels in different live cells, such as HeLa and NIH-3T3 cells. It is found that the concentration of endogenous MAO-A in HeLa cells is approximately 1.8 times higher than that in NIH-3T3 cells, which is validated by the result from an ELISA kit. Additionally, the proposed probes may find more uses in the specific detection of MAO-A between the two isoforms of MAO

  14. A Sensitive Secondary Users Selection Algorithm for Cognitive Radio Ad Hoc Networks

    PubMed Central

    Li, Aohan; Han, Guangjie; Wan, Liangtian; Shu, Lei

    2016-01-01

    Secondary Users (SUs) are allowed to use the temporarily unused licensed spectrum without disturbing Primary Users (PUs) in Cognitive Radio Ad Hoc Networks (CRAHNs). Existing architectures for CRAHNs impose energy-consuming Cognitive Radios (CRs) on SUs. However, the advanced CRs will increase energy cost for their cognitive functionalities, which is undesirable for the battery powered devices. A new architecture referred to as spectral Requirement-based CRAHN (RCRAHN) is proposed to enhance energy efficiency for CRAHNs in this paper. In RCRAHNs, only parts of SUs are equipped with CRs. SUs equipped with CRs are referred to as Cognitive Radio Users (CRUs). To further enhance energy efficiency of CRAHNs, we aim to select minimum CRUs to sense available spectrum. A non-linear programming problem is mathematically formulated under the constraints of energy efficiency and real-time. Considering the NP-hardness of the problem, a framework of a heuristic algorithm referred to as Sensitive Secondary Users Selection (SSUS) was designed to compute the near-optimal solutions. The simulation results demonstrate that SSUS not only improves the energy efficiency, but also achieves satisfied performances in end-to-end delay and communication reliability. PMID:27023562

  15. A Sensitive Secondary Users Selection Algorithm for Cognitive Radio Ad Hoc Networks.

    PubMed

    Li, Aohan; Han, Guangjie; Wan, Liangtian; Shu, Lei

    2016-01-01

    Secondary Users (SUs) are allowed to use the temporarily unused licensed spectrum without disturbing Primary Users (PUs) in Cognitive Radio Ad Hoc Networks (CRAHNs). Existing architectures for CRAHNs impose energy-consuming Cognitive Radios (CRs) on SUs. However, the advanced CRs will increase energy cost for their cognitive functionalities, which is undesirable for the battery powered devices. A new architecture referred to as spectral Requirement-based CRAHN (RCRAHN) is proposed to enhance energy efficiency for CRAHNs in this paper. In RCRAHNs, only parts of SUs are equipped with CRs. SUs equipped with CRs are referred to as Cognitive Radio Users (CRUs). To further enhance energy efficiency of CRAHNs, we aim to select minimum CRUs to sense available spectrum. A non-linear programming problem is mathematically formulated under the constraints of energy efficiency and real-time. Considering the NP-hardness of the problem, a framework of a heuristic algorithm referred to as Sensitive Secondary Users Selection (SSUS) was designed to compute the near-optimal solutions. The simulation results demonstrate that SSUS not only improves the energy efficiency, but also achieves satisfied performances in end-to-end delay and communication reliability. PMID:27023562

  16. Purifying Selection Maintains Dosage-Sensitive Genes during Degeneration of the Threespine Stickleback Y Chromosome.

    PubMed

    White, Michael A; Kitano, Jun; Peichel, Catherine L

    2015-08-01

    Sex chromosomes are subject to unique evolutionary forces that cause suppression of recombination, leading to sequence degeneration and the formation of heteromorphic chromosome pairs (i.e., XY or ZW). Although progress has been made in characterizing the outcomes of these evolutionary processes on vertebrate sex chromosomes, it is still unclear how recombination suppression and sequence divergence typically occur and how gene dosage imbalances are resolved in the heterogametic sex. The threespine stickleback fish (Gasterosteus aculeatus) is a powerful model system to explore vertebrate sex chromosome evolution, as it possesses an XY sex chromosome pair at relatively early stages of differentiation. Using a combination of whole-genome and transcriptome sequencing, we characterized sequence evolution and gene expression across the sex chromosomes. We uncovered two distinct evolutionary strata that correspond with known structural rearrangements on the Y chromosome. In the oldest stratum, only a handful of genes remain, and these genes are under strong purifying selection. By comparing sex-linked gene expression with expression of autosomal orthologs in an outgroup, we show that dosage compensation has not evolved in threespine sticklebacks through upregulation of the X chromosome in males. Instead, in the oldest stratum, the genes that still possess a Y chromosome allele are enriched for genes predicted to be dosage sensitive in mammals and yeast. Our results suggest that dosage imbalances may have been avoided at haploinsufficient genes by retaining function of the Y chromosome allele through strong purifying selection.

  17. FTIR gas analysis with improved sensitivity and selectivity for CWA and TIC detection

    NASA Astrophysics Data System (ADS)

    Phillips, Charles M.; Tan, Huwei

    2010-04-01

    This presentation describes the use of an FTIR (Fourier Transform Infrared)-based spectrometer designed to continuously monitor ambient air for the presence of chemical warfare agents (CWAs) and toxic industrial chemicals (TICs). The necessity of a reliable system capable of quickly and accurately detecting very low levels of CWAs and TICs while simultaneously retaining a negligible false alarm rate will be explored. Technological advancements in FTIR sensing have reduced noise while increasing selectivity and speed of detection. These novel analyzer design characteristics are discussed in detail and descriptions are provided which show how optical throughput, gas cell form factor, and detector response are optimized. The hardware and algorithms described here will explain why this FTIR system is very effective for the simultaneous detection and speciation of a wide variety of toxic compounds at ppb concentrations. Analytical test data will be reviewed demonstrating the system's sensitivity to and selectivity for specific CWAs and TICs; this will include recent data acquired as part of the DHS ARFCAM (Autonomous Rapid Facility Chemical Agent Monitor) project. These results include analyses of the data from live agent testing for the determination of CWA detection limits, immunity to interferences, detection times, residual noise analysis and false alarm rates. Sensing systems such as this are critical for effective chemical hazard identification which is directly relevant to the CBRNE community.

  18. Highly sensitive and selective WO3 nanoparticle gas sensor operating in thermally modulated dynamic mode

    NASA Astrophysics Data System (ADS)

    Ionescu, Radu; Hoel, Anders; Granqvist, Claes-Goran; Llobet, Eduard; Heszler, Peter

    2004-05-01

    Nanoparticle films of crystalline WO3, designed for gas sensing applications, were deposited on alumina substrates by reactive gas deposition. H2S, ethanol vapour, and binary mixtures of ethanol/H2S, ethanol/NO2 and H2S/NO2 were used in different concentrations for testing the performance of the sensor device. The sensor was operated in dynamic mode by modulating its temperature between 150 and 250 °C. Coefficients were extracted by applying Fast Fourier Transform (FFT) and Discrete Wavelet Transform (DWT) methods to the dynamic resistance response of the sensor. These coefficients were then used as inputs for pattern recognition methods to extract both quantitative (concentration) and qualitative (chemical selectivity) information about the test gases. After sensor calibration, it was possible to detect as little as 200 ppb of ethanol and 20 ppb of H2S with good accuracy. Furthermore, ethanol and H2S could be detected with good sensitivity and selectivity in the presence of both reducing and oxidising gases.

  19. A Sensitive Secondary Users Selection Algorithm for Cognitive Radio Ad Hoc Networks.

    PubMed

    Li, Aohan; Han, Guangjie; Wan, Liangtian; Shu, Lei

    2016-03-26

    Secondary Users (SUs) are allowed to use the temporarily unused licensed spectrum without disturbing Primary Users (PUs) in Cognitive Radio Ad Hoc Networks (CRAHNs). Existing architectures for CRAHNs impose energy-consuming Cognitive Radios (CRs) on SUs. However, the advanced CRs will increase energy cost for their cognitive functionalities, which is undesirable for the battery powered devices. A new architecture referred to as spectral Requirement-based CRAHN (RCRAHN) is proposed to enhance energy efficiency for CRAHNs in this paper. In RCRAHNs, only parts of SUs are equipped with CRs. SUs equipped with CRs are referred to as Cognitive Radio Users (CRUs). To further enhance energy efficiency of CRAHNs, we aim to select minimum CRUs to sense available spectrum. A non-linear programming problem is mathematically formulated under the constraints of energy efficiency and real-time. Considering the NP-hardness of the problem, a framework of a heuristic algorithm referred to as Sensitive Secondary Users Selection (SSUS) was designed to compute the near-optimal solutions. The simulation results demonstrate that SSUS not only improves the energy efficiency, but also achieves satisfied performances in end-to-end delay and communication reliability.

  20. Selective allergy to lobster in a case of primary sensitization to house dust mites.

    PubMed

    Iparraguirre, A; Rodríguez-Pérez, R; Juste, S; Ledesma, A; Moneo, I; Caballero, M L

    2009-01-01

    Allergy to only 1 kind of seafood is uncommon. We report a case of selective allergy to lobster. We studied a 30-year-old man who suffered generalized urticaria, facial erythema, and pharyngeal pruritus after eating lobster. He had a more than 10-year history of mild persistent asthma and sensitization to house dust mites. The study was performed by skin prick test, and prick-prick test, oral food challenge, specific immunoglobulin (Ig) E determinations by CAP (Phadia, Uppsala, Sweden) and ADVIA-Centaur (ALK-Abelló, Madrid, Spain), and IgE-immunoblotting. The patient's serum recognized 2 allergens of around 198 kDa and 2 allergens of around 65 kDa from the lobster extract, allergens of around 15, 90, and 120 kDa from Dermatophagoides pteronyssinus extract, and allergens of around 15 and 65 kDa from Dermatophagoides farinae extract. Serum did not recognize purified shrimp tropomyosin. Immunoblot-inhibition assay results indicated cross-reactivity between lobster and mite allergens. This is the first report of selective allergy to lobster.

  1. A high-sensitivity 2x2 multi-aperture color camera based on selective averaging

    NASA Astrophysics Data System (ADS)

    Zhang, Bo; Kagawa, Keiichiro; Takasawa, Taishi; Seo, Min-Woong; Yasutomi, Keita; Kawahito, Shoji

    2015-03-01

    To demonstrate the low-noise performance of the multi-aperture imaging system using a selective averaging method, an ultra-high-sensitivity multi-aperture color camera with 2×2 apertures is being developed. In low-light conditions, random telegraph signal (RTS) noise and dark current white defects become visible, which greatly degrades the quality of the image. To reduce these kinds of noise as well as to increase the number of incident photons, the multi-aperture imaging system composed of an array of lens and CMOS image sensor (CIS), and the selective averaging for minimizing the synthetic sensor noise at every pixel is utilized. It is verified by simulation that the effective noise at the peak of noise histogram is reduced from 1.44 e- to 0.73 e- in a 2×2-aperture system, where RTS noise and dark current white defects have been successfully removed. In this work, a prototype based on low-noise color sensors with 1280×1024 pixels fabricated in 0.18um CIS technology is considered. The pixel pitch is 7.1μm×7.1μm. The noise of the sensor is around 1e- based on the folding-integration and cyclic column ADCs, and the low voltage differential signaling (LVDS) is used to improve the noise immunity. The synthetic F-number of the prototype is 0.6.

  2. Sensitive and selective magnetoimmunosensing platform for determination of the food allergen Ara h 1.

    PubMed

    Ruiz-Valdepeñas Montiel, V; Campuzano, S; Pellicanò, A; Torrente-Rodríguez, R M; Reviejo, A J; Cosio, M S; Pingarrón, J M

    2015-06-23

    A highly sensitive disposable amperometric immunosensor based on the use of magnetic beads (MBs) is described for determination of Ara h 1, the major peanut allergen, in only 2h. The approach uses a sandwich configuration involving selective capture and biotinylated detector antibodies and carboxylic acid-modified MBs (HOOC-MBs). The MBs bearing the immunoconjugates are captured by a magnet placed under the surface of a disposable screen-printed carbon electrode (SPCE) and the affinity reactions are monitored amperometrically at -0.20 V (vs a Ag pseudo-reference electrode) in the presence of hydroquinone (HQ) as electron transfer mediator and upon addition of H2O2 as the enzyme substrate. The developed immunosensor exhibits a wide range of linearity between 20.8 and 1000.0 ng mL(-1) Ara h 1, a detection limit of 6.3 ng mL(-1), a great selectivity, a good reproducibility with a RSD of 6.3% for six different immunosensors and a useful lifetime of 25 days. The usefulness of the immunosensor was demonstrated by determining Ara h 1 in different matrices (food extracts and saliva). The results correlated properly with those provided by a commercial ELISA method offering a reliable and promising analytical screening tool in the development of user-friendly devices for on-site determination of Ara h 1. PMID:26092337

  3. Sensitive indoor air monitoring of monoterpenes using different adsorbents and thermal desorption gas chromatography with mass-selective detection.

    PubMed

    Hollender, Juliane; Sandner, Frank; Möller, Manfred; Dott, Wolfgang

    2002-07-12

    A simple method using active trapping on adsorbents and thermal desorption followed by GC-MS analysis was developed for the indoor air monitoring of monoterpenes. The study was carried out using a dynamically generated atmosphere consisting of 11 monoterpenes: camphene, camphor, delta 3-carene, 1,8-cineol, limonene, linalool, alpha-pinene, beta-pinene, alpha-terpinene, gamma-terpinene, fenchyl alcohol. The influence of the different adsorbents Tenax TA, Tenax GR, Carbosieve SIII, Chromosorb 106 on the yield of six selected monoterpenes at indoor air concentrations was studied. The adsorbent Tenax GR gave relatively the best yields followed by Tenax TA. Detection limits of approximately 1 microgram m3 were determined with Tenax GR for most of the monoterpenes.

  4. Selection and Characterization of [alpha]-Methyltryptophan-Resistant Lines of Lemna gibba Showing a Rapid Rate of Indole-3-Acetic Acid Turnover.

    PubMed Central

    Tam, Y. Y.; Slovin, J. P.; Cohen, J. D.

    1995-01-01

    Turnover rate is an important aspect of the regulation of plant processes by plant growth substances. To study turnover of indole-3-acetic acid (IAA), two [alpha]-methyltryptophan-resistant lines (MTR1 and MTR2) of Lemna gibba were generated by nitrosomethyl urea treatment of an inbred line derived from L. gibba G-3. In this report we describe: (a) the development of a selection system using this near isogenic line of L. gibba; (b) techniques for chemical mutation of the lines and selection for [alpha]-methyltryptophan resistance; and (c) the partial characterization of the selected lines. MTR lines contained 3-fold higher levels of anthranilate synthase activity. The enzyme in the MTR lines required higher levels of tryptophan for feedback inhibition. MTR lines also contained 8-fold higher levels of tryptophan, 3-fold higher levels of free IAA, and similar levels of total IAA compared to the inbred line. Turnover rates in the inbred and selected lines were calculated, using the first-order rate equation, based on the decrease over time in isotopic enrichment of I3C6-IAA introduced into L. gibba during a 1-h pulse period. Isotope enrichment in IAA was determined by using gas chromatography-mass spectrometry. Both MTR lines had an approximately 10-fold higher rate of IAA turnover than the parent inbred line. PMID:12228344

  5. Pattern of retinoid-induced teratogenic effects: possible relationship with relative selectivity for nuclear retinoid receptors RAR alpha, RAR beta, and RAR gamma.

    PubMed

    Elmazar, M M; Reichert, U; Shroot, B; Nau, H

    1996-03-01

    Retinoic acid, an oxidative metabolite of vitamin A, is involved in the control of many biological processes including embryonic development. Excess as well as deficiency of retinoids were found to be teratogenic. The effects of retinoids in normal as well as abnormal development may be mediated by two members of retinoid receptors, the RAR's and RXR's, which exhibit a specific temporal and spatial expression during development. The significance of the retinoid receptors was investigated here by studying the teratogenic effects of retinoid ligands with relative selectivity for binding and transactivation of the retinoic acid receptors RAR alpha, RAR beta and RAR gamma. Pregnant NMRI mice were administered 5 or 15 mg/kg of CD 336 (Am 580) (alpha-ligand), CD 2019 (beta-ligand), CD 437 (gamma-ligand) or 37.5 mg/kg all-trans-retinoic acid in 25% Cremophor EL on day 8.25 or day 11 of gestation by gastric intubation. External, visceral and skeletal malformations were observed on day 18 of gestation. The order of teratogenic potency was: alpha-ligand > beta-ligand > gamma-ligand. In addition, these retinoids also produced a different spectrum of defects. The alpha-ligand induced the most varied defects including severe ear, mandible, and limb malformations. The beta-ligand induced defects of the urinary system and liver in greater frequency than expected from its relative potency. The gamma-ligand preferentially induced ossification deficiencies and defects of the sternebrae and vertebral body. Our results show that these three retinoids, which were previously demonstrated to exhibit retinoid-like activities in several systems, exert differing teratogenic activities, in regard to both potency and regioselectivity: we hypothesize that the relative selectivity for binding and transactivation of the three retinoic acid receptors could possibly be related to the differences of teratogenic effects observed in this study. The low potency of the gamma-ligand may lead the way to

  6. Fatty acid specificity of hormone-sensitive lipase. Implication in the selective hydrolysis of triacylglycerols.

    PubMed

    Raclot, T; Holm, C; Langin, D

    2001-12-01

    The selective mobilization of fatty acids from white fat cells depends on their molecular structure, in particular the degree of unsaturation. The present study was designed to examine if the release of fatty acids by hormone-sensitive lipase (HSL) in vitro i) is influenced by the amount of unsaturation, ii) depends on the temperature, and iii) could explain the selective pattern of fatty acid mobilization and notably the preferential mobilization of certain highly unsaturated fatty acids. Recombinant rat and human HSL were incubated with a lipid emulsion. The hydrolysis of 35 individual fatty acids, ranging in chain length from 12 to 24 carbon atoms and in unsaturation from 0 to 6 double bonds was measured. Fatty acid composition of in vitro released NEFA was compared with that of fat cell triacylglycerols (TAG), the ratio % NEFA/% TAG being defined as the relative hydrolysis. The relative hydrolysis of individual fatty acids differed widely, ranging from 0.44 (24:1n-9) to 1.49 (18:1n-7) with rat HSL, and from 0.38 (24:1n-9) to 1.67 (18:1n-7) with human HSL. No major difference was observed between rat and human HSL. The relative release was dependent on the number of double bonds according to chain length. The amount of fatty acid released by recombinant rat HSL was decreased but remained robust at 4 degrees C compared with 37 degrees C, and the relative hydrolysis of some individual fatty acids was affected. The relative hydrolysis of fatty acids moderately, weakly, and highly mobilized by adipose tissue in vivo was similar and close to unity in vitro. We conclude that i) the release of fatty acids by HSL is only slightly affected by their degree of unsaturation, ii) the ability of HSL to efficiently and selectively release fatty acids at low temperature could reflect a cold adaptability for poikilotherms or hibernators when endogenous lipids are needed, and iii) the selectivity of fatty acid hydrolysis by HSL does not fully account for the selective pattern of

  7. Design and synthesis of novel alpha(1)(a) adrenoceptor-selective antagonists. 2. Approaches to eliminate opioid agonist metabolites via modification of linker and 4-methoxycarbonyl-4-phenylpiperidine moiety.

    PubMed

    Murali Dhar, T G; Nagarathnam, D; Marzabadi, M R; Lagu, B; Wong, W C; Chiu, G; Tyagarajan, S; Miao, S W; Zhang, F; Sun, W; Tian, D; Shen, Q; Zhang, J; Wetzel, J M; Forray, C; Chang, R S; Broten, T P; Schorn, T W; Chen, T B; O'Malley, S; Ransom, R; Schneck, K; Bendesky, R; Harrell, C M; Vyas, K P

    1999-11-18

    We have previously described compound 1a as a high-affinity subtype selective alpha(1a) antagonist. In vitro and in vivo evaluation of compound 1a showed its major metabolite to be a mu-opioid agonist, 4-methoxycarbonyl-4-phenylpiperidine (3). Several dihydropyrimidinone analogues were synthesized with the goal of either minimizing the formation of 3 by modification of the linker or finding alternative piperidine moieties which when cleaved as a consequence of metabolism would not give rise to mu-opioid activity. Modification of the linker gave several compounds with good alpha(1a) binding affinity (K(i) = < 1 nM) and selectivity (>300-fold over alpha(1b) and alpha(1d)). In vitro analysis in the microsomal assay revealed these modifications did not significantly affect N-dealkylation and the formation of the piperidine 3. The second approach, however, yielded several piperidine replacements for 3, which did not show significant mu-opioid activity. Several of these compounds maintained good affinity at the alpha(1a) adrenoceptor and selectivity over alpha(1b) and alpha(1d). For example, the piperidine fragments of (+)-73 and (+)-83, viz. 4-cyano-4-phenylpiperidine and 4-methyl-4-phenylpiperidine, were essentially inactive at the mu-opioid receptor (IC(50) > 30 microM vs 3 microM for 3). Compounds (+)-73 and (+)-83 were subjected to detailed in vitro and in vivo characterization. Both these compounds, in addition to their excellent selectivity (>880-fold) over alpha(1b) and alpha(1d), also showed good selectivity over several other recombinant human G-protein coupled receptors. Compounds (+)-73 and (+)-83 showed good functional potency in isolated human prostate tissues, with K(b)s comparable to their in vitro alpha(1a) binding data. In addition, compound (+)-73 also exhibited good uroselectivity (DBP K(b)/IUP K(b) > 20-fold) in the in vivo experiments in dogs, similar to 1a.

  8. Aspartic acid-promoted highly selective and sensitive colorimetric sensing of cysteine in rat brain.

    PubMed

    Qian, Qin; Deng, Jingjing; Wang, Dalei; Yang, Lifen; Yu, Ping; Mao, Lanqun

    2012-11-01

    Direct selective determination of cysteine in the cerebral system is of great importance because of the crucial roles of cysteine in physiological and pathological processes. In this study, we report a sensitive and selective colorimetric assay for cysteine in the rat brain with gold nanoparticles (Au-NPs) as the signal readout. Initially, Au-NPs synthesized with citrate as the stabilizer are red in color and exhibit absorption at 520 nm. The addition of an aqueous solution (20 μL) of cysteine or aspartic acid alone to a 200 μL Au-NP dispersion causes no aggregation, while the addition of an aqueous solution of cysteine into a Au-NP dispersion containing aspartic acid (1.8 mM) causes the aggregation of Au-NPs and thus results in the color change of the colloid from wine red to blue. These changes are ascribed to the ion pair interaction between aspartic acid and cysteine on the interface between Au-NPs and solution. The concentration of cysteine can be visualized with the naked eye and determined by UV-vis spectroscopy. The signal output shows a linear relationship for cysteine within the concentration range from 0.166 to 1.67 μM with a detection limit of 100 nM. The assay demonstrated here is highly selective and is free from the interference of other natural amino acids and other thiol-containing species as well as the species commonly existing in the brain such as lactate, ascorbic acid, and glucose. The basal dialysate level of cysteine in the microdialysate from the striatum of adult male Sprague-Dawley rats is determined to be around 9.6 ± 2.1 μM. The method demonstrated here is facile but reliable and durable and is envisaged to be applicable to understanding the chemical essence involved in physiological and pathological events associated with cysteine. PMID:23025476

  9. Cortical Granule Exocytosis Is Mediated by Alpha-SNAP and N-Ethilmaleimide Sensitive Factor in Mouse Oocytes

    PubMed Central

    de Paola, Matilde; Bello, Oscar Daniel; Michaut, Marcela Alejandra

    2015-01-01

    Cortical granule exocytosis (CGE), also known as cortical reaction, is a calcium- regulated secretion that represents a membrane fusion process during meiotic cell division of oocytes. The molecular mechanism of membrane fusion during CGE is still poorly understood and is thought to be mediated by the SNARE pathway; nevertheless, it is unkown if SNAP (acronym for soluble NSF attachment protein) and NSF (acronym for N-ethilmaleimide sensitive factor), two key proteins in the SNARE pathway, mediate CGE in any oocyte model. In this paper, we documented the gene expression of α-SNAP, γ-SNAP and NSF in mouse oocytes. Western blot analysis showed that the expression of these proteins maintains a similar level during oocyte maturation and early activation. Their localization was mainly observed at the cortical region of metaphase II oocytes, which is enriched in cortical granules. To evaluate the function of these proteins in CGE we set up a functional assay based on the quantification of cortical granules metaphase II oocytes activated parthenogenetically with strontium. Endogenous α-SNAP and NSF proteins were perturbed by microinjection of recombinant proteins or antibodies prior to CGE activation. The microinjection of wild type α-SNAP and the negative mutant of α-SNAP L294A in metaphase II oocytes inhibited CGE stimulated by strontium. NEM, an irreversibly inhibitor of NSF, and the microinjection of the negative mutant NSF D1EQ inhibited cortical reaction. The microinjection of anti-α-SNAP and anti-NSF antibodies was able to abolish CGE in activated metaphase II oocytes. The microinjection of anti-γ SNAP antibody had no effect on CGE. Our findings indicate, for the first time in any oocyte model, that α-SNAP, γ-SNAP, and NSF are expressed in mouse oocytes. We demonstrate that α-SNAP and NSF have an active role in CGE and propose a working model. PMID:26267363

  10. Cortical Granule Exocytosis Is Mediated by Alpha-SNAP and N-Ethilmaleimide Sensitive Factor in Mouse Oocytes.

    PubMed

    de Paola, Matilde; Bello, Oscar Daniel; Michaut, Marcela Alejandra

    2015-01-01

    Cortical granule exocytosis (CGE), also known as cortical reaction, is a calcium- regulated secretion that represents a membrane fusion process during meiotic cell division of oocytes. The molecular mechanism of membrane fusion during CGE is still poorly understood and is thought to be mediated by the SNARE pathway; nevertheless, it is unkown if SNAP (acronym for soluble NSF attachment protein) and NSF (acronym for N-ethilmaleimide sensitive factor), two key proteins in the SNARE pathway, mediate CGE in any oocyte model. In this paper, we documented the gene expression of α-SNAP, γ-SNAP and NSF in mouse oocytes. Western blot analysis showed that the expression of these proteins maintains a similar level during oocyte maturation and early activation. Their localization was mainly observed at the cortical region of metaphase II oocytes, which is enriched in cortical granules. To evaluate the function of these proteins in CGE we set up a functional assay based on the quantification of cortical granules metaphase II oocytes activated parthenogenetically with strontium. Endogenous α-SNAP and NSF proteins were perturbed by microinjection of recombinant proteins or antibodies prior to CGE activation. The microinjection of wild type α-SNAP and the negative mutant of α-SNAP L294A in metaphase II oocytes inhibited CGE stimulated by strontium. NEM, an irreversibly inhibitor of NSF, and the microinjection of the negative mutant NSF D1EQ inhibited cortical reaction. The microinjection of anti-α-SNAP and anti-NSF antibodies was able to abolish CGE in activated metaphase II oocytes. The microinjection of anti-γ SNAP antibody had no effect on CGE. Our findings indicate, for the first time in any oocyte model, that α-SNAP, γ-SNAP, and NSF are expressed in mouse oocytes. We demonstrate that α-SNAP and NSF have an active role in CGE and propose a working model.

  11. Expression of protein phosphatase 2A mutants and silencing of the regulatory B alpha subunit induce a selective loss of acetylated and detyrosinated microtubules.

    PubMed

    Nunbhakdi-Craig, Viyada; Schuechner, Stefan; Sontag, Jean-Marie; Montgomery, Lisa; Pallas, David C; Juno, Claudia; Mudrak, Ingrid; Ogris, Egon; Sontag, Estelle

    2007-05-01

    Carboxymethylation and phosphorylation of protein phosphatase 2A (PP2A) catalytic C subunit are evolutionary conserved mechanisms that critically control PP2A holoenzyme assembly and substrate specificity. Down-regulation of PP2A methylation and PP2A enzymes containing the B alpha regulatory subunit occur in Alzheimer's disease. In this study, we show that expressed wild-type and methylation- (L309 Delta) and phosphorylation- (T304D, T304A, Y307F, and Y307E) site mutants of PP2A C subunit differentially bind to B, B', and B''-type regulatory subunits in NIH 3T3 fibroblasts and neuro-2a (N2a) neuroblastoma cells. They also display distinct binding affinity for microtubules (MTs). Relative to controls, expression of the wild-type, T304A and Y307F C subunits in N2a cells promotes the accumulation of acetylated and detyrosinated MTs. However, expression of the Y307E, L309 Delta, and T304D mutants, which are impaired in their ability to associate with the B alpha subunit, induces their loss. Silencing of B alpha subunit in N2a and NIH 3T3 cells is sufficient to induce a similar breakdown of acetylated and detyrosinated MTs. It also confers increased sensitivity to nocodazole-induced MT depolymerization. Our findings suggest that changes in intracellular PP2A subunit composition can modulate MT dynamics. They support the hypothesis that reduced amounts of neuronal B alpha-containing PP2A heterotrimers contribute to MT destabilization in Alzheimer's disease.

  12. Sex-selective effects of neonatal isolation on fear conditioning and foot shock sensitivity.

    PubMed

    Kosten, Therese A; Miserendino, Mindy J D; Bombace, Joan C; Lee, Hongjoo J; Kim, Jeansok J

    2005-02-28

    Our previous work demonstrates enduring effects of the early life stress of neonatal isolation (ISO). ISO facilitates appetitive response learning in adult female, but not male rats, and enhances corticosterone levels and stress responsivity in infant and juvenile rats of both sexes. Corticosterone acts at brain areas such as hippocampus that are rich in glucocorticoid receptors, differentiate postnatally, are sexually dimorphic, and involved in learning. Thus, ISO is hypothesized to alter aversive learning in adult rats in a sex-specific manner. This study tests this hypothesis using context and cue fear conditioning. Pups were isolated for 1h a day on postnatal (PN) days 2-9 or were non-handled and were then tested in adulthood (PN70-90). In Experiment 1, context- and cue-elicited freezing and ultrasonic vocalizations (USVs; 22 kHz range) were measured. Experiments 2-4, respectively, examined three unconditioned foot shock responses (flinch, jump, vocalization), unconditioned fear (time in center of an open, novel arena), and appetitive (stroking-induced; 50 kHz range) USVs. ISO had a sex-selective effect on context-induced USVs that may reflect changes in foot shock sensitivity. ISO increases foot shock sensitivity and tends to enhance context-induced fear in female rats, whereas ISO tends to impair context-induced fear in male rats. Overall, male rats show greater conditioned fear, and female rats show greater unconditioned fear as well as enhanced responses to the aversive and appetitive stimuli. The sex-specific effect of ISO on context fear may reflect neuronal reorganization in stress responsive areas and/or sex differences in some unconditioned responses.

