bluuebunny/crossref_metadata_2025 huggingface dataset:
A novel fluorescent sensor for measurement of CFTR function by flow cytometry
Authors: Vijftigschild Lodewijk A. W. , van der Ent Cornelis K. , Beekman Jeffrey M.
Published: March 2013

Mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene cause cystic fibrosis. CFTR‐dependent iodide transport measured by fluorescent quenching of ectopically expressed halide‐sensitive yellow fluorescent protein (YFP) is widely being used to study CFTR function by microscopy or plate readers. Since YFP fluorescence in these systems is dependent on YFP expression levels and iodide concentration, differences in sensor expression level between experimental units are normalized at the start of each experiment. To allow accurate measurement of CFTR function by flow cytometry, we reasoned that co‐expression of an iodide insensitive fluorescent protein would allow for normalization of sensor expression levels and more accurate quantification of CFTR function. Our data indicated that dsRed and mKate fluorescence are iodide insensitive, and we determined an optimal format for co‐expression of these fluorescent proteins with halide‐sensitive YFP. We showed using microscopy that ratiometric measurement (YFP/mKate) corrects for differences in sensor expression levels. Ratiometric measurements were essential to accurately measure CFTR function by flow cytometry that we here describe for the first time. Mixing of wild type or mutant CFTR expressing cells indicated that addition of approximately 10% of wild type CFTR expressing cells could be distinguished by ratiometric YFP quenching. Flow cytometric ratiometric YFP quenching also allowed us to study CFTR mutants associated with differential residual function upon ectopic expression. Compared with conventional plate‐bound CFTR function assays, the flow cytometric approach described here can be used to study CFTR function in suspension cells. It may be further adapted to study CFTR function in heterologous cell populations using cell surface markers and selection of cells that display high CFTR function by cell sorting. © 2013 International Society for Advancement of Cytometry
Sex and litter effects on anxiety and DNA methylation levels of stress and neurotrophin genes in adolescent rats
Authors: Kosten Therese A. , Huang Wen , Nielsen David A.
Published: March 2013

Maternal care variations associate with DNA methylation of the glucocorticoid receptor gene, Nr3c1, in hippocampus at a nerve‐growth factor‐inducible protein 1 binding site. Epigenetic regulation of brain‐derived neurotrophin factor is affected by early stress. These systems contribute to anxiety and fear. Early stress has sex‐dependent effects perhaps reflecting sex differences in maternal care. Altering litter gender composition affects maternal behavior and DNA methylation levels of another gene in hippocampus and nucleus accumbens (NAc). We now test if DNA methylation levels of Nr3c1, Egr1, and Bdnf differ by litter composition or sex. Rats from mixed‐ or single‐sex litters were tested for anxiety and fear on postnatal day 35. Brain tissues were collected and analyzed using direct sequencing methods. Females showed hypermethylation of Nr3c1 of hippocampal DNA and litter composition modified sex effects on methylation of Egr1 in NAc. Few differences were seen for Bdnf. LGC modified some sex differences in behavior. © 2013 Wiley Periodicals, Inc. © 2013 Wiley Periodicals, Inc. Dev Psychobiol 56: 392–406, 2014.
Analysis of new classes of recreational drugs in sewage: Synthetic cannabinoids and amphetamine‐like substances
Authors: Reid Malcolm J. , Derry Lisa , Thomas Kevin V.
Published: March 2013

The analysis of sewage for the residues of commonly used illicit drugs has successfully been applied as a suitable approach for estimating community illicit drug use. The drug market is increasingly dynamic with new substances continually being marketed for recreational purposes. In this study, ultra‐high pressure liquid chromatography coupled to tandem mass spectrometry (UHPLC‐MS/MS) was used to simultaneously and quantitatively detect the exogenous biomarkers of new classes of recreational drugs in sewage collected from three different Norwegian cities (Oslo, Bergen, Hamar). The samples were screened for the presence of khat (d‐norpseudoephedrine and cathinone), mephedrone, pseudoephedrine, 7‐aminoflunitrazepam, para‐methoxyamphetamine (PMA), para‐methoxy‐N‐methylamphetamine (PMMA) and a selection of urinary metabolites of synthetic cannabinoids collectively termed ´Spice´ (5‐3‐1‐naphthoyl‐1H‐indol‐1‐yl‐pentanoic acid (JWH 018 N‐pentanoic acid), 1‐5‐hydroxypentyl‐1H‐indol‐3‐ylnaphthalen‐1‐yl‐methanone (JWH 018 N‐5‐hydroxypentyl), 4‐3‐1‐naphthoyl‐1H‐indol‐1‐yl‐butanoic acid (JWH 073 N‐butanoic acid), 1‐4‐hydroxybutyl‐1H‐indol‐3‐ylnaphthalen‐1‐yl‐methanone (JWH 073 N‐4‐hydroxybutyl), 1‐5‐hydroxypentyl‐1H‐indol‐3‐yl4‐methylnaphthalen‐1‐yl‐methanone (JWH 122 N‐5‐hydroxypentyl), 1‐5‐fluoro‐4‐hydroxypentyl‐1H‐indol‐3‐ylnaphthalen‐1‐ylmethanone (AM2201 N‐4‐hydroxypentyl), and 1‐5‐hydroxypentyl‐1H‐indol‐3‐yl4‐methoxyphenyl‐methanone (RCS‐4 N‐5‐hydroxypentyl)). Limits of detection were 1 ng/L for amphetamine like compounds and 5 ng/L for the metabolites of synthetic cannabinoids while the limits of quantification were 3 and 15 ng/L, respectively. Three of the fourteen selected biomarkers (cathine, pseudoephedrine and the synthetic cannabinoid metabolite JWH‐018 N‐5‐hydroxypentyl) were detected in sewage, alongside the illicit drugs (and/or metabolites) typically found in sewage (cocaine, benzoylecognine, methamphetamine, MDMA, and THC‐COOH). The khat biomarker d‐norpseudoephedrine was detected in Oslo sewage at a mean concentration of 93 ng/L that represents a daily load of 54 mg/day/1000 inhabitants. Pseudoephedrine was present at mean concentrations of between 27 and 67 ng/L representing normalized daily loads of between 10 (Hamar) and 24 mg/day/1000 inhabitants (Bergen). The daily normalized loads of JWH‐018 N‐5‐hydroxypentyl were between 49 (Oslo) and 62 mg/day/1000 inhabitants (Hamar). This study demonstrates for the first time that sewage biomarker analysis can be applied to evaluate not only the use the traditional illicit drugs (cocaine, cannabis and amphetamines), but also the use of certain new synthetic drugs. Copyright © 2013 John Wiley & Sons, Ltd.
Genetic diversity and phylogenetic analysis of native mountain ponies of Britain and Ireland reveals a novel rare population
Authors: Winton Clare L. , Hegarty Matthew J. , McMahon Robert , Slavov Gancho T. , McEwan Neil R. , Davies‐Morel Mina C.G. , Morgan Charly M. , Powell Wayne , Nash Deborah M.
Published: March 2013

