Sample records for cdta

  1. Current Mode Full-Wave Rectifier Based on a Single MZC-CDTA

    Directory of Open Access Journals (Sweden)

    Neeta Pandey


    Full Text Available This paper presents a current mode full-wave rectifier based on single modified Z copy current difference transconductance amplifier (MZC-CDTA and two switches. The circuit is simple and is suitable for IC implementation. The functionality of the circuit is verified with SPICE simulation using 0.35 μm TSMC CMOS technology parameters.

  2. A new functional site W115 in CdtA is critical for Aggregatibacter actinomycetemcomitans cytolethal distending toxin.

    Directory of Open Access Journals (Sweden)

    Lu Li

    Full Text Available Aggregatibacter actinomycetemcomitans, a specific pathogen of localized aggressive periodontitis, produces a cytolethal distending toxin (CDT that arrests eukaryotic cells irreversibly in G0/G1 or G2/M phase of the cell cycle. Although structural studies show that the aromatic patch region of CdtA plays an important role in its biological activity, the functional sites of CdtA have not been firmly established. In this study, site-specific mutagenesis strategy was employed for cdtA point mutations construction so as to examine the contributions of individual amino acids to receptor binding and the biological activity of holotoxin. The binding ability was reduced in CdtA(Y181ABC holotoxin and the biological function of CDT was not weaken in CdtA(Y105ABC, CdtA(Y125ABC, CdtA(F109ABC and CdtA(S106NBC holotoxin suggesting that these sites were not critical to CDT. But the binding activity and cell cycle arrest ability of holotoxin complexes were inhibited in CdtA(W115GBC. And this site did not affect the holotoxin assembly by size exclusion chromatography. Therefore, W115 might be a critical site of CdtA binding ability. These findings suggest that the functional sites of CdtA are not only in the aromatic patch region. W115, the new functional site is critical for receptor binding and cell cycle arrest, which provides potential targets for pharmacological disruption of CDT activity.

  3. Distinct Roles for CdtA and CdtC during Intoxication by Cytolethal Distending Toxins.

    Directory of Open Access Journals (Sweden)

    Shandee D Dixon

    Full Text Available Cytolethal distending toxins (CDTs are heterotrimeric protein exotoxins produced by a diverse array of Gram-negative pathogens. The enzymatic subunit, CdtB, possesses DNase and phosphatidylinositol 3-4-5 trisphosphate phosphatase activities that induce host cell cycle arrest, cellular distension and apoptosis. To exert cyclomodulatory and cytotoxic effects CDTs must be taken up from the host cell surface and transported intracellularly in a manner that ultimately results in localization of CdtB to the nucleus. However, the molecular details and mechanism by which CDTs bind to host cells and exploit existing uptake and transport pathways to gain access to the nucleus are poorly understood. Here, we report that CdtA and CdtC subunits of CDTs derived from Haemophilus ducreyi (Hd-CDT and enteropathogenic E. coli (Ec-CDT are independently sufficient to support intoxication by their respective CdtB subunits. CdtA supported CdtB-mediated killing of T-cells and epithelial cells that was nearly as efficient as that observed with holotoxin. In contrast, the efficiency by which CdtC supported intoxication was dependent on the source of the toxin as well as the target cell type. Further, CdtC was found to alter the subcellular trafficking of Ec-CDT as determined by sensitivity to EGA, an inhibitor of endosomal trafficking, colocalization with markers of early and late endosomes, and the kinetics of DNA damage response. Finally, host cellular cholesterol was found to influence sensitivity to intoxication mediated by Ec-CdtA, revealing a role for cholesterol or cholesterol-rich membrane domains in intoxication mediated by this subunit. In summary, data presented here support a model in which CdtA and CdtC each bind distinct receptors on host cell surfaces that direct alternate intracellular uptake and/or trafficking pathways.

  4. The Moessbauer effect in Fe(III) HEDTA, Fe(III) EDTA, and Fe(III) CDTA compounds

    International Nuclear Information System (INIS)

    Prado, F.R.


    The dependence of Moessbauer spectra with pH value of Fe(III)HEDTA and Fe(III)CDTA compounds is studied. Informations on formation processes of LFe-O-FeL (L=ligand) type dimers by the relation of titration curves of Fe(III)EDTA, Fe(III)HEDTA and Fe(III)CDTA compounds with the series of Moessbauer spectra, are obtained. Some informations on Fe-O-Fe bond structure are also obtained. Comparing the titration curves with the series of Moessbauer spectra, it is concluded that the dimerization process begins when a specie of the form FeXOH α (X = EDTA, HEDTA, CDTA; α = -1, -2) arises. (M.C.K.) [pt

  5. The CDTA-soluble pectic substances from soybean meal are composed of rhamnogalacturonan and xylogalacturonan but not homogalacturonan

    NARCIS (Netherlands)

    Vliegenthart, J.F.G.; Huisman, M.M.H.; Kamerling, J.P.; Schols, H.A.; Voragen, A.G.J.