  13. SynLethDB: synthetic lethality database toward discovery of selective and sensitive anticancer drug targets.

    PubMed

    Guo, Jing; Liu, Hui; Zheng, Jie

    2016-01-01

    Synthetic lethality (SL) is a type of genetic interaction between two genes such that simultaneous perturbations of the two genes result in cell death or a dramatic decrease of cell viability, while a perturbation of either gene alone is not lethal. SL reflects the biologically endogenous difference between cancer cells and normal cells, and thus the inhibition of SL partners of genes with cancer-specific mutations could selectively kill cancer cells but spare normal cells. Therefore, SL is emerging as a promising anticancer strategy that could potentially overcome the drawbacks of traditional chemotherapies by reducing severe side effects. Researchers have developed experimental technologies and computational prediction methods to identify SL gene pairs on human and a few model species. However, there has not been a comprehensive database dedicated to collecting SL pairs and related knowledge. In this paper, we propose a comprehensive database, SynLethDB (http://histone.sce.ntu.edu.sg/SynLethDB/), which contains SL pairs collected from biochemical assays, other related databases, computational predictions and text mining results on human and four model species, i.e. mouse, fruit fly, worm and yeast. For each SL pair, a confidence score was calculated by integrating individual scores derived from different evidence sources. We also developed a statistical analysis module to estimate the druggability and sensitivity of cancer cells upon drug treatments targeting human SL partners, based on large-scale genomic data, gene expression profiles and drug sensitivity profiles on more than 1000 cancer cell lines. To help users access and mine the wealth of the data, we developed other practical functionalities, such as search and filtering, orthology search, gene set enrichment analysis. Furthermore, a user-friendly web interface has been implemented to facilitate data analysis and interpretation. With the integrated data sets and analytics functionalities, SynLethDB would

  14. SynLethDB: synthetic lethality database toward discovery of selective and sensitive anticancer drug targets.

    PubMed

    Guo, Jing; Liu, Hui; Zheng, Jie

    2016-01-01

    Synthetic lethality (SL) is a type of genetic interaction between two genes such that simultaneous perturbations of the two genes result in cell death or a dramatic decrease of cell viability, while a perturbation of either gene alone is not lethal. SL reflects the biologically endogenous difference between cancer cells and normal cells, and thus the inhibition of SL partners of genes with cancer-specific mutations could selectively kill cancer cells but spare normal cells. Therefore, SL is emerging as a promising anticancer strategy that could potentially overcome the drawbacks of traditional chemotherapies by reducing severe side effects. Researchers have developed experimental technologies and computational prediction methods to identify SL gene pairs on human and a few model species. However, there has not been a comprehensive database dedicated to collecting SL pairs and related knowledge. In this paper, we propose a comprehensive database, SynLethDB (http://histone.sce.ntu.edu.sg/SynLethDB/), which contains SL pairs collected from biochemical assays, other related databases, computational predictions and text mining results on human and four model species, i.e. mouse, fruit fly, worm and yeast. For each SL pair, a confidence score was calculated by integrating individual scores derived from different evidence sources. We also developed a statistical analysis module to estimate the druggability and sensitivity of cancer cells upon drug treatments targeting human SL partners, based on large-scale genomic data, gene expression profiles and drug sensitivity profiles on more than 1000 cancer cell lines. To help users access and mine the wealth of the data, we developed other practical functionalities, such as search and filtering, orthology search, gene set enrichment analysis. Furthermore, a user-friendly web interface has been implemented to facilitate data analysis and interpretation. With the integrated data sets and analytics functionalities, SynLethDB would

  15. Elucidating the Complex Lineshapes Resulting from the Highly Sensitive, Ion Selective, Technique Nice-Ohvms

    NASA Astrophysics Data System (ADS)

    Hodges, James N.; Siller, Brian; McCall, Benjamin J.

    2015-06-01

    The technique Noise Immune Cavity Enhanced Optical Heterodyne Velocity Modulation Spectroscopy, or NICE-OHVMS, has been used to great effect to precisely and accurately measure a variety of molecular ion transitions from species such as H_3^+, CH_5^+, HeH^+, and HCO^+, achieving MHz or in some cases sub-MHz uncertainty. It is a powerful technique, but a complete theoretical understanding of the complex NICE-OHVMS lineshape is needed to fully unlock its potential. NICE-OHVMS is the direct result of the combination of the highly sensitive spectroscopic technique Noise Immune Cavity Enhanced Optical Heterodyne Molecular Spectroscopy(NICE-OHMS) with Velocity Modulation Spectroscopy(VMS), applying the most sensitive optical detection method with ion species selectivity. The theoretical underpinnings of NICE-OHMS lineshapes are well established, as are those of VMS. This presentation is the logical extension of those two preceding bodies of work. Simulations of NICE-OHVMS lineshapes under a variety of conditions and fits of experimental data to the model are presented. The significance and accuracy of the various inferred parameters, along with the prospect of using them to extract additional information from observed transitions, are discussed. J.~N. Hodges, et al. J. Chem. Phys. (2013), 139, 164201. A.~J. Perry, et al. J. Chem. Phys. (2014), 141, 101101. K.~N. Crabtree, et al. Chem. Phys. Lett. (2012), 551, 1-6. F.~M. Schmidt, et al. J. Opt. Soc. Amer. A (2008), 24, 1392--1405. J.~W. Farley, J. Chem. Phys. (1991), 95, 5590--5602.

  16. Sensitivity of Lower Stratospheric Assimilated Ozone on Error Covariance Modeling and Data Selection

    NASA Technical Reports Server (NTRS)

    Stajner, Ivanka; Rood, Richard B.; Winslow, Nathan; Wargan, Krzysztof; Pawson, Steven

    2002-01-01

    Assimilated ozone is produced at the NASA/Goddard Data Assimilation Office by blending ozone retrieved from the Solar Backscatter UltraViolet/2 (SBUV/2) instrument and the Earth Probe Total Ozone Mapping Spectrometer (EP TOMS) measurements into an off-line transport model. The current system tends to overestimate the amount of lower stratospheric ozone. This is a region where ozone plays a key role in the forcing of climate. A biased ozone field in this region will adversely impact calculations of the stratosphere-troposphere exchange and, when used as a first guess in retrievals, the values determined from satellite observations. Since these are all important applications of assimilated ozone products, effort is being directed towards reducing this bias. The SBUV ozone data have a coarse vertical resolution with increased uncertainty below the ozone maximum, and TOMS provides only total ozone columns. Thus, the assimilated ozone in the lower stratosphere, and its vertical distribution in particular, are only weakly constrained by the incoming SBUV and TOMS data. Consequently, the assimilated ozone distribution should be sensitive to changes in inputs to the statistical analysis scheme. Accordingly, the sensitivity of the assimilated lower stratospheric ozone fields to changes in the TOMS error-covariance modeling and the SBUV data selection has been investigated. The use of a spatially correlated TOMS error covariance model led to improvements in the product. However, withholding the SBUV/2 data for the layer between 63 and 126 hPa typically degraded the product, a result which vindicates the use of this layer ozone product, despite its known errors. These efforts to improve the lower stratospheric distribution will be extended to include a more advanced forecast error covariance model, and by assimilating ozone products from new instruments on Envisat and EOS Aura.

  17. Caging of plumbagin on silver nanoparticles imparts selectivity and sensitivity to plumbagin for targeted cancer cell apoptosis.

    PubMed

    Duraipandy, N; Lakra, Rachita; Kunnavakkam Vinjimur, Srivatsan; Samanta, Debasis; K, Purna Sai; Kiran, Manikantan Syamala

    2014-11-01

    Plumbagin is a nutraceutical with potent anti-cancer activity. However, the therapeutic efficacy of plumbagin is overshadowed by the lack of sensitivity and selectivity towards cancer cells. The present study evaluated the use of nano-biotechnological intervention to cage plumbagin in silver nanoparticles for selective targeting of its biological effects towards cancerous cells. Caging of plumbagin in silver nanoparticles imparted selectivity and sensitivity to plumbagin for selective killing of cancer cells by altering the redox signalling events in the cancer cells. The selectivity and sensitivity of plumbagin towards cancer cells was due to the cumulative expression of the properties of plumbagin and nanoparticles which specifically affected the differential cancer cell microenvironment by altering the pyruvate kinase activity that regulates the ROS challenge in cancerous cells. The positive surface charge of plumbagin caged silver nanoparticles (PCSN) aids in getting them targeted towards anionic cancerous cells due to the exposed terminal carboxyl group of sialic acid residues. Furthermore, we observed that the effective concentration of the drug required to induce apoptosis was brought down to 50% upon caging of plumbagin on silver nanoparticles. We observed no such effect with the individual compound alone. The results indicated that the physico-chemical and biochemical properties of plumbagin significantly changed after conjugation with nanomaterials that facilitated "adding-in" therapeutical values to plumbagin which would otherwise be overshadowed by its lack of sensitivity and selectivity towards cancer cells. PMID:25188862

  18. Laser selective microablation of sensitized intracellular components within auditory receptor cells

    NASA Astrophysics Data System (ADS)

    Harris, David M.; Evans, Burt N.; Santos-Sacchi, Joseph

    1995-05-01

    A laser system can be coupled to a light microscope for laser microbeam ablation and trapping of single cells in vitro. We have extended this technology by sensitization of target structures with vital dyes to provide selective ablation of specific subcellular components. Isolated auditory receptor cells (outer hair cells, OHCs) are known to elongate and contract in response to electrical, chemical and mechanical stimulation. Various intracellular structures are candidate components mediating motility of OHCs, but the exact mechanism(s) is currently unknown. In ongoing studies of OHC motility, we have used the microbeam for selective ablation of lateral wall components and of an axial cytoskeletal core that extends from the nucleus to the cell apex. Both the area beneath the subsurface cistemae of the lateral wall and the core are rich in mitochondria. OHCs isolated from guinea pig cochlea are suspended in L- 15 medium containing 2.0 (mu) M Rhodamine 123, a porphyrin with an affinity for mitochondria. A spark-pumped nitrogen laser pumping a dye cell (Coumarin 500) was aligned on the optical axis of a Nikon Optiphot-2 to produce a 3 ns, 0.5 - 10 micrometers spot (diameter above ablation threshold w/50X water immersion, N.A. 0.8), and energy at the target approximately equals 10 (mu) J/pulse. At short incubation times in Rh123 irradiation caused local blebbing or bulging of cytoplastic membrane and thus loss of the OHC's cylindrical shape. At longer Rh123 incubation times when the central axis of the cell was targeted we observed cytoplasmic clearing, immediate cell elongation (approximately equals 5%) and clumping of core material at nuclear and apical attachments. Experiments are underway to examine the significance of these preliminary observations.

  19. A highly selective biosensor with nanomolar sensitivity based on cytokinin dehydrogenase.

    PubMed

    Tian, Faming; Greplová, Marta; Frébort, Ivo; Dale, Nicholas; Napier, Richard

    2014-01-01

    We have developed a N6-dimethylallyladenine (cytokinin) dehydrogenase-based microbiosensor for real-time determination of the family of hormones known as cytokinins. Cytokinin dehydrogenase from Zea mays (ZmCKX1) was immobilised concurrently with electrodeposition of a silica gel film on the surface of a Pt microelectrode, which was further functionalized by free electron mediator 2,6-dichlorophenolindophenol (DCPIP) in supporting electrolyte to give a bioactive film capable of selective oxidative cleavage of the N6- side chain of cytokinins. The rapid electron shuffling between freely diffusible DCPIP and the FAD redox group in ZmCKX1 endowed the microbiosensor with a fast response time of less than 10 s. The immobilised ZmCKX1 retained a high affinity for its preferred substrate N6-(Δ2-isopentenyl) adenine (iP), and gave the miniaturized biosensor a large linear dynamic range from 10 nM to 10 µM, a detection limit of 3.9 nM and a high sensitivity to iP of 603.3 µAmM-1cm-2 (n = 4, R2 = 0.9999). Excellent selectivity was displayed for several other aliphatic cytokinins and their ribosides, including N6-(Δ2-isopentenyl) adenine, N6-(Δ2-isopentenyl) adenosine, cis-zeatin, trans-zeatin and trans-zeatin riboside. Aromatic cytokinins and metabolites such as cytokinin glucosides were generally poor substrates. The microbiosensors exhibited excellent stability in terms of pH and long-term storage and have been used successfully to determine low nanomolar cytokinin concentrations in tomato xylem sap exudates.

  20. A Highly Selective Biosensor with Nanomolar Sensitivity Based on Cytokinin Dehydrogenase

    PubMed Central

    Tian, Faming; Greplová, Marta; Frébort, Ivo; Dale, Nicholas; Napier, Richard

    2014-01-01

    We have developed a N6-dimethylallyladenine (cytokinin) dehydrogenase-based microbiosensor for real-time determination of the family of hormones known as cytokinins. Cytokinin dehydrogenase from Zea mays (ZmCKX1) was immobilised concurrently with electrodeposition of a silica gel film on the surface of a Pt microelectrode, which was further functionalized by free electron mediator 2,6-dichlorophenolindophenol (DCPIP) in supporting electrolyte to give a bioactive film capable of selective oxidative cleavage of the N6- side chain of cytokinins. The rapid electron shuffling between freely diffusible DCPIP and the FAD redox group in ZmCKX1 endowed the microbiosensor with a fast response time of less than 10 s. The immobilised ZmCKX1 retained a high affinity for its preferred substrate N6-(Δ2-isopentenyl) adenine (iP), and gave the miniaturized biosensor a large linear dynamic range from 10 nM to 10 µM, a detection limit of 3.9 nM and a high sensitivity to iP of 603.3 µAmM−1cm−2 (n = 4, R2 = 0.9999). Excellent selectivity was displayed for several other aliphatic cytokinins and their ribosides, including N6-(Δ2-isopentenyl) adenine, N6-(Δ2-isopentenyl) adenosine, cis-zeatin, trans-zeatin and trans-zeatin riboside. Aromatic cytokinins and metabolites such as cytokinin glucosides were generally poor substrates. The microbiosensors exhibited excellent stability in terms of pH and long-term storage and have been used successfully to determine low nanomolar cytokinin concentrations in tomato xylem sap exudates. PMID:24595403

  1. Synthesis of Water-Dispersible Molecularly Imprinted Electroactive Nanoparticles for the Sensitive and Selective Paracetamol Detection.

    PubMed

    Luo, Jing; Ma, Qiang; Wei, Wei; Zhu, Ye; Liu, Ren; Liu, Xiaoya

    2016-08-17

    A novel kind of water-dispersible molecularly imprinted electroactive nanoparticles was prepared combining macromolecular self-assembly with molecularly imprinting technique employing paracetamol (PCM) as template molecule. An amphiphilic electroactive copolymer (P(NVC-EHA-AA), PNEA) containing carbazole group was first synthesized through a one-pot free radical copolymerization. The coassembly of the electroactive copolymers with the template molecules (PCM) in aqueous solution generated nanoparticles embedded with PCM, leading to the formation of molecularly imprinted electroactive nanoparticles (MIENPs). A robust MIP film was formed on the surface of electrode by electrodeposition of MIENPs and subsequent electropolymerization of the carbazole units in MIENPs. After the extraction of PCM molecules, a MIP sensor was successfully constructed. It should be noted that electropolymerization of the electroactive units in MIENPs creates cross-conjugated polymer network, which not only locks the recognition sites but also significantly accelerates the electron transfer and thus enhances the response signal of the MIP sensor. These advantages endowed the MIP sensor with good selectivity and high sensitivity for PCM detection. The MIP sensor could recognize PCM from its possible interfering substances with good selectivity. Under the optimal conditions, two linear ranges from 1 μM to 0.1 mM and 0.1 to 10 mM with a detection limit of 0.3 μM were obtained for PCM detection. The MIP sensor also showed good stability and repeatability, which has been successfully used to analyze PCM in tablets and human urine samples with satisfactory results.

  2. Synthesis of Water-Dispersible Molecularly Imprinted Electroactive Nanoparticles for the Sensitive and Selective Paracetamol Detection.

    PubMed

    Luo, Jing; Ma, Qiang; Wei, Wei; Zhu, Ye; Liu, Ren; Liu, Xiaoya

    2016-08-17

    A novel kind of water-dispersible molecularly imprinted electroactive nanoparticles was prepared combining macromolecular self-assembly with molecularly imprinting technique employing paracetamol (PCM) as template molecule. An amphiphilic electroactive copolymer (P(NVC-EHA-AA), PNEA) containing carbazole group was first synthesized through a one-pot free radical copolymerization. The coassembly of the electroactive copolymers with the template molecules (PCM) in aqueous solution generated nanoparticles embedded with PCM, leading to the formation of molecularly imprinted electroactive nanoparticles (MIENPs). A robust MIP film was formed on the surface of electrode by electrodeposition of MIENPs and subsequent electropolymerization of the carbazole units in MIENPs. After the extraction of PCM molecules, a MIP sensor was successfully constructed. It should be noted that electropolymerization of the electroactive units in MIENPs creates cross-conjugated polymer network, which not only locks the recognition sites but also significantly accelerates the electron transfer and thus enhances the response signal of the MIP sensor. These advantages endowed the MIP sensor with good selectivity and high sensitivity for PCM detection. The MIP sensor could recognize PCM from its possible interfering substances with good selectivity. Under the optimal conditions, two linear ranges from 1 μM to 0.1 mM and 0.1 to 10 mM with a detection limit of 0.3 μM were obtained for PCM detection. The MIP sensor also showed good stability and repeatability, which has been successfully used to analyze PCM in tablets and human urine samples with satisfactory results. PMID:27463123

  3. Metabolites related to gut bacterial metabolism, peroxisome proliferator-activated receptor-alpha activation, and insulin sensitivity are associated with physical function in functionally-limited older adults.

    PubMed

    Lustgarten, Michael S; Price, Lori L; Chalé, Angela; Fielding, Roger A

    2014-10-01

    Identification of mechanisms underlying physical function will be important for addressing the growing challenge that health care will face with physical disablement in the expanding aging population. Therefore, the goals of the current study were to use metabolic profiling to provide insight into biologic mechanisms that may underlie physical function by examining the association between baseline and the 6-month change in serum mass spectrometry-obtained amino acids, fatty acids, and acylcarnitines with baseline and the 6-month change in muscle strength (leg press one repetition maximum divided by total lean mass, LP/Lean), lower extremity function [short physical performance battery (SPPB)], and mobility (400 m gait speed, 400-m), in response to 6 months of a combined resistance exercise and nutritional supplementation (whey protein or placebo) intervention in functionally-limited older adults (SPPB ≤ 10; 70-85 years, N = 73). Metabolites related to gut bacterial metabolism (cinnamoylglycine, phenol sulfate, p-cresol sulfate, 3-indoxyl sulfate, serotonin, N-methylproline, hydrocinnamate, dimethylglycine, trans-urocanate, valerate) that are altered in response to peroxisome proliferator-activated receptor-alpha (PPAR-α) activation (α-hydroxyisocaproate, α-hydroxyisovalerate, 2-hydroxy-3-methylvalerate, indolelactate, serotonin, 2-hydroxypalmitate, glutarylcarnitine, isobutyrylcarnitine, cinnamoylglycine) and that are related to insulin sensitivity (monounsaturated fatty acids: 5-dodecenoate, myristoleate, palmitoleate; γ-glutamylamino acids: γ-glutamylglutamine, γ-glutamylalanine, γ-glutamylmethionine, γ-glutamyltyrosine; branched-chain amino acids: leucine, isoleucine, valine) were associated with function at baseline, with the 6-month change in function or were identified in backward elimination regression predictive models. Collectively, these data suggest that gut microbial metabolism, PPAR-α activation, and insulin sensitivity may be involved in

  4. Rapid and sensitive lateral flow immunoassay method for determining alpha fetoprotein in serum using europium (III) chelate microparticles-based lateral flow test strips.

    PubMed

    Liang, Rong-Liang; Xu, Xu-Ping; Liu, Tian-Cai; Zhou, Jian-Wei; Wang, Xian-Guo; Ren, Zhi-Qi; Hao, Fen; Wu, Ying-Song

    2015-09-01

    Alpha-fetoprotein (AFP), a primary marker for many diseases including various cancers, is important in clinical tumor diagnosis and antenatal screening. Most immunoassays provide high sensitivity and accuracy for determining AFP, but they are expensive, often complex, time-consuming procedures. A simple and rapid point-of-care system that integrates Eu (III) chelate microparticles with lateral flow immunoassay (LFIA) has been developed to determine AFP in serum with an assay time of 15 min. The approach is based on a sandwich immunoassay performed on lateral flow test strips. A fluorescence strip reader was used to measure the fluorescence peak heights of the test line (HT) and the control line (HC); the HT/HC ratio was used for quantitation. The Eu (III) chelate microparticles-based LFIA assay exhibited a wide linear range (1.0-1000 IU mL(-1)) for AFP with a low limit of detection (0.1 IU mL(-1)) based on 5ul of serum. Satisfactory specificity and accuracy were demonstrated and the intra- and inter-assay coefficients of variation (CV) for AFP were both <10%. Furthermore, in the analysis of human serum samples, excellent correlation (n = 284, r = 0.9860, p < 0.0001) was obtained between the proposed method and a commercially available CLIA kit. Results indicated that the Eu (III) chelate microparticles-based LFIA system provided a rapid, sensitive and reliable method for determining AFP in serum, indicating that it would be suitable for development in point-of-care testing.

  5. Rapid and sensitive lateral flow immunoassay method for determining alpha fetoprotein in serum using europium (III) chelate microparticles-based lateral flow test strips.

    PubMed

    Liang, Rong-Liang; Xu, Xu-Ping; Liu, Tian-Cai; Zhou, Jian-Wei; Wang, Xian-Guo; Ren, Zhi-Qi; Hao, Fen; Wu, Ying-Song

    2015-09-01

    Alpha-fetoprotein (AFP), a primary marker for many diseases including various cancers, is important in clinical tumor diagnosis and antenatal screening. Most immunoassays provide high sensitivity and accuracy for determining AFP, but they are expensive, often complex, time-consuming procedures. A simple and rapid point-of-care system that integrates Eu (III) chelate microparticles with lateral flow immunoassay (LFIA) has been developed to determine AFP in serum with an assay time of 15 min. The approach is based on a sandwich immunoassay performed on lateral flow test strips. A fluorescence strip reader was used to measure the fluorescence peak heights of the test line (HT) and the control line (HC); the HT/HC ratio was used for quantitation. The Eu (III) chelate microparticles-based LFIA assay exhibited a wide linear range (1.0-1000 IU mL(-1)) for AFP with a low limit of detection (0.1 IU mL(-1)) based on 5ul of serum. Satisfactory specificity and accuracy were demonstrated and the intra- and inter-assay coefficients of variation (CV) for AFP were both <10%. Furthermore, in the analysis of human serum samples, excellent correlation (n = 284, r = 0.9860, p < 0.0001) was obtained between the proposed method and a commercially available CLIA kit. Results indicated that the Eu (III) chelate microparticles-based LFIA system provided a rapid, sensitive and reliable method for determining AFP in serum, indicating that it would be suitable for development in point-of-care testing. PMID:26388387

  6. Enhanced sensitivity and selectivity in a dual cell ion mobility spectrometer

    NASA Astrophysics Data System (ADS)

    Griffin, Matthew T.; Fulton, Jack E., Jr.; McAtee, Robert F.; Gao, Rong; Tsoukalas, Lefteri H.

    2003-08-01

    While ion mobility spectrometry (IMS) has been used as a portable trace vapor detector, these handheld systems suffer from poor selectivity. Their low resolution makes confident identification of chemical species difficult. One major application for these IMS systems is in Homeland Defense. IMS systems are fielded for the detection of chemical warfare agents, explosives, narcotics, and other hazardous chemicals. Recently, a novel signal processing methodology using wavelet filtering, statistical evaluators, and genetic algorithms was demonstrated to improve sensitivity and specificity of an ion mobility spectrometer. Previous work involved a single (single polarity) IMS cell. Since both positive and negative ions are created in the same environment and a common sample interface is used for the dual IMS system, there is cross talk between the positive and negative cell. Typically, this cross talk provides little information on the identity of the chemical species present. However, using this new methodology, valuable sample information is obtained. Moreover, ion beam modulation has been incorporated to allow for the ion beam to be broken up into discrete packets. The modulation allows the rejection of common background interferents. This paper will present the process of using cell cross talk, ion beam modulation, and application and extension of the signal processing methodology. The application to field instrumentation will also be discussed.

  7. A novel bi-enzyme electrochemical biosensor for selective and sensitive determination of methyl salicylate.

    PubMed

    Fang, Yi; Umasankar, Yogeswaran; Ramasamy, Ramaraja P

    2016-07-15

    An amperometric sensor based on a bi-enzyme modified electrode was fabricated to detect methyl salicylate, a volatile organic compound released by pathogen-infected plants via systemic response. The detection is based on cascadic conversion reactions that result in an amperometric electrochemical signal. The bi-enzyme electrode is made of alcohol oxidase and horseradish peroxidase enzymes immobilized on to a carbon nanotube matrix through a molecular tethering method. Methyl salicylate undergoes hydrolysis to form methanol, which is consumed by alcohol oxidase to form formaldehyde while simultaneously reducing oxygen to hydrogen peroxide. The hydrogen peroxide will be further reduced to water by horseradish peroxidase, which results in an amperometric signal via direct electron transfer. The bi-enzyme biosensor was evaluated by cyclic voltammetry and constant potential amperometry using hydrolyzed methyl salicylate as the analyte. The sensitivity of the bi-enzyme biosensor as determined by cyclic voltammetry and constant potential amperometry were 112.37 and 282.82μAcm(-2)mM(-1) respectively, and the corresponding limits of detection were 22.95 and 0.98μM respectively. Constant potential amperometry was also used to evaluate durability, repeatability and interference from other compounds. Wintergreen oil was used for real sample study to establish the application of the bi-enzyme sensor for selective determination of plant pathogen infections.

  8. A Sensitive Ratiometric Long-Wavelength Fluorescent Probe for Selective Determination of Cysteine/Homocysteine.

    PubMed

    Manibalan, Kesavan; Chen, Sin-Ming; Mani, Veerappan; Huang, Tsung-Tao; Huang, Sheng-Tung

    2016-07-01

    The development of sensitive fluorescence probes to detect biothiols such as cysteine and homocysteine has attracted great attention in recent times. Herein, we described the design and synthesis of coumarin based long-wavelength fluorescence probe, Bromo-2-benzothiazolyl-3-cyano-7-hydroxy coumarin (BBCH, 2) for selective detections of cysteine and homocysteine. The probe is rationally designed in such a way that both sulfhydryl and adjacent amino groups of thiols are involved in sensing process. Only cysteine/homocysteine able to react with BBCH to release fluorescence reporter (BCH, 1); while, glutathione and other amino acids unable to react with BBCH due to the absence of adjacent amino groups. In presence of cysteine, the color of BBCH is turns from colorless to red and thus BBCH is a naked eye fluorescence indicator for cysteine. Besides, BBCH can discriminate cysteine and homocysteine based on color changes and different reaction rates. The described sensing platform showed good sensing performances to detect cysteine and homocysteine with detection limits of 0.87 and 0.19 μM, respectively. Practical applicability was verified in biological and pharmaceutical samples. PMID:27290640

  9. Tanshinone IIA induces TRAIL sensitization of human lung cancer cells through selective ER stress induction.

    PubMed

    Kim, Eun-Ok; Kang, Shi Eun; Im, Chang Rak; Lee, Jun-Hee; Ahn, Kwang Seok; Yang, Woong Mo; Um, Jae-Young; Lee, Seok-Geun; Yun, Miyong

    2016-05-01

    Although tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promised anticancer medicine targeting only the tumor, most cancers show resistance to TRAIL-induced apoptosis. For this reason, new therapeutic strategies to overcome the TRAIL resistance are required for more effective tumor treatment. In the present study, potential of tanshinone IIA as a TRAIL sensitizer was evaluated in human non-small cell lung cancer (NSCLC) cells. NSCLC cells showed resistance to TRAIL-mediated cell death, but combination treatment of Tanshinone IIA and TRAIL synergistically decreased cell viability and increased apoptosis in TRAIL-resistant NSCLC cells. Tanshinone IIA greatly induced death receptor 5 (DR5), but not death receptor 4 (DR4). Furthermore, DR5 knockdown attenuated the combination treatment of tanshinone IIA with TRAIL-mediated cell death in human NSCLC cells. Tanshinone IIA also increased CHOP and activated the PERK-ATF4 pathway suggesting that tanshinone IIA increased DR5 and CHOP by activating the PERK-ATF4 pathway. Tanshinone IIA also downregulated phosphorylation of STAT3 and expression of survivin. Taken together, these results indicate that tanshinone IIA increases TRAIL-induced cell death via upregulating DR5 and downregulating survivin mediated by, respectively, selective activation of PERK/ATF4 and inhibition of STAT3, suggesting combinatorial intervention of tanshinone IIA and TRAIL as a new therapeutic strategy for human NSCLC. PMID:26983803

  10. Protein MRI contrast agent with unprecedented metal selectivity and sensitivity for liver cancer imaging.

    PubMed

    Xue, Shenghui; Yang, Hua; Qiao, Jingjuan; Pu, Fan; Jiang, Jie; Hubbard, Kendra; Hekmatyar, Khan; Langley, Jason; Salarian, Mani; Long, Robert C; Bryant, Robert G; Hu, Xiaoping Philip; Grossniklaus, Hans E; Liu, Zhi-Ren; Yang, Jenny J

    2015-05-26

    With available MRI techniques, primary and metastatic liver cancers that are associated with high mortality rates and poor treatment responses are only diagnosed at late stages, due to the lack of highly sensitive contrast agents without Gd(3+) toxicity. We have developed a protein contrast agent (ProCA32) that exhibits high stability for Gd(3+) and a 10(11)-fold greater selectivity for Gd(3+) over Zn(2+) compared with existing contrast agents. ProCA32, modified from parvalbumin, possesses high relaxivities (r1/r2: 66.8 mmol(-1)⋅s(-1)/89.2 mmol(-1)⋅s(-1) per particle). Using T1- and T2-weighted, as well as T2/T1 ratio imaging, we have achieved, for the first time (to our knowledge), robust MRI detection of early liver metastases as small as ∼0.24 mm in diameter, much smaller than the current detection limit of 10-20 mm. Furthermore, ProCA32 exhibits appropriate in vivo preference for liver sinusoidal spaces and pharmacokinetics for high-quality imaging. ProCA32 will be invaluable for noninvasive early detection of primary and metastatic liver cancers as well as for monitoring treatment and guiding therapeutic interventions, including drug delivery.

  11. Golgi Phosphoprotein 4 (GPP130) is a Sensitive and Selective Cellular Target of Manganese Exposure

    PubMed Central

    Masuda, Melisa; Braun-sommargren, Michelle; Crooks, Dan; Smith, Donald R.

    2014-01-01

    Chronic elevated exposure to manganese (Mn) is associated with neurocognitive and fine motor deficits in children. However, relatively little is understood about cellular responses to Mn spanning the transition between physiologic to toxic levels of exposure. Here, we investigated the specificity, sensitivity, and time course of the Golgi Phosphoprotein 4 (GPP130) response to Mn exposure in AF5 GABAergic neuronal cells, and we determined the extent to which GPP130 degradation occurs in brain cells in vivo in rats subchronically exposed to Mn. Our results show that GPP130 degradation in AF5 cells was specific to Mn, and did not occur following exposure to cobalt, copper, iron, nickel, or zinc. GPP130 degradation occurred without measurable increases in intracellular Mn levels and at Mn exposures as low as 0.54 µM. GPP130 protein was detectable by immunofluorescence in only ~15–30% of cells in striatal and cortical rat brain slices, and Mn-exposed animals exhibited a significant reduction in both the number of GPP130-positive cells, and the overall levels of GPP130 protein, demonstrating the in vivo relevance of this Mn-specific response within the primary target organ of Mn toxicity. These results provide insight into specific mechanism(s) of cellular Mn regulation and toxicity within the brain, including the selective susceptibility of cells to Mn cytotoxicity. PMID:23280773

  12. A novel OFET-based biosensor for the selective and sensitive detection of lactate levels.

    PubMed

    Minami, Tsuyoshi; Sato, Tsubasa; Minamiki, Tsukuru; Fukuda, Kenjiro; Kumaki, Daisuke; Tokito, Shizuo

    2015-12-15

    Biosensors based on organic field effect transistors (OFETs) are one of the more promising device applications in organic electronics. However, OFET-based biosensors are still in their early stages of development compared to other electrochemical biosensors. This study is the first to report on an extended-gate type organic field effect transistor (OFET) for lactate detection in aqueous media. Here, the extended-gate electrode of the OFET was modified with layers of a lactate oxidase and a horseradish peroxidase osmium-redox polymer on a flexible plastic film substrate for an enzymatic redox reaction of lactate. The device exhibited both high selectivity and sensitivity. The limit of detection (LOD) and the limit of quantification (LOQ) were estimated to be 66 nM and 220 nM, respectively, which are the sufficient detection limit for practical sensor applications. The obtained results confirm that extended-gate type OFET devices are applicable to enzyme-based biosensors for detecting lactate levels. PMID:26101795

  13. Highly sensitive and selective glucose sensor based on ultraviolet-treated nematic liquid crystals.

    PubMed

    Zhong, Shenghong; Jang, Chang-Hyun

    2014-09-15

    Glucose is an extremely important biomolecule, and the ability to sense it has played a significant role in facilitating the understanding of many biological processes. Here, we report a novel glucose sensor based on ultraviolet (UV)-treated nematic liquid crystals. Submerging UV-treated 4-cyano-4'-pentylbiphenyl (5CB) in a glucose solution (while carefully adjusting its pH to 7.5 with NaOH and HCl) triggered an optical response, from dark to bright, observed with a polarized microscope. Notably, 5CB was located inside a glucose oxidase (GOx)-modified gold grid. We exploited this pH-driven phenomenon to design a new glucose sensor. This device could detect as little as 1 pM analyte, which is 3 orders of magnitude lower than the detection limit of the most sensitive glucose sensor currently available. It also exhibits high selectivity due to GOx modification. Thus, this is a promising technique for glucose detection, not only for clinical diagnostics, but also for sensing low levels of glucose in a biological environment (e.g., single cells and bacterial cultures).