The conservation of unique populations of animals is critical in order to preserve valuable genetic diversity and, where populations are free‐living, maintain their irreplaceable influence upon habitat ecology. An accurate assessment of genetic diversity and structure within and between populations is crucial in order to design and implement conservation strategies in natural and domesticated species. Moreover, where it is possible to identify relic populations that are related to a structured breed an ideal opportunity presents itself to model processes that reveal historical factors that have shaped genetic diversity. The origins of native UK mountain and moorland ponies are uncertain, but they may have directly descended from prehistoric populations and potentially harbour specific adaptations to the uplands of Britain and Ireland. To date, there have been no studies of population structure and genetic diversity present within a free‐living group of ponies in the Carneddau mountain range of North Wales. Herein, we describe the use of microsatellites and SNPs together with analysis of the mitochondrial control region to quantify the extent and magnitude of genetic diversity present in the feral Carneddau pony and relate this to several recognised British and Irish pony breeds. Our results establish that the feral Carneddau ponies represent a unique and distinctive population that merits recognition as a defined population and conservation priority. We discuss the implications for conservation of this population as a unique pool of genetic diversity adapted to the British uplands and potentially of particular value in maintaining the biodiversity of these habitats.
A novel Lyn‐protein kinase Cδ/ε‐protein kinase D axis is activated in B cells by signalosome‐independent alternate pathway BCR signaling
Authors: Guo Benchang , Rothstein Thomas L.
Published: March 2013

BCR signaling initiates multiple activities critical for B‐cell function. Recently, we identified an alternate BCR signaling pathway, induced by IL‐4, that is signalosome‐independent, unlike the classical signalosome‐dependent pathway, and that leads to activation of the MAP kinase, ERK. Here we questioned whether alternate pathway signaling extends to other key downstream events, especially protein kinase D (PKD) activation. We found that in murine spleen‐derived B cells the IL‐4‐induced alternate pathway for BCR signaling results in PKD and PKD substrate phosphorylation, and that alternate pathway phosphorylation of HDAC5/7 and other key substrates requires PKD. Furthermore, we found that tyrosine phosphorylation of PKCδ/ε occurs as a result of alternate but not classical pathway signaling and is required for phosphorylation of PKD and PKD substrates. This result identifies PKCδ/ε tyrosine phosphorylation as a unique outcome of the alternate pathway. The alternate pathway is mediated by Lyn that is not required for classical pathway signaling and we found that Lyn associates directly with PKCδ/ε and is required for phosphorylation of PKCδ/ε and of PKD. These findings indicate that IL‐4 influences B‐cell activation by inducing a novel signaling pathway from BCR to Lyn to PKCδ/ε to PKD.
Mycobacterium tuberculosis‐specific CD8+ T cells are functionally and phenotypically different between latent infection and active disease
Authors: Rozot Virginie , Vigano Selena , Mazza‐Stalder Jesica , Idrizi Elita , Day Cheryl L. , Perreau Matthieu , Lazor‐Blanchet Catherine , Petruccioli Elisa , Hanekom Willem , Goletti Delia , Bart Pierre‐Alexandre , Nicod Laurent , Pantaleo Giuseppe , Harari Alexandre
Published: March 2013

Protective immunity to Mycobacterium tuberculosis (Mtb) remains poorly understood and the role of Mtb‐specific CD8+ T cells is controversial. Here we performed a broad phenotypic and functional characterization of Mtb‐specific CD8+ T cells in 326 subjects with latent Mtb infection (LTBI) or active TB disease (TB). Mtb‐specific CD8+ T cells were detected in most (60%) TB patients and few (15%) LTBI subjects but were of similar magnitude. Mtb‐specific CD8+ T cells in LTBI subjects were mostly TEMRA cells (CD45RA+CCR7−), coexpressing 2B4 and CD160, and in TB patients were mostly TEM cells (CD45RA−CCR7−), expressing 2B4 but lacking PD‐1 and CD160. The cytokine profile was not significantly different in both groups. Furthermore, Mtb‐specific CD8+ T cells expressed low levels of perforin and granulysin but contained granzymes A and B. However, in vitro‐expanded Mtb‐specific CD8+ T cells expressed perforin and granulysin. Finally, Mtb‐specific CD8+ T‐cell responses were less frequently detected in extrapulmonary TB compared with pulmonary TB patients. Mtb‐specific CD8+ T‐cell proliferation was also greater in patients with extrapulmonary compared with pulmonary TB. Thus, the activity of Mtb infection and clinical presentation are associated with distinct profiles of Mtb‐specific CD8+ T‐cell responses. These results provide new insights in the interaction between Mtb and the host immune response.
Preparation of Mesoporous Titania Thin Films with Well‐Crystallized Frameworks by Using Thermally Stable Triblock Copolymers
Authors: Zakaria Mohamed B. , Suzuki Norihiro , Torad Nagy L. , Matsuura Mikiya , Maekawa Kazuhiko , Tanabe Hirofumi , Yamauchi Yusuke
Published: March 2013