    Structural characteristics of pectic substances extracted from soybean meal cell walls (water unextractable solids) with a chelating agent-containing buffer (0.05M 1,2-diaminocyclohexane-N,N,N,N-tetraacetic acid (CDTA) and 0.05M NH4-oxalate in 0.05M NaOAc buffer) were studied. The arabinogalactans

  6. 1HN, 13C, and 15N Resonance Assignments of the CDTb-Interacting Domain (CDTaBID) from the Clostridium difficile Binary Toxin Catalytic Component (CDTa, residues 1–221) (United States)

    Roth, Braden M.; Varney, Kristen M.; Rustandi, Richard R.; Weber, David J.


    Once considered a relatively harmless bacterium, Clostridium difficile has become a major concern for healthcare facilities, now the most commonly reported hospital-acquired pathogen. C. difficile infection (CDI) is usually contracted when the normal gut microbiome is compromised by antibiotic therapy, allowing the opportunistic pathogen to grow and produce its toxins. The severity of infection ranges from watery diarrhea and abdominal cramping to pseudomembranous colitis, sepsis, or death. The past decade has seen a marked increase in the frequency and severity of CDI among industrialized nations owing directly to the emergence of a highly virulent C. difficile strain, NAP1. Along with the large Clostridial toxins expressed by non-epidemic strains, C. difficile NAP1 produces a binary toxin, CDT (C. difficile transferase). As the name suggests, CDT is a two-component toxin comprised of an ADP-ribosyltransferase (ART) component (CDTa) and a cell-binding/translocation component (CDTb) that function to destabilize the host cytoskeleton by covalent modification of actin monomers. Central to the mechanism of binary toxin-induced pathogenicity is the formation of CDTa/CDTb complexes at the cell surface. From the perspective of CDTa, this interaction is mediated by the N-terminal domain (residues 1–215) and is spatially and functionally independent of ART activity, which is located in the C-terminal domain (residues 216–420). Here we report the 1HN, 13C, and 15N backbone resonance assignments of a 212 amino acid, ~23 kDa N-terminal CDTb-interacting domain (CDTaBID) construct by heteronuclear NMR spectroscopy. These NMR assignments represent the first component coordination domain for a family of Clostridium or Bacillus species harboring ART activity. Our assignments lay the foundation for detailed solution state characterization of structure-function relationships, toxin complex formation, and NMR-based drug discovery efforts. PMID:27351891

  7. A novel ±0.8 V high-performance voltage-tunable CDTA with enhanced bandwidth (United States)

    Xia, Zanming; Wang, Chunhua; Kuang, Jun; Jin, Jie


    In this article, we propose a novel high-performance complementary metal oxide semiconductor (CMOS) current differencing transconductance amplifier (CDTA) with a transconductance gain (GM) that can be linearly tuned by a voltage. By using a high-speed, low-voltage, cascaded current mirror active resistance compensation technique, the proposed CDTA circuit exhibits wide frequency bandwidths, high current tracking precisions as well as large output impedances. The linear-tunable GM of the CDTA is designed with the use of linear composite metal oxide semiconductor field-effect transistor as basic cells in the circuit. Combining these two approaches, several design concerns are studied, including: impedance characteristic, tracking errors, offset and linearity and noise. The prototype chip with a 0.25 mm2 area is fabricated in a GlobalFoundries'0.18 μm CMOS process. The simulated results and measured results with ±0.8 V DC supply voltages are presented, and show extremely wide bandwidths and wide linear tuning range. In addition, a fully differential band-pass filter for a high-speed system is also given as an example to confirm the high performance of the proposed circuit.

  8. Evaluation of the efect of EDTA, EGTA and CDTA on dentin adhesiveness and microleakage with different root canal sealers

    Directory of Open Access Journals (Sweden)

    Sousa-Neto Manoel D.