  14. Highly sensitive and selective fluorescent assay for guanine based on the Cu2 +/eosin Y system

    NASA Astrophysics Data System (ADS)

    Shi, Huimin; Cui, Yi; Gong, Yijun; Feng, Suling

    2016-05-01

    A fluorescent probe has been developed for the determination of guanine based on the quenched fluorescence signal of Cu2 +/eosin Y. Cu2 + interacted with eosin Y, resulting in fluorescence quenching. Subsequently, with the addition of guanine to the Cu2 +/eosin Y system, guanine reacted with Cu2 + to form 1:1 chelate cation, which further combined with eosin Y to form a 1:1 ternary ion-association complex by electrostatic attraction and hydrophobic interaction, resulting in significant decrease of the fluorescence. Hence, a fluorescent system was constructed for rapid, sensitive and selective detection of guanine with a detection limit as low as 1.5 nmol L- 1 and a linear range of 3.3-116 nmol L- 1. The method has been applied satisfactorily to the determination of guanine in DNA and urine samples with the recoveries from 98.7% to 105%. This study significantly expands the realm of application of ternary ion-association complex in fluorescence probe.

  15. A novel bi-enzyme electrochemical biosensor for selective and sensitive determination of methyl salicylate.

    PubMed

    Fang, Yi; Umasankar, Yogeswaran; Ramasamy, Ramaraja P

    2016-07-15

    An amperometric sensor based on a bi-enzyme modified electrode was fabricated to detect methyl salicylate, a volatile organic compound released by pathogen-infected plants via systemic response. The detection is based on cascadic conversion reactions that result in an amperometric electrochemical signal. The bi-enzyme electrode is made of alcohol oxidase and horseradish peroxidase enzymes immobilized on to a carbon nanotube matrix through a molecular tethering method. Methyl salicylate undergoes hydrolysis to form methanol, which is consumed by alcohol oxidase to form formaldehyde while simultaneously reducing oxygen to hydrogen peroxide. The hydrogen peroxide will be further reduced to water by horseradish peroxidase, which results in an amperometric signal via direct electron transfer. The bi-enzyme biosensor was evaluated by cyclic voltammetry and constant potential amperometry using hydrolyzed methyl salicylate as the analyte. The sensitivity of the bi-enzyme biosensor as determined by cyclic voltammetry and constant potential amperometry were 112.37 and 282.82μAcm(-2)mM(-1) respectively, and the corresponding limits of detection were 22.95 and 0.98μM respectively. Constant potential amperometry was also used to evaluate durability, repeatability and interference from other compounds. Wintergreen oil was used for real sample study to establish the application of the bi-enzyme sensor for selective determination of plant pathogen infections. PMID:26918616

  16. Sensitive and selective electrochemical detection of artemisinin based on its reaction with p-aminophenylboronic acid.

    PubMed

    Wang, Chao; Zholudov, Yuriy T; Nsabimana, Anaclet; Xu, Guobao; Li, Jianping

    2016-09-21

    The electrochemical detection of artemisinin generally requires high oxidation potential or the use of complex electrode modification. We find that artemisinin can react with p-aminophenylboronic acid to produce easily electrochemically detectable aminophenol for the first time. By making use of the new reaction, we report an alternative method to detect artemisinin through the determination of p-aminophenol. The calibration curve for the determination of artemisinin is linear in the range of 2 μmol L(-1) to 200 μmol L(-1) with the detection limit of 0.8 μmol L(-1), which is more sensitive than other reported electrochemical methods. The relative standard deviation is 4.83% for the determination of 10 μM artemisinin. Because the oxidation potential of p-aminophenol is around 0 V, the present method is high selective. When 40 μM, 90 μM and 140 μM of artemisinin were spiked to compound naphthoquine phosphate tablet samples, the recoveries are 107.6%, 105.4% and 101.7%, respectively. This detection strategy is attractive for the detection of artemisinin and its derivatives. The finding that artemisinin can react with aromatic boronic acid has the potential to be exploited for the development of other sensors, such as fluorescence artemisinin sensors. PMID:27590543

  17. Determination of the biotin content of select foods using accurate and sensitive HPLC/avidin binding

    PubMed Central

    Staggs, C.G.; Sealey, W.M.; McCabe, B.J.; Teague, A.M.; Mock, D.M.

    2006-01-01

    Assessing dietary biotin content, biotin bioavailability, and resulting biotin status are crucial in determining whether biotin deficiency is teratogenic in humans. Accuracy in estimating dietary biotin is limited both by data gaps in food composition tables and by inaccuracies in published data. The present study applied sensitive and specific analytical techniques to determine values for biotin content in a select group of foods. Total biotin content of 87 foods was determined using acid hydrolysis and the HPLC/avidin-binding assay. These values are consistent with published values in that meat, fish, poultry, egg, dairy, and some vegetables are relatively rich sources of biotin. However, these biotin values disagreed substantially with published values for many foods. Assay values varied between 247 times greater than published values for a given food to as much as 36% less than the published biotin value. Among 51 foods assayed for which published values were available, only seven agreed within analytical variability (720%). We conclude that published values for biotin content of foods are likely to be inaccurate. PMID:16648879

  18. Contaminant tailing in highly heterogeneous porous formations: Sensitivity on model selection and material properties

    NASA Astrophysics Data System (ADS)

    Maghrebi, Mahdi; Jankovic, Igor; Weissmann, Gary S.; Matott, L. Shawn; Allen-King, Richelle M.; Rabideau, Alan J.

    2015-12-01

    Coupled impacts of slow advection, diffusion and sorption were investigated using two heterogeneity models that differ in structure and in the mathematical framework that was used to simulate flow and transport and to quantify contaminant tailing. Both models were built using data from a highly heterogeneous exposure of the Borden Aquifer at a site located 2 km north-west of the Stanford-Waterloo experimental site at Canadian Forces Base Borden, Ontario, Canada. The inclusions-based model used a simplified representation of the different materials found at the site, while the second model was based on transitional probability geostatistics of the formation. These two models were used to investigate sensitivity of contaminant tailing on model selection and on geometric and material properties. While simulations were based on data collected at Borden, models were exercised beyond the geometric and material properties that characterize the site. Various realizations have identified very low conductive silty clay, found at volume fraction of 23.4%, as the material with dominant influence on tailing, and vertical diffusion in and out of low conductive units, affected by sorption, as the dominant transport mechanism causing tailing. The two models yielded almost identical transport results when vertical correlation lengths of silty clay were matched. Several practical implications relevant for characterization of low conductive units were identified and briefly discussed.

  19. Simple, Selective and Sensitive Determination of CH₃Hg⁺ Using Gold Nanocluster.

    PubMed

    Chen, Xiaoxia; Zhao, Jiating; Cong, Wenshu; Li, Xixuan; Fan, Hongyi; Sun, Jinzhe; Lin, Jing; Li, Bai; Gao, Yuxi; Qin, Chuan; Li, Yu-Feng

    2016-01-01

    Mercury is a potent neurotoxin, which causes serious and permanent damage to biota including human beings. Nanomaterials like gold have been used to detect mercury, especially the mercuric ions in recent years, but few have been done on the determination of methylmercury (CH₃Hg⁺). In this study, we present a simple, selective and sensitive method for the determination of CH₃Hg⁺ co-existing with Hg²⁺ based on the fluorescence quenching of bovine serum albumin (BSA) stabilized gold nanoclusters (BSA-Au NCs). It was found that the fluorescence quenching of BSA-Au NCs by Hg²⁺ is stronger than CH₃Hg⁺ at the same concentration. Hg²⁺ can be masked by EDTA while CH₃Hg⁺ is less affected. Therefore, the determination of CH₃Hg⁺ was achieved through EDTA masking. The detection limit for CH₃Hg⁺ is 35 nmol/L after masking Hg²⁺ with EDTA. This method has been successfully applied to quantify CH₃Hg⁺ in rice paddy water from Qingzhen, Guizhou and tap water from Beijing. PMID:27398521

  20. Selective and Sensitive Chemiluminescence Determination of MCPB: Flow Injection and Liquid Chromatography.

    PubMed

    Meseguer-Lloret, Susana; Torres-Cartas, Sagrario; Catalá-Icardo, Mónica; Gómez-Benito, Carmen

    2016-02-01

    Two new chemiluminescence (CL) methods are described for the determination of the herbicide 4-(4-chloro-o-tolyloxy) butyric acid (MCPB). First, a flow injection chemiluminescence (FI-CL) method is proposed. In this method, MCPB is photodegraded with an ultraviolet (UV) lamp and the photoproducts formed provide a great CL signal when they react with ferricyanide in basic medium. Second, a high-performance liquid chromatography chemiluminescence (HPLC-CL) method is proposed. In this method, before the photodegradation and CL reaction, the MCPB and other phenoxyacid herbicides are separated in a C18 column. The experimental conditions for the FI-CL and HPLC-CL methods are optimized. Both methods present good sensitivity, the detection limits being 0.12 µg L(-1) and 0.1 µg L(-1) (for FI-CL and HPLC-CL, respectively) when solid phase extraction (SPE) is applied. Intra- and interday relative standard deviations are below 9.9%. The methods have been satisfactorily applied to the analysis of natural water samples. FI-CL method can be employed for the determination of MCPB in simple water samples and for the screening of complex water samples in a fast, economic, and simple way. The HPLC-CL method is more selective, and allows samples that have not been resolved with the FI-CL method to be solved.

  1. Increasing the selectivity and sensitivity of gas sensors for the detection of explosives

    NASA Astrophysics Data System (ADS)

    Mallin, Daniel

    Over the past decade, the use of improvised explosive devices (IEDs) has increased, domestically and internationally, highlighting a growing need for a method to quickly and reliably detect explosive devices in both military and civilian environments before the explosive can cause damage. Conventional techniques have been successful in explosive detection, however they typically suffer from enormous costs in capital equipment and maintenance, costs in energy consumption, sampling, operational related expenses, and lack of continuous and real-time monitoring. The goal was thus to produce an inexpensive, portable sensor that continuously monitors the environment, quickly detects the presence of explosive compounds and alerts the user. In 2012, here at URI, a sensor design was proposed for the detection of triacetone triperoxide (TATP). The design entailed a thermodynamic gas sensor that measures the heat of decomposition between trace TATP vapor and a metal oxide catalyst film. The sensor was able to detect TATP vapor at the part per million level (ppm) and showed great promise for eventual commercial use, however, the sensor lacked selectivity. Thus, the specific objective of this work was to take the original sensor design proposed in 2012 and to make several key improvements to advance the sensor towards commercialization. It was demonstrated that a sensor can be engineered to detect TATP and ignore the effects of interferent H2O2 molecules by doping SnO2 films with varying amounts of Pd. Compared with a pure SnO2 catalyst, a SnO2, film doped with 8 wt. % Pd had the highest selectivity between TATP and H2O2. Also, at 12 wt. % Pd, the response to TATP and H2O2 was enhanced, indicating that sensitivity, not only selectivity, can be increased by modifying the composition of the catalyst. An orthogonal detection system was demonstrated. The platform consists of two independent sensing mechanisms, one thermodynamic and one conductometric, which take measurements from

  2. A Sensitivity Study of Liric Algorithm to User-defined Input Parameters, Using Selected Cases from Thessaloniki's Measurements

    NASA Astrophysics Data System (ADS)

    Filioglou, M.; Balis, D.; Siomos, N.; Poupkou, A.; Dimopoulos, S.; Chaikovsky, A.

    2016-06-01

    A targeted sensitivity study of the LIRIC algorithm was considered necessary to estimate the uncertainty introduced to the volume concentration profiles, due to the arbitrary selection of user-defined input parameters. For this purpose three different tests were performed using Thessaloniki's Lidar data. Overall, tests in the selection of the regularization parameters, an upper and a lower limit test were performed. The different sensitivity tests were applied on two cases with different predominant aerosol types, a dust episode and a typical urban case.

  3. Selective central alpha-2 adrenoceptor control of regional haemodynamic responses to air jet stress in conscious spontaneously hypertensive rats.

    PubMed

    Kapusta, D R; Knardahl, S; Koepke, J P; Johnson, A K; DiBona, G F

    1989-03-01

    The role of central nervous system alpha 2-adrenoceptors in the regulation of peripheral sympathetic outflow to regional vascular resistance beds during environmental stress was examined in conscious chronically instrumented spontaneously hypertensive rats (SHR). SHR were instrumented with pulsed Doppler flow probes on the renal and mesenteric arteries and the lower abdominal aorta. The mean arterial pressure (MAP), heart rate and regional vascular resistance responses to air jet stress were determined before and after cumulative administration of the alpha 2-adrenoceptor agonist, guanabenz, into the lateral cerebral ventricle in doses of 5 and 25 micrograms. Compared with intracerebroventricular (i.c.v.) administration of isotonic saline vehicle which did not affect baseline systemic and regional haemodynamic measurements, guanabenz produced significant decreases in baseline MAP and heart rate but did not affect regional vascular resistances. During air jet stress, the characteristic pattern of the classic defense reaction with an increase in MAP, heart rate, renal and mesenteric vascular resistances and a decrease in hindquarters vascular resistance was observed and was not affected by i.c.v. administration of isotonic saline vehicle. Guanabenz did not affect the MAP, heart rate or renal vascular resistance responses to air jet stress. The air jet stress-induced increase in mesenteric vascular resistance was reduced by 25 micrograms guanabenz. The air jet stress-induced decrease in hindquarters vascular resistance was converted to an increase by 25 micrograms guanabenz. These results demonstrate that the regulation of environmental stress-stimulated sympathetic neural outflow to different vascular beds may be independently controlled by central nervous system alpha 2-adrenoceptors.

  4. In silico selection of an aptamer to estrogen receptor alpha using computational docking employing estrogen response elements as aptamer-alike molecules

    PubMed Central

    Ahirwar, Rajesh; Nahar, Smita; Aggarwal, Shikha; Ramachandran, Srinivasan; Maiti, Souvik; Nahar, Pradip

    2016-01-01

    Aptamers, the chemical-antibody substitute to conventional antibodies, are primarily discovered through SELEX technology involving multi-round selections and enrichment. Circumventing conventional methodology, here we report an in silico selection of aptamers to estrogen receptor alpha (ERα) using RNA analogs of human estrogen response elements (EREs). The inverted repeat nature of ERE and the ability to form stable hairpins were used as criteria to obtain aptamer-alike sequences. Near-native RNA analogs of selected single stranded EREs were modelled and their likelihood to emerge as ERα aptamer was examined using AutoDock Vina, HADDOCK and PatchDock docking. These in silico predictions were validated by measuring the thermodynamic parameters of ERα -RNA interactions using isothermal titration calorimetry. Based on the in silico and in vitro results, we selected a candidate RNA (ERaptR4; 5′-GGGGUCAAGGUGACCCC-3′) having a binding constant (Ka) of 1.02 ± 0.1 × 108 M−1 as an ERα-aptamer. Target-specificity of the selected ERaptR4 aptamer was confirmed through cytochemistry and solid-phase immunoassays. Furthermore, stability analyses identified ERaptR4 resistant to serum and RNase A degradation in presence of ERα. Taken together, an efficient ERα-RNA aptamer is identified using a non-SELEX procedure of aptamer selection. The high-affinity and specificity can be utilized in detection of ERα in breast cancer and related diseases. PMID:26899418

  5. Selective ablation of dorsal horn NK1 expressing cells reveals a modulation of spinal alpha2-adrenergic inhibition of dorsal horn neurones.

    PubMed

    Rahman, Wahida; Suzuki, Rie; Hunt, Stephen P; Dickenson, Anthony H

    2008-06-01

    Activity in descending systems from the brainstem modulates nociceptive transmission through the dorsal horn. Intrathecal injection of the neurotoxin saporin conjugated to SP (SP-SAP) into the lumbar spinal cord results in the selective ablation of NK(1) receptor expressing (NK(1)+ve) neurones in the superficial dorsal horn (lamina I/III). Loss of these NK(1)+ve neurones attenuates excitability of deep dorsal horn neurones due to a disruption of both intrinsic spinal circuits and a spino-bulbo-spinal loop, which activates a descending excitatory drive, mediated through spinal 5HT(3) receptors. Descending inhibitory pathways also modulate spinal activity and hence control the level of nociceptive transmission relayed to higher centres. To ascertain the spinal origins of the major descending noradrenergic inhibitory pathway we studied the effects of a selective alpha2-adrenoceptor antagonist, atipamezole, on neuronal activity in animals pre-treated with SP-SAP. Intrathecal application of atipamezole dose dependently facilitated the mechanically evoked neuronal responses of deep dorsal horn neurones to low intensity von Frey hairs (5-15 g) and noxious thermal (45-50 degrees C) evoked responses in SAP control animals indicating a physiological alpha2-adrenoceptor control. This facilitatory effect of atipamezole was lost in the SP-SAP treated group. These data suggest that activity within noradrenergic pathways have a dependence on dorsal horn NK(1)+ve cells. Further, noradrenergic descending inhibition may in part be driven by lamina I/III (NK(1)+ve) cells, and mediated via spinal alpha2-adrenoceptor activation. Since the same neuronal population drives descending facilitation and inhibition, the reduced excitability of lamina V/VI WDR neurones seen after loss of these NK(1)+ve neurones indicates a dominant role of descending facilitation. PMID:18462764

  6. Sensitive and Selective Plasmon Ruler Nanosensors for Monitoring the Apoptotic Drug Response in Leukemia

    PubMed Central

    2015-01-01

    Caspases are proteases involved in cell death, where caspase-3 is the chief executioner that produces an irreversible cutting event in downstream protein substrates and whose activity is desired in the management of cancer. To determine such activity in clinically relevant samples with high signal-to-noise, plasmon rulers are ideal because they are sensitively affected by their interparticle separation without ambiguity from photobleaching or blinking effects. A plasmon ruler is a noble metal nanoparticle pair, tethered in close proximity to one another via a biomolecule, that acts through dipole–dipole interactions and results in the light scattering to increase exponentially. In contrast, a sharp decrease in intensity is observed when the pair is confronted by a large interparticle distance. To align the mechanism of protease activity with building a sensor that can report a binary signal in the presence or absence of caspase-3, we present a caspase-3 selective plasmon ruler (C3SPR) composed of a pair of Zn0.4Fe2.6O4@SiO2@Au core–shell nanoparticles connected by a caspase-3 cleavage sequence. The dielectric core (Zn0.4Fe2.6O4@SiO2)-shell (Au) geometry provided a brighter scattering intensity versus solid Au nanoparticles, and the magnetic core additionally acted as a purification handle during the plasmon ruler assembly. By monitoring the decrease in light scattering intensity per plasmon ruler, we detected caspase-3 activity at single molecule resolution across a broad dynamic range. This was observed to be as low as 100 fM of recombinant material or 10 ng of total protein from cellular lysate. By thorough analyses of single molecule trajectories, we show caspase-3 activation in a drug-treated chronic myeloid leukemia (K562) cancer system as early as 4 and 8 h with greater sensitivity (2- and 4-fold, respectively) than conventional reagents. This study provides future implications for monitoring caspase-3 as a biomarker and efficacy of drugs. PMID:25166742

  7. Sensitive and selective plasmon ruler nanosensors for monitoring the apoptotic drug response in leukemia.

    PubMed

    Tajon, Cheryl A; Seo, Daeha; Asmussen, Jennifer; Shah, Neil; Jun, Young-wook; Craik, Charles S

    2014-09-23

    Caspases are proteases involved in cell death, where caspase-3 is the chief executioner that produces an irreversible cutting event in downstream protein substrates and whose activity is desired in the management of cancer. To determine such activity in clinically relevant samples with high signal-to-noise, plasmon rulers are ideal because they are sensitively affected by their interparticle separation without ambiguity from photobleaching or blinking effects. A plasmon ruler is a noble metal nanoparticle pair, tethered in close proximity to one another via a biomolecule, that acts through dipole-dipole interactions and results in the light scattering to increase exponentially. In contrast, a sharp decrease in intensity is observed when the pair is confronted by a large interparticle distance. To align the mechanism of protease activity with building a sensor that can report a binary signal in the presence or absence of caspase-3, we present a caspase-3 selective plasmon ruler (C3SPR) composed of a pair of Zn0.4Fe2.6O4@SiO2@Au core-shell nanoparticles connected by a caspase-3 cleavage sequence. The dielectric core (Zn0.4Fe2.6O4@SiO2)-shell (Au) geometry provided a brighter scattering intensity versus solid Au nanoparticles, and the magnetic core additionally acted as a purification handle during the plasmon ruler assembly. By monitoring the decrease in light scattering intensity per plasmon ruler, we detected caspase-3 activity at single molecule resolution across a broad dynamic range. This was observed to be as low as 100 fM of recombinant material or 10 ng of total protein from cellular lysate. By thorough analyses of single molecule trajectories, we show caspase-3 activation in a drug-treated chronic myeloid leukemia (K562) cancer system as early as 4 and 8 h with greater sensitivity (2- and 4-fold, respectively) than conventional reagents. This study provides future implications for monitoring caspase-3 as a biomarker and efficacy of drugs.

  8. Optical Near Field Studies of Plasmonic and Optical Antennas For Sensitive and Selective Biosensing Applications

    NASA Astrophysics Data System (ADS)

    Gelfand, Ryan M.

    For biosensing applications a useful device needs at least two properties: high sensitivity and high selectivity. Optical spectroscopy offers unique advantages over other sensing techniques however one big challenge to overcome is the mismatch between wavelength and the size of biologically relevant molecules. In order to have high enough sensitivity to approach the single-molecule limit, the interaction between the light and the molecule should be strong. However, the diffraction limit of light is approximately half the incidence wavelength, on the order of 100 nm for the smallest nondestructive wavelengths. This presents a significant mismatch between the size of the molecule and the smallest focus spot of the light. The photo-excitation should be compressed more than 100 fold to interact strongly. We must use metallic antennas that convert the incidence radiation into plasmonic modes which can then be compressed well below the wavelength diffraction limit. Studying the near field characteristics of these metallic nanostructures will help us gain insight into this emerging field and allow us to better use them in developing next generation devices. We have developed different geometries of these antennas and simulated their performance using Finite Difference Time Domain software. We have concentrated our efforts in the mid-infrared because that is the natural molecular vibration frequency region and also the near infrared because at these frequencies there exists a mature industry for compact sources, detectors, and fiber optic components. Our simulations show a 6,000 fold mode compression for a bowtie antenna and a million fold compression for a plasmonic photonic crystal (ppc) antenna. The bull's-eye antenna does not have as a high a mode compression but it has a natural geometry for molecular sensing due to the central metallic disc. Experimentally, we have measured the near field of these antennas with a custom back reflection apertureless NSOM setup in both

  9. 21-(/sup 18/F)fluoro-16 alpha-ethyl-19-norprogesterone: synthesis and target tissue selective uptake of a progestin receptor based radiotracer for positron emission tomography

    SciTech Connect

    Pomper, M.G.; Katzenellenbogen, J.A.; Welch, M.J.; Brodack, J.W.; Mathias, C.J.

    1988-07-01

    We have synthesized 21-(/sup 18/F)fluoro-16 alpha-ethyl-19-norprogesterone (FENP), a high affinity ligand for the progesterone receptor, labeled with the positron-emitting radionuclide fluorine-18 (t1/2 = 110 min). The synthesis proceeds in two steps from 21-hydroxy-16 alpha-ethyl-19-norprogesterone and involves (/sup 18/F)fluoride ion displacement of the 21-trifluoromethanesulfonate (21-triflate). This material is purified by HPLC and is obtained in 4-30% overall yield (decay corrected) within 40 min after the end of bombardment to produce (/sup 18/F)fluoride ion. The effective specific activity, determined by competitive radioreceptor binding assays, is 700-1400 Ci/mmol. In vivo, (/sup 18/F)FENP demonstrates highly selective, receptor-mediated uptake by the uterus of estrogen-primed rats; the uterus to blood and uterus to muscle ratios were respectively 26 and 16 at 1 h and 71 and 41 at 3 h after injection. The high target tissue selectivity of this uptake suggests that this compound may be useful for the in vivo imaging of progestin target tissues and receptor-rich tumors (such as human breast tumors) by positron emission tomography.

  10. Melibiose permease and alpha-galactosidase of Escherichia coli: Identification by selective labeling using a T7 RNA polymerase/promoter expression system

    SciTech Connect

    Pourcher, T.; Bassilana, M.; Sarkar, H.K.; Kaback, H.R.; Leblanc, G. )

    1990-01-23

    Identification and selective labeling of the melibiose permease and alpha-galactosidase in Escherichia coli, which are encoded by the melB and melA genes, respectively, have been accomplished by selectively labeling the two gene products with a T7 RNA polymerase expression system. Following generation of a novel EcoRI restriction site in the intergenic sequence between the two genes of the mel operon by oligonucleotide-directed, site-specific mutagenesis, melA and melB were separately inserted into plasmid pT7-6 of the T7 expression system. Expression of melB was markedly enhanced by placing a strong, synthetic ribosome binding site at an optimal distance upstream from the initiation codon of melB. Expression of cloned gene products was characterized functionally and by performing autoradiographic analysis on total cell, inner membrane, and cytoplasmic proteins from cells pulse labeled with (35S)methionine in the presence of rifampicin and resolved by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. The results first confirm that alpha-galactosidase is a cytoplasmic protein with an Mr of 50K; in contrast, the membrane-bound melibiose permease is identified as a protein with an apparent Mr of 39K, a value significantly higher than that of 30K previously suggested.

  11. A phosphatidylinositol 4,5-bisphosphate-sensitive casein kinase I alpha associates with synaptic vesicles and phosphorylates a subset of vesicle proteins

    PubMed Central

    1995-01-01

    In interphase cells, alpha-casein kinase I (alpha-CKI) is found associated with cytosolic vesicular structures, the centrosome, and within the nucleus. To identify the specific vesicular structures with which alpha-CKI is associated, established cell lines and primary rat neurons were immunofluorescently labeled with an antibody raised to alpha-CKI. In nonneuronal cells, alpha-CKI colocalizes with vesicular structures which align with microtubules and are partially coincident with both Golgi and endoplasmic reticulum markers. In neurons, alpha- CKI colocalizes with synaptic vesicle markers. When synaptic vesicles were purified from rat brain, they were highly enriched in a CKI, based on activity and immunoreactivity. The synaptic vesicle-associated CKI is an extrinsic kinase and was eluted from synaptic vesicles and purified. This purified CKI has properties most similar to alpha-CKI. When the activities of casein kinase I or II were specifically inhibited on isolated synaptic vesicles, CKI was shown to phosphorylate a specific subset of vesicle proteins, one of which was identified as the synaptic vesicle-specific protein SV2. As with alpha-CKI, the synaptic vesicle CKI is inhibited by phosphatidylinositol 4,5- bisphosphate (PIP2). However, synthesis of PIP2 was detected only in plasma membrane-containing fractions. Therefore, PIP2 may spatially regulate CKI. Since PIP2 synthesis is required for secretion, this inhibition of CKI may be important for the regulation of secretion. PMID:7622570

  12. STAR FORMATION RATES AND STELLAR MASSES OF H{alpha} SELECTED STAR-FORMING GALAXIES AT z = 0.84: A QUANTIFICATION OF THE DOWNSIZING

    SciTech Connect

    Villar, Victor; Gallego, Jesus; Perez-Gonzalez, Pablo G.; Barro, Guillermo; Zamorano, Jaime; Noeske, Kai; Koo, David C. E-mail: j.gallego@fis.ucm.es E-mail: gbarro@fis.ucm.es E-mail: noeske@stsci.edu

    2011-10-10

    In this work we analyze the physical properties of a sample of 153 star-forming galaxies at z {approx} 0.84, selected by their H{alpha} flux with a narrowband filter. B-band luminosities of the objects are higher than those of local star-forming galaxies. Most of the galaxies are located in the blue cloud, though some objects are detected in the green valley and in the red sequence. After the extinction correction is applied, virtually all these red galaxies move to the blue sequence, unveiling their dusty nature. A check on the extinction law reveals that the typical extinction law for local starbursts is well suited for our sample but with E(B - V){sub stars} = 0.55 E(B - V){sub gas}. We compare star formation rates (SFRs) measured with different tracers (H{alpha}, far-ultraviolet, and infrared), finding that they agree within a factor of three after extinction correction. We find a correlation between the ratios SFR{sub FUV}/SFR{sub H{alpha}}, SFR{sub IR}/SFR{sub H{alpha}}, and the EW(H{alpha}) (i.e., weighted age), which accounts for part of the scatter. We obtain stellar mass estimations by fitting templates to multi-wavelength photometry. The typical stellar mass of a galaxy within our sample is {approx}10{sup 10} M{sub sun}. The SFR is correlated with stellar mass and the specific SFR decreases with it, indicating that massive galaxies are less affected by star formation processes than less massive ones. This result is consistent with the downsizing scenario. To quantify this downsizing we estimated the quenching mass M{sub Q} for our sample at z {approx} 0.84, finding that it declines from M{sub Q} {approx} 10{sup 12} M{sub sun} at z {approx} 0.84 to M{sub Q} {approx} 8 x 10{sup 10} M{sub sun} at the local universe.

  13. Fluorescent Chemosensors for Selective and Sensitive Detection of Phosmet/Chlorpyrifos with Octahedral Ni(2+) Complexes.

    PubMed

    Raj, Pushap; Singh, Amanpreet; Kaur, Kamalpreet; Aree, Thammarat; Singh, Ajnesh; Singh, Narinder

    2016-05-16

    The hexadentate ligands H2L1-L3 with mixed S, N, O donor sites and possessing substituents having either "no" or electron-releasing/withdrawing nature at terminal ends are synthesized. The ligands H2L1-L3 were tested for binding with library of metal ions, wherein maximum efficiency was observed with Ni(2+), and it motivated us to prepare the Ni(2+) complexes. The ligand H2L1 underwent deprotonation and formed binuclear complex when interacted with Ni(2+) as evident from its crystal structure. The H2L2 and H2L3 having electron-withdrawing/electron releasing groups, respectively, were also deprotonated; however, they afforded mononuclear complexes with Ni(2+) ion. This signifies the importance of steric parameters instead of electronic factors in these particular cases. Impressed by differential behavior of complexes of H2L1 and H2L2/H2L3 with Ni(2+) and their photophysical and electrochemical properties, all the metal complexes were studied for their chemosensing ability. Nowadays with increased use of organophosphate, there is alarming increase of these agents in the environment, and thus we require efficient technique to estimate the level of these agents with high sensitivity and selectivity in aqueous medium. The Ni(2+) complexes with hydrophobic nature were suspended into aqueous medium for testing them as sensor for organophosphate. The (L1)2.(Ni(2+))2 could sense phosmet with detection limit of 44 nM, whereas L2.Ni(2+) and L3.Ni(2+) exhibited the detection limits of 62 and 71 nM, respectively, for chlorpyrifos.

  14. Using Impact-Relevant Sensitivities to Efficiently Evaluate and Select Climate Change Scenarios

    NASA Astrophysics Data System (ADS)

    Vano, J. A.; Kim, J. B.; Rupp, D. E.; Mote, P.

    2014-12-01

    We outline an efficient approach to help researchers and natural resource managers more effectively use global climate model information in their long-term planning. The approach provides an estimate of the magnitude of change of a particular impact (e.g., summertime streamflow) from a large ensemble of climate change projections prior to detailed analysis. These estimates provide both qualitative information as an end unto itself (e.g., the distribution of future changes between emissions scenarios for the specific impact) and a judicious, defensible evaluation structure that can be used to qualitatively select a sub-set of climate models for further analysis. More specifically, the evaluation identifies global climate model scenarios that both (1) span the range of possible futures for the variable/s most important to the impact under investigation, and (2) come from global climate models that adequately simulate historical climate, providing plausible results for the future climate in the region of interest. To identify how an ecosystem process responds to projected future changes, we methodically sample, using a simple sensitivity analysis, how an impact variable (e.g., streamflow magnitude, vegetation carbon) responds locally to projected regional temperature and precipitation changes. We demonstrate our technique over the Pacific Northwest, focusing on two types of impacts each in three distinct geographic settings: (a) changes in streamflow magnitudes in critical seasons for water management in the Willamette, Yakima, and Upper Columbia River basins; and (b) changes in annual vegetation carbon in the Oregon and Washington Coast Ranges, Western Cascades, and Columbia Basin ecoregions.