Mesoporous titania (TiO2) thin films are prepared by a spin‐coating method using new laboratory‐made polystyrene‐block‐polybutadiene‐block‐polystyrene‐based triblock copolymers in which the polystyrene block is functionalized with hydrophilic sulfonic groups (abbreviated as sulfonated HmSEBmS). The polymer used in this study is more thermally stable than commercial Pluronic‐type block copolymers (e.g. P123 and F127). As‐prepared TiO2 thin films are calcined under various temperatures. The calcined films are systematically characterized by low‐angle and wide‐angle XRD, scanning electron microscopy (SEM), and nitrogen gas adsorption–desorption isotherms. As calcination temperatures increase, crystal growth proceeds in the pore walls and the phase transformation from anatase to rutile occurs. Even after calcination at 550 and 600 °C, the thin films nicely preserve high surface areas. On the other hand, the surface areas of mesoporous TiO2 prepared from P123 are markedly decreased after calcination at 550 °C.
Thermally Reversible, Flexible, Transparent, and Luminescent Ionic Organosilica Gels
Authors: Ru Qiaorong , Wang Yige , Zhang Wenjun , Yu Xiaoyan , Li Huanrong
Published: March 2013

Herein, we report a facile method to prepare thermally reversible, flexible, transparent, and luminescent ionic organosilica gels with tunable emission colors. The reaction of an amino‐functionalized ionic liquid ([NH2‐Bim]Br) with 3‐(triethoxysilyl)propyl isocyanate results in an ionic‐liquid‐modified alkoxysilane, the hydrolysis and condensation of which lead to thermally reversible, flexible, and luminescent ionic gels in the presence of lanthanide species. The luminescence color can be fine‐tuned by changing the Ln3+ ions (Eu3+, Tb3+, or a mixture of Eu3+ and Tb3+) and by changing the excitation wavelength. The absolute quantum yields of the flexible luminescent gels with europium(III) and terbium(III) complexes are determined to be 15 and 21 %, respectively, by using an integrating sphere. The thermal reversibility of the organosilicas makes them attractive for the development of flexible optical devices and flexible luminescent devices such as organic light‐emitting diodes (OLEDs).
Glycosynthase with Broad Substrate Specificity – an Efficient Biocatalyst for the Construction of Oligosaccharide Library
Authors: Wei Jinhua , Lv Xun , Lü Yang , Yang Gangzhu , Fu Lifeng , Yang Liu , Wang Jianjun , Gao Jianhui , Cheng Shuihong , Duan Qian , Jin Cheng , Li Xuebing
Published: March 2013

A versatile glycosynthase (TnG‐E338A) with strikingly broad substrate scope has been developed from Thermus nonproteolyticus β‐glycosidase (TnG) by using site‐directed mutagenesis. The practical utility of this biocatalyst has been demonstrated by the facile generation of a small library containing various oligosaccharides and a steroidal glycoside (total 25 compounds) in up to 100 % isolated yield. Moreover, an array of eight gluco‐oligosaccharides has been readily synthesized by the enzyme in a one‐pot, parallel reaction, which highlights its potential in the combinatorial construction of a carbohydrate library that will assist glycomic and glycotherapeutic research. Significantly, the enzyme provides a means by which glycosynthase technology may be extended to combinatorial chemistry.
Synthesis of Nopinone from β‐Pinene – A Journey Revisiting Methods for Oxidative Cleavage of C=C Bonds in Terpenoid Chemistry
Authors: Stolle Achim
Published: March 2013

The synthesis of nopinone from β‐pinene is an interesting model reaction for the investigation of methods for the oxidative cleavage of C=C bonds. Nopinone is used as chiral component for the total synthesis of different ligands or natural compounds such as di‐ or sesquiterpenes, as well as for the production of pharmaceutically active components. The cleavage of the C(2)=C(10) bond in β‐pinene by suitable oxidants leads to the replacement of the exo‐methylene functionality by a carbonyl group, which is more suitable for subsequent synthetic steps and the construction of complex condensed ring systems. Oxidants such as ozone, potassium permanganate, or sodium periodate have been applied successfully for this reaction. Ozonolysis of the double bond follows the classical Criegee mechanism, after which the secondary intermediate is isolated and reductive workup yields the carbonyl products. Dihydroxylation of the double bond can be initiated by permanganate or periodate. Further oxidation leads to the same products known from ozonolysis. Periodate oxidations in particular can be supported by catalysts such as RuCl3 or OsO4, because the oxidant is generally too weak to dihydroxylate the double bond. OsO4‐catalyzed reactions, for example, can take advantage of a co‐oxidant such as trimethylammonium N‐oxide for the dihydroxylation step. Other oxidants such as hydrogen peroxide or dioxygen have also been tested but with minor success with respect to conversion or selectivity.
Synthesis of 1,2‐Diaryl‐1H‐indol‐4‐ols and 1,2‐Diaryl‐7‐ethoxy‐1,5,6,7‐tetrahydroindol‐4‐ones from Arylglyoxals and Enamines through Domino Reactions
Authors: Maity Subhendu , Pathak Sudipta , Pramanik Animesh
Published: March 2013

A series of 1,2‐diaryl‐ and 1‐alkyl‐2‐aryl‐1H‐indol‐4‐ols and 1,2‐diaryl‐7‐ethoxy‐1,5,6,7‐tetrahydroindol‐4‐ones have been prepared by the domino reactions of arylglyoxals and enamines at reflux in non‐nucleophilic and nucleophilic solvents such as acetonitrile and ethanol, respectively. The transformation occurs by annulation followed by aromatization without the use of any metal or catalyst.
Synthesis of Photoreactive 2‐Phenethylamine Derivatives – Synthesis of Adenosine Derivatives Enabling Functional Analysis of Adenosine Receptors by Photoaffinity Labeling
Authors: Murai Yuta , Masuda Katsuyoshi , Ogasawara Yui , Wang Lei , Hashidoko Yasuyuki , Hatanaka Yasumaru , Iwata So , Kobayashi Takuya , Hashimoto Makoto
Published: March 2013

2‐Phenylethylamine is well known as a substructure of many biologically active compounds, and the synthesis of its photoreactive derivatives to allow the analysis of biological functions is reported. This allowed us to synthesise ligands for adenosine receptors, which have many functional roles in biology and have been extensively studied for their many roles in maintaining homeostasis. Adenosine is one of the most common biochemical compounds, but photoaffinity labeling has not yet been used to study adenosine receptors. Synthetic methods for producing photoreactive adenosine derivatives that can be used with adenosine receptors were established for the photophores phenyl azide and benzophenone. The effect of the introduction of the photoreactive components was determined using an adenosine receptor assay.
Synthesis of (+)‐Centrolobine and Its Analogues by Using Acyl Anion Chemistry
Authors: Sudarshan Kasireddy , Aidhen Indrapal Singh
Published: March 2013

A new route based on the use of acyl anion chemistry was developed for the synthesis of (+)‐centrolobine and its analogues. Acid‐catalyzed benzylic cation initiated cyclization was the key step in the stereoselective formation of the cis‐2,6‐disubstituted tetrahydropyran ring. The developed methodology was applied to the synthesis of (+)‐centrolobine analogues containing electron‐donating substituents in the aryl rings.
Electrochemical Sensing of Dissolved Hydrogen in Aqueous Solutions as a Tool to Monitor Magnesium Alloy Corrosion
Authors: Kuhlmann Julia , Witte Frank , Heineman William R.
Published: March 2013

Magnesium and its alloys have been the focus of the development of biodegradable metallic implant materials for years. Since water is reduced to form hydrogen gas during their corrosion, the amount and rate of hydrogen evolution, and therefore the dissolved hydrogen, could be used as an indicator to monitor and compare the corrosion. Here we report on a commercially available Clark‐Type amperometric microsensor and a simple potentiometric sensor for hydrogen to monitor the corrosion of a magnesium alloy in aqueous solutions. The sensors were compared using rare‐earth containing Mg alloy discs (Mg with 4 % Y, 2 % Nd, 0.5 % Ga, 0.5 % Dy) immersed in phosphate buffered saline (pH 7.4) and 3.5 % NaCl.
A Sensitive Microchip‐Based Immunosensor for Electrochemical Detection of Low‐Level Biomarker S100B
Authors: Liu Yun , Wang Huixiang , Chen Jiayao , Liu Chengcheng , Li Wenxian , Kong Jilie , Yang Pengyuan , Liu Baohong
Published: March 2013

A sensitive microchip‐based electrochemical immunosensor has been developed for quantification of low level S100B, which is an important biomarker of neurological diseases. Poly(ethyleneimine) (PEI), containing NH2 groups, was firstly modified on the surface of the poly(methyl methacrylate) (PMMA) microchannel to capture monoclonal anti‐S100B. Afterward, S100B and second polyclonal anti‐S100 were sequentially coupled by antigen‐antibody specific interaction, and the alkaline phosphatase conjugates (ALP‐conjugates) were bound to the second antibodies in a PMMA microchannnel. Using differential pulse voltammetry (DPV), the immunosensor achieved the detection limit of 0.1 pg mL−1 with the linear concentration range of 0.1 pg  mL−1–100 pg mL−1. The results indicate that the proposed strategy not only has high specificity and sensitivity, but also resists nonspecific protein adsorption. The disposable microchip immunosensor shows good promise for clinical screening of important biomarkers and convenient point‐of‐care diagnostics.
Electrochemical Pseudo‐Titration of Water‐Soluble Antioxidants
Authors: Tacchini Philippe , Lesch Andreas , Neequaye Alice , Lagger Grégoire , Liu Jifeng , Cortés‐Salazar Fernando , Girault Hubert H.
Published: March 2013

An amperometric test for the antioxidant power (AOP) of biological and food samples is presented. The gist of the method is to measure by linear sweep voltammetry the anodic current produced during the oxidation of the labile species present in the sample and then rationalizing this signal with a mathematical treatment that allows the pseudo‐titration of antioxidants (AOs) around a given threshold potential. As a result, the AOP of the sample is calculated. This method allows the discrimination of the most biologically relevant AOs that react rapidly and at low oxidation potentials, from less reactive AOs that are oxidised slowly and at much higher oxidation potentials. This methodology was applied for measuring the AOP of blood, saliva, and natural drinks like orange juice.
Adsorption and Electrooxidation of Nucleic Acids at Glassy Carbon Electrodes Modified with Multiwalled Carbon Nanotubes Dispersed In Polylysine
Authors: Jalit Yamile , Moreno Mónica , Gutierrez Fabiana A. , Sanchez Arribas Alberto , Chicharro Manuel , Bermejo Esperanza , Zapardiel Antonio , Parrado Concepción , Rivas Gustavo A. , Rodríguez Marcela C.
Published: March 2013