  15. The lathyrus toxin, {beta}-N-oxalyl-L-{alpha},{beta}-diaminopropionic acid (ODAP), and homocysteic acid sensitize CA1 pyramidal neurons to cystine and L-2-amino-6-phosphonohexanoic acid

    SciTech Connect

    Chase, L.A. . E-mail: chase@hope.edu; Peterson, N.L. . E-mail: nlpeterson@noctrl.edu; Koerner, J.F. . E-mail: koern003@umn.edu

    2007-02-15

    A brief exposure of hippocampal slices to L-quisqualic acid (QUIS) sensitizes CA1 pyramidal neurons 30- to 250-fold to depolarization by certain excitatory amino acids analogues, e.g., L-2-amino-6-phosphonohexanoic acid (L-AP6), and by the endogenous compound, L-cystine. This phenomenon has been termed QUIS sensitization. A mechanism similar to that previously described for QUIS neurotoxicity has been proposed to describe QUIS sensitization. Specifically, QUIS has been shown to be sequestered into GABAergic interneurons by the System x{sub c} {sup -} and subsequently released by heteroexchange with cystine or L-AP6, resulting in activation of non-NMDA receptors. We now report two additional neurotoxins, the Lathyrus excitotoxin, {beta}-N-oxalyl-L-{alpha},{beta}-diaminopropionic acid (ODAP), and the endogenous compound, L-homocysteic acid (HCA), sensitize CA1 hippocampal neurons > 50-fold to L-AP6 and > 10-fold to cystine in a manner similar to QUIS. While the cystine- or L-AP6-mediated depolarization can be inhibited by the non-NMDA receptor antagonist CNQX in ODAP- or QUIS-sensitized slices, the NMDA antagonist D-AP5 inhibits depolarization by cystine or L-AP6 in HCA-sensitized slices. Thus, HCA is the first identified NMDA agonist that induces phosphonate or cystine sensitization. Like QUIS sensitization, the sensitization evoked by either ODAP or HCA can be reversed by a subsequent exposure to 2 mM {alpha}-aminoadipic acid. Finally, we have demonstrated that there is a correlation between the potency of inducers for triggering phosphonate or cystine sensitivity and their affinities for System x{sub c} {sup -} and either the non-NMDA or NMDA receptor. Thus, the results of this study support our previous model of QUIS sensitization and have important implications for the mechanisms of neurotoxicity, neurolathyrism and hyperhomocystinemia.

  16. Effect of Alpha-Hederin, the active constituent of Nigella sativa, on miRNA-126, IL-13 mRNA levels and inflammation of lungs in ovalbumin-sensitized male rats

    PubMed Central

    Fallahi, Maryam; Keyhanmanesh, Rana; Khamaneh, Amir Mahdi; Ebrahimi Saadatlou, Mohammad Ali; Saadat, Saeideh; Ebrahimi, Hadi

    2016-01-01

    Objective: In previous studies the therapeutic effects of Nigella sativa have been demonstrated on asthmatic animals. In the present study, the preventive effect of single dose of alpha-hederin, its active constituent, has been evaluated on lung inflammation and some inflammatory mediators in lungs of ovalbumin sensitized rat in order to elicit its mechanism. Materials and Methods: Forty rats were randomly grouped in 4 groups; control (C), sensitized (S), sensitized pretreated groups with thymoquinone (3 mg/kg i.p., S+TQ) and alpha-hederin (0.02 mg/kg i.p., S+AH). Levels of IL-13 mRNA and miRNA-126 in lung tissue and its pathological changes in each group were assessed. Results: Elevated levels of miRNA-126, IL-13 mRNA and pathological changes were observed in the sensitized group compared to the control group (p<0.001 to p<0.05). All of these factors were significantly reduced in S+TQ and S+AH groups in comparison to S group (p<0.001 to p<0.05). Although alpha-hederin decreased the levels of miRNA-126, IL-13 mRNA and pathological changes in comparison with thymoquinone, the results were statistically not significant. Conclusion: The results suggested that alpha-hederin had preventive effect on sensitized rats like thymoquinone. It may intervene in miRNA-126 expression, which consequently could interfere with IL-13 secretion pathway leading to a reduction in inflammatory responses. PMID:27247924

  17. Nitric oxide enhances the sensitivity of alpaca melanocytes to respond to {alpha}-melanocyte-stimulating hormone by up-regulating melanocortin-1 receptor

    SciTech Connect

    Dong, Yanjun; Cao, Jing; Wang, Haidong; Zhang, Jie; Zhu, Zhiwei; Bai, Rui; Hao, HuanQing; He, Xiaoyan; Fan, Ruiwen; Dong, Changsheng

    2010-06-11

    Nitric oxide (NO) and {alpha}-melanocyte-stimulating hormone ({alpha}-MSH) have been correlated with the synthesis of melanin. The NO-dependent signaling of cellular response to activate the hypothalamopituitary proopiomelanocortin system, thereby enhances the hypophysial secretion of {alpha}-MSH to stimulate {alpha}-MSH-receptor responsive cells. In this study we investigated whether an NO-induced pathway can enhance the ability of the melanocyte to respond to {alpha}-MSH on melanogenesis in alpaca skin melanocytes in vitro. It is important for us to know how to enhance the coat color of alpaca. We set up three groups for experiments using the third passage number of alpaca melanocytes: the control cultures were allowed a total of 5 days growth; the UV group cultures like the control group but the melanocytes were then irradiated everyday (once) with 312 mJ/cm{sup 2} of UVB; the UV + L-NAME group is the same as group UV but has the addition of 300 {mu}M L-NAME (every 6 h). To determine the inhibited effect of NO produce, NO produces were measured. To determine the effect of the NO to the key protein and gene of {alpha}-MSH pathway on melanogenesis, the key gene and protein of the {alpha}-MSH pathway were measured by quantitative real-time PCR and Western immunoblotting. The results provide exciting new evidence that NO can enhance {alpha}-MSH pathway in alpaca skin melanocytes by elevated MC1R. And we suggest that the NO pathway may more rapidly cause the synthesis of melanin in alpaca skin under UV, which at that time elevates the expression of MC1R and stimulates the keratinocytes to secrete {alpha}-MSH to enhance the {alpha}-MSH pathway on melanogenesis. This process will be of considerable interest in future studies.

  18. Nitric oxide enhances the sensitivity of alpaca melanocytes to respond to alpha-melanocyte-stimulating hormone by up-regulating melanocortin-1 receptor.

    PubMed

    Dong, Yanjun; Cao, Jing; Wang, Haidong; Zhang, Jie; Zhu, Zhiwei; Bai, Rui; Hao, HuanQing; He, Xiaoyan; Fan, Ruiwen; Dong, Changsheng

    2010-06-11

    Nitric oxide (NO) and alpha-melanocyte-stimulating hormone (alpha-MSH) have been correlated with the synthesis of melanin. The NO-dependent signaling of cellular response to activate the hypothalamopituitary proopiomelanocortin system, thereby enhances the hypophysial secretion of alpha-MSH to stimulate alpha-MSH-receptor responsive cells. In this study we investigated whether an NO-induced pathway can enhance the ability of the melanocyte to respond to alpha-MSH on melanogenesis in alpaca skin melanocytes in vitro. It is important for us to know how to enhance the coat color of alpaca. We set up three groups for experiments using the third passage number of alpaca melanocytes: the control cultures were allowed a total of 5 days growth; the UV group cultures like the control group but the melanocytes were then irradiated everyday (once) with 312 mJ/cm(2) of UVB; the UV+L-NAME group is the same as group UV but has the addition of 300 microM L-NAME (every 6h). To determine the inhibited effect of NO produce, NO produces were measured. To determine the effect of the NO to the key protein and gene of alpha-MSH pathway on melanogenesis, the key gene and protein of the alpha-MSH pathway were measured by quantitative real-time PCR and Western immunoblotting. The results provide exciting new evidence that NO can enhance alpha-MSH pathway in alpaca skin melanocytes by elevated MC1R. And we suggest that the NO pathway may more rapidly cause the synthesis of melanin in alpaca skin under UV, which at that time elevates the expression of MC1R and stimulates the keratinocytes to secrete alpha-MSH to enhance the alpha-MSH pathway on melanogenesis. This process will be of considerable interest in future studies.

  19. Boronic Acid: A Bio-Inspired Strategy To Increase the Sensitivity and Selectivity of Fluorescent NADH Probe.

    PubMed

    Wang, Lu; Zhang, Jingye; Kim, Beomsue; Peng, Juanjuan; Berry, Stuart N; Ni, Yong; Su, Dongdong; Lee, Jungyeol; Yuan, Lin; Chang, Young-Tae

    2016-08-24

    Fluorescent probes have emerged as an essential tool in the molecular recognition events in biological systems; however, due to the complex structures of certain biomolecules, it remains a challenge to design small-molecule fluorescent probes with high sensitivity and selectivity. Inspired by the enzyme-catalyzed reaction between biomolecule and probe, we present a novel combination-reaction two-step sensing strategy to improve sensitivity and selectivity. Based on this strategy, we successfully prepared a turn-on fluorescent reduced nicotinamide adenine dinucleotide (NADH) probe, in which boronic acid was introduced to bind with NADH and subsequently accelerate the sensing process. This probe shows remarkably improved sensitivity (detection limit: 0.084 μM) and selectivity to NADH in the absence of any enzymes. In order to improve the practicality, the boronic acid was further modified to change the measurement conditions from alkalescent (pH 9.5) to physiological environment (pH 7.4). Utilizing these probes, we not only accurately quantified the NADH weight in a health care product but also evaluated intracellular NADH levels in live cell imaging. Thus, these bio-inspired fluorescent probes offer excellent tools for elucidating the roles of NADH in biological systems as well as a practical strategy to develop future sensitive and selective probes for complicated biomolecules. PMID:27500425

  20. A microporous anionic metal-organic framework for a highly selective and sensitive electrochemical sensor of Cu(2+) ions.

    PubMed

    Jin, Jun-Cheng; Wu, Ju; Yang, Guo-Ping; Wu, Yun-Long; Wang, Yao-Yu

    2016-06-28

    We first reported an anionic metal-organic framework for electrode material for the electrochemical detection of Cu(2+). The modified electrode shows an excellent selectivity, high stability and sensitivity, wide linear range and lower detection limit. This strategy for generating new electrode materials will be useful for preparing new sensors and reporters for biological systems.

  1. Oncogenic transformation of C3H 10T1/2 cells exposed to alpha particles: Sensitivity through the cell cycle

    SciTech Connect

    Bettega, D.; Calzolari, P.; Chiorda, G.N.

    1995-06-01

    Oncogenic transformation of synchronized C3H 10T1/2 cells was determined after exposure to 4.3 MeV {alpha} particles (LET = 101 keV/{mu}m). Two synchronization techniques were tested using basic and modified protocols: one based on the release of cells from contact inhibition and the second on the mitotic shake-off method. Progression of cells through the cycle was followed as a function of time by flow cytometric analysis, DNA labeling for passage through S phase, the growth curve for the cell number and mitotic index measurements. The conclusion is that, although the release of cells from confluence provides higher yields of synchronized cells, mitotic shake-off proved to be the best way of collecting a synchronized population of minimally perturbed cells. Cells synchronized by mitotic shake-off were irradiated with 0.30 Gy in the interval between 2 and 10 h corresponding to G{sub 1} and early S phases. For comparison asynchronous populations were irradiated in parallel. Oncogenic transformation frequency, corrected for background, in mid-G{sub 1} phase was (18 {+-} 4) x 10{sup {minus}5} (average values of frequencies at 4 and 6 h) compared with the value of (8 {+-} 4) x 10{sup {minus}5} for the asynchronous population. While these data are suggestive of a trend toward a slightly increased sensitivity in mid-G{sub 1} phase, it is not statistically significant. The surviving fraction is constant in G{sub 1} phase. 14 refs., 2 figs., 2 tabs.

  2. Mitogen-activated protein kinase kinase 1/extracellular signal-regulated kinase (MEK-1/ERK) inhibitors sensitize reduced glucocorticoid response mediated by TNF{alpha} in human epidermal keratinocytes (HaCaT)

    SciTech Connect

    Onda, Kenji . E-mail: knjond@ps.toyaku.ac.jp; Nagashima, Masahiro; Kawakubo, Yo; Inoue, Shota; Hirano, Toshihiko; Oka, Kitaro

    2006-12-08

    Glucocorticoids (GCs) are essential drugs administered topically or systematically for the treatment of autoimmune skin diseases such as pemphigus. However, a certain proportion of patients does not respond well to GCs. Although studies on the relationship between cytokines and GC insensitivity in local tissues have attracted attention recently, little is known about the underlying mechanism(s) for GC insensitivity in epidermal keratinocytes. Here, we report that tumor necrosis factor (TNF) {alpha} reduces GC-induced transactivation of endogenous genes as well as a reporter plasmid which contains GC responsive element (GRE) in human epidermal keratinocyte cells (HaCaT). The GC insensitivity by TNF{alpha} was not accompanied by changes in mRNA expressions of GR isoforms ({alpha} or {beta}). However, we observed that mitogen-activated protein kinase kinase-1/extracellular signal-regulated kinase (MEK-1/ERK) inhibitors (PD98059 and U0126) significantly sensitized the GC-induced transactivation of anti-inflammatory genes (glucocorticoid-induced leucine zipper (GILZ) and mitogen-activated protein kinase phosphatase (MKP)-1) and FK506 binding protein (FKBP) 51 gene in the presence of TNF{alpha}. Additionally, we observed that TNF{alpha} reduced prednisolone (PSL)-dependent nuclear translocation of GR, which was restored by pre-treatment of MEK-1 inhibitors. This is the first study demonstrating a role of the MEK-1/ERK cascade in TNF{alpha}-mediated GC insensitivity. Our data suggest that overexpression of TNF{alpha} leads to topical GC insensitivity by reducing GR nuclear translocation in keratinocytes, and our findings also suggest that inhibiting the MEK-1/ERK cascade may offer a therapeutic potential for increasing GC efficacy in epidermis where sufficient inflammatory suppression is required.

  3. Strongly coupled hybrid nanostructures for selective hydrogen detection - understanding the role of noble metals in reducing cross-sensitivity

    NASA Astrophysics Data System (ADS)

    Liu, Bin; Cai, Daoping; Liu, Yuan; Wang, Dandan; Wang, Lingling; Xie, Wuyuan; Li, Qiuhong; Wang, Taihong

    2014-04-01

    Noble metal-semiconductor hybrid nanostructures can offer outperformance to gas sensors in terms of sensitivity and selectivity. In this work, a catalytically activated (CA) hydrogen sensor is realized based on strongly coupled Pt/Pd-WO3 hybrid nanostructures constructed by a galvanic replacement participated solvothermal procedure. The room-temperature operation and high selectivity distinguish this sensor from the traditional ones. It is capable of detecting dozens of parts per million (ppm) hydrogen in the presence of thousands of ppm methane gas. An insight into the role of noble metals in reducing cross-sensitivity is provided by comparing the sensing properties of this sensor with a traditional thermally activated (TA) one made from the same pristine WO3. Based on both experimental and density functional theory (DFT) calculation results, the cross-sensitivity of the TA sensor is found to have a strong dependence on the highest occupied molecular orbital (HOMO) level of the hydrocarbon molecules. The high selectivity of the CA sensor comes from the reduced impact of gas frontier orbitals on the charge transfer process by the nano-scaled metal-semiconductor (MS) interface. The methodology demonstrated in this work indicates that rational design of MS hybrid nanostructures can be a promising strategy for highly selective gas sensing applications.Noble metal-semiconductor hybrid nanostructures can offer outperformance to gas sensors in terms of sensitivity and selectivity. In this work, a catalytically activated (CA) hydrogen sensor is realized based on strongly coupled Pt/Pd-WO3 hybrid nanostructures constructed by a galvanic replacement participated solvothermal procedure. The room-temperature operation and high selectivity distinguish this sensor from the traditional ones. It is capable of detecting dozens of parts per million (ppm) hydrogen in the presence of thousands of ppm methane gas. An insight into the role of noble metals in reducing cross-sensitivity

  4. (+/-)-1-[[2-(3,4-Dimethoxphenyl)ethyl]amino]-3-(3-methylphenoxy)-2- propanol hydrochloride (bevantolol, NC-1400) as a beta 1-selective adrenoceptor blocker with alpha 1-adrenoceptor blocking activity.

    PubMed

    Takayanagi, I; Kizawa, Y; Iwasaki, S; Nakagoshi, A

    1987-01-01

    Blocking activities of alpha- and beta-adrenoceptors by (+/-)-1-[[2-(3,4-dimethoxyphenyl)-ethyl]amino]-3-(3-methylphenoxy)-2 -propanol hydrochloride (NC-1400) were tested on the isolated muscles, comparing with those of labetalol and atenolol. In blocking the beta 1-adrenoceptor, NC-1400 was slightly more potent than labetalol and atenolol. NC-1400 was about 1/10th as potent as labetalol and about ten times as potent as atenolol in blocking the beta 2-receptor. NC-1400 was beta 1-adrenoceptor selective. NC-1400 was about 1/30th as potent as labetalol in blocking the alpha 1-receptor. NC-1400 did not interact with the alpha 2-adrenoceptor in concentrations up to 10(-5) M. The present results indicate that NC-1400 is the selective beta 1-adrenoceptor blocker with some blocking activity of alpha 1-adrenoceptors.

  5. Highly selective and sensitive detection of neurotransmitters using receptor-modified single-walled carbon nanotube sensors

    NASA Astrophysics Data System (ADS)

    Kim, Byeongju; Song, Hyun Seok; Jin, Hye Jun; Park, Eun Jin; Lee, Sang Hun; Lee, Byung Yang; Park, Tai Hyun; Hong, Seunghun

    2013-07-01

    We present receptor-modified carbon nanotube sensors for the highly selective and sensitive detection of acetylcholine (ACh), one kind of neurotransmitter. Here, we successfully expressed the M1 muscarinic acetylcholine receptor (M1 mAChR), a family of G protein-coupled receptors (GPCRs), in E. coli and coated single-walled carbon nanotube (swCNT)-field effect transistors (FETs) with lipid membrane including the receptor, enabling highly selective and sensitive ACh detection. Using this sensor, we could detect ACh at 100 pM concentration. Moreover, we showed that this sensor could selectively detect ACh among other neurotransmitters. This is the first demonstration of the real-time detection of ACh using specific binding between ACh and M1 mAChR, and it may lead to breakthroughs for various applications such as disease diagnosis and drug screening.

  6. FA-SIFT study of reactions of protonated water and ethanol clusters with [alpha]-pinene and linalool in view of their selective detection by CIMS

    NASA Astrophysics Data System (ADS)

    Dhooghe, F.; Amelynck, C.; Rimetz-Planchon, J.; Schoon, N.; Vanhaecke, F.

    2010-02-01

    The use of protonated water clusters and protonated ethanol clusters as reagent ions has been evaluated for the resolution of an interference encountered in CIMS when measuring monoterpenes (C10H16) and linalool (C10H18O) simultaneously. To this end, the reactions of H3O+.(H2O)n (n = 1-3), (C2H5OH)mH+ (m = 1-3) and (C2H5OH.H2O)H+ with [alpha]-pinene and linalool have been characterized in a flowing afterglow-selected ion flow tube (FA-SIFT) instrument at a SIFT He buffer gas pressure of 1.43 hPa and a temperature of 298 K. All reactions with linalool were found to occur at the collision limit. The reaction of (C2H5OH)2H+ with [alpha]-pinene proceeds at half the collision rate and both the reactions of (C2H5OH)3H+ and H3O+.(H2O)3 with [alpha]-pinene have a very low rate constant. All other reactions involving [alpha]-pinene proceed at the collision rate. The reactions of H3O+.H2O, H3O+.(H2O)2, C2H5OH2+, (C2H5OH.H2O)H+ and (C2H5OH)2H+ with [alpha]-pinene mainly proceed by proton transfer. Additionally, ligand switching channels have been observed for the reactions of (C2H5OH)2H+ and H3O+.(H2O)2 with [alpha]-pinene. Protonated linalool was observed as a minor product for the reactions of (C2H5OH.H2O)H+ and H3O+.(H2O)n (n = 1-3) with linalool. For all linalool reactions, a contribution of the dissociative proton transfer product at m/z 137 was found and this ion was the main product ion for the reactions with H3O+.H2O, C2H5OH2+ and (C2H5OH.H2O)H+. For the (C2H5OH.H2O)H+/linalool reaction, ligand switching with both water and ethanol has been observed. Major ligand switching channels were observed for the reactions of (C2H5OH)2H+, (C2H5OH)3H+ and H3O+.(H2O)2 with linalool. Also, for the H3O+.(H2O)3/linalool reaction, several ligand switching channels have been observed. These results are discussed in view of their applicability for the selective detection of monoterpenes and linalool with CIMS instrumentation such as SIFT-MS, PTR-MS and APCI-MS.

  7. Selective activation of p38alpha and p38gamma by hypoxia. Role in regulation of cyclin D1 by hypoxia in PC12 cells.

    PubMed

    Conrad, P W; Rust, R T; Han, J; Millhorn, D E; Beitner-Johnson, D

    1999-08-13

    Hypoxic/ischemic trauma is a primary factor in the pathology of a multitude of disease states. The effects of hypoxia on the stress- and mitogen-activated protein kinase signaling pathways were studied in PC12 cells. Exposure to moderate hypoxia (5% O(2)) progressively stimulated phosphorylation and activation of p38gamma in particular, and also p38alpha, two stress-activated protein kinases. In contrast, hypoxia had no effect on enzyme activity of p38beta, p38beta(2), p38delta, or on c-Jun N-terminal kinase, another stress-activated protein kinase. Prolonged hypoxia also induced phosphorylation and activation of p42/p44 mitogen-activated protein kinase, although this activation was modest compared with nerve growth factor- and ultraviolet light-induced activation. Hypoxia also dramatically down-regulated immunoreactivity of cyclin D1, a gene that is known to be regulated negatively by p38 at the level of gene expression (Lavoie, J. N., L'Allemain, G., Brunet, A., Muller, R., and Pouyssegur, J. (1996) J. Biol. Chem. 271, 20608-20616). This effect was partially blocked by SB203580, an inhibitor of p38alpha but not p38gamma. Overexpression of a kinase-inactive form of p38gamma was also able to reverse in part the effect of hypoxia on cyclin D1 levels, suggesting that p38alpha and p38gamma converge to regulate cyclin D1 during hypoxia. These studies demonstrate that an extremely typical physiological stress (hypoxia) causes selective activation of specific p38 signaling elements; and they also identify a downstream target of these pathways. PMID:10438538

  8. Selective Enhancement of Dopamine Release in the Ventral Pallidum of Methamphetamine-Sensitized Mice

    PubMed Central

    2016-01-01

    Drugs of abuse induce sensitization, which is defined as enhanced response to additional drug following a period of withdrawal. Sensitization occurs in both humans and animal models of drug reinforcement and contributes substantially to the addictive nature of drugs of abuse, because it is thought to represent enhanced motivational wanting for drug. The ventral pallidum, a key member of the reward pathway, contributes to behaviors associated with reward, such as sensitization. Dopamine inputs to the ventral pallidum have not been directly characterized. Here we provide anatomical, neurochemical, and behavioral evidence demonstrating that dopamine terminals in the ventral pallidum contribute to reward in mice. We report subregional differences in dopamine release, measured by ex vivo fast-scan cyclic voltammetry: rostral ventral pallidum exhibits increased dopamine release and uptake compared with caudal ventral pallidum, which is correlated with tissue expression of dopaminergic proteins. We then subjected mice to a methamphetamine-sensitization protocol to investigate the contribution of dopaminergic projections to the region in reward related behavior. Methamphetamine-sensitized animals displayed a 508% and 307% increase in baseline dopamine release in the rostral and caudal ventral pallidum, respectively. Augmented dopamine release in the rostral ventral pallidum was significantly correlated with sensitized locomotor activity. Moreover, this presynaptic dopaminergic plasticity occurred only in the ventral pallidum and not in the ventral or dorsal striatum, suggesting that dopamine release in the ventral pallidum may be integrally important to drug-induced sensitization. PMID:27501345

  9. Vibration isolation technology: Sensitivity of selected classes of experiments to residual accelerations

    NASA Technical Reports Server (NTRS)

    Alexander, J. Iwan D.

    1990-01-01

    The solution was sought of a 2-D axisymmetric moving boundary problem for the sensitivity of isothermal and nonisothermal liquid columns and the sensitivity of thermo-capillary flows to buoyancy driven convection caused by residual accelerations. The sensitivity of a variety of space experiments to residual accelerations are examined. In all the cases discussed, the sensitivity is related to the dynamic response of a fluid. In some cases the sensitivity can be defined by the magnitude of the response of the velocity field. This response may involve motion of the fluid associated with internal density gradients, or the motion of a free liquid surface. For fluids with internal density gradients, the type of acceleration to which the experiment is sensitive will depend on whether buoyancy driven convection must be small in comparison to other types of fluid motion (such as thermocapillary flow), or fluid motion must be suppressed or eliminated (such as in diffusion studies, or directional solidification experiments). The effect of the velocity on the composition and temperature field must be considered, particularly in the vicinity of the melt crystal interface. As far as the response to transient disturbances is concerned the sensitivity is determined by both the magnitude and frequency the acceleration and the characteristic momentum and solute diffusion times.

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

    PubMed

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

    2016-03-01

    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. PMID:26619046

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

    PubMed

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

    2016-03-01

    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.

  12. Vibration isolation technology: Sensitivity of selected classes of space experiments to residual accelerations

    NASA Technical Reports Server (NTRS)

    Alexander, J. Iwan D.; Zhang, Y. Q.; Adebiyi, Adebimpe

    1989-01-01

    Progress performed on each task is described. Order of magnitude analyses related to liquid zone sensitivity and thermo-capillary flow sensitivity are covered. Progress with numerical models of the sensitivity of isothermal liquid zones is described. Progress towards a numerical model of coupled buoyancy-driven and thermo-capillary convection experiments is also described. Interaction with NASA personnel is covered. Results to date are summarized and they are discussed in terms of the predicted space station acceleration environment. Work planned for the second year is also discussed.

  13. UV-induced endonuclease III-sensitive sites at the mating type loci in Saccharomyces cerevisiae are repaired by nucleotide excision repair: RAD7 and RAD16 are not required for their removal from HML alpha.

    PubMed

    Reed, S H; Boiteux, S; Waters, R

    1996-03-01

    Ultraviolet irradiation of DNA induces cyclobutane pyrimidine dimers (CPDs) 6-4'-(pyrimidine 2'-one) pyrimidines and pyrimidine hydrates. The dimer is the major photoproduct, and is specifically recognized by endonuclease V of phage T4. Pyrimidine hydrates represent a small fraction of the total photoproducts, and are substrates for endonuclease III of Escherichia coli. We used these enzymes to follow the fate of their substrates in the mating type loci of Saccharomyces cerevisiae. In a RAD strain, CPSs in the transcriptionally active MAT alpha locus are preferentially repaired relative to the inactive HML alpha locus, whilst repair of endonuclease III-sensitive sites is not preferential. The rad1, 2, 3 and 4 mutants, which lack factors that are essential for the incision step of nucleotide excision repair (NER), repair neither CPDs nor endonuclease III-sensitive sites, clearly showing that these lesions are repaired by by NER pathway. Previously it had been shown that the products of the RAD7 and RAD16 genes are required for the NER of CPDs from the HML alpha locus. We show that, in the same locus, these gene products are not needed for removal of endonuclease III-sensitive sites by the same mechanism. This indicates that the components required for NER differ depending on either the type of lesion encountered or on the specific location of the lesion within the genome.

  14. Ligation of retinoic acid receptor alpha regulates negative selection of thymocytes by inhibiting both DNA binding of nur77 and synthesis of bim.

    PubMed

    Szegezdi, Eva; Kiss, Ildikó; Simon, Agnes; Blaskó, Bernadett; Reichert, Uwe; Michel, Serge; Sándor, Mátyás; Fésüs, László; Szondy, Zsuzsa

    2003-04-01

    Negative selection refers to the selective deletion of autoreactive thymocytes. Its molecular mechanisms have not been well defined. Previous studies in our laboratory have demonstrated that retinoic acids, physiological ligands for the nuclear retinoid receptors, selectively inhibit TCR-mediated death under in vitro conditions, and the inhibition is mediated via the retinoic acid receptor (RAR) alpha. The present studies were undertaken to investigate whether ligation of RARalpha leads to inhibition of TCR-mediated death in vivo and to identify the molecular mechanisms involved. Three models of TCR-mediated death were studied: anti-CD3-mediated death of thymocytes in wild-type mice, and Ag- and bacterial superantigen-driven thymocyte death in TCR-transgenic mice expressing a receptor specific for a fragment of pigeon cytochrome c in the context of the E(k) (class II MHC) molecule. Our data demonstrate that the molecular program of both anti-CD3- and Ag-driven, but not that of superantigen-mediated apoptosis involves up-regulation of nur77, an orphan nuclear receptor, and bim, a BH3-only member of the proapoptotic bcl-2 protein family, proteins previously implicated to participate in the negative selection. Ligation of RARalpha by the synthetic agonist CD336 inhibited apoptosis, DNA binding of nur77, and synthesis of bim induced by anti-CD3 or the specific Ag, but had no effect on the superantigen-driven cell death. Our data imply that retinoids are able to inhibit negative selection in vivo as well, and they interfere with multiple steps of the T cell selection signal pathway.

  15. Shared receptor components but distinct complexes for alpha and beta interferons.

    PubMed

    Lewerenz, M; Mogensen, K E; Uzé, G

    1998-09-25

    The type I interferon family includes 13 alpha, one omega and one beta subtypes recognized by a complex containing the receptor subunits ifnar1 and ifnar2 and their associated Janus tyrosine kinases, Tyk2 and Jak1. To investigate the reported differences in the way that alpha and beta interferons signal through the receptor, we introduced alanine-substitutions in the ifnar2 extracellular domain, and expressed the mutants in U5A cells, lacking endogenous ifnar2. A selection, designed to recover mutants that responded preferentially to alpha or beta interferon yielded three groups: I, neutral; II, sensitive to alpha interferon, partially resistant to beta interferon; III, resistant to alpha interferon, partially sensitive to beta interferon. A mutant clone, TMK, fully resistant to alpha interferon with good sensitivity to beta interferon, was characterized in detail and compared with U5A cells complemented with wild-type ifnar2 and also with Tyk2-deficient 11.1 cells, which exhibit a similar alpha-unresponsive phenotype with a partial beta interferon response. Using anti-receptor antibodies and mutant forms of beta interferon, three distinct modes of ligand interaction could be discerned: (i) alpha interferon with ifnar1 and ifnar2; (ii) beta interferon with ifnar1 and ifnar2; (iii) beta interferon with ifnar2 alone. We conclude that alpha and beta interferons signal differently through their receptors because the two ligand subtypes interact with the receptor subunits ifnar 1 and ifnar2 in entirely different ways.

  16. Genomic organization and differential signature of positive selection in the alpha and beta globin gene clusters in two cetacean species.

    PubMed

    Nery, Mariana F; Arroyo, José Ignacio; Opazo, Juan C

    2013-01-01

    The hemoglobin of jawed vertebrates is a heterotetramer protein that contains two α- and two β-chains, which are encoded by members of α- and β-globin gene families. Given the hemoglobin role in mediating an adaptive response to chronic hypoxia, it is likely that this molecule may have experienced a selective pressure during the evolution of cetaceans, which have to deal with hypoxia tolerance during prolonged diving. This selective pressure could have generated a complex history of gene turnover in these clusters and/or changes in protein structure themselves. Accordingly, we aimed to characterize the genomic organization of α- and β-globin gene clusters in two cetacean species and to detect a possible role of positive selection on them using a phylogenetic framework. Maximum likelihood and Bayesian phylogeny reconstructions revealed that both cetacean species had retained a similar complement of putatively functional genes. For the α-globin gene cluster, the killer whale presents a complement of genes composed of HBZ, HBK, and two functional copies of HBA and HBQ genes, whereas the dolphin possesses HBZ, HBK, HBA and HBQ genes, and one HBA pseudogene. For the β-globin gene cluster, both species retained a complement of four genes, two early expressed genes-HBE and HBH-and two adult expressed genes-HBD and HBB. Our natural selection analysis detected two positively selected sites in the HBB gene (56 and 62) and four in HBA (15, 21, 49, 120). Interestingly, only the genes that are expressed during the adulthood showed the signature of positive selection. PMID:24259315

  17. Relative sensitivity of three endangered fishes, Colorado squawfish, bonytail, and razorback sucker, to selected metal pollutants

    USGS Publications Warehouse

    Buhl, K.J.