We report the advantages of the adsorption and electrooxidation of oligonucleotides and calf‐thymus double stranded DNA (dsDNA) at glassy carbon electrodes (GCE) modified with a dispersion of multiwalled carbon nanotubes (MWCNT) in poly‐L‐lysine (Plys) (GCE/MWCNT‐Plys). Important enhancement in the guanine oxidation signal was obtained by adsorptive stripping voltammetry (AdSV) due to a most favorable interaction between the negatively charged DNAs and the positively charged Plys that support the MWCNT. The layer of oligo(dG)11 immobilized at GCE/MWCNT‐Plys was successfully used for the selective detection of the hybridization event using oligo(dG)11/oligo(dC)11 as model.
Electrochemical Behaviour of Isoflavones Genistein and Biochanin A at a Glassy Carbon Electrode
Authors: Popa Oana M. , Diculescu Victor C.
Published: March 2013

The electrochemical behaviour of genistein and biochanin A was studied at a glassy carbon electrode by cyclic, differential pulse and square wave voltammetry. Genistein undergoes three irreversible, pH dependent oxidation reactions with the transfer of one electron and one proton from each hydroxyl group. The formation of two electroactive products that undergo reversible redox reactions was observed. Biochanin A undergoes two irreversible, pH dependent reactions due to the oxidation of the two hydroxyl groups. The electrochemical behaviour of the chemical analogue daidzein was also investigated. The electroactive centres of genistein and biochanin A were identified and their oxidation mechanisms discussed.
Label‐Free Amperometric Magnetoimmunosensors for Direct Determination of Lactoperoxidase in Milk
Authors: Torrente‐Rodríguez Rebeca M. , Campuzano Susana , Gamella María , Conzuelo Felipe , Reviejo A. Julio , Pingarrón José M.
Published: March 2013

Novel label free electrochemical magnetoimmunosensors for the determination of lactoperoxidase (LPO) were developed based on the use of streptavidin‐magnetic beads (Strep‐MBs) and graphite‐Teflon composite or screen‐printed carbon electrodes (SPCEs) as electrochemical transducers. Biotinylated antiLPO antibodies were immobilized onto the Strep‐MBs and a direct‐type immunoassay by detecting the enzymatic activity of the captured target enzyme using amperometric detection at −0.20 V and the H2O2/hydroquinone (HQ) system was employed. The SPCEs‐based magnetoimmunosensors allowed a detection limit (LOD) of 0.12 μg mL−1 and a good selectivity against nontarget proteins. Moreover, their analytical usefulness was successfully demonstrated by analyzing different types of milk samples.
Simple and Sensitive Electrochemical DNA Detection of Primer Generation‐Rolling Circle Amplification
Authors: Lee Derek Chun , Yip Shea Ping , Lee Thomas M. H.
Published: March 2013

A new electrochemical sequence‐specific DNA detection platform based on primer generation‐rolling circle amplification (PG‐RCA), methylene blue (MB) redox indicator, and indium tin oxide (ITO) electrode is reported. In the presence of a specific target sequence, PG‐RCA, an isothermal DNA amplification technique, produced large amounts of amplicons in an exponential manner. In addition to the standard components, the reaction mixture contained MB, which bound with the PG‐RCA amplicons. End‐point electrochemical measurement by differential pulse voltammetry (DPV) was performed using ITO electrode. The amplicon‐bound MB resulted in a lower DPV signal than free MB due to a smaller diffusion coefficient as well as electrostatic repulsion between the negatively charged amplicon‐bound MB and ITO electrode. With simple assay design (recognition probe) and instrumentation (operating temperature at 37 °C and ITO electrode without the need for probe immobilization), this detection platform is well suited for point‐of‐care and on‐site testing. Real‐time measurement was also achieved by pretreating the ITO electrode with bovine serum albumin.
Preparation and application of trimethylamine amination polychloromethyl styrene nanolatex coated capillary column for the determination of bromate by field‐amplified sample stacking open‐tubular capillary electrochromatography
Authors: Guo Yaxiao , Xu Feifei , Meng Lei , Tang Wei , Xia Yan , Wu Yangjie , Zhang Shusheng
Published: March 2013

A new trimethylamine amination polychloromethyl styrene nanolatex (TMAPL) and TMAPL coated capillary column (ccc‐TMAPL) were successfully prepared. The TMAPL coating was characterized with reversed steady EOF values of ca. ‒16.8 × 10−5 cm2 V−1 s−1. It was applied to establish open‐tubular (OT) CEC and field‐amplified sample stacking (FASS) OT‐CEC methods for the determination of bromate in tap water. Compared to OT‐CEC, the LOD with FASS‐OT‐CEC was improved from 80 to 8 ng/mL. The developed FASS‐OT‐CEC method was practically used for the analysis of bromate in tap water samples with recoveries ranging from 93.6 to 103.5%.
Preparation of poly(m‐phenylenediamine)/ZnO composites and their photocatalytic activities for degradation of C.I. acid red 249 under UV and visible light irradiations
Authors: Peng Yong‐Gang , Ji Jun‐Ling , Zhang Yun‐Long , Wan Huai‐Xin , Chen Da‐Jun
Published: March 2013

Poly(m‐phenylenediamine)/ZnO (PMPD/ZnO) composites were successfully synthesized by chemical oxidative polymerization method. Powder X‐ray diffraction (XRD), Fourier‐transform infrared spectra (FTIR), transmission electron microscope, and UV–vis spectra (UV–vis) were used to characterize the prepared PMPD/ZnO composites. Their photocatalytic activities were evaluated by the degradation of C.I. Acid Red 249 (AR249) aqueous solutions under UV and visible light irradiations. Results showed that PMPD/ZnO composites exhibited good photocatalytic activities under UV and visible light irradiations, whereas neat ZnO had only ultraviolet light photocatalytic activity. Moreover, PMPD/ZnO composites had better photocatalytic stabilities. After five successive cycles, their photocatalytic activities for degradation of AR249 were still over 75% of that for the first cycling run under UV and visible light irradiations. The inhibition of AR249 degradation by adding isopropyl alcohol (IPA) and 1‐butanol (1‐BuOH) to the UV/PMPD/ZnO system revealed that both photogenerated holes (h+) and hydroxyl radicals (OH·) were the main oxidants in the photodegradation reactions. However, in the visible light/PMPD/ZnO system, the main oxidant was hydroxyl radicals (OH·). © 2013 American Institute of Chemical Engineers Environ Prog, 33: 123–130, 2014
GPT2: Empirical slant delay model for radio space geodetic techniques
Authors: Lagler K. , Schindelegger M. , Böhm J. , Krásná H. , Nilsson T.
Published: March 2013

Up to now, state‐of‐the‐art empirical slant delay modeling for processing observations from radio space geodetic techniques has been provided by a combination of two empirical models. These are GPT (Global Pressure and Temperature) and GMF (Global Mapping Function), both operating on the basis of long‐term averages of surface values from numerical weather models. Weaknesses in GPT/GMF, specifically their limited spatial and temporal variability, are largely eradicated by a new, combined model GPT2, which provides pressure, temperature, lapse rate, water vapor pressure, and mapping function coefficients at any site, resting upon a global 5° grid of mean values, annual, and semi‐annual variations in all parameters. Built on ERA‐Interim data, GPT2 brings forth improved empirical slant delays for geophysical studies. Compared to GPT/GMF, GPT2 yields a 40% reduction of annual and semi‐annual amplitude differences in station heights with respect to a solution based on instantaneous local pressure values and the Vienna mapping functions 1, as shown with a series of global VLBI (Very Long Baseline Interferometry) solutions.
How systematic is organized tropical convection within the MJO?
Authors: Dias J. , Leroux S. , Tulich S. N. , Kiladis G. N.
Published: March 2013

Recent theories of the dynamics of the Madden‐Julian Oscillation (MJO) have been building on its multiscale structure. While it is observed that much of the convection within the MJO envelope is related to higher frequency equatorial waves, it is not known to what extent this organization is systematic from one MJO event to another. For instance, it is of interest to know whether MJO favors waves of particular types and propagation direction, and how this variability changes seasonally. This work presents a wavenumber‐frequency spectral analysis of the MJO convective envelope, which indicates that there is no strong preferred scale of high frequency organization that is ubiquitous to the MJO. This null finding supports the use of convective parametrizations in modeling the MJO since, while moist convection is likely critical to its initiation and maintenance, the particular details of that organization may not be critical for its prediction and simulation.
Triple check: Observations verify structural realism of an ocean ecosystem model
Authors: Santora Jarrod A. , Sydeman William J. , Messié Monique , Chai Fei , Chao Yi , Thompson Sarah Ann , Wells Brian K. , Chavez Francisco P.
Published: March 2013

Improvements in fisheries and ecosystem management could be made if the prediction of key zooplankton, such as krill, were possible using ocean ecosystem models. To examine structural realism, hence the validity of a coupled physical‐biogeochemical model, we compared measured spatiotemporal dynamics of krill and seabird abundance off California to hindcasted mesozooplankton derived from an independently designed model. Observed krill and modeled mesozooplankton (Z2) displayed latitudinal coherence but distinct longitudinal offsets, possibly related to unrealistic bathymetry in the model. Temporally, Z2, Thysanoessa spinifera (a neritic krill species) and seabird density and reproductive performance were well correlated, indicating that quantitative prediction regarding marine predators in upwelling ecosystems is within reach. Despite its basin‐scale framework, the ROMS‐CoSiNE model captures zooplankton and top predator dynamics regionally in the central California region, suggesting its utility for management of marine ecosystems and highlighting rapid advances that can be made through collaboration between empirical scientists and ecosystem modelers.
The influence of fracture anisotropy on CO2 flow
Authors: Bond Clare E. , Wightman Ruth , Ringrose Philip S.
Published: March 2013

Greenhouse gas mitigation through geological storage of carbon dioxide is dependent on rock formations storing CO2 effectively. Secure containment for periods of 100 × 105 years nee7ds to be verifiable. The effectiveness of geological storage is reliant on the chemical and physical properties of the geological storage complex and its ability to inhibit migration of CO2. Petroleum reservoir data and field evidence show that fracture networks often act as pathways for fluid movement, potentially allowing fluids to migrate to the surface within the time scale of interest. We demonstrate the importance of predicting the effects of fracture networks on flow, using a case study from the In Salah CO2 storage site, and show how fracture permeability is closely controlled by the stress regime determining the conductive fracture network. Our results demonstrate that fracture network prediction combined with present‐day stress analysis can be used to successfully predict CO2 movement in the subsurface.
Effects of the GABA‐uptake blocker NNC‐711 on spontaneous sharp wave–ripple complexes in mouse hippocampal slices
Authors: Viereckel Thomas , Kostic Milos , Bähner Florian , Draguhn Andreas , Both Martin
Published: March 2013

The precise temporal and spatial activity patterns of neurons in cortical networks are organized by different state‐dependent types of network oscillations. GABAergic inhibition plays a key role in the underlying mechanisms of such oscillations and it has been suggested that the duration of widely distributed phasic inhibitory postsynaptic potentials (IPSPs) determines the frequency of the resulting network oscillation. Here, we test this hypothesis in an in vitro model of sharp wave–ripple (SPW‐R) complexes, a particularly fast pattern of network oscillations at ∼200 Hz which is involved in memory consolidation. We recorded SPW‐R in mouse hippocampal slices in the absence and presence of NCC‐711, an inhibitor of GABA uptake. The resulting prolongation of IPSP resulted in reduced occurrence of SPW‐R, whereas the superimposed fast oscillations as well as the precision of rhythmic cell synchronization remained stable. Application of Diazepam which is a positive modulator of the GABAA receptor led to consistent results. We conclude that phasic inhibition is a major regulator of network excitability in CA3 (where SPW‐Rs are generated), but does not set the frequency of hippocampal ripples. © 2013 Wiley Periodicals, Inc.
Differential roles of the dorsal and ventral hippocampus in predator odor contextual fear conditioning
Authors: Wang Melissa E. , Fraize Nicolas P. , Yin Linda , Yuan Robin K. , Petsagourakis Despina , Wann Ellen G. , Muzzio Isabel A.
Published: March 2013