    1997-01-01

    The acute toxicity of four metal pollutants to larval and juvenile stages of endangered Colorado squawfish (Ptychocheilus lucius), bonytail (Gila elegans), and razorback sucker (Xyrauchen texanus) were determined in a water quality representative of that in the Green River, Utah. The rank order of toxicity (96-hr LC50) of the metals to all species and life stages from most toxic to least toxic was mercury (57-168 ??G/liter) > cadmium (78-168 pg/liter) > hexavalent chromium (32,000-123,000 ??g/liter) > lead (>170,000 ??G/liter). In tests with lead, a precipitate formed in all test solutions and no mortalities occurred in these treatments. The larvae of each species were as sensitive or more sensitive than the juveniles to cadmium, hexavalent chromium, and mercury. Overall, the three species exhibited similar sensitivities to cadmium, hexavalent chromium, and mercury. Comparison of test results for the juveniles with toxicity values reported for other freshwater fishes tested in different water qualities indicates that the endangered fishes are more sensitive to cadmium than other cyprinids and centrarchids and less sensitive than salmonids, whereas their sensitivity to hexavalent chromium and mercury is similar to that of other cyprinids, centrarchids, and salmonids.

  18. Relative sensitivity of three endangered fishes, Colorado squawfish, bonytail, and razorback sucker, to selected metal pollutants.

    PubMed

    Buhl, K J

    1997-07-01

    The acute toxicity of four metal pollutants to larval and juvenile stages of endangered Colorado squawfish (Ptychocheilus lucius), bonytail (Gila elegans), and razorback sucker (Xyrauchen texanus) were determined in a water quality representative of that in the Green River, Utah. The rank order of toxicity (96-hr LC50) of the metals to all species and life stages from most toxic to least toxic was mercury (57-168 micrograms/liter) > cadmium (78-168 micrograms/liter) > hexavalent chromium (32,000-123,000 micrograms/liter) > lead (> 170,000 micrograms/liter). In tests with lead, a precipitate formed in all test solutions and no mortalities occurred in these treatments. The larvae of each species were as sensitive or more sensitive than the juveniles to cadmium, hexavalent chromium, and mercury. Overall, the three species exhibited similar sensitivities to cadmium, hexavalent chromium, and mercury. Comparison of test results for the juveniles with toxicity values reported for other freshwater fishes tested in different water qualities indicates that the endangered fishes are more sensitive to cadmium than other cyprinids and centrarchids and less sensitive than salmonids, whereas their sensitivity to hexavalent chromium and mercury is similar to that of other cyprinids, centrarchids, and salmonids.

  19. Effects of hippocampal injections of a novel ligand selective for the alpha 5 beta 2 gamma 2 subunits of the GABA/benzodiazepine receptor on Pavlovian conditioning.

    PubMed

    Bailey, David J; Tetzlaff, Julie E; Cook, James M; He, Xiaohui; Helmstetter, Fred J

    2002-07-01

    Benzodiazepine pharmacology has led to greater insight into the neural mechanisms underlying learning and anxiety. The synthesis of new compounds capable of modulating responses produced by these receptors has been made possible by the development of an isoform model of the GABA(A)/benzodiazepine receptor complex. In the current experiment, rats were pretreated with several concentrations of the novel ligand RY024 (an alpha 5 beta 2 gamma 2 -selective benzodiazepine receptor inverse agonist) in the hippocampus and were trained in a Pavlovian fear conditioning paradigm. RY024 independently produced fear-related behavior prior to training and, at the highest concentration, decreased the strength of conditioning observed 24 h after training. These data provide further evidence for the involvement of hippocampal GABA(A)/benzodiazepine receptors in learning and anxiety.

  20. Fabrication of highly sensitive and selective H₂ gas sensor based on SnO₂ thin film sensitized with microsized Pd islands.

    PubMed

    Nguyen, Van Toan; Nguyen, Viet Chien; Nguyen, Van Duy; Hoang, Si Hong; Hugo, Nguyen; Nguyen, Duc Hoa; Nguyen, Van Hieu

    2016-01-15

    Ultrasensitive and selective hydrogen gas sensor is vital component in safe use of hydrogen that requires a detection and alarm of leakage. Herein, we fabricated a H2 sensing devices by adopting a simple design of planar-type structure sensor in which the heater, electrode, and sensing layer were patterned on the front side of a silicon wafer. The SnO2 thin film-based sensors that were sensitized with microsized Pd islands were fabricated at a wafer-scale by using a sputtering system combined with micro-electronic techniques. The thicknesses of SnO2 thin film and microsized Pd islands were optimized to maximize the sensing performance of the devices. The optimized sensor could be used for monitoring hydrogen gas at low concentrations of 25-250 ppm, with a linear dependence to H2 concentration and a fast response and recovery time. The sensor also showed excellent selectivity for monitoring H2 among other gases, such as CO, NH3, and LPG, and satisfactory characteristics for ensuring safety in handling hydrogen. The hydrogen sensing characteristics of the sensors sensitized with Pt and Au islands were also studied to clarify the sensing mechanisms.

  1. Melanoma prevention by MC1R selective small peptide analogs of alpha MSH | Division of Cancer Prevention

    Cancer.gov

    DESCRIPTION (provided by applicant): This revised application will test the hypothesis that small peptide analogs of α-melanocortin (α-MSH) that are selective agonists of the melanocortin 1 receptor (MC1R) will prevent melanoma tumor formation in transgenic mouse melanoma models by enhancing repair of ultraviolet radiation (UV)-induced DNA damage and stimulating melanogenesis. |

  2. Sensitivity and Specificity Improvement in Abdominal Obesity Diagnosis Using Cluster Analysis during Waist Circumference Cut-Off Point Selection

    PubMed Central

    Bermúdez, Valmore; Añez, Roberto; Toledo, Alexandra; Bello, Luis; Apruzzese, Vanessa; González, Robys; Chacín, Maricarmen; Cabrera, Mayela; Cano, Clímaco; Velasco, Manuel; López-Miranda, José

    2015-01-01

    Introduction. The purpose of this study was to analyze the influence of metabolic phenotypes during the construction of ROC curves for waist circumference (WC) cutpoint selection. Materials and Methods. A total of 1,902 subjects of both genders were selected from the Maracaibo City Metabolic Syndrome Prevalence Study database. Two-Step Cluster Analysis (TSCA) was applied to select metabolically healthy and sick men and women. ROC curves were constructed to determine WC cutoff points by gender. Results. Through TSCA, metabolic phenotype predictive variables were selected: HOMA2-IR and HOMA2-βcell for women and HOMA2-IR, HOMA2-βcell, and TAG for men. Subjects were classified as healthy normal weight, metabolically obese normal weight, healthy and metabolically disturbed overweight, and healthy and metabolically disturbed obese. Final WC cutpoints were 91.50 cm for women (93.4% sensitivity, 93.7% specificity) and 98.15 cm for men (96% sensitivity, 99.5% specificity). Conclusions. TSCA in the selection of the groups used in ROC curves construction proved to be an important tool, aiding in the detection of MOWN and MHO which cannot be identified with WC alone. The resulting WC cutpoints were <91.00 cm for women and <98.00 cm for men. Furthermore, anthropometry is insufficient to determine healthiness, and, biochemical analysis is needed to properly filter subjects during classification. PMID:25945356

  3. Depletion of OLFM4 gene inhibits cell growth and increases sensitization to hydrogen peroxide and tumor necrosis factor-alpha induced-apoptosis in gastric cancer cells

    PubMed Central

    2012-01-01

    Background Human olfactomedin 4 (OLFM4) gene is a secreted glycoprotein more commonly known as the anti-apoptotic molecule GW112. OLFM4 is found to be frequently up-regulated in many types of human tumors including gastric cancer and it was believed to play significant role in the progression of gastric cancer. Although the function of OLFM4 has been indicated in many studies, recent evidence strongly suggests a cell or tissue type-dependent role of OLFM4 in cell growth and apoptosis. The aim of this study is to examine the role of gastric cancer-specific expression of OLFM4 in cell growth and apoptosis resistance. Methods OLFM4 expression was eliminated by RNA interference in SGC-7901 and MKN45 cells. Cell proliferation, anchorage-independent growth, cell cycle and apoptosis were characterized in vitro. Tumorigenicity was analyzed in vivo. The apoptosis and caspase-3 activation in response to hydrogen peroxide (H2O2) or tumor necrosis factor-alpha (TNF α) were assessed in the presence or absence of caspase inhibitor Z-VAD-fmk. Results The elimination of OLFM4 protein by RNA interference in SGC-7901 and MKN45 cells significantly inhibits tumorigenicity both in vitro and in vivo by induction of cell G1 arrest (all P < 0.01). OLFM4 knockdown did not trigger obvious cell apoptosis but increased H2O2 or TNF α-induced apoptosis and caspase-3 activity (all P < 0.01). Treatment of Z-VAD-fmk attenuated caspase-3 activity and significantly reversed the H2O2 or TNF α-induced apoptosis in OLFM4 knockdown cells (all P < 0.01). Conclusion Our study suggests that depletion of OLFM4 significantly inhibits tumorigenicity of the gastric cancer SGC-7901 and MKN45 cells. Blocking OLFM4 expression can sensitize gastric cancer cells to H2O2 or TNF α treatment by increasing caspase-3 dependent apoptosis. A combination strategy based on OLFM4 inhibition and anticancer drugs treatment may provide therapeutic potential in gastric cancer intervention. PMID:22471589

  4. Highly selective water and fat imaging applying multislice sequences without sensitivity to B1 field inhomogeneities.

    PubMed

    Schick, F; Forster, J; Machann, J; Huppert, P; Claussen, C D

    1997-08-01

    Improved selectivity to one chemical shift component was obtained using simultaneous slice-selective and chemical shift-selective excitation in sequences with usual spin-echo refocusing. The new type of sequences can be applied on modern whole-body units and permits multislice operation. Spatial-spectral excitation is based on prior research in this field, but the proposed improved version provides off-center slice excitation by the usual processing of the RF pulse envelopes. In addition, no irregular gradient shapes are necessary. The required B0 homogeneity of the new method is similar to conventional "fat-sat" techniques. In contrast to fat-sat methods, selectivity to water is not reduced by unavoidable misadjustments of the transmitter or B1 field inhomogeneities in the newly developed approach. Thus, the reported method has the potential to replace standard frequency selective fat-sat sequences for most applications. PMID:9256107

  5. Redox-sensitivity and mobility of selected pharmaceutical compounds in a laboratory column experiment

    NASA Astrophysics Data System (ADS)

    Banzhaf, S.; Nödler, K.; Licha, T.; Krein, A.; Scheytt, T.

    2012-04-01

    Laboratory column experiments are suitable to investigate the sediment water interaction and to study the transport behaviour of solutes. Processes like retardation and degradation can be identified and quantified. The conducted experiment, which is closely connected to a field study in Luxembourg, investigated the transport behaviour of selected pharmaceutical compounds and their redox-dependent metabolism under water saturated conditions. Fine-grained natural sediment with a low hydraulic conductivity from a study site in Luxembourg was filled into the column. The water for the experiment was taken from a small stream at the same fieldsite. It was spiked with four pharmaceutical compounds (carbamazepine, diclofenac, ibuprofen, sulfamethoxazole) with concentrations between 170 and 300 ng/L for the different substances. The chosen pharmaceuticals were also detected in groundwater and surface water samples at the study site and used to qualify exchange/mixing of surface water and groundwater (BANZHAF et al., 2011). As some of the substances are known to exhibit redox-sensitive degradation, the redox-conditions were systematically varied throughout the experiment. This was realised by adding nitrate at the inflow of the column. During the experiment, which lasted for 2.5 months, four different nitrate concentrations (20-130 mg/L) were applied, beginning with the highest concentration. During the experiment water from the reservoir tank was sampled daily in order to detect a potential degradation of the pharmaceutical compounds before they enter the column. The effluent water was sampled every three hours to guarantee a maximum resolution for the analysis of the pharmaceuticals where necessary. In addition, major ions were analysed in the influent and effluent samples. Throughout the experiment physicochemical parameters (oxidation reduction potential (ORP), dissolved oxygen, electrical conductivity, and pH-value) were measured and logged at the outflow of the column

  6. Two Dimensional Host-Guest Metal-Organic Framework Sensor with High Selectivity and Sensitivity to Picric Acid.

    PubMed

    Bagheri, Minoo; Masoomi, Mohammad Yaser; Morsali, Ali; Schoedel, Alexander

    2016-08-24

    A dye-sensitized metal-organic framework, TMU-5S, was synthesized based on introducing the laser dye Rhodamine B into the porous framework TMU-5. TMU-5S was investigated as a ratiometric fluorescent sensor for the detection of explosive nitro aromatic compounds and showed four times greater selectivity to picric acid than any state-of-the-art luminescent-based sensor. Moreover, it can selectively discriminate picric acid concentrations in the presence of other nitro aromatics and volatile organic compounds. Our findings indicate that using this sensor in two dimensions leads to a greatly reduced environmental interference response and thus creates exceptional sensitivity toward explosive molecules with a fast response. PMID:27479837

  7. Increasing the sensitivity and single-base mismatch selectivity of the molecular beacon using graphene oxide as the "nanoquencher".

    PubMed

    Lu, Chun-Hua; Li, Juan; Liu, Jing-Jing; Yang, Huang-Hao; Chen, Xi; Chen, Guo-Nan

    2010-04-26

    Here, we report a novel, highly sensitive, selective and economical molecular beacon using graphene oxide as the "nanoquencher". This novel molecular beacon system contains a hairpin-structured fluorophore-labeled oligonucleotide and a graphene oxide sheet. The strong interaction between hairpin-structured oligonucleotide and graphene oxide keep them in close proximity, facilitating the fluorescence quenching of the fluorophore by graphene oxide. In the presence of a complementary target DNA, the binding between hairpin-structured oligonucleotide and target DNA will disturb the interaction between hairpin-structured oligonucleotide and graphene oxide, and release the oligonucleotide from graphene oxide, resulting in restoration of fluorophore fluorescence. In the present study, we show that this novel graphene oxide quenched molecular beacon can be used to detect target DNA with higher sensitivity and single-base mismatch selectivity compared to the conventional molecular beacon.

  8. A Post-Synthetically Modified MOF for Selective and Sensitive Aqueous-Phase Detection of Highly Toxic Cyanide Ions.

    PubMed

    Karmakar, Avishek; Kumar, Naveen; Samanta, Partha; Desai, Aamod V; Ghosh, Sujit K

    2016-01-18

    Selective and sensitive detection of toxic cyanide (CN(-) ) by a post-synthetically altered metal-organic framework (MOF) has been achieved. A post-synthetic modification was employed in the MOF to incorporate the specific recognition site with the CN(-) ion over all other anions, such as Cl(-) , Br(-) , and SCN(-) . The aqueous-phase sensing and very low detection limit, the essential prerequisites for an effective sensory material, have been fulfilled by the MOF. Moreover, the present detection level meets the standard set by the World Health Organization (WHO) for the permissible limit of cyanide concentration in drinking water. The utilization of MOF-based materials as the fluorometric probes for selective and sensitive detection of CN(-) ions has not been explored till now.

  9. FRET-based dimeric aptamer probe for selective and sensitive Lup an 1 allergen detection.

    PubMed

    Mairal, T; Nadal, P; Svobodova, M; O'Sullivan, C K

    2014-04-15

    A sensitive method for the rapid and sensitive detection of the anaphylactic food allergen Lup an 1 (β-conglutin) exploiting fluorescence resonance energy transfer (FRET) has been developed. A high affinity dimeric form of a truncated 11-mer aptamer against β-conglutin was used, with each monomeric aptamer being flanked by donor/acceptor moieties. The dimeric form in the absence of target yields fluorescence emission due to the FRET from the excited fluorophore to the proximal second fluorophore. However, upon addition of β-conglutin, the specific interaction induces a change in the bi-aptameric structure resulting in an increase in fluorescence emission. The method is highly specific and sensitive, with a detection limit of 150 pM, providing an effective tool for the direct detection of the toxic β-conglutin subunit in foodstuffs in just 1 min at room temperature. PMID:24280051

  10. Sensitivity analysis of add-on price estimate for select silicon wafering technologies

    NASA Technical Reports Server (NTRS)

    Mokashi, A. R.

    1982-01-01

    The cost of producing wafers from silicon ingots is a major component of the add-on price of silicon sheet. Economic analyses of the add-on price estimates and their sensitivity internal-diameter (ID) sawing, multiblade slurry (MBS) sawing and fixed-abrasive slicing technique (FAST) are presented. Interim price estimation guidelines (IPEG) are used for estimating a process add-on price. Sensitivity analysis of price is performed with respect to cost parameters such as equipment, space, direct labor, materials (blade life) and utilities, and the production parameters such as slicing rate, slices per centimeter and process yield, using a computer program specifically developed to do sensitivity analysis with IPEG. The results aid in identifying the important cost parameters and assist in deciding the direction of technology development efforts.

  11. Selective skin sensitivity changes and sensory reweighting following short-duration space flight.

    PubMed

    Lowrey, Catherine R; Perry, Stephen D; Strzalkowski, Nicholas D J; Williams, David R; Wood, Scott J; Bent, Leah R

    2014-03-15

    Skin sensory input from the foot soles is coupled with vestibular input to facilitate body orientation in a gravitational environment. Anecdotal observations suggest that foot sole skin becomes hypersensitive following space flight. The veritable level of skin sensitivity and its impact on postural disequilibrium observed post space flight have not been documented. Skin sensitivity of astronauts (n = 11) was measured as vibration perception at the great toe, fifth metatarsal and heel. Frequencies targeted four classes of receptors: 3 and 25 Hz for slow-adapting (SA) receptors and 60 and 250 Hz for fast-adapting (FA) receptors. Data were collected pre- and post-space flight. We hypothesized that skin sensitivity would increase post-space flight and correlate to balance measures. Decreased skin sensitivity was found on landing day at 3 and 25 Hz on the great toe. Hypersensitivity was found for a subset of astronauts (n = 6) with significantly increased sensitivity to 250 Hz at the heel. This subset displayed a greater reduction in computerized dynamic posturography (CDP) equilibrium (EQ) scores (-54%) on landing vs. non-hypersensitive participants (-11%). Observed hyposensitivity of SA (pressure) receptors may indicate a strategy to reduce pressure input during periods of unloading. Hypersensitivity of FAs coupled with reduced EQ scores may reflect targeted sensory reweighting. Altered gravito-inertial environments reduce vestibular function in balance control which may trigger increased weighting of FAs (that signal foot contact, slips). Understanding modulations to skin sensitivity has translational implications for mitigating postural disequilibrium following space flight and for on-Earth preventative strategies for imbalance in older adults.

  12. Homomers of alpha 8 and alpha 7 subunits of nicotinic receptors exhibit similar channel but contrasting binding site properties.

    PubMed

    Gerzanich, V; Anand, R; Lindstrom, J

    1994-02-01

    alpha 8 subunits of alpha-bungarotoxin-sensitive chick neuronal nicotinic acetylcholine receptors expressed in Xenopus oocytes from cRNA are shown to form homomeric, acetylcholine-gated, rapidly desensitizing, inwardly rectifying, Ca(2+)-permeable cation channels similar to those of alpha 7 homomers. alpha 8 forms oligomers of several sizes, of which < 14% are expressed on the oocyte surface, which is less efficient than for alpha 7 homomers. alpha 8 homomers are more sensitive to agonists but less sensitive to antagonists than are alpha 7 homomers, and some agonists for alpha 8 homomers are partial agonists or antagonists for alpha 7 homomers. The pharmacological properties of homomers of alpha 8 and alpha 7 subunits generally reflect those of native alpha 8 and alpha 7 receptors.

  13. In vitro sensitivity of Hungarian Actinobaculum suis strains to selected antimicrobials.

    PubMed

    Biksi, I; Major, Andrea; Fodor, L; Szenci, O; Vetési, F

    2003-01-01

    In vitro antimicrobial sensitivity of 12 Hungarian isolates and the type strain ATCC 33144 of Actinobaculum suis to different antimicrobial compounds was determined both by the agar dilution and by the disc diffusion method. By agar dilution, MIC50 values in the range of 0.05-3.125 micrograms/ml were determined for penicillin, ampicillin, ceftiofur, doxycycline, tylosin, pleuromutilins, chloramphenicol, florfenicol, enrofloxacin and lincomycin. The MIC50 value of oxytetracycline and spectinomycin was 6.25 and 12.5 micrograms/ml, respectively. For ofloxacin, flumequine, neomycin, streptomycin, gentamicin, nalidixic acid, nitrofurantoin and sulphamethoxazole + trimethoprim MIC50 values were in the range of 25-100 micrograms/ml. With the disc diffusion method, all strains were sensitive to penicillin, cephalosporins examined, chloramphenicol and florfenicol, tetracyclines examined, pleuromutilins, lincomycin and tylosin. Variable sensitivity was observed for fluoroquinolones (flumequine, enrofloxacin, ofloxacin), most of the strains were susceptible to marbofloxacin. Almost all strains were resistant to aminoglycosides but most of them were sensitive to spectinomycin. A strong correlation was determined for disc diffusion and MIC results (Spearman's rho 0.789, p < 0001). MIC values of the type strain and MIC50 values of other tested strains did not differ significantly. Few strains showed a partially distinct resistance pattern for erythromycin, lincomycin and ampicillin in both methods. PMID:12688126

  14. Co-dominant effect of selected natural dye sensitizers in DSSC performance

    NASA Astrophysics Data System (ADS)

    Lim, Andery; Ekanayake, Piyasiri; Lim, Linda Biaw Leng; Bandara, J. M. R. Sarath

    2016-10-01

    Natural dyes from Ixora coccinea (RX) and Bougainvillea sp. (BG) were extracted and these individual dyes as well as the cocktail dye of RX and BG (1:1 v/v ratio) were investigated as dye sensitizers in dye sensitized solar cells (DSSCs). To analyze the capability of the dyes, various characterization methods were deployed such as incident photon-to-current efficiency (IPCE), current-to-voltage (I-V) characteristics, cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). It was found that DSSC sensitized with RX performed the best with η = 0.76%, followed by the cocktail dye (η = 0.40%) and BG (η = 0.21%). This best overall performance of RX was attributed to the charge transport resistance (Rtr = 3.89 Ω), with the highest electron density (ɳs = 9.45 × 1028) and effective diffusion coefficient (Deff = 2.43 × 10- 8 m2/s), which accounted for high Jsc. Co-dominance performance of DSSC sensitized with cocktail dye was observed where the cocktail dye has demonstrated an improved Voc as compared to RX and improved Jsc against BG, suggesting individual influence from both dyes.

  15. Vibration isolation technology: Sensitivity of selected classes of experiments to residual accelerations

    NASA Technical Reports Server (NTRS)

    Alexander, J. Iwan D.

    1991-01-01

    Work was completed on all aspects of the following tasks: order of magnitude estimates; thermo-capillary convection - two-dimensional (fixed planar surface); thermo-capillary convection - three-dimensional and axisymmetric; liquid bridge/floating zone sensitivity; transport in closed containers; interaction: design and development stages; interaction: testing flight hardware; and reporting. Results are included in the Appendices.

  16. A Highly Sensitive Diketopyrrolopyrrole-Based Ambipolar Transistor for Selective Detection and Discrimination of Xylene Isomers.

    PubMed

    Wang, Bin; Huynh, Tan-Phat; Wu, Weiwei; Hayek, Naseem; Do, Thu Trang; Cancilla, John C; Torrecilla, Jose S; Nahid, Masrur Morshed; Colwell, John M; Gazit, Oz M; Puniredd, Sreenivasa Reddy; McNeill, Christopher R; Sonar, Prashant; Haick, Hossam

    2016-06-01

    An ambipolar poly(diketopyrrolopyrrole-terthiophene)-based field-effect transistor (FET) sensitively detects xylene isomers at low ppm levels with multiple sensing features. Combined with pattern-recognition algorithms, a sole ambipolar FET sensor, rather than arrays of sensors, can discriminate highly similar xylene structural isomers from one another. PMID:26996398

  17. A Highly Sensitive Diketopyrrolopyrrole-Based Ambipolar Transistor for Selective Detection and Discrimination of Xylene Isomers.

    PubMed

    Wang, Bin; Huynh, Tan-Phat; Wu, Weiwei; Hayek, Naseem; Do, Thu Trang; Cancilla, John C; Torrecilla, Jose S; Nahid, Masrur Morshed; Colwell, John M; Gazit, Oz M; Puniredd, Sreenivasa Reddy; McNeill, Christopher R; Sonar, Prashant; Haick, Hossam

    2016-06-01

    An ambipolar poly(diketopyrrolopyrrole-terthiophene)-based field-effect transistor (FET) sensitively detects xylene isomers at low ppm levels with multiple sensing features. Combined with pattern-recognition algorithms, a sole ambipolar FET sensor, rather than arrays of sensors, can discriminate highly similar xylene structural isomers from one another.

  18. The Usefulness of Selected Physicochemical Indices, Cell Membrane Integrity and Sperm Chromatin Structure in Assessments of Boar Semen Sensitivity

    PubMed Central

    Wysokińska, A.; Kondracki, S.; Iwanina, M.

    2015-01-01

    The present work describes experiments undertaken to evaluate the usefulness of selected physicochemical indices of semen, cell membrane integrity and sperm chromatin structure for the assessment of boar semen sensitivity to processes connected with pre-insemination procedures. The experiments were carried out on 30 boars: including 15 regarded as providers of sensitive semen and 15 regarded as providers of semen that is little sensitive to laboratory processing. The selection of boars for both groups was based on sperm morphology analyses, assuming secondary morphological change incidence in spermatozoa as the criterion. Two ejaculates were manually collected from each boar at an interval of 3 to 4 months. The following analyses were carried out for each ejaculate: sperm motility assessment, sperm pH measurement, sperm morphology assessment, sperm chromatin structure evaluation and cell membrane integrity assessment. The analyses were performed three times. Semen storage did not cause an increase in the incidence of secondary morphological changes in the group of boars considered to provide sperm of low sensitivity. On the other hand, with continued storage there was a marked increase in the incidence of spermatozoa with secondary morphological changes in the group of boars regarded as producing more sensitive semen. Ejaculates of group I boars evaluated directly after collection had an approximately 6% smaller share of spermatozoa with undamaged cell membranes than the ejaculates of boars in group II (p≤0.05). In the process of time the percentage of spermatozoa with undamaged cell membranes decreased. The sperm of group I boars was characterised with a lower sperm motility than the semen of group II boars. After 1 hour of storing diluted semen, the sperm motility of boars producing highly sensitive semen was already 4% lower (p≤0.05), and after 24 hours of storage it was 6.33% lower than that of the boars that produced semen with a low sensitivity. Factors

  19. A Novel Strategy for Gene Selection of Microarray Data Based on Gene-to-Class Sensitivity Information

    PubMed Central

    Han, Fei; Sun, Wei; Ling, Qing-Hua

    2014-01-01

    To obtain predictive genes with lower redundancy and better interpretability, a hybrid gene selection method encoding prior information is proposed in this paper. To begin with, the prior information referred to as gene-to-class sensitivity (GCS) of all genes from microarray data is exploited by a single hidden layered feedforward neural network (SLFN). Then, to select more representative and lower redundant genes, all genes are grouped into some clusters by K-means method, and some low sensitive genes are filtered out according to their GCS values. Finally, a modified binary particle swarm optimization (BPSO) encoding the GCS information is proposed to perform further gene selection from the remainder genes. For considering the GCS information, the proposed method selects those genes highly correlated to sample classes. Thus, the low redundant gene subsets obtained by the proposed method also contribute to improve classification accuracy on microarray data. The experiments results on some open microarray data verify the effectiveness and efficiency of the proposed approach. PMID:24844313

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

    SciTech Connect

    Yang Yongmin; Barankiewicz, Teresa J.; He Mingyue; Taussig, Michael J.; Chen, Swey-Shen . E-mail: alexchen.IGE.com@gmail.com

    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.

  1. Selective T-cell Ablation with Bismuth-213 Labeled Anti-TCR Alpha Beta as Nonmyeloablative Conditionaing for Allogeneic Canine Marrow Transplantion

    SciTech Connect

    Bethge, W. A.; Wilbur, D. Scott; Storb, R.; Hamlin, Donald K.; Santos, E. B.; Brechbiel, M. W.; Fisher, Darrell R.; Sandmaier, B. M.

    2003-06-15

    Two major immunological barriers, the host versus graft (HVG) and the graft versus host (GVH) reaction, must be overcome for successful allogeneic hematopoietic stem cell transplantation. T-cells are involved in these barriers in the major histocompatibility complex-identical settings. We hypothesized that selective ablation of T-cells using radioimmunotherapy, together with postgrafting immunosuppression, would ensure stable allogeneic engraftment. We developed a canine model of nonmyeloablative marrow transplantation in which host immune reactions are impaired by a single dose of 2 Gy total body irradiation (TBI), and where both GVH and residual HVG reactions are controlled by postgrafting immunosuppression with mycophenolate mofetil (MMF) and cyclosporine (CSP). We substituted the alpha-emitter bismuth-213 linked to a monoclonal antibody against TCR(alpha,beta)using the metal-binding chelate CHX-A”-DTPA, for 2 Gy TBI. Biodistribution studies using a gamma-emitting indium-111-labeled anti-TCR mAb showed uptake primarily in blood, marrow, lymph nodes, spleen and liver. In a dosimetry study, 4 dogs were treated with 0.13-0.46 mg/kg TCR mAb labeled with 3.7-5.6 mCi/kg (137-207 MBq/kg) Bi-213. The treatment was administered in 6 injections on days -3 and -2 followed by transplantion of dog leukocyte antigen-identical marrow on day 0 and postgrafting immunosuppression with MMF and CSP. Therapy was well tolerated except for elevations of transaminases, which were transient in all but one dog. No other organ toxicities or signs of graft-versus-host-disease were noted. The dogs had prompt allogeneic hematopoietic engraftment and achieved stable mixed donor-host hematopoietic chimerism with donor contributions ranging from 5-55 % with >30 weeks follow up.

  2. Carbon-fiber microelectrodes modified with 4-sulfobenzene have increased sensitivity and selectivity for catecholamines

    PubMed Central

    Hermans, Andre; Seipel, Andrew T.; Miller, Charles E.; Wightman, R. Mark

    2008-01-01

    Elliptical and cylindrical geometries of carbon-fiber microelectrodes were modified by covalent attachment of 4-sulfobenzenediazonium tetrafluoroborate following its electroreduction. Elliptical electrodes fabricated from Thornel P-55 carbon fibers show the highest amount of 4-sulfobenzene attached to the electrode. Fast-scan cyclic voltammetry was used to compare the response to dopamine and other neurochemicals at these modified carbon-fiber microelectrodes. The grafted layer causes an increased sensitivity to dopamine and other positively charged analytes that is due to increased adsorption of analyte in the grafted layer. However, this layer remains permeable to negatively charged compounds. Modified electrodes retain the increased sensitivity for dopamine during measurements in mouse brain tissue. PMID:16489775

  3. Prior exposure to chemotherapy: a marker of sensitivity and selection for antiangiogenic therapy in breast cancer?

    PubMed

    Xynos, Ioannis D; Tanna, Nikhil; Patten, Darren K; Palmieri, Carlo

    2014-02-01

    The identification of patients who are more likely to derive benefit from antiangiogenic therapy is a key to refine patient selection and so maximize clinical benefit, and reduce unnecessary treatment costs. Improved patient selection will equally be effective in minimizing the exposure of non-eligible patients to ineffectual treatment which could be associated with adverse effects as well as delaying effective treatment. Herein, we review the literature from clinical trials suggesting that the addition of antiangiogenic agents to chemotherapy for the treatment of HER-2 negative metastatic breast cancer in patients previously exposed to chemotherapy may deliver differential therapeutic benefit and may serve as a selection criteria in the current absence of a robust biomarker.

  4. The effect of selective photosuppression of sensitized pathogenic microflora: Part II. Experimental validation on animals

    NASA Astrophysics Data System (ADS)

    Masychev, Viktor I.; Risovannaya, Olga N.

    2005-03-01

    Results of in vivo experiments have shown the maximum effectiveness of combined use of photo sensitizer 0,1% gel Radachlorine simultaneously with continuous and super pulse low energy irradiation of the diode laser with energy density 400 J/cm2, and power 1W. Given parameters have lead to complete elimination of Streptococcus pyogenes from inflammation foci in oral cavity of experimental animals.

  5. Vanilloids selectively sensitize thermal glutamate release from TRPV1 expressing solitary tract afferents.

    PubMed

    Hofmann, Mackenzie E; Andresen, Michael C

    2016-02-01

    Vanilloids, high temperature, and low pH activate the transient receptor potential vanilloid type 1 (TRPV1) receptor. In spinal dorsal root ganglia, co-activation of one of these gating sites on TRPV1 sensitized receptor gating by other modes. Here in rat brainstem slices, we examined glutamate synaptic transmission in nucleus of the solitary tract (NTS) neurons where most cranial primary afferents express TRPV1, but TRPV1 sensitization is unknown. Electrical shocks to the solitary tract (ST) evoked EPSCs (ST-EPSCs). Activation of TRPV1 with capsaicin (100 nM) increased spontaneous EPSCs (sEPSCs) but inhibited ST-EPSCs. High concentrations of the ultra-potent vanilloid resiniferatoxin (RTX, 1 nM) similarly increased sEPSC rates but blocked ST-EPSCs. Lowering the RTX concentration to 150 pM modestly increased the frequency of the sEPSCs without causing failures in the evoked ST-EPSCs. The sEPSC rate increased with raising bath temperature to 36 °C. Such thermal responses were larger in 150 pM RTX, while the ST-EPSCs remained unaffected. Vanilloid sensitization of thermal responses persisted in TTX but was blocked by the TRPV1 antagonist capsazepine. Our results demonstrate that multimodal activation of TRPV1 facilitates sEPSC responses in more than the arithmetic sum of the two activators, i.e. co-activation sensitizes TRPV1 control of spontaneous glutamate release. Since action potential evoked glutamate release is unaltered, the work provides evidence for cooperativity in gating TRPV1 plus a remarkable separation of calcium mechanisms governing the independent vesicle pools responsible for spontaneous and evoked release at primary afferents in the NTS.

  6. Temperature-Sensitive Mutants of a Chinese Hamster Cell Line. I. Selection of Clones with Defective Macromolecular Biosynthesis

    PubMed Central

    Roufa, Donald J.; Reed, Susan J.

    1975-01-01

    Temperature-sensitive clones have been selected from a mutagenized culture of Chinese hamster lung cells by a procedure involving bromodeoxy-uridine (BrdU) incorporation and irradiation with black light. The selection procedure used in these studies was adapted from methods developed by others to yield mutants that cease DNA replication within a short time after they are transferred to nonpermissive temperature. After mutagenesis with ethyl methanosulfonate ten clones survived the selection procedure. Three of the clones (mutants) were temperature-sensitive as measured by growth properties. Two mutants ceased DNA synthesis within six hours of being shifted to 39° and the third mutant continued to synthesize DNA at nonpermissive temperature at a reduced rate for at least 24 hours. Thus, all three mutants survived the selection procedure for understandable reasons, since each was unable to incorporate sufficient BrdU at 39° to lethally protosensitize its DNA during the standard exposure period. The two mutants that cease DNA synthesis at high temperature (clones 115–47 and 115–53) also stop incorporating radioactive amino acids and uridine within six hours at 39°. Their complex phenotype, i.e. defective DNA, RNA and protein biosynthesis, is reversible. When these mutants were returned to 33° after 8 hours at 39°, both resumed DNA synthesis immediately (< 1 hour). Reversal of defective DNA synthesis in both mutants was sensitive to drugs that inhibit protein biosynthesis specifically. Those same drugs, as well as toxic amino acids analogs, also effected a striking mutant phenocopy in wild-type cells. The phenocopy produced by amino acid analogs that are incorporated into mammalian proteins suggested that one or more proteins must be synthesized continuously to support mammalian cells engaged in programmed DNA replication. PMID:1232024

  7. MASS MEASUREMENTS BY AN ACCURATE AND SENSITIVE SELECTED ION RECORDING TECHNIQUE

    EPA Science Inventory

    Trace-level components of mixtures were successfully identified or confirmed by mass spectrometric accurate mass measurements, made at high resolution with selected ion recording, using GC and LC sample introduction. Measurements were made at 20 000 or 10 000 resolution, respecti...

  8. Brief Report: Sensitivity of Children with Autism Spectrum Disorders to Face Appearance in Selective Trust

    ERIC Educational Resources Information Center

    Li, Pengli; Zhang, Chunhua; Yi, Li

    2016-01-01

    The current study examined how children with Autism Spectrum Disorders (ASD) could selectively trust others based on three facial cues: the face race, attractiveness, and trustworthiness. In a computer-based hide-and-seek game, two face images, which differed significantly in one of the three facial cues, were presented as two cues for selective…

  9. Mesoporous carbon nitride based biosensor for highly sensitive and selective analysis of phenol and catechol in compost bioremediation.

    PubMed

    Zhou, Yaoyu; Tang, Lin; Zeng, Guangming; Chen, Jun; Cai, Ye; Zhang, Yi; Yang, Guide; Liu, Yuanyuan; Zhang, Chen; Tang, Wangwang

    2014-11-15

    Herein, we reported here a promising biosensor by taking advantage of the unique ordered mesoporous carbon nitride material (MCN) to convert the recognition information into a detectable signal with enzyme firstly, which could realize the sensitive, especially, selective detection of catechol and phenol in compost bioremediation samples. The mechanism including the MCN based on electrochemical, biosensor assembly, enzyme immobilization, and enzyme kinetics (elucidating the lower detection limit, different linear range and sensitivity) was discussed in detail. Under optimal conditions, GCE/MCN/Tyr biosensor was evaluated by chronoamperometry measurements and the reduction current of phenol and catechol was proportional to their concentration in the range of 5.00 × 10(-8)-9.50 × 10(-6)M and 5.00 × 10(-8)-1.25 × 10(-5)M with a correlation coefficient of 0.9991 and 0.9881, respectively. The detection limits of catechol and phenol were 10.24 nM and 15.00 nM (S/N=3), respectively. Besides, the data obtained from interference experiments indicated that the biosensor had good specificity. All the results showed that this material is suitable for load enzyme and applied to the biosensor due to the proposed biosensor exhibited improved analytical performances in terms of the detection limit and specificity, provided a powerful tool for rapid, sensitive, especially, selective monitoring of catechol and phenol simultaneously. Moreover, the obtained results may open the way to other MCN-enzyme applications in the environmental field.

  10. Highly sensitive and selective determination of methylergometrine maleate using carbon nanofibers/silver nanoparticles composite modified carbon paste electrode.

    PubMed

    Kalambate, Pramod K; Rawool, Chaitali R; Karna, Shashi P; Srivastava, Ashwini K

    2016-12-01

    A highly sensitive and selective voltammetric method for determination of Methylergometrine maleate (MM) in pharmaceutical formulations, urine and blood serum samples has been developed based on enhanced electrochemical response of MM at carbon nanofibers and silver nanoparticles modified carbon paste electrode (CNF-AgNP-CPE). The electrode material was characterized by various techniques viz., X-ray diffraction, scanning electron microscopy and energy dispersive X-ray spectroscopy. The electrocatalytic response of MM at CNF-AgNP-CPE was studied by cyclic voltammetry (CV), differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). Under optimized conditions, the proposed sensor exhibits excellent electrochemical response towards MM. The DPV study shows greatly enhanced electrochemical signal for MM at CNF-AgNP-CPE lending high sensitivity to the proposed sensor for MM detection. The peak (Ip) current for MM is found to be rectilinear in the range 4.0×10(-8)-2.0×10(-5)M with a detection limit of 7.1×10(-9)M using DPV. The feasibility of the proposed sensor in analytical applications was investigated by conducting experiments on commercial pharmaceutical formulations, human urine and blood serum samples, which yielded satisfactory recoveries of MM. The proposed electrochemical sensor offers high sensitivity, selectivity, reproducibility and practical utility. We recommend it as an authentic and productive electrochemical sensor for successful determination of MM. PMID:27612735

  11. Single-walled carbon nanotube field-effect transistors with graphene oxide passivation for fast, sensitive, and selective protein detection.

    PubMed

    Chang, Jingbo; Mao, Shun; Zhang, Yang; Cui, Shumao; Steeber, Douglas A; Chen, Junhong

    2013-04-15

    We report a novel technique to design an insulating membrane with attachment sites on top of single-walled carbon nanotubes (SWNTs) for achieving high sensitivity and selectivity in an SWNT field-effect transistor (FET) biosensor. Because electronic properties of SWNTs are extremely sensitive to the surface state, direct immobilization of proteins or DNAs onto SWNTs will generate surface defects through chemical reactions or physical adsorption, resulting in degradation of performance and instability of SWNT-FET biosensor devices. Here we demonstrate fabrication of novel FET biosensor devices using SWNTs as semiconducting channels, and a monolayer of graphene oxide (GO) membrane covered on the SWNTs as a passivating layer to avoid direct attachment of biomaterials on SWNTs, thereby preserving intrinsic electrical properties of SWNTs. Gold nanoparticles (Au NPs) are decorated on the GO layer for the covalent attachment of biotin, which is then used to selectively detect the target avidin. The passivation with GO layers can effectively lead to enhanced sensitivity of biosensor devices through increasing the on/off ratio of FET sensors.

  12. Highly sensitive and selective determination of methylergometrine maleate using carbon nanofibers/silver nanoparticles composite modified carbon paste electrode.

    PubMed

    Kalambate, Pramod K; Rawool, Chaitali R; Karna, Shashi P; Srivastava, Ashwini K

    2016-12-01

    A highly sensitive and selective voltammetric method for determination of Methylergometrine maleate (MM) in pharmaceutical formulations, urine and blood serum samples has been developed based on enhanced electrochemical response of MM at carbon nanofibers and silver nanoparticles modified carbon paste electrode (CNF-AgNP-CPE). The electrode material was characterized by various techniques viz., X-ray diffraction, scanning electron microscopy and energy dispersive X-ray spectroscopy. The electrocatalytic response of MM at CNF-AgNP-CPE was studied by cyclic voltammetry (CV), differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). Under optimized conditions, the proposed sensor exhibits excellent electrochemical response towards MM. The DPV study shows greatly enhanced electrochemical signal for MM at CNF-AgNP-CPE lending high sensitivity to the proposed sensor for MM detection. The peak (Ip) current for MM is found to be rectilinear in the range 4.0×10(-8)-2.0×10(-5)M with a detection limit of 7.1×10(-9)M using DPV. The feasibility of the proposed sensor in analytical applications was investigated by conducting experiments on commercial pharmaceutical formulations, human urine and blood serum samples, which yielded satisfactory recoveries of MM. The proposed electrochemical sensor offers high sensitivity, selectivity, reproducibility and practical utility. We recommend it as an authentic and productive electrochemical sensor for successful determination of MM.

  13. Single-walled carbon nanotube field-effect transistors with graphene oxide passivation for fast, sensitive, and selective protein detection.

    PubMed

    Chang, Jingbo; Mao, Shun; Zhang, Yang; Cui, Shumao; Steeber, Douglas A; Chen, Junhong

    2013-04-15

    We report a novel technique to design an insulating membrane with attachment sites on top of single-walled carbon nanotubes (SWNTs) for achieving high sensitivity and selectivity in an SWNT field-effect transistor (FET) biosensor. Because electronic properties of SWNTs are extremely sensitive to the surface state, direct immobilization of proteins or DNAs onto SWNTs will generate surface defects through chemical reactions or physical adsorption, resulting in degradation of performance and instability of SWNT-FET biosensor devices. Here we demonstrate fabrication of novel FET biosensor devices using SWNTs as semiconducting channels, and a monolayer of graphene oxide (GO) membrane covered on the SWNTs as a passivating layer to avoid direct attachment of biomaterials on SWNTs, thereby preserving intrinsic electrical properties of SWNTs. Gold nanoparticles (Au NPs) are decorated on the GO layer for the covalent attachment of biotin, which is then used to selectively detect the target avidin. The passivation with GO layers can effectively lead to enhanced sensitivity of biosensor devices through increasing the on/off ratio of FET sensors. PMID:23202350

  14. Prevalence of -alpha(3.7) and alpha alpha alpha(anti3.7) alleles in sickle cell trait and beta-thalassemia patients in Mexico.

    PubMed

    Nava, María Paulina; Ibarra, Bertha; Magaña, María Teresa; de la Luz Chávez, María; Perea, F Javier

    2006-01-01

    The aim of this study was to determine the frequency of alpha-globin gene mutations in three groups of Mexican unrelated individuals. The first two groups were normal and sickle cell trait individuals from the Costa Chica region, a place with a 12.8% frequency of HbS carriers, and the third group comprised of Mexican mestizo patients with beta-thalassemia. We searched for -alpha(3.7) and -alpha(4.2) alpha(+)-thalassemia deletion alleles, as well as the alpha alpha alpha(anti3.7) triplication through long-gap PCR. The alleles -alpha(3.7) and alpha alpha alpha(anti3.7) were found in the heterozygote state only; 19% of the normal subjects had the -alpha(3.7) allele, and 2% showed the alpha alpha alpha(anti3.7) allele. In individuals with the sickle cell trait, 17% had the -alpha(3.7) deletion, and the alpha alpha alpha(anti3.7) triplication was observed in 3% of these individuals. We revealed that 16% of the subjects with beta-thalassemia showed the -alpha(3.7) deletion and 28% the alpha alpha alpha(anti3.7) triplication. The -alpha(4.2) deletion was not detected in any individual. The frequency of the -alpha(3.7) allele was roughly the same in the three groups studied; this can be explained by the fact that the three groups have common genes from Africa and the Mediterranean, where a high prevalence of alpha(+)-thalassemia has been observed. To our knowledge, the frequency of alpha alpha alpha(anti3.7) triplication observed in the Mexican beta-thalassemia patients is the highest reported. As the -alpha(3.7) and alpha alpha alpha(anti3.7) alleles are very common in our selected populations, we believe that there is a need to investigate systematically the alpha-globin gene mutations in all hemoglobinopathies in the Mexican population.

  15. Disperse fine equiaxed alpha alumina nanoparticles with narrow size distribution synthesised by selective corrosion and coagulation separation

    PubMed Central

    Pu, Sanxu; Li, Lu; Ma, Ji; Lu, Fuliang; Li, Jiangong

    2015-01-01

    Disperse fine equiaxed α-Al2O3 nanoparticles with narrow size distribution are important materials in nanotechnology and nanomaterials, but syntheses of disperse fine equiaxed α-Al2O3 nanoparticles usually result in fine γ-Al2O3 nanoparticles or large α-Al2O3 nanoparticles larger than 15 nm. α-Al2O3 has a higher surface energy than γ-Al2O3 and becomes thermodynamically not stable with respect to γ-Al2O3 at specific surface areas larger than 100 m2/g (at sizes smaller than 15 nm for spherical particles) at room temperature. So disperse fine equiaxed α-Al2O3 nanoparticles smaller than 15 nm with narrow size distribution are extremely difficult to synthesise. Here we show the successful synthesis of disperse fine equiaxed α-Al2O3 nanoparticles with average sizes below 10 nm and narrow size distribution by selective corrosion and refined fractionated coagulation separation. An almost fully dense nanocrystalline α-Al2O3 ceramic with a relative density of 99.5% and an average grain size of 60 nm can be sintered from disperse fine equiaxed α-Al2O3 nanoparticles with narrow size distribution. PMID:26166455

  16. Disperse fine equiaxed alpha alumina nanoparticles with narrow size distribution synthesised by selective corrosion and coagulation separation

    NASA Astrophysics Data System (ADS)

    Pu, Sanxu; Li, Lu; Ma, Ji; Lu, Fuliang; Li, Jiangong

    2015-07-01

    Disperse fine equiaxed α-Al2O3 nanoparticles with narrow size distribution are important materials in nanotechnology and nanomaterials, but syntheses of disperse fine equiaxed α-Al2O3 nanoparticles usually result in fine γ-Al2O3 nanoparticles or large α-Al2O3 nanoparticles larger than 15 nm. α-Al2O3 has a higher surface energy than γ-Al2O3 and becomes thermodynamically not stable with respect to γ-Al2O3 at specific surface areas larger than 100 m2/g (at sizes smaller than 15 nm for spherical particles) at room temperature. So disperse fine equiaxed α-Al2O3 nanoparticles smaller than 15 nm with narrow size distribution are extremely difficult to synthesise. Here we show the successful synthesis of disperse fine equiaxed α-Al2O3 nanoparticles with average sizes below 10 nm and narrow size distribution by selective corrosion and refined fractionated coagulation separation. An almost fully dense nanocrystalline α-Al2O3 ceramic with a relative density of 99.5% and an average grain size of 60 nm can be sintered from disperse fine equiaxed α-Al2O3 nanoparticles with narrow size distribution.

  17. IL-6 mediated isotype specific suppression of hapten specific IgE in serum of BPO-KLH sensitized mice: role of IFN alpha in maintainance of hapten specific IgE responses.

    PubMed

    Auci, D L; Miller, H; Chice, S M; Durkin, H G

    1994-04-01

    The ability of IL-6 or IFN alpha or antibodies to these cytokines to regulate serum levels of hapten specific immunoglobulins (IgM, IgG1, IgE, IgA) was studied in BPO-KLH (benzylpenicilloyl-keyhole limpet hemocyanin) sensitized BALB/c mice at the peak of a hapten specific IgE antibody forming cell (AFC) response. To induce peak IgE responses, mice were injected intraperitonealy (i.p.) with BPO-KLH (10 micrograms) in aluminum hydroxide gel (alum) on days 0, 21, and 42. On day 44, mice were injected s.c. with IL-6 (100-1000 U), IFN alpha (1000-10,000 U), anti-IL-6 (100-1000 neutralizing units [NU]), or anti-IFN alpha (1000-10,000 NU). On day 46, levels of BPO specific IgM, IgG1, IgE and IgA in serum were determined (ELISA). Data are expressed as micrograms/ml. IL-6 suppressed BPO specific IgE in serum in isotype specific fashion (to > 90%), increasing IgA (approximately 3 fold), and leaving IgM and IgG1 unchanged. Since removal of endogenous IL-6 with anti-IL-6 increased serum IgE, and suppressed IgG1 (approximately 50%), with IgM and IgA unchanged, this suggests that IL-6 is an isotype specific suppressor of peak IgE responses and as such may be useful in the therapeutic management of atopic disease. IFN alpha treatment increased serum IgE levels (60%), and potentiated IgA responses (> 30 fold), with IgM and IgG1 unchanged. Since removal of endogenous IFN alpha with anti-IFN alpha decreased IgE levels (approximately 50%), increasing IgA, with IgM and IgG1 unchanged, this suggests a role for IFN alpha as an isotype specific helper of peak IgE responses and in maintenance of IgA responses.

  18. AMPA receptors and motivation for drug: effect of the selective antagonist NBQX on behavioural sensitization and on self-administration in mice.

    PubMed

    Jackson, A; Mead, A N; Rocha, B A; Stephens, D N

    1998-09-01

    A series of experiments was carried out in which the potency of the selective alpha-amino-3-hydroxy-5-methyl-4-isoxazole proprionate (AMPA)-receptor antagonist 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)quinoxaline (NBQX) (10-100 mg/kg) on locomotor activity was investigated, in mice. NBQX reduced all forms of activity studied, but its potency to do so varied according to the conditions of the experiment. The smallest dose of NBQX significantly reducing spontaneous or cocaine-induced activity was 100 mg/kg. Mice that had been repeatedly treated with 16 mg/kg cocaine once per week, for 7 weeks, showed a sensitized locomotor response to a challenge dose of cocaine (16 mg/kg). NBQX reversed the sensitized response at 30 and 100 mg/kg. The pattern of results obtained leaves open the role that AMPA-receptors may have in the expression of behavioural sensitization. In two further experiments, mice were trained to self-administer cocaine (30 micrograms per reinforcer) via intravenous catheters, using an operant lever pressing technique. When the amount of cocaine per reinforcer was doubled (to 60 micrograms) or halved (to 15 micrograms) the mice adapted lever pressing rates to maintain some constancy of self-dosing (but not at 7.5 micrograms per reinforcer) and when saline was substituted for cocaine, response rates increased considerably (extinction bursting). NBQX (10 and 30 mg/kg) reduced the self-administration of 30 micrograms reinforcers of cocaine, but only during the first 30 min of the test session. There was no evidence that NBQX specifically antagonized the reinforcing effect of cocaine, as responding was similarly reduced on both the reinforced and the non-reinforced lever, nor did the response to NBQX mimic behaviour seen following changes in the concentration of the reinforcer. The results of the locomotor experiments and the self-administration experiments are discussed together, in terms of current hypotheses about glutamatergic mechanisms involved in

  19. Error modeling and sensitivity analysis of a parallel robot with SCARA(selective compliance assembly robot arm) motions

    NASA Astrophysics Data System (ADS)

    Chen, Yuzhen; Xie, Fugui; Liu, Xinjun; Zhou, Yanhua

    2014-07-01

    Parallel robots with SCARA(selective compliance assembly robot arm) motions are utilized widely in the field of high speed pick-and-place manipulation. Error modeling for these robots generally simplifies the parallelogram structures included by the robots as a link. As the established error model fails to reflect the error feature of the parallelogram structures, the effect of accuracy design and kinematic calibration based on the error model come to be undermined. An error modeling methodology is proposed to establish an error model of parallel robots with parallelogram structures. The error model can embody the geometric errors of all joints, including the joints of parallelogram structures. Thus it can contain more exhaustively the factors that reduce the accuracy of the robot. Based on the error model and some sensitivity indices defined in the sense of statistics, sensitivity analysis is carried out. Accordingly, some atlases are depicted to express each geometric error's influence on the moving platform's pose errors. From these atlases, the geometric errors that have greater impact on the accuracy of the moving platform are identified, and some sensitive areas where the pose errors of the moving platform are extremely sensitive to the geometric errors are also figured out. By taking into account the error factors which are generally neglected in all existing modeling methods, the proposed modeling method can thoroughly disclose the process of error transmission and enhance the efficacy of accuracy design and calibration.

  20. Electrosprayed Metal Oxide Semiconductor Films for Sensitive and Selective Detection of Hydrogen Sulfide

    PubMed Central

    Ghimbeu, Camelia Matei; Lumbreras, Martine; Schoonman, Joop; Siadat, Maryam

    2009-01-01

    Semiconductor metal oxide films of copper-doped tin oxide (Cu-SnO2), tungsten oxide (WO3) and indium oxide (In2O3) were deposited on a platinum coated alumina substrate employing the electrostatic spray deposition technique (ESD). The morphology studied with scanning electron microscopy (SEM) and atomic force microscopy (AFM) shows porous homogeneous films comprising uniformly distributed aggregates of nano particles. The X-ray diffraction technique (XRD) proves the formation of crystalline phases with no impurities. Besides, the Raman cartographies provided information about the structural homogeneity. Some of the films are highly sensitive to low concentrations of H2S (10 ppm) at low operating temperatures (100 and 200 °C) and the best response in terms of Rair/Rgas is given by Cu-SnO2 films (2500) followed by WO3 (1200) and In2O3 (75). Moreover, all the films exhibit no cross-sensitivity to other reducing (SO2) or oxidizing (NO2) gases. PMID:22291557

  1. Electrosprayed metal oxide semiconductor films for sensitive and selective detection of hydrogen sulfide.

    PubMed

    Ghimbeu, Camelia Matei; Lumbreras, Martine; Schoonman, Joop; Siadat, Maryam

    2009-01-01

    Semiconductor metal oxide films of copper-doped tin oxide (Cu-SnO(2)), tungsten oxide (WO(3)) and indium oxide (In(2)O(3)) were deposited on a platinum coated alumina substrate employing the electrostatic spray deposition technique (ESD). The morphology studied with scanning electron microscopy (SEM) and atomic force microscopy (AFM) shows porous homogeneous films comprising uniformly distributed aggregates of nano particles. The X-ray diffraction technique (XRD) proves the formation of crystalline phases with no impurities. Besides, the Raman cartographies provided information about the structural homogeneity. Some of the films are highly sensitive to low concentrations of H(2)S (10 ppm) at low operating temperatures (100 and 200 °C) and the best response in terms of R(air)/R(gas) is given by Cu-SnO(2) films (2500) followed by WO(3) (1200) and In(2)O(3) (75). Moreover, all the films exhibit no cross-sensitivity to other reducing (SO(2)) or oxidizing (NO(2)) gases. PMID:22291557

  2. RNA aptamer based electrochemical biosensor for sensitive and selective detection of cAMP.

    PubMed

    Zhao, Fulin; Xie, Qingyun; Xu, Mingfei; Wang, Shouyu; Zhou, Jiyong; Liu, Fei

    2015-04-15

    Cyclic adenosine monophosphate (cAMP) is an important small biological molecule associated with the healthy state of living organism. In order to realize highly sensitive and specific detection of cAMP, here an RNA aptamer and electrochemical impedance spectroscopy (EIS) based biosensor enhanced by gold nanoparticles electrodeposited on the surface of gold electrode is designed. The designed aptasensor has a wide effective measuring range from 50pM to 250pM with a detection limit of 50pM in PBS buffer, and an effective measuring range from 50nM to 1μM with a detection limit of 50nM in serum. The designed biosensor is also able to detect cAMP with high sensitivity, specificity, and stability. Since the biosensor can be easily fabricated with low cost and repeatedly used for at least two times, it owns great potential in wide application fields such as clinical test and food inspection, etc.

  3. Sensitive and selective amplified fluorescence DNA detection based on exonuclease III-aided target recycling.

    PubMed

    Zuo, Xiaolei; Xia, Fan; Xiao, Yi; Plaxco, Kevin W

    2010-02-17

    A limitation of many traditional approaches to the detection of specific oligonucleotide sequences, such as molecular beacons, is that each target strand hybridizes with (and thus activates) only a single copy of the relevant probe sequence. This 1:1 hybridization ratio limits the gain of most approaches and thus their sensitivity. Here we demonstrate a nuclease-amplified DNA detection scheme in which exonuclease III is used to "recycle" target molecules, thus leading to greatly improved sensitivity relative to, for example, traditional molecular beacons without any significant restriction in the choice of target sequences. The exonuclease-amplified assay can detect target DNA at concentrations as low as 10 pM when performed at 37 degrees C, which represents a significant improvement over the equivalent molecular beacon alone. Moreover, at 4 degrees C we can obtain a detection limit as low as 20 aM, albeit at the cost of a 24 h incubation period. Finally, our assay can be easily interrogated with the naked eye and is thus amenable to deployment in the developing world, where fluorometric detection is more problematic.

  4. A highly sensitive and selective diagnostic assay based on virus nanoparticles

    NASA Astrophysics Data System (ADS)

    Park, Jin-Seung; Cho, Moon Kyu; Lee, Eun Jung; Ahn, Keum-Young; Lee, Kyung Eun; Jung, Jae Hun; Cho, Yunjung; Han, Sung-Sik; Kim, Young Keun; Lee, Jeewon

    2009-04-01

    Early detection of the protein marker troponin I in patients with a higher risk of acute myocardial infarction can reduce the risk of death from heart attacks. Most troponin assays are currently based on the conventional enzyme linked immunosorbent assay and have detection limits in the nano- and picomolar range. Here, we show that by combining viral nanoparticles, which are engineered to have dual affinity for troponin antibodies and nickel, with three-dimensional nanostructures including nickel nanohairs, we can detect troponin levels in human serum samples that are six to seven orders of magnitude lower than those detectable using conventional enzyme linked immunosorbent assays. The viral nanoparticle helps to orient the antibodies for maximum capture of the troponin markers. High densities of antibodies on the surfaces of the nanoparticles and nanohairs lead to greater binding of the troponin markers, which significantly enhances detection sensitivities. The nickel nanohairs are re-useable and can reproducibly differentiate healthy serum from unhealthy ones. We expect other viral nanoparticles to form similar highly sensitive diagnostic assays for a variety of other protein markers.

  5. Ion imprinted polymeric nanoparticles for selective separation and sensitive determination of zinc ions in different matrices.

    PubMed

    Shamsipur, Mojtaba; Rajabi, Hamid Reza; Pourmortazavi, Seied Mahdi; Roushani, Mahmoud

    2014-01-01

    Preparation of Zn(2+) ion-imprinted polymer (Zn-IIP) nanoparticles is presented in this report. The Zn-IIP nanoparticles are prepared by dissolving stoichiometric amounts of zinc nitrate and selected chelating ligand, 3,5,7,20,40-pentahydroxyflavone, in 15 mL ethanol-acetonitrile (2:1; v/v) mixture as a porogen solvent in the presence of ethylene glycol-dimethacrylate (EGDMA) as cross-linking, methacrylic acid (MAA) as functional monomer, and 2,2-azobisisobutyronitrile (AIBN) as initiator. After polymerization, Cavities in the polymer particles corresponding to the Zn(2+) ions were created by leaching the polymer in HCl aqueous solution. The synthesized IIPs were characterized by scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy, fluorescence spectroscopy and thermal analysis techniques. Also, the pH range for rebinding of Zn(2+) ion on the IIP and equilibrium binding time were optimized, using flame atomic absorption spectrometry. In selectivity study, it was found that imprinting results increased affinity of the material toward Zn(2+) ion over other competitor metal ions with the same charge and close ionic radius. The prepared IIPs were repeatedly used and regenerated for six times without any significant decrease in polymer binding affinities. Finally, the prepared sorbent was successfully applied to the selective recognition and determination of zinc ion in different real samples.

  6. Using Temporal Modulation Sensitivity to Select Stimulation Sites for Processor MAPs in Cochlear Implant Listeners

    PubMed Central

    Garadat, Soha N.; Zwolan, Teresa A.; Pfingst, Bryan E.

    2013-01-01

    Previous studies in our laboratory showed that temporal acuity as assessed by modulation detection thresholds (MDTs) varied across activation sites and that this site-to-site variability was subject specific. Using two 10-channel MAPs, the previous experiments showed that processor MAPs that had better across-site mean (ASM) MDTs yielded better speech recognition than MAPs with poorer ASM MDTs tested in the same subject. The current study extends our earlier work on developing more optimal fitting strategies to test the feasibility of using a site-selection approach in the clinical domain. This study examined the hypothesis that revising the clinical speech processor MAP for cochlear implant (CI) recipients by turning off selected sites that have poorer temporal acuity and reallocating frequencies to the remaining electrodes would lead to improved speech recognition. Twelve CI recipients participated in the experiments. We found that site selection procedure based on MDTs in the presence of a masker resulted in improved performance on consonant recognition and recognition of sentences in noise. In contrast, vowel recognition was poorer with the experimental MAP than with the clinical MAP, possibly due to reduced spectral resolution when sites were removed from the experimental MAP. Overall, these results suggest a promising path for improving recipient outcomes using personalized processor-fitting strategies based on a psychophysical measure of temporal acuity. PMID:23881208

  7. Boronic acid functionalized N-doped carbon quantum dots as fluorescent probe for selective and sensitive glucose determination

    NASA Astrophysics Data System (ADS)

    Jiang, Guohua; Jiang, Tengteng; Li, Xia; Wei, Zheng; Du, Xiangxiang; Wang, Xiaohong

    2014-04-01

    Nitrogen doped carbon quantum dots (NCQDs) of about 10 nm in diameter have been obtained by hydrothermal reaction from collagen. Because of the superiority of water dispersion, low toxicity and ease of functionlization, the NCQDs were designed as a glucose sensor after covalent grafting by 3-aminophenylboronic (APBA) (APBA-NCQDs). The as-prepared APBA-NCQDs were imparted with glucose sensitivity and selectivity from other saccharides via fluorescence (FL) quenching effect at physiological pH and at room temperature, which show high sensitivity and specificity for glucose determination with a wide range from 1 mM to 14 mM. FL quenching mechanism of APBA-NCQDs was also investigated by adding an external quencher. The APBA-NCQDs-based platform is an environmentally friendly way to substitute inorganic quantum dots containing heavy metals which offer a facile and low cost detection method.

  8. A sensitive and selective immuno-nanogold resonance-scattering spectral method for the determination of trace penicillin G.