The study of fear memory is important for understanding various anxiety disorders in which patients experience persistent recollections of traumatic events. These memories often involve associations of contextual cues with aversive events; consequently, Pavlovian classical conditioning is commonly used to study contextual fear learning. The use of predator odor as a fearful stimulus in contextual fear conditioning has become increasingly important as an animal model of anxiety disorders. Innate fear responses to predator odors are well characterized and reliable; however, attempts to use these odors as unconditioned stimuli in fear conditioning paradigms have proven inconsistent. Here we characterize a contextual fear conditioning paradigm using coyote urine as the unconditioned stimulus. We found that contextual conditioning induced by exposure to coyote urine produces long‐term freezing, a stereotypic response to fear observed in mice. This paradigm is context‐specific and parallels shock‐induced contextual conditioning in that it is responsive to extinction training and manipulations of predator odor intensity. Region‐specific lesions of the dorsal and ventral hippocampus indicate that both areas are independently required for the long‐term expression of learned fear. These results in conjunction with c‐fos immunostaining data suggest that while both the dorsal and ventral hippocampus are required for forming a contextual representation, the ventral region also modulates defensive behaviors associated with predators. This study provides information about the individual contributions of the dorsal and ventral hippocampus to ethologically relevant fear learning. © 2013 Wiley Periodicals, Inc.
OPTIMAL WATER SCHEDULING IN IRRIGATION CANAL NETWORK USING PARTICLE SWARM OPTIMIZATION
Authors: Pawde Anil W. , Mathur Yogesh P. , Kumar Rajesh
Published: March 2013

Rotational water scheduling in the irrigation canal network is a problem of preparing the operation schedule of lateral canals to deliver the required quantity of water to the irrigation fields under it within the rotational period as per the crop water requirement. The important objectives of a rotational canal schedule are to minimize capacity of canal and operational stages of the head gate of supply canal; subject to canal capacity constraint. In the present paper, to achieve these objectives, the decision variables considered are rate of water delivery and start time of delivery points, which are determined by the model within the specified range. Particle swarm optimization has been used to solve the problem and applied on the two water scheduling problems for generating the optimal water schedule. The schedules generated by the model shows that the inflow hydrographs of supply canals are approximately constant throughout the rotation period with least operation of canal head gate which could results in minimum conveyance losses. This model could be used for the preparation of a rotational schedule for the existing irrigation canal system and a more economic design of the supply canals for a new irrigation canal system. Copyright © 2013 John Wiley & Sons, Ltd.
COMBINATION OF TRADITION AND THE MODERN AGE: NEW METHODS FOR EFFICIENT WATER MANAGEMENT
Authors: Röttcher Klaus
Published: March 2013

The focus of this paper is on whether and which traditional structures and methods, perhaps in combination with modern approaches and elements, could lead to a better and more sustainable management of water resources. Arid and semi‐arid regions are at the centre of this paper. Some subject areas and methods in which the consideration of traditional methods and structures appears particularly promising are described. The examples listed show that in the various areas there are technically interesting aspects of historical water structures that still have relevance today. In addition to this, however, there is another overriding benefit of water historic research, which is independent of any single structure. In some cases these benefits exist even when the system itself has no further use today or its traces have largely disappeared. Copyright © 2013 John Wiley & Sons, Ltd.
Structures and reaction mechanisms of GTP cyclohydrolases
Authors: Gräwert Tobias , Fischer Markus , Bacher Adelbert
Published: March 2013

GTP cyclohydrolases generate the first committed intermediates for the biosynthesis of certain vitamins/cofactors (folic acid, riboflavin, deazaflavin, and tetrahydrobiopterin), deazapurine antibiotics, some t‐RNA bases (queuosine, archaeosine), and the phytotoxin, toxoflavin. They depend on divalent cations for hydrolytic opening of the imidazole ring of the substrate, guanosine triphosphate (GTP). Surprisingly, the ring opening reaction is not the rate‐limiting step for GTP cyclohydrolases I and II whose mechanism have been studied in some detail. GTP cyclohydrolase I, Ib, and II are potential targets for novel anti‐infectives. Genetic factors modulating the activity of human GTP cyclohydrolase are highly pleiotropic, since the signal transponders whose biosyntheses require their participation (nitric oxide, catecholamines) impact a very wide range of physiological phenomena. Recent studies suggest that human GTP cyclohydrolase may become an oncology target. © 2013 IUBMB Life 65(4):310–322, 2013.
Preservice Counselors' Knowledge of Classmates' Problems of Professional Competency
Authors: Brown‐Rice Kathleen A. , Furr Susan
Published: March 2013

Master's‐level students (N= 389) in programs accredited by the Council for Accreditation of Counseling and Related Educational Programs were surveyed to determine their knowledge of classmates’ problems of professional competency (PPC). Descriptive statistics were used to analyze the data, and a principal components analysis and Cronbach's alpha coefficients were applied to reduce and investigate the pattern of correlations among participants’ responses. Findings suggest that the majority of preservice counselors are aware of classmates with PPC and report frustration with faculty for not addressing problematic peers.
Counseling in Australia: Past, Present, and Future
Authors: Schofield Margot J.
Published: March 2013

The author describes the evolution of the counseling profession in Australia within the broader historic and sociocultural context. The process of professionalization is outlined, particularly over the past 15 years, and data on the demographic, professional, and practice characteristics of counselors are reviewed. Future professional directions and challenges are explored.
Outplacement services for displaced employees: attitudes of human resource managers based on differences in internal and external delivery
Authors: Kilcrease Kelly M.
Published: March 2013

The results from a survey of 238 human resources executives from organizations that offer outplacement counseling (OPC) internally and 168 that offer it externally suggest that internal OPC delivery is inferior to external OPC delivery. The author found that most internal OPC organizations did not offer the 13 traditional OPC services, even when they were viewed as important. However, both groups had deficiencies in measuring their OPC relative to gender, age, and effectiveness. Internal OPC organizations rated the quality of their OPC service as 4.77 on a scale of 10, whereas external OPC organizations rated their services as 7.96.
Roles of I(f) and intracellular Ca2+ release in spontaneous activity of ventricular cardiomyocytes during murine embryonic development
Authors: Wang Peng , Tang Ming , Gao Linlin , Luo Hongyan , Wang Guoping , Ma Xue , Duan Yaqi
Published: March 2013

In recent years, the contribution of I(f), an important pacemaker current, and intracellular Ca2+ release (ICR) from sarcoplasmic reticulum to pacemaking and arrhythmia has been intensively studied. However, their functional roles in embryonic heart remain uncertain. Using patch clamp, Ca2+ imaging, and RT‐PCR, we found that I(f) regulated the firing rate in early and late stage embryonic ventricular cells, as ivabradine (30 µM), a specific blocker of I(f), slowed down action potential (AP) frequency. This inhibitory effect was even stronger in late stage cells, though I(f) was down‐regulated. In contrast to I(f), ICR was found to be indispensable for the occurrence of APs in ventricular cells of different stages, because abolishment of ICR with ryanodine and 2‐aminoethoxydiphenyl borate (2‐APB), specific blockers of ryanodine receptors (RyRs) and inositol trisphosphate receptors (IP3Rs), completely abolished APs. In addition, we noticed that RyR‐ and IP3R‐mediated ICR coexisted in early‐stage ventricular cells and RyRs functionally dominated. While at late stage RyRs, but not IP3Rs, mediated ICR. In both early and late stage ventricular cells, Na‐Ca exchanger current (INa/Ca) mediated ICR‐triggered depolarization of membrane potential and resulted in the initiation of APs. We also observed that different from I(f), which presented as the substantial component of the earlier diastolic depolarization current, application of ryanodine, and/or 2‐APB slowed the late phase of diastolic depolarization. Thus, we conclude that in murine embryonic ventricular cells I(f) regulates firing rate, while RyRs and IP3Rs (early stage) or RyRs (late stage)‐mediated ICR determines the occurrence of APs. J. Cell. Biochem. 114: 1852–1862, 2013. © 2013 Wiley Periodicals, Inc.
Cadmium affects focal adhesion kinase (FAK) in mesangial cells: Involvement of CaMK‐II and the actin cytoskeleton
Authors: Choong Grace , Liu Ying , Templeton Douglas M.
Published: March 2013

The toxic metal ion cadmium (Cd2+) induces pleiotropic effects on cell death and survival, in part through effects on cell signaling mechanisms and cytoskeletal dynamics. Linking these phenomena appears to be calmodulin‐dependent activation of the Ca2+/calmodulin‐dependent protein kinase II (CaMK‐II). Here we show that interference with the dynamics of the filamentous actin cytoskeleton, either by stabilization or destabilization, results in disruption of focal adhesions at the ends of organized actin structures, and in particular the loss of vinculin and focal adhesion kinase (FAK) from the contacts is a result. Low‐level exposure of renal mesangial cells to CdCl2 disrupts the actin cytoskeleton and recapitulates the effects of manipulation of cytoskeletal dynamics with biological agents. Specifically, Cd2+ treatment causes loss of vinculin and FAK from focal contacts, concomitant with cytoskeletal disruption, and preservation of cytoskeletal integrity with either a calmodulin antagonist or a CaMK‐II inhibitor abrogates these effects of Cd2+. Notably, inhibition of CaMK‐II decreases the migration of FAK‐phosphoTyr925 to a membrane‐associated compartment where it is otherwise sequestered from focal adhesions in a Cd2+‐dependent manner. These results add further insight into the mechanism of the CaMK‐II‐dependent effects of Cd2+ on cellular function. J. Cell. Biochem. 114: 1832–1842, 2013. © 2013 Wiley Periodicals, Inc.
Snail mediates PDGF‐BB‐induced invasion of rat bone marrow mesenchymal stem cells in 3D collagen and chick chorioallantoic membrane
Authors: Lu Changlian , Sun Xiaojiao , Sun Lijun , Sun Jianping , Lu Yan , Yu Xin , Zhou Lingyun , Gao Xu
Published: March 2013

We previously reported that membrane type‐1 matrix metalloproteinase (MT1‐MMP) enables mesenchymal stem cells (MSCs) to move through both three‐dimensional (3D) type I collagen and basement membrane barriers; however, its upstream regulating factors were unidentified. Here, we report that PDGF‐BB upregulates both mRNA and protein expression of snail in rat bone marrow MSCs (rBMMSCs). PDGF‐BB enhances rBMMSC invasion in 3D collagen, which is blocked by snail specific siRNA transfection. Snail overexpression induced by plasmid transfection results in increased rBMMSC invasion in 3D collagen. Snail expression induced by PDGF‐BB in MSCs is inhibited by LY294002 and PD98059, which are inhibitors of the PI3K/AKT and MAPK1/2/ERK1/2 signaling pathways, respectively. MT1‐MMP expression in rBMMSCs, both as mRNA and protein, is decreased by snail siRNA transfection, but increased by snail overexpression, indicating that they are controlled by snail. Finally, snail controls MSC transmigration through chorioallantoic membrane of 11‐day‐old chick embryos. Taken together, these in vitro and in vivo data identify snail as a critical mediator for rBMMSC invasion induced by PDGF‐BB. J. Cell. Physiol. 228: 1827–1833, 2013. © 2013 Wiley Periodicals, Inc.