    PubMed

    Jiang, Zhiliang; Li, Yan; Liang, Aihui; Qin, Aimiao

    2008-01-01

    A sensitive and selective immuno-nanogold resonance scattering spectral assay was developed for the determination of trace hapten penicillin G, based on the resonance scattering (RS) effect of the nanogold at 560 nm, and the nanogold-labelled immunoreaction took place in pH 5.4 phosphate citric acid buffer solutions and in the presence of polythylene glycol (PEG). The nanogold-labelled immunocomplex formed more and more with addition of penicillin G. The enhanced RS intensity at 560 nm Delta I(RS) was linear to the penicillin G concentration in the range 7.5-1700 ng/mL, with a detection limit of 0.78 ng/mL. The results indicate that the immunonanogold-labelled RS spectral assay has a high specificity and sensitivity for quantitative determination of penicillin G in raw milk samples.

  9. Highly selective and sensitive detection of Cu2+ with lysine enhancing bovine serum albumin modified-carbon dots fluorescent probe.

    PubMed

    Liu, Jia-Ming; Lin, Li-ping; Wang, Xin-Xing; Lin, Shao-Qin; Cai, Wen-Lian; Zhang, Li-Hong; Zheng, Zhi-Yong

    2012-06-01

    Based on the ability of lysine (Lys) to enhance the fluorescence intensity of bovine serum albumin modified-carbon dots (CDs-BSA) to decrease surface defects and quench fluorescence of the CDs-BSA-Lys system in the presence of Cu(2+) under conditions of phosphate buffer (PBS, pH = 5.0) at 45 °C for 10 min, a sensitive Lys enhancing CDs-BSA fluorescent probe was designed. The environment-friendly, simple, rapid, selective and sensitive fluorescent probe has been utilized to detect Cu(2+) in hair and tap water samples and it achieved consistent results with those obtained by inductively coupled plasma mass spectroscopy (ICP-MS). The mechanism of the proposed assay for the detection of Cu(2+) is discussed. PMID:22531278

  10. Self-locked aptamer probe mediated cascade amplification strategy for highly sensitive and selective detection of protein and small molecule.

    PubMed

    Li, Wei; Jiang, Wei; Wang, Lei

    2016-10-12

    In this work, a novel self-locked aptamer probe mediated cascade amplification strategy has been constructed for highly sensitive and specific detection of protein. First, the self-locked aptamer probe was designed with three functions: one was specific molecular recognition attributed to the aptamer sequence, the second was signal transduction owing to the transduction sequence, and the third was self-locking through the hybridization of the transduction sequence and part of the aptamer sequence. Then, the aptamer sequence specific recognized the target and folded into a three-way helix junction, leading to the release of the transduction sequence. Next, the 3'-end of this three-way junction acted as primer to trigger the strand displacement amplification (SDA), yielding a large amount of primers. Finally, the primers initiated the dual-exponential rolling circle amplification (DE-RCA) and generated numerous G-quadruples sequences. By inserting the fluorescent dye N-methyl mesoporphyrin IX (NMM), enhanced fluorescence signal was achieved. In this strategy, the self-locked aptamer probe was more stable to reduce the interference signals generated by the uncontrollable folding in unbounded state. Through the cascade amplification of SDA and DE-RCA, the sensitivity was further improved with a detection limit of 3.8 × 10(-16) mol/L for protein detection. Furthermore, by changing the aptamer sequence of the probe, sensitive and selective detection of adenosine has been also achieved, suggesting that the proposed strategy has good versatility and can be widely used in sensitive and selective detection of biomolecules.

  11. Self-locked aptamer probe mediated cascade amplification strategy for highly sensitive and selective detection of protein and small molecule.

    PubMed

    Li, Wei; Jiang, Wei; Wang, Lei

    2016-10-12

    In this work, a novel self-locked aptamer probe mediated cascade amplification strategy has been constructed for highly sensitive and specific detection of protein. First, the self-locked aptamer probe was designed with three functions: one was specific molecular recognition attributed to the aptamer sequence, the second was signal transduction owing to the transduction sequence, and the third was self-locking through the hybridization of the transduction sequence and part of the aptamer sequence. Then, the aptamer sequence specific recognized the target and folded into a three-way helix junction, leading to the release of the transduction sequence. Next, the 3'-end of this three-way junction acted as primer to trigger the strand displacement amplification (SDA), yielding a large amount of primers. Finally, the primers initiated the dual-exponential rolling circle amplification (DE-RCA) and generated numerous G-quadruples sequences. By inserting the fluorescent dye N-methyl mesoporphyrin IX (NMM), enhanced fluorescence signal was achieved. In this strategy, the self-locked aptamer probe was more stable to reduce the interference signals generated by the uncontrollable folding in unbounded state. Through the cascade amplification of SDA and DE-RCA, the sensitivity was further improved with a detection limit of 3.8 × 10(-16) mol/L for protein detection. Furthermore, by changing the aptamer sequence of the probe, sensitive and selective detection of adenosine has been also achieved, suggesting that the proposed strategy has good versatility and can be widely used in sensitive and selective detection of biomolecules. PMID:27662754

  12. Recent approaches to improving selectivity and sensitivity of enzyme-based biosensors for organophosphorus pesticides: A review.

    PubMed

    Songa, Everlyne A; Okonkwo, Jonathan O

    2016-08-01

    Pesticide determination has attracted great attention due to the fact that they exhibit high acute toxicity and can cause long-term damage to the environment and human lives even at trace levels. Although classical analytical methods (including gas chromatography, high performance liquid chromatography, capillary electrophoresis and mass spectrometry) have been effectively used for analysis of pesticides in contaminated samples, they present certain limitations such as time-consuming sample preparation, complexity, and the requirement of expensive instrumentation and highly skilled personnel. For these reasons, there is an expanding need for analytical methods able to provide simple, rapid, sensitive, selective, low cost and reliable detection of pesticides at trace levels. Over the past decades, acetylcholinesterase (AChE) biosensors have emerged as simple, rapid and ultra-sensitive tools for toxicity detection of pesticides in the environment and food. These biosensors have the potential to complement or replace the classical analytical methods by simplifying or eliminating sample preparation and making field-testing easier and faster with significant decrease in cost per analysis. With the recent engineering of more sensitive AChE enzymes, the development of more reliable immobilization matrices and the progress in the area of microelectronics, AChE biosensors could become competitive for multi-analyte screening and soon be used for the development of portable instrumentation for rapid toxicity testing of samples. The enzymes organophosphorus hydrolase (OPH) and organophosphorus acid anhydrolase (OPAA) have also shown considerable potential in OP biosensor applications and they have been used for direct detection of OPs. This review presents the recent advances in the fabrication of enzyme biosensors for organophosphorus pesticides (OPs) and their possible applications for toxicity monitoring of organophosphorus pesticide residues in real samples. The focus will

  13. Recent approaches to improving selectivity and sensitivity of enzyme-based biosensors for organophosphorus pesticides: A review.

    PubMed

    Songa, Everlyne A; Okonkwo, Jonathan O

    2016-08-01

    Pesticide determination has attracted great attention due to the fact that they exhibit high acute toxicity and can cause long-term damage to the environment and human lives even at trace levels. Although classical analytical methods (including gas chromatography, high performance liquid chromatography, capillary electrophoresis and mass spectrometry) have been effectively used for analysis of pesticides in contaminated samples, they present certain limitations such as time-consuming sample preparation, complexity, and the requirement of expensive instrumentation and highly skilled personnel. For these reasons, there is an expanding need for analytical methods able to provide simple, rapid, sensitive, selective, low cost and reliable detection of pesticides at trace levels. Over the past decades, acetylcholinesterase (AChE) biosensors have emerged as simple, rapid and ultra-sensitive tools for toxicity detection of pesticides in the environment and food. These biosensors have the potential to complement or replace the classical analytical methods by simplifying or eliminating sample preparation and making field-testing easier and faster with significant decrease in cost per analysis. With the recent engineering of more sensitive AChE enzymes, the development of more reliable immobilization matrices and the progress in the area of microelectronics, AChE biosensors could become competitive for multi-analyte screening and soon be used for the development of portable instrumentation for rapid toxicity testing of samples. The enzymes organophosphorus hydrolase (OPH) and organophosphorus acid anhydrolase (OPAA) have also shown considerable potential in OP biosensor applications and they have been used for direct detection of OPs. This review presents the recent advances in the fabrication of enzyme biosensors for organophosphorus pesticides (OPs) and their possible applications for toxicity monitoring of organophosphorus pesticide residues in real samples. The focus will

  14. Intermittent Preventive Treatment with Dihydroartemisinin-Piperaquine in Ugandan Schoolchildren Selects for Plasmodium falciparum Transporter Polymorphisms That Modify Drug Sensitivity.

    PubMed

    Nankabirwa, Joaniter I; Conrad, Melissa D; Legac, Jennifer; Tukwasibwe, Stephen; Tumwebaze, Patrick; Wandera, Bonnie; Brooker, Simon J; Staedke, Sarah G; Kamya, Moses R; Nsobya, Sam L; Dorsey, Grant; Rosenthal, Philip J

    2016-10-01

    Dihydroartemisinin-piperaquine (DP) offers prolonged protection against malaria, but its impact on Plasmodium falciparum drug sensitivity is uncertain. In a trial of intermittent preventive treatment in schoolchildren in Tororo, Uganda, in 2011 to 2012, monthly DP for 1 year decreased the incidence of malaria by 96% compared to placebo; DP once per school term offered protection primarily during the first month after therapy. To assess the impact of DP on selection of drug resistance, we compared the prevalence of key polymorphisms in isolates that emerged at different intervals after treatment with DP. Blood obtained monthly and at each episode of fever was assessed for P. falciparum parasitemia by microscopy. Samples from 160 symptomatic and 650 asymptomatic episodes of parasitemia were assessed at 4 loci (N86Y, Y184F, and D1246Y in pfmdr1 and K76T in pfcrt) that modulate sensitivity to aminoquinoline antimalarials, utilizing a ligase detection reaction-fluorescent microsphere assay. For pfmdr1 N86Y and pfcrt K76T, but not the other studied polymorphisms, the prevalences of mutant genotypes were significantly greater in children who had received DP within the past 30 days than in those not treated within 60 days (86Y, 18.0% versus 8.3% [P = 0.03]; 76T, 96.0% versus 86.1% [P = 0.05]), suggesting selective pressure of DP. Full sequencing of pfcrt in a subset of samples did not identify additional polymorphisms selected by DP. In summary, parasites that emerged soon after treatment with DP were more likely than parasites not under drug pressure to harbor pfmdr1 and pfcrt polymorphisms associated with decreased sensitivity to aminoquinoline antimalarials. (This study has been registered at ClinicalTrials.gov under no. NCT01231880.). PMID:27401569

  15. Fabrication of bimetallic Cu/Au nanotubes and their sensitive, selective, reproducible and reusable electrochemical sensing of glucose

    NASA Astrophysics Data System (ADS)

    Tee, Si Yin; Ye, Enyi; Pan, Pei Hua; Lee, Coryl Jing Jun; Hui, Hui Kim; Zhang, Shuang-Yuan; Koh, Leng Duei; Dong, Zhili; Han, Ming-Yong

    2015-06-01

    Herein, we report a facile two-step approach to produce gold-incorporated copper (Cu/Au) nanostructures through controlled disproportionation of the Cu+-oleylamine complex at 220 °C to form copper nanowires and the subsequent reaction with Au3+ at different temperatures of 140, 220 and 300 °C. In comparison with copper nanowires, these bimetallic Cu/Au nanostructures exhibit their synergistic effect to greatly enhance glucose oxidation. Among them, the shape-controlled Cu/Au nanotubes prepared at 140 °C show the highest electrocatalytic activity for non-enzymatic glucose sensing in alkaline solution. In addition to high sensitivity and fast response, the Cu/Au nanotubes possess high selectivity against interferences from other potential interfering species and excellent reproducibility with long-term stability. By introducing gold into copper nanostructures at a low level of 3, 1 and 0.1 mol% relative to the initial copper precursor, a significant electrocatalytic enhancement of the resulting bimetallic Cu/Au nanostructures starts to occur at 1 mol%. Overall, the present fabrication of stable Cu/Au nanostructures offers a promising low-cost platform for sensitive, selective, reproducible and reusable electrochemical sensing of glucose.Herein, we report a facile two-step approach to produce gold-incorporated copper (Cu/Au) nanostructures through controlled disproportionation of the Cu+-oleylamine complex at 220 °C to form copper nanowires and the subsequent reaction with Au3+ at different temperatures of 140, 220 and 300 °C. In comparison with copper nanowires, these bimetallic Cu/Au nanostructures exhibit their synergistic effect to greatly enhance glucose oxidation. Among them, the shape-controlled Cu/Au nanotubes prepared at 140 °C show the highest electrocatalytic activity for non-enzymatic glucose sensing in alkaline solution. In addition to high sensitivity and fast response, the Cu/Au nanotubes possess high selectivity against interferences from other

  16. A naphthalene-based two-photon fluorescent probe for selective and sensitive detection of endogenous hypochlorous acid.

    PubMed

    Zhou, Xiao-Hong; Jiang, Yu-Ren; Zhao, Xiong-Jie; Guo, Dong

    2016-11-01

    An efficient naphthalene-based two-photon fluorescent probe for endogenous HClO has been reported in the present study, which consists of a 6-(2-benzothiazolyl)-2-naphthalenol fluorophore connected with a 4-aminophenol (the fluorescence quenching and response group). This probe exhibits a high selectivity and excellent sensitivity with a detection limit of 7.6nM over other reactive oxygen species and analyte species, and the fluorescence intensity enhanced 103-fold when responsed. Furthermore, it was successfully used for two-photon imaging of endogenous HClO in live cells with high-resolution. PMID:27591640

  17. Highly selective and sensitive fluorescence chemosensor for the detection of palladium species based on Tsuji-Trost reaction.

    PubMed

    Xu, Zhong-Yong; Li, Jing; Guan, Su; Zhang, Lei; Dong, Chang-Zhi

    2015-09-01

    A new chemosensor 7-nitro-2,1,3-benzoxadiazole-4-allyl-N-(thiophen-2-ylmethyl)carbamate (NBDTC) was synthesized and utilized for palladium detection based on the Tsuji-Trost reaction. NBDTC displayed specific and ratiometric fluorescent responses toward palladium species. The chemosensor showed more than 50-fold enhancement in fluorescence intensity with the presence of PEG400 and palladium because NBDTC can be transformed to NBDT under palladium-catalyzing Tsuji-Trost reaction. NBDTC displayed high selectivity and sensitivity for palladium species with the detection limit of 1.13×10(-9) M.

  18. Rhodamine 6G hydrazone bearing thiophene unit: A highly sensitive and selective off-on fluorescent chemosensor for Al3+

    NASA Astrophysics Data System (ADS)

    Wu, Wei-Na; Mao, Pan-Dong; Wang, Yuan; Zhao, Xiao-Lei; Jia, Lei; Xu, Zhou-Qing

    2016-10-01

    A rhodamine derivative (R1) has been synthesized by a hydrazone formation of rhodamine 6G hydrazide with 3-methylthiophene-2-carbaldehyde, which exhibits high selectivity and sensitivity as an "off-on" fluorescent sensor toward Al3+ in water containing media. The binding process was confirmed by UV-vis absorption, fluorescence measurements, mass spectroscopy and DFT calculation. The probe functions by Al3+ induced hydrolytic cleavage of the imine-bond to produce an intense rhodamine-based emission. To test the practical use of the probe, the determination of Al3+ in real water samples was also evaluated.

  19. A naphthalene-based two-photon fluorescent probe for selective and sensitive detection of endogenous hypochlorous acid.

    PubMed

    Zhou, Xiao-Hong; Jiang, Yu-Ren; Zhao, Xiong-Jie; Guo, Dong

    2016-11-01

    An efficient naphthalene-based two-photon fluorescent probe for endogenous HClO has been reported in the present study, which consists of a 6-(2-benzothiazolyl)-2-naphthalenol fluorophore connected with a 4-aminophenol (the fluorescence quenching and response group). This probe exhibits a high selectivity and excellent sensitivity with a detection limit of 7.6nM over other reactive oxygen species and analyte species, and the fluorescence intensity enhanced 103-fold when responsed. Furthermore, it was successfully used for two-photon imaging of endogenous HClO in live cells with high-resolution.

  20. Bioluminescence assay platform for selective and sensitive detection of Ub/Ubl proteases.

    PubMed

    Orcutt, Steven J; Wu, Jian; Eddins, Michael J; Leach, Craig A; Strickler, James E

    2012-11-01

    As the importance of ubiquitylation in certain disease states becomes increasingly apparent, the enzymes responsible for removal of ubiquitin (Ub) from target proteins, deubiquitylases (DUBs), are becoming attractive targets for drug discovery. For rapid identification of compounds that alter DUB function, in vitro assays must be able to provide statistically robust data over a wide dynamic range of both substrate and enzyme concentrations during high throughput screening (HTS). The most established reagents for HTS are Ubs with a quenched fluorophore conjugated to the C-terminus; however, a luciferase-based strategy for detecting DUB activity (DUB-Glo™, Promega) provides a wider dynamic range than traditional fluorogenic reagents. Unfortunately, this assay requires high enzyme concentrations and lacks specificity for DUBs over other isopeptidases (e.g. desumoylases), as it is based on an aminoluciferin (AML) derivative of a peptide derived from the C-terminus of Ub (Z-RLRGG-). Conjugation of aminoluciferin to a full-length Ub (Ub-AML) yields a substrate that has a wide dynamic range, yet displays detection limits for DUBs 100- to 1000-fold lower than observed with DUB-Glo™. Ub-AML was even a sensitive substrate for DUBs (e.g. JosD1 and USP14) that do not show appreciable activity with DUB-Glo™. Aminoluciferin derivatives of hSUMO2 and NEDD8 were also shown to be sensitive substrates for desumoylases and deneddylases, respectively. Ub/Ubl-AML substrates are amenable to HTS (Z'=0.67) yielding robust signal, and providing an alternative drug discovery platform for Ub/Ubl isopeptidases. This article is part of a Special Issue entitled: Ubiquitin Drug Discovery and Diagnostics.

  1. Highly selective and sensitive nucleic acid detection based on polysaccharide-functionalized silver nanoparticles

    NASA Astrophysics Data System (ADS)

    Yan, Jing-Kun; Ma, Hai-Le; Cai, Pan-Fu; Wu, Jian-Yong

    2015-01-01

    Polysaccharide-functionalized silver nanoparticles (Oc-AgNPs) with a mean diameter of 15 nm were utilized as a novel and effective fluorescence-sensing platform for nucleic acid detection. Tests on the oligonucleotide sequences associated with the human immunodeficiency virus as a model system showed that the Oc-AgNPs effectively absorbed and quenched dye-labeled single-stranded DNA through strong hydrogen bonding interactions and slight electrostatic attractive interactions. The proposed system efficiently differentiated between complementary and mismatched nucleic acid sequences with high selectivity and good reproducibility at room temperature.

  2. Tailoring dye-sensitized upconversion nanoparticle excitation bands towards excitation wavelength selective imaging

    SciTech Connect

    Wu, Xiang; Lee, Hyungseok; Bilsel, Osman; Zhang, Yuanwei; Li, Zhanjun; Chen, Teresa; Liu, Yi; Duan, Chunying; Shen, Jie; Punjabi, Amol; Han, Gang

    2015-01-01

    One of the key roadblocks in UCNP development is its extremely limited choices of excitation wavelengths. We report a generic design to program UCNPs to possess highly tunable dye characteristic excitation bands. Using such distinctive properties, we were able to develop a new excitation wavelength selective security imaging. Finally, this work unleashed the greater freedom of the excitation wavelengths of the upconversion nanoparticles and we believe it is a game-changer in the field and this method will enable numerous applications that are currently limited by existing UCNPs.

  3. Tailoring Dye-sensitized Upconversion Nanoparticles Excitation Bands towards Excitation Wavelength Selective Imaging

    PubMed Central

    Wu, Xiang; Lee, Hyungseok; Bilsel, Osman; Zhang, Yuanwei; Li, Zhanjun; Chen, Teresa; Liu, Yi; Duan, Chunying; Shen, Jie; Punjabi, Amol; Han, Gang

    2015-01-01

    One of key roadblocks in UCNP development is its extremely limited choices of excitation wavelengths. We report a generic design to program UCNPs to possess highly tunable dye characteristic excitation bands. Using such distinctive properties, we were able to develop a new excitation wavelength selective security imaging. This work unleashed the greater freedom of the excitation wavelengths of the upconversion nanoparticles and we believe it is a game-changer in the field and this method will enable numerous applications that are currently limited by existing UCNPs. PMID:26499208

  4. Fast, sensitive point of care electrochemical molecular system for point mutation and select agent detection.

    PubMed

    MacLeod, J A; Nemeth, A C; Dicke, W C; Wang, D; Manalili Wheeler, S; Hannis, J C; Collier, G B; Drader, J J

    2016-07-01

    Point of care molecular diagnostics benefits from a portable battery-operated device capable of performing a fast turnaround using reliable inexpensive cartridges. We describe a prototype device for performing a molecular diagnostics test for clinical and biodefense samples in 16 minutes using a prototype capable of an 8 minute PCR reaction, followed by hybridization and detection on an electrochemical microarray based on the i-STAT® system. We used human buccal swabs for hemochromatosis testing including in-device DNA extraction. Additional clinical and biodefense samples included influenza A and bacterial select agents Bacillus anthracis, Yersinia pestis and Francisella tularensis. PMID:27280174

  5. Highly sensitive and selective fluoride detection in water through fluorophore release from a metal-organic framework

    PubMed Central

    Hinterholzinger, Florian M.; Rühle, Bastian; Wuttke, Stefan; Karaghiosoff, Konstantin; Bein, Thomas

    2013-01-01

    The detection, differentiation and visualization of compounds such as gases, liquids or ions are key challenges for the design of selective optical chemosensors. Optical chemical sensors employ a transduction mechanism that converts a specific analyte recognition event into an optical signal. Here we report a novel concept for fluoride ion sensing where a porous crystalline framework serves as a host for a fluorescent reporter molecule. The detection is based on the decomposition of the host scaffold which induces the release of the fluorescent dye molecule. Specifically, the hybrid composite of the metal-organic framework NH2-MIL-101(Al) and fluorescein acting as reporter shows an exceptional turn-on fluorescence in aqueous fluoride-containing solutions. Using this novel strategy, the optical detection of fluoride is extremely sensitive and highly selective in the presence of many other anions. PMID:24008779

  6. Highly sensitive and selective fluoride detection in water through fluorophore release from a metal-organic framework.

    PubMed

    Hinterholzinger, Florian M; Rühle, Bastian; Wuttke, Stefan; Karaghiosoff, Konstantin; Bein, Thomas

    2013-01-01

    The detection, differentiation and visualization of compounds such as gases, liquids or ions are key challenges for the design of selective optical chemosensors. Optical chemical sensors employ a transduction mechanism that converts a specific analyte recognition event into an optical signal. Here we report a novel concept for fluoride ion sensing where a porous crystalline framework serves as a host for a fluorescent reporter molecule. The detection is based on the decomposition of the host scaffold which induces the release of the fluorescent dye molecule. Specifically, the hybrid composite of the metal-organic framework NH2-MIL-101(Al) and fluorescein acting as reporter shows an exceptional turn-on fluorescence in aqueous fluoride-containing solutions. Using this novel strategy, the optical detection of fluoride is extremely sensitive and highly selective in the presence of many other anions.

  7. An aptasensor for selective, sensitive and fast detection of lead(II) based on polyethyleneimine and gold nanoparticles.

    PubMed

    Taghdisi, Seyed Mohammad; Danesh, Noor Mohammad; Lavaee, Parirokh; Ramezani, Mohammad; Abnous, Khalil

    2015-05-01

    Lead (Pb), as a major environmental contaminant, could be harmful to humans when inhaled or ingested. In this study, we developed a sensitive, selective and fast colorimetric aptasensor for Pb(+2) based on polyethylenimine (PEI) and gold nanoparticles (AuNPs). In the absence of Pb(+2), aptamer binds to PEI. So the well-dispersed AuNPs remain stable with a wine-red color. Upon the addition of Pb(+2), a conformational change happens and a G-quadruplex aptamer/Pb(+2) complex is formed, leading to the aggregation of AuNPs and a color change to blue. This sensor showed a high selectivity toward Pb(+2) with a limit of detection (LOD) as low as 702pM. Moreover, our fabricated sensor was successfully applied for Pb(+2) detection in rat serum and tap water. PMID:25989533

  8. Improved Selectivity and Sensitivity of Gas Sensing Using a 3D Reduced Graphene Oxide Hydrogel with an Integrated Microheater.

    PubMed

    Wu, Jin; Tao, Kai; Miao, Jianmin; Norford, Leslie K

    2015-12-16

    Low-cost, one-step, and hydrothermal synthesized 3D reduced graphene oxide hydrogel (RGOH) is exploited to fabricate a high performance NO2 and NH3 sensor with an integrated microheater. The sensor can experimentally detect NO2 and NH3 at low concentrations of 200 ppb and 20 ppm, respectively, at room temperature. In addition to accelerating the signal recovery rate by elevating the local silicon substrate temperature, the microheater is exploited for the first time to improve the selectivity of NO2 sensing. Specifically, the sensor response from NH3 can be effectively suppressed by a locally increased temperature, while the sensitivity of detecting NO2 is not significantly affected. This leads to good discrimination between NO2 and NH3. This strategy paves a new avenue to improve the selectivity of gas sensing by using the microheater to raise substrate temperature. PMID:26630364

  9. Highly sensitive and selective fluorescence sensor based on functional SBA-15 for detection of Hg2+ in aqueous media.

    PubMed

    Song, Chunxia; Zhang, Xiaolin; Jia, Cuiying; Zhou, Peng; Quan, Xie; Duan, Chunying

    2010-04-15

    A new Rhodamine-based chemosensor RBSN designed for the selective detection of Hg(2+) in aqueous media is synthesized and structural characterized. It features brightness blue emission in the presence of Hg(2+) with the fluorescent detection limit for Hg(2+) in aqueous media being ppb levels and exhibits excellent Hg(2+)-specific luminescence enhancement over various competitive cations, including alkali and alkaliearth, the first-row transition metals and heavy metals. By immobilizing RBSN to the mesoporous SBA-15, a highly selective and sensitive chemosensor toward mercury cations in aqueous solution was achieved. RBSN/SBA-15 could quantitatively determinate mercury cations at ppb level in the practical environmental media, suggesting the possibility for real-time quantitative detection of Hg(2+) and the convenience for potential application in toxicological and environmental science.

  10. A sensitive and selective resonance Rayleigh scattering method for quick detection of avidin using affinity labeling Au nanoparticles

    NASA Astrophysics Data System (ADS)

    Wang, Qi; Huang, Xi; Fu, Xuan; Deng, Huan; Ma, Meihu; Cai, Zhaoxia

    2016-06-01

    Avidin is a glycoprotein with antinutritional property, which should be limited in daily food. We developed an affinity biosensor system based on resonance Rayleigh scattering (RRS) and using affinity biotin labeling Au nanoparticles (AuNPs). This method was selective and sensitive for quick avidin detection due to the avidin-biotin affinitive interaction. Under optimal conditions, RRS intensity of biotin-AuNPs increase linearly with an increasing concentration of avidin from 5 to 160 ng/mL. The lower limit of detection was 0.59 ng/mL. This rapid and selective avidin detection method was used in synthetic samples and egg products with recoveries of between 102.97 and 107.92%, thereby demonstrating the feasible and practical application of this assay.

  11. Gold nanoparticle-sensitized quartz crystal microbalance sensor for rapid and highly selective determination of Cu(II) ions.

    PubMed

    Jin, Yulong; Huang, Yanyan; Liu, Guoquan; Zhao, Rui

    2013-09-21

    A novel quartz crystal microbalance (QCM) sensor for rapid, highly selective and sensitive detection of copper ions was developed. As a signal amplifier, gold nanoparticles (Au NPs) were self-assembled onto the surface of the sensor. A simple dip-and-dry method enabled the whole detection procedure to be accomplished within 20 min. High selectivity of the sensor towards copper ions is demonstrated by both individual and coexisting assays with interference ions. This gold nanoparticle mediated amplification allowed a detection limit down to 3.1 μM. Together with good repeatability and regeneration, the QCM sensor was also applied to the analysis of copper contamination in drinking water. This work provides a flexible method for fabricating QCM sensors for the analysis of important small molecules in environmental and biological samples. PMID:23888301

  12. Audit Report on "Management Controls over the Department's Excess Weapons Inventories and Selected Sensitive Equipment used by Protective Forces"

    SciTech Connect

    2009-01-01

    Since September 11, 2001, the Department of Energy has, on several occasions, revised its security posture based on identified threats and adversaries. These revisions in security posture have driven Departmental sites to upgrade their defensive and tactical equipment. Subsequent changes in the perceived threats have, in some cases, led to a reduction in the need for certain types of weapons, thus creating a pool of surplus equipment. These surplus weapons could potentially be used by other Department sites and Federal law enforcement agencies. Recent Office of Inspector General reports have raised concerns with the adequacy of controls related to defensive and tactical equipment. For example, our report on Management Controls Over Defense Related High Risk Property (OAS-M-08-06, April 2008) found that administrative controls over certain defense related high risk property were not sufficient for providing accountability over these items. Because of prior reported weaknesses in controls over defensive and tactical equipment, we initiated this audit to determine whether the Department and its contractors were properly managing excess weapons inventories and selected sensitive equipment used by protective forces. Our review disclosed that the Department was not always properly managing its inventories of excess weapons and selected sensitive equipment. We identified issues with the retention of unneeded weapons at many locations and with the identification and tracking of sensitive items. More specifically: Sites maintained large inventories of weapons that were no longer needed but had not been made available for use by either other Departmental sites or other Federal law enforcement agencies. For instance, at six of the locations included in our review we identified a total of 2,635 unneeded weapons with a total acquisition value of over $2.8 million that had not been officially declared as excess - an action that would have made them available for others to use

  13. A ratiometric fluorescence nanosensor for highly selective and sensitive detection of selenite.

    PubMed

    Chen, Linfeng; Tian, Xike; Zhao, Yuan; Li, Yong; Yang, Chao; Zhou, Zhaoxin; Liu, Xiangwen

    2016-08-01

    The instant and on-site detection of selenium still remains a challenge for environmental monitoring and medical prevention. We herein developed a ratiometric fluorescent nanosensor for accurate and on-site sensing of SeO3(2-) by linking the recognition molecule 3,3'-diaminobenzidine (DAB) onto the surface of carboxyl group modified CdTe@SiO2. The fluorescence of DAB on the surface of silica nanospheres could be selectively and efficiently enhanced by SeO3(2-) through a surface chelating reaction between DAB and SeO3(2-). Thus, in the presence of SeO3(2-), the nanosensor would show two characteristic fluorescence emissions of Se-DAB and CdTe QDs under a single excitation wavelength. The selectivity and the optimal conditions for the detection of SeO3(2-) were carefully investigated. The ratio of F530/F635 linearly increased with increasing SeO3(2-) concentration in the range of 0 to 2.5 μM and the detection limit reaches as low as 6.68 nM (0.53 ppb). This developed nanosensor has the capability of on-site detection in an aqueous system without any separation step. The Se concentrations in selenium-rich food were detected and the results were consistent with the values determined by ICP-AES. PMID:27241591

  14. Molecularly imprinted polymer decorated nanoporous gold for highly selective and sensitive electrochemical sensors

    PubMed Central

    Li, Yingchun; Liu, Yuan; Liu, Jie; Liu, Jiang; Tang, Hui; Cao, Cong; Zhao, Dongsheng; Ding, Yi

    2015-01-01

    Electrochemical nanosensors based on nanoporous gold leaf (NPGL) and molecularly imprinted polymer (MIP) are developed for pharmaceutical analysis by using metronidazole (MNZ) as a model analyte. NPGL, serving as the loading platform for MIP immobilization, possesses large accessible surface area with superb electric conductivity, while electrochemically synthesized MIP thin layer affords selectivity for specific recognition of MNZ molecules. For MNZ determination, the hybrid electrode shows two dynamic linear range of 5 × 10−11 to 1 × 10−9 mol L−1 and 1 × 10−9 to 1.4 × 10−6 mol L−1 with a remarkably low detection limit of 1.8 × 10−11 mol L−1 (S/N = 3). In addition, the sensor exhibits high binding affinity and selectivity towards MNZ with excellent reproducibility and stability. Finally, the reliability of MIP-NPGL for MNZ detection is proved in real fish tissue samples, demonstrating the potential for the proposed electrochemical sensors in monitoring drug and biological samples. PMID:25572290

  15. Selectively Sensitizing Malignant Cells to Photothermal Therapy Using a CD44-Targeting Heat Shock Protein 72 Depletion Nanosystem.

    PubMed

    Wang, Shouju; Tian, Ying; Tian, Wei; Sun, Jing; Zhao, Shuang; Liu, Ying; Wang, Chunyan; Tang, Yuxia; Ma, Xingqun; Teng, Zhaogang; Lu, Guangming

    2016-09-27

    Selectively enhance the therapeutic efficacy to malignancy is one of the most important issues for photothermal therapy (PTT). However, most solid tumors, such as triple negative breast cancer (TNBC), do not have identifiable surface markers to distinguish themselves from normal cells, thus it is challenging to selectively identify and eliminate those malignances by PTT. In this report, we hypothesized that, by targeting CD44 (one TNBC-overexpressed surface molecule) and depleting heat shock protein 72 (HSP72, one malignancy-specific-overexpressed thermotolerance-related chaperone) subsequently, the TNBC could be selectively sensitized to PTT and improve the accuracy of treatment. To this end, a rationally designed nanosystem gold nanostar (GNS)/siRNA against HSP72 (siHSP72)/hyaluronic acid (HA) was successfully constructed using a layer-by-layer method. Hydrodynamic diameter and zeta potential analysis demonstrated the formation of GNS/siHSP72/HA having a particle size of 73.2 ± 3.8 nm and a negative surface charge of -18.3 ± 1.6 mV. The CD44-targeting ability of GNS/siHSP72/HA was confirmed by the flow cytometer, confocal microscopic imaging, and competitive binding analysis. The HSP72 silencing efficacy of GNS/siHSP72/HA was ∼95% in complete culture medium. By targeting CD44 and depleting HSP72 sequentially, GNS/siHSP72/HA could selectively sensitize TNBC cells to hyperthermia and enhance the therapeutic efficacy to TNBC with minimal side effect both in vitro and in vivo. Other advantages of GNS/siHSP72/HA included easy synthesis, robust siRNA loading capacity, endosome/lysosome escaping ability, high photothermal conversion efficacy and superior hemo- and biocompatibility.

  16. A fast response hafnium selective polymeric membrane electrode based on N,N'-bis(alpha-methyl-salicylidene)-dipropylenetriamine as a neutral carrier.

    PubMed

    Rezaei, B; Meghdadi, S; Zarandi, R Fazel

    2008-08-30

    In this study a new hafnium selective sensor was fabricated from polyvinylchloride (PVC) matrix membrane containing neutral carrier N,N'-bis(alpha-methyl-salicylidene)-dipropylenetriamine (Mesaldpt) as a new ionophore, sodium tetraphenyl borate (NaTPB) as anionic discriminator and dioctyl phthalate (DOP) as plasticizing solvent mediator in tetrahydrofuran solvent. The electrode exhibits Nernstian response for Hf(4+) (Hafnium(IV)) over a wide concentration range (2.0 x 10(-7) to 1.0 x 10(-1)M) with the determination coefficient of 0.9966 and slope of 15.1+/-0.1 mVdecades(-1). The limit of detection is 1.9 x 10(-7)M. The electrode has a fast response time of 18s and a working pH range of 4-8. The proposed membrane shows excellent discriminating ability towards Hf(4+) ion with regard to several alkali, alkaline earth transition and heavy metal ions. It can be used over a period of 1.5 months with good reproducibility. It is successfully applied for direct determination of Hf(4+) in solutions by standard addition method for real sample analysis.

  17. A simple procedure for expression and purification of selected non-structural (alpha and beta) herpes simplex virus 1 (HSV-1) proteins.

    PubMed

    Kosovský, J; Durmanová, V; Kúdelová, M; Rezuchová, I; Tkáciková, L; Rajcáni, J

    2001-04-01

    The expression and isolation of herpes simplex virus 1 (HSV-1) immediate early (alpha) IE63 (ICP27) and of the early (beta) thymidine kinase (Tk) polypeptides in Escherichia coli JM 109 cells transformed with the PinPoint Xa-1 (Promega) plasmid construct carrying either the HSV-1 UL54 or UL23 genes are described. The resulting biotinylated fusion protein(s) could be easily induced and were purified in appropriate amounts by means of a monomeric avidin-conjugated resin (SoftLink Soft Release Avidin Resin, Promega) provided that: (1) the exponential growth of the selected transformed cells was monitored carefully; (2) the post-induction harvest interval was properly chosen; and (3) the period for adsorption to the avidin resin suitably adjusted. The isolated protein(s), although partially digested in the case of the IE63 polypeptide, were suitable antigen(s) for immunization of various animal species. Co-purification of trace amounts of endogenous biotinylated protein(s) produced in E. coli was eliminated by shortening the duration of adsorption to the avidin resin.

  18. Solid-state (17)O NMR as a sensitive probe of keto and gem-diol forms of alpha-keto acid derivatives.

    PubMed

    Zhu, Jianfeng; Geris, Amanda J; Wu, Gang

    2009-08-28

    We have used solid-state (17)O NMR experiments to measure the (17)O quadrupole coupling (QC) and chemical shift (CS) tensors for two alpha-keto acids: sodium [2-(17)O]pyruvate and lithium [2,2'-(17)O(2)]pyruvate. In the solid state, sodium [2-(17)O]pyruvate is in the keto form (-C(double bond (17)O)-) whereas lithium [2,2'-(17)O(2)]pyruvate takes the gem-diol form (-C((17)OH)(2)-). This study represents the first time that a full set of (17)O NMR tensors are experimentally determined for alpha-keto acids in these two different tautomeric forms. We have found that the two forms exhibit drastically different (17)O QC and CS tensors: for the keto form, delta(iso) = 543 +/- 1 ppm, C(Q) = 10.8 +/- 0.2 MHz, eta(Q) = 0.48 +/- 0.05, delta(11) = 1020 +/- 10, delta(22) = 640 +/- 10, delta(33) = -40 +/- 10 ppm, alpha = 80 +/- 5 degrees, beta = 90 +/- 2 degrees, and gamma = 83 +/- 2 degrees; for the gem-diol form, delta(iso) = 62 +/- 1 ppm, C(Q) = 8.5 +/- 0.5 MHz, eta(Q) = 1.0 +/- 0.05, delta(11) = 140 +/- 5, delta(22) = 45 +/- 5, delta(33) = 0 +/- 5 ppm, alpha = 55 +/- 5 degrees, beta = 90 +/- 5 degrees, and gamma = 80 +/- 2 degrees. The (17)O chemical shift tensor observed for the gem-diol functional group also represents the first such measurement for any -ol functional group (e.g., alcohols, phenols, carbohydrates, etc.) Using these accurate experimental (17)O NMR tensors, we were able to evaluate the accuracy of quantum chemical calculations. Our results showed that quantum chemical calculations using the crystal lattice approach are in much better agreement with the experimental solid-state (17)O NMR data than those calculated using the molecular cluster approach. Quantum chemical calculations have also provided information about the sign of the (17)O quadrupolar coupling constants and about the (17)O NMR tensor orientations in the molecular frame of reference. Our findings suggest that solid-state (17)O NMR may be useful in probing the tautomeric form of the alpha

  19. Criteria for Sample Selection to Maximize Planet Sensitivity and Yield from Space-Based Microlens Parallax Surveys

    NASA Astrophysics Data System (ADS)

    Yee, Jennifer C.; Gould, Andrew; Beichman, Charles; Calchi Novati, Sebastiano; Carey, Sean; Gaudi, B. Scott; Henderson, Calen B.; Nataf, David; Penny, Matthew; Shvartzvald, Yossi; Zhu, Wei

    2015-09-01

    Space-based microlens parallax measurements are a powerful tool for understanding planet populations, especially their distribution throughout the Galaxy. However, if space-based observations of the microlensing events must be specifically targeted, it is crucial that microlensing events enter the parallax sample without reference to the known presence or absence of planets. Hence, it is vital to define objective criteria for selecting events where possible and to carefully consider and minimize the selection biases where not possible so that the final sample represents a controlled experiment. We present objective criteria for initiating observations and determining their cadence for a subset of events, and we define procedures for isolating subjective decision making from information about detected planets for the remainder of events. We also define procedures to resolve conflicts between subjective and objective selections. These procedures maximize the planet sensitivity of the sample as a whole by allowing for planet detections even if they occur before satellite observations for objectively selected events and by helping to trigger fruitful follow-up observations for subjectively chosen events. This paper represents our public commitment to these procedures, which is a necessary component of enforcing objectivity on the experimental protocol. They will be implemented for the 2015 Spitzer microlensing campaign.

  20. Discovery of an Oxybenzylglycine Based Peroxisome Proliferator Activated Receptor [alpha] Selective Agonist 2-((3-((2-(4-Chlorophenyl)-5-methyloxazol-4-yl)methoxy)benzyl)(methoxycarbonyl)amino)acetic Acid (BMS-687453)

    SciTech Connect

    Li, Jun; Kennedy, Lawrence J.; Shi, Yan; Tao, Shiwei; Ye, Xiang-Yang; Chen, Stephanie Y.; Wang, Ying; Hernndez, Andrs S.; Wang, Wei; Devasthale, Pratik V.; Chen, Sean; Lai, Zhi; Zhang, Hao; Wu, Shung; Smirk, Rebecca A.; Bolton, Scott A.; Ryono, Denis E.; Zhang, Huiping; Lim, Ngiap-Kie; Chen, Bang-Chi; Locke, Kenneth T.; O’Malley, Kevin M.; Zhang, Litao; Srivastava, Rai Ajit; Miao, Bowman; Meyers, Daniel S.; Monshizadegan, Hossain; Search, Debra; Grimm, Denise; Zhang, Rongan; Harrity, Thomas; Kunselman, Lori K.; Cap, Michael; Kadiyala, Pathanjali; Hosagrahara, Vinayak; Zhang, Lisa; Xu, Carrie; Li, Yi-Xin; Muckelbauer, Jodi K.; Chang, Chiehying; An, Yongmi; Krystek, Stanley R.; Blanar, Michael A.; Zahler, Robert; Mukherjee, Ranjan; Cheng, Peter T.W.; Tino, Joseph A.

    2010-04-12

    An 1,3-oxybenzylglycine based compound 2 (BMS-687453) was discovered to be a potent and selective peroxisome proliferator activated receptor (PPAR) {alpha} agonist, with an EC{sub 50} of 10 nM for human PPAR{alpha} and 410-fold selectivity vs human PPAR{gamma} in PPAR-GAL4 transactivation assays. Similar potencies and selectivity were also observed in the full length receptor co-transfection assays. Compound 2 has negligible cross-reactivity against a panel of human nuclear hormone receptors including PPAR{delta}. Compound 2 demonstrated an excellent pharmacological and safety profile in preclinical studies and thus was chosen as a development candidate for the treatment of atherosclerosis and dyslipidemia. The X-ray cocrystal structures of the early lead compound 12 and compound 2 in complex with PPAR{alpha} ligand binding domain (LBD) were determined. The role of the crystal structure of compound 12 with PPAR{alpha} in the development of the SAR that ultimately resulted in the discovery of compound 2 is discussed.

  1. Environmental sensitivity differs between rabbit lines selected for reproductive intensity and longevity.

    PubMed

    Savietto, D; Cervera, C; Blas, E; Baselga, M; Larsen, T; Friggens, N C; Pascual, J J

    2013-12-01

    To better understand the mechanisms that allow some animals to sustain their productive effort in harsh environmental conditions, rabbit does from two selection lines (LP and V) were housed in normal (NC), nutritional (NF) or heat (HC) challenging environmental conditions from first to third partum. The LP line (n=85) was founded on reproductive longevity criteria by selecting does from commercial farms that had a minimum of 25 partum with more than 7.5 kits born alive per parity. Line V (n=79) was constituted from four specialised maternal lines into a composite synthetic line and then selected by litter size at weaning for 36 generations. Female rabbits in NC and NF environments were housed at normal room temperature (18°C to 24°C) and fed with control [11.6 MJ digestible energy (DE)/kg dry matter (DM)] or low-energy diets (9.1 MJ DE/kg DM). HC does were housed at high room temperatures (25°C to 35°C) and fed the control diet. Female rabbits in the HC and NF environments ingested 11.5% and 6% less DE than NC does, respectively (P<0.05). These differences between environments occurred in both lines, with the differences being higher for LP than for V does (+6%; P<0.05). Milk yield responses followed those of energy intake also being higher for LP does (+21.3 g/day; P<0.05). The environmental conditions did not affect the perirenal fat thickness (PFT), but a genotype by environment interaction was observed. In NC and HC, the PFT was higher for line V (+0.23 and +0.35 mm, respectively; P<0.05) than for LP does, but this was not the case at NF (-0.01 mm). Moreover, the PFT evolution was different between them. In the NC environment, LP does used the accreted PFT in late lactation (-0.29 mm), whereas V does did not (-0.08 mm). Conversely, in the HC environment, LP does showed a flat PFT evolution in late lactation, whereas V does accumulated PFT. In the NF environment, LP and V does had a similar PFT evolution. There was also a litter size reduction for V does of

  2. Utility-based criteria for selecting patients with hepatocellular carcinoma for liver transplantation: A multicenter cohort study using the alpha-fetoprotein model as a survival predictor.

    PubMed

    Vitale, Alessandro; Farinati, Fabio; Burra, Patrizia; Trevisani, Franco; Giannini, Edoardo G; Ciccarese, Francesca; Piscaglia, Fabio; Rapaccini, Gian Lodovico; Di Marco, Mariella; Caturelli, Eugenio; Zoli, Marco; Borzio, Franco; Cabibbo, Giuseppe; Felder, Martina; Sacco, Rodolfo; Morisco, Filomena; Missale, Gabriele; Foschi, Francesco Giuseppe; Gasbarrini, Antonio; Svegliati Baroni, Gianluca; Virdone, Roberto; Chiaramonte, Maria; Spolverato, Gaya; Cillo, Umberto

    2015-10-01

    The lifetime utility of liver transplantation (LT) in patients with hepatocellular carcinoma (HCC) is still controversial. The aim of this study was to ascertain when LT is cost-effective for HCC patients, with a view to proposing new transplant selection criteria. The study involved a real cohort of potentially transplantable Italian HCC patients (n = 2419 selected from the Italian Liver Cancer group database) who received nontransplant therapies. A non-LT survival analysis was conducted, the direct costs of therapies were calculated, and a Markov model was used to compute the cost utility of LT over non-LT therapies in Italian and US cost scenarios. Post-LT survival was calculated using the alpha-fetoprotein (AFP) model on the basis of AFP values and radiological size and number of nodules. The primary endpoint was the net health benefit (NHB), defined as LT survival benefit in quality-adjusted life years minus incremental costs (US $)/willingness to pay. The calculated median cost of non-LT therapies per patient was US $53,042 in Italy and US $62,827 in the United States. On Monte Carlo simulation, the NHB of LT was always positive for AFP model values ≤ 3 and always negative for values > 7 in both countries. A multivariate model showed that nontumor variables (patient's age, Child-Turcotte-Pugh [CTP] class, and alternative therapies) had the potential to shift the AFP model threshold of LT cost-ineffectiveness from 3 to 7. LT proved always cost-effective for HCC patients with AFP model values ≤ 3, whereas the cost-ineffectiveness threshold ranged between 3 and 7 using nontumor variables. PMID:26183802

  3. Utility-based criteria for selecting patients with hepatocellular carcinoma for liver transplantation: A multicenter cohort study using the alpha-fetoprotein model as a survival predictor.

    PubMed

    Vitale, Alessandro; Farinati, Fabio; Burra, Patrizia; Trevisani, Franco; Giannini, Edoardo G; Ciccarese, Francesca; Piscaglia, Fabio; Rapaccini, Gian Lodovico; Di Marco, Mariella; Caturelli, Eugenio; Zoli, Marco; Borzio, Franco; Cabibbo, Giuseppe; Felder, Martina; Sacco, Rodolfo; Morisco, Filomena; Missale, Gabriele; Foschi, Francesco Giuseppe; Gasbarrini, Antonio; Svegliati Baroni, Gianluca; Virdone, Roberto; Chiaramonte, Maria; Spolverato, Gaya; Cillo, Umberto

    2015-10-01

    The lifetime utility of liver transplantation (LT) in patients with hepatocellular carcinoma (HCC) is still controversial. The aim of this study was to ascertain when LT is cost-effective for HCC patients, with a view to proposing new transplant selection criteria. The study involved a real cohort of potentially transplantable Italian HCC patients (n = 2419 selected from the Italian Liver Cancer group database) who received nontransplant therapies. A non-LT survival analysis was conducted, the direct costs of therapies were calculated, and a Markov model was used to compute the cost utility of LT over non-LT therapies in Italian and US cost scenarios. Post-LT survival was calculated using the alpha-fetoprotein (AFP) model on the basis of AFP values and radiological size and number of nodules. The primary endpoint was the net health benefit (NHB), defined as LT survival benefit in quality-adjusted life years minus incremental costs (US $)/willingness to pay. The calculated median cost of non-LT therapies per patient was US $53,042 in Italy and US $62,827 in the United States. On Monte Carlo simulation, the NHB of LT was always positive for AFP model values ≤ 3 and always negative for values > 7 in both countries. A multivariate model showed that nontumor variables (patient's age, Child-Turcotte-Pugh [CTP] class, and alternative therapies) had the potential to shift the AFP model threshold of LT cost-ineffectiveness from 3 to 7. LT proved always cost-effective for HCC patients with AFP model values ≤ 3, whereas the cost-ineffectiveness threshold ranged between 3 and 7 using nontumor variables.

  4. Recent Results on the CKM Angle Alpha

    SciTech Connect

    Mihalyi, A.; /Wisconsin U., Madison

    2005-10-18

    The method to measure the CKM angle {alpha} and the modes sensitive to it are discussed. It is shown that the B {yields} {rho}{rho} decays provide the most stringent constraint on {alpha}, which is found to be {alpha} = 96{sup o} {+-} 10{sup o}(stat) {+-} 4{sup o}(syst){+-} 13{sup o}(penguin).

  5. Oxygen-evoked changes in transcriptional activity of the 5'-flanking region of the human amiloride-sensitive sodium channel (alphaENaC) gene: role of nuclear factor kappaB.

    PubMed Central

    Baines, Deborah L; Janes, Mandy; Newman, David J; Best, Oliver G

    2002-01-01

    Expression of the alpha-subunit of the amiloride-sensitive sodium channel (alphaENaC) is regulated by a number of factors in the lung, including oxygen partial pressure (PO2). As transcriptional activation is a mechanism for raising cellular mRNA levels, we investigated the effect of physiological changes in PO2 on the activity of the redox-sensitive transcription factor nuclear factor kappaB (NF-kappaB) and transcriptional activity of 5'-flanking regions of the human alphaENaC gene using luciferase reporter-gene vectors transiently transfected into human adult alveolar carcinoma A549 cells. By Western blotting we confirmed the presence of NF-kappaB p65 but not p50 in these cells. Transiently increasing PO2 from 23 to 42 mmHg for 24 h evoked a significant increase in NF-kappaB DNA-binding activity and transactivation of a NF-kappaB-driven luciferase construct (pGLNF-kappaBpro), which was blocked by the NF-kappaB activation inhibitor sulphasalazine (5 mM). Transcriptional activity of alphaENaC-luciferase constructs containing 5'-flanking sequences (including the NF-kappaB consensus) were increased by raising PO2 from 23 to 142 mmHg if they contained transcriptional initiation sites (TIS) for exons 1A and 1B (pGL3E2.2) or the 3' TIS of exon 1B alone (pGL3E0.8). Sulphasalazine had no significant effect on the activity of these constructs, suggesting that the PO2-evoked rise in activity was not a direct consequence of NF-kappaB activation. Conversely, the relative luciferase activity of a construct that lacked the 3' TIS, a 3' intron and splice site but still retained the 5' TIS and NF-kappaB consensus sequence was suppressed significantly by raising PO2. This effect was reversed by sulphasalazine, suggesting that activation of NF-kappaB mediated PO2-evoked suppression of transcription from the exon 1A TIS of alphaENaC. PMID:12023897

  6. Functionalized polymeric magnetic nanoconstructs for selective capturing and sensitive detection of Salmonella typhimurium.

    PubMed

    Chattopadhyay, Sruti; Kaur, Avneet; Jain, Swati; Sabharwal, Prabhjot K; Singh, Harpal

    2016-09-21

    Rapid detection and enumeration of pathogens is essential for monitoring contamination and spoilage of food products to ensure improved quality control management. Functionalized polymeric magnetic nanoconstructs (FPMNCs) were developed as an effective immunomagnetic separator and sensing platform for the selective capturing of Salmonella typhimurium. Novel FPMNCs were prepared in three stages involving synthesis of iron oxide (IO) dispersion, capping with sodium oleate and encapsulation of preformed IO nanoparticles by in-situ free radical emulsion polymerization of styrene (St), methyl methacrylate (MMA) and acetoacetoxy ethylmethacrylate (AAEM). PMMA improves the stability of FPMNCs by bridging extremely hydrophobic PS and hydrophilic PAAEM. Core-shell morphology of hydrophobic core of IO, PS & PMMA and hydrophilic shell of PAAEM was demonstrated by SEM, TEM and FTIR studies. FPMNCs with surface functionalized acetoacetoxy groups were covalently attached with polyclonal antibodies against Salmonella common structural antigen (CSA-1-Ab) without using any linker and catalyst. Colorimetric readout signal was acquired using CSA-1-Ab-HRP as secondary antibody after formation of sandwich immunocomplex with bacteria where the optical density of the samples were recorded using ELISA plate reader at 450 nm. The developed immunoassay was specific and selective which captures only targeted S. typhimurium with a detection limit of 10 cells/mL lower than infectious dose of salmonellosis infection. Minimal interference of food matrix with high signal to noise ratio was shown by various food samples. In addition, the performance of developed FPMNC based immunoassay was superior to commercially available immunomagnetic microbeads demonstrating undisputed advantage for capturing and detecting specific bacteria without any pre-enrichment of sample. PMID:27590554

  7. SIMPLE, SENSITIVE AND SELECTIVE SPECTROPHOTOMETRIC ASSAY OF NAPROXEN IN PURE, PHARMACEUTICAL PREPARATION AND HUMAN SERUM SAMPLES.

    PubMed

    Alizadeh, Nina; Keyhanian, Fereshteh

    2015-01-01

    Two simple, rapid and sensitive spectrophotometric methods have been developed for the determination of naproxen in pure, pharmaceutical preparation and human serum samples. These methods are based on the formation of yellow ion-pair complexes between naproxen and two sulfophthalein acid dyes, namely bromocresol green (BCG method) and bromothymol blue (BTB method). The resulting complexes were measured at 424 nm (BCG method) and at 422 nm (BTB method). The effects of variables such as reagent concentration and reaction time were investigated to optimize the procedure. Beer's law was obeyed in the concentration range of 10-105 µg/mL and 5-85 µg/mL and the detection limits were found to be 0.347 and 0.31 µg/mL for BCG and BTB methods, respectively. The developed methods have been successfully applied for the determination of naproxen in bulk drugs, pharmaceutical formulations and human serum samples with good accuracy and precision. The results are comparable to those of reference methods, and hence are recommended for quality control and routine analysis.

  8. Based on magnetic graphene oxide highly sensitive and selective imprinted sensor for determination of sunset yellow.

    PubMed

    Li, Jianbo; Wang, Xiaojiao; Duan, Huimin; Wang, Yanhui; Bu, Yanan; Luo, Chuannan

    2016-01-15

    A new imprinted material based on β-cyclodextrin/ionic liquid/gold nanoparticles functionalized magnetic graphene oxide has been successfully synthesized and modified to the glassy carbon electrode surface to constructed imprinted electrochemical sensor to detect sunset yellow. The sensitivity and electrochemical response of the electrode can be improved by nanomaterials. The surface morphology and crystal structure of the hybrid nanomaterial has been characterized by scanning electron microscopy, X-ray diffraction, Fourier transform infrared spectroscopy. The electrochemical behaviors of the hybrid nanomaterials based sensor were evaluated through cyclic voltammetry and electrochemical impedance spectroscopy. Under the optimized conditions, the proposed electrochemical sensor showed a fast rebinding dynamics, which was successfully applied to sunset yellow detection with a wide linear range from 5.0×10(-9) to 2.0×10(-6)mol L(-1) and a detection limit of 2.0×10(-9)mol L(-1). The electrochemical sensor has been successfully applied in the determination of SY in spiked water samples, mirinda drink and minute maid, and the recoveries for the standards added are 97-105%.

  9. Microglial brain region-dependent diversity and selective regional sensitivities to ageing

    PubMed Central

    Grabert, Kathleen; Michoel, Tom; Karavolos, Michail H; Clohisey, Sara; Baillie, J Kenneth; Stevens, Mark P; Freeman, Tom C; Summers, Kim M; McColl, Barry W

    2015-01-01

    Microglia play critical roles in neural development, homeostasis and neuroinflammation and are increasingly implicated in age-related neurological dysfunction. Neurodegeneration often occurs in disease-specific spatially-restricted patterns, the origins of which are unknown. We performed the first genome-wide analysis of microglia from discrete brain regions across the adult lifespan of the mouse and reveal that microglia have distinct region-dependent transcriptional identities and age in a regionally variable manner. In the young adult brain, differences in bioenergetic and immunoregulatory pathways were the major sources of heterogeneity and suggested that cerebellar and hippocampal microglia exist in a more immune vigilant state. Immune function correlated with regional transcriptional patterns. Augmentation of the distinct cerebellar immunophenotype and a contrasting loss in distinction of the hippocampal phenotype among forebrain regions were key features during ageing. Microglial diversity may enable regionally localised homeostatic functions but could also underlie region-specific sensitivities to microglial dysregulation and involvement in age-related neurodegeneration. PMID:26780511

  10. Microglial brain region-dependent diversity and selective regional sensitivities to aging.

    PubMed

    Grabert, Kathleen; Michoel, Tom; Karavolos, Michail H; Clohisey, Sara; Baillie, J Kenneth; Stevens, Mark P; Freeman, Tom C; Summers, Kim M; McColl, Barry W

    2016-03-01

    Microglia have critical roles in neural development, homeostasis and neuroinflammation and are increasingly implicated in age-related neurological dysfunction. Neurodegeneration often occurs in disease-specific, spatially restricted patterns, the origins of which are unknown. We performed to our knowledge the first genome-wide analysis of microglia from discrete brain regions across the adult lifespan of the mouse, and found that microglia have distinct region-dependent transcriptional identities and age in a regionally variable manner. In the young adult brain, differences in bioenergetic and immunoregulatory pathways were the major sources of heterogeneity and suggested that cerebellar and hippocampal microglia exist in a more immune-vigilant state. Immune function correlated with regional transcriptional patterns. Augmentation of the distinct cerebellar immunophenotype and a contrasting loss in distinction of the hippocampal phenotype among forebrain regions were key features during aging. Microglial diversity may enable regionally localized homeostatic functions but could also underlie region-specific sensitivities to microglial dysregulation and involvement in age-related neurodegeneration.

  11. High-sensitivity high-selectivity detection of CWAs and TICs using tunable laser photoacoustic spectroscopy

    NASA Astrophysics Data System (ADS)

    Pushkarsky, Michael; Webber, Michael; Patel, C. Kumar N.

    2005-03-01

    We provide a general technique for evaluating the performance of an optical sensor for the detection of chemical warfare agents (CWAs) in realistic environments and present data from a simulation model based on a field deployed discretely tunable 13CO2 laser photoacoustic spectrometer (L-PAS). Results of our calculations show the sensor performance in terms of usable sensor sensitivity as a function of probability of false positives (PFP). The false positives arise from the presence of many other gases in the ambient air that could be interferents. Using the L-PAS as it exists today, we can achieve a detection threshold of about 4 ppb for the CWAs while maintaining a PFP of less than 1:106. Our simulation permits us to vary a number of parameters in the model to provide guidance for performance improvement. We find that by using a larger density of laser lines (such as those obtained through the use of tunable semiconductor lasers), improving the detector noise and maintaining the accuracy of laser frequency determination, optical detection schemes can make possible CWA sensors having sub-ppb detection capability with <1:108 PFP. We also describe the results of a preliminary experiment that verifies the results of the simulation model. Finally, we discuss the use of continuously tunable quantum cascade lasers in L-PAS for CWA and TIC detection.

  12. CHEF induced highly selective and sensitive turn-on fluorogenic and colorimetric sensor for Fe3+.

    PubMed

    Goswami, Shyamaprosad; Das, Sangita; Aich, Krishnendu; Sarkar, Deblina; Mondal, Tapan Kumar; Quah, Ching Kheng; Fun, Hoong-Kun

    2013-11-14

    A new fluorescent probe was synthesized from rhodamine-6G and 6-(hydroxymethyl) picolinohydrazide for the sensing of Fe(3+) in an aqueous environment. The structure of the sensor was confirmed through its single crystal X-ray study. It exhibits a high specificity and sensitivity towards Fe(3+) over other interfering heavy and transition metal ions (HTM). The turn-on greenish-yellow fluorescence and a colorimetric change from colourless to pink were observed upon addition of Fe(3+) which evokes almost 116- and 23-fold enhancement in absorbance and emission intensity respectively in an acetonitrile-water (1:1, v/v, 25 °C) solution at a neutral pH (pH = 7.2). The Fe(3+) promoted ring opening of the spirolactam framework to the open chain amide platform of the sensor is responsible for its visible colour change and turn-on fluorescence activity. It also exhibits an excellent performance in the "dip stick" method. Moreover the limit of detection of the probe is in the 10(-8) M range.

  13. Selective delivery of doxorubicin by novel stimuli-sensitive nano-ferritins overcomes tumor refractoriness.

    PubMed

    Fracasso, Giulio; Falvo, Elisabetta; Colotti, Gianni; Fazi, Francesco; Ingegnere, Tiziano; Amalfitano, Adriana; Doglietto, Giovanni Battista; Alfieri, Sergio; Boffi, Alberto; Morea, Veronica; Conti, Giamaica; Tremante, Elisa; Giacomini, Patrizio; Arcovito, Alessandro; Ceci, Pierpaolo

    2016-10-10

    Human ferritin heavy chain (HFt) has been demonstrated to possess considerable potential for targeted delivery of drugs and diagnostic agents to cancer cells. Here, we report the development of a novel HFt-based genetic construct (HFt-MP-PAS) containing a short peptide linker (MP) between each HFt subunit and an outer shielding polypeptide sequence rich in proline (P), serine (S) and alanine (A) residues (PAS). The peptide linker contains a matrix-metalloproteinases (MMPs) cleavage site that permits the protective PAS shield to be removed by tumor-driven proteolytic cleavage within the tumor microenvironment. For the first time HFt-MP-PAS ability to deliver doxorubicin to cancer cells, subcellular localization, and therapeutic efficacy on a xenogeneic mouse model of a highly refractory to conventional chemotherapeutics type of cancer were evaluated. HFt-MP-PAS-DOXO performance was compared with the novel albumin-based drug delivery system INNO-206, currently in phase III clinical trials. The results of this work provide solid evidence indicating that the stimuli-sensitive, long-circulating HFt-MP-PAS nanocarriers described herein have the potential to be exploited in cancer therapy. PMID:27524282

  14. CHEF induced highly selective and sensitive turn-on fluorogenic and colorimetric sensor for Fe3+.

    PubMed

    Goswami, Shyamaprosad; Das, Sangita; Aich, Krishnendu; Sarkar, Deblina; Mondal, Tapan Kumar; Quah, Ching Kheng; Fun, Hoong-Kun

    2013-11-14

    A new fluorescent probe was synthesized from rhodamine-6G and 6-(hydroxymethyl) picolinohydrazide for the sensing of Fe(3+) in an aqueous environment. The structure of the sensor was confirmed through its single crystal X-ray study. It exhibits a high specificity and sensitivity towards Fe(3+) over other interfering heavy and transition metal ions (HTM). The turn-on greenish-yellow fluorescence and a colorimetric change from colourless to pink were observed upon addition of Fe(3+) which evokes almost 116- and 23-fold enhancement in absorbance and emission intensity respectively in an acetonitrile-water (1:1, v/v, 25 °C) solution at a neutral pH (pH = 7.2). The Fe(3+) promoted ring opening of the spirolactam framework to the open chain amide platform of the sensor is responsible for its visible colour change and turn-on fluorescence activity. It also exhibits an excellent performance in the "dip stick" method. Moreover the limit of detection of the probe is in the 10(-8) M range. PMID:24000084