    Full Text Available The effect of chelating solutions EDTA, EGTA and CDTA on human dentin adhesiveness and microleakage with 4 sealers (Sealer 26, Sealapex, N-Rickert and Endofill was evaluated in vitro. Whether or not there was a mathematical correlation between the tests of adhesiveness and microleakage was also evaluated. A total of eighty maxillary and mandibular molars were used to test adhesiveness. After wearing of the occlusal surface to obtain a flat surface, the sealer was placed with an aluminum cylinder (10 mm x 6 mm. Adhesiveness was evaluated with a 4444 Instron universal testing machine. Microleakage was evaluated in 160 maxillary canines after root canal instrumentation, obturation and clearing. The penetration of India ink in the apical region was measured with a measurescope. The teeth were divided into 4 groups: group 1, distilled water; group 2, EDTA; group 3, EGTA; group 4, CDTA. Sealer 26 and EDTA had the best results (p<0.01 for adhesiveness and microleakage. There was no correlation between the test for adhesiveness and microleakage.

  9. 1H, 13C, and 15N resonance assignments of an enzymatically active domain from the catalytic component (CDTa, residues 216-420) of a binary toxin from Clostridium difficile. (United States)

    Roth, Braden M; Godoy-Ruiz, Raquel; Varney, Kristen M; Rustandi, Richard R; Weber, David J


    Clostridium difficile is a bacterial pathogen and is the most commonly reported source of nosocomial infection in industrialized nations. Symptoms of C. difficile infection (CDI) include antibiotic-associated diarrhea, pseudomembranous colitis, sepsis and death. Over the last decade, rates and severity of hospital infections in North America and Europe have increased dramatically and correlate with the emergence of a hypervirulent strain of C. difficile characterized by the presence of a binary toxin, CDT (C. difficile toxin). The binary toxin consists of an enzymatic component (CDTa) and a cellular binding component (CDTb) that together form the active binary toxin complex. CDTa harbors a pair of structurally similar but functionally distinct domains, an N-terminal domain (residues 1-215; (1-215)CDTa) that interacts with CDTb and a C-terminal domain (residues 216-420; (216-420)CDTa) that harbors the intact ADP-ribosyltransferase (ART) active site. Reported here are the (1)H, (13)C, and (15)N backbone resonance assignments of the 23 kDa, 205 amino acid C-terminal enzymatic domain of CDTa, termed (216-420)CDTa. These NMR resonance assignments for (216-420)CDTa represent the first for a family of ART binary toxins and provide the framework for detailed characterization of the solution-state protein structure determination, dynamic studies of this domain, as well as NMR-based drug discovery efforts.

  10. Využití prvků CDTA a CFTA v elektronických obvodech a kmitočtových filtrech


    Kotlán, Petr


    Práce pojednává o aktivních prvcích v proudovém módu CDTA (current differencing transconductance amplifier) a CFTA (current follower transconductance amplifier). CDTA je prvek, jenž porovnává dva vstupní proudy. Výsledný proud vystupuje přes přídavnou svorku Z na zátěž. Vzniklé napětí je přes transkonduktanci g převedeno na výstupní proud, který může být kladný (X+ výstup) nebo záporný (X- výstup). CFTA využívá narozdíl od CDTA jen jeden proudový vstup. Je ukázáno aplikační využití prvků v km...

  11. Rates of nickel(II) capture from complexes with NTA, EDDA, and related tetradentate chelating agents by the hexadentate chelating agents EDTA and CDTA: Evidence of a "semijunctive" ligand exchange pathway (United States)

    Boland, Nathan E.; Stone, Alan T.


    Many siderophores and metallophores produced by soil organisms, as well as anthropogenic chelating agent soil amendments, rely upon amine and carboxylate Lewis base groups for metal ion binding. UV-visible spectra of metal ion-chelating agent complexes are often similar and, as a consequence, whole-sample absorbance measurements are an unreliable means of monitoring the progress of exchange reactions. In the present work, we employ capillary electrophoresis to physically separate Ni(II)-tetradentate chelating agent complexes (NiL) from Ni(II)-hexadentate chelating agent complexes (NiY) prior to UV detection, such that progress of the reaction NiL + Y → NiY + L can be conveniently monitored. Rates of ligand exchange for Ni(II) are lower than for other +II transition metal ions. Ni(II) speciation in environmental media is often under kinetic rather than equilibrium control. Nitrilotriacetic acid (NTA), with three carboxylate groups all tethered to a central amine Lewis base group, ethylenediamine-N,N‧-diacetic acid (EDDA), with carboxylate-amine-amine-carboxylate groups arranged linearly, plus four structurally related compounds, are used as tetradentate chelating agents. Ethylenediaminetetraacetic acid (EDTA) and the structurally more rigid analog trans-cyclohexaneethylenediaminetetraacetic acid (CDTA) are used as hexadentate chelating agents. Effects of pH and reactant concentration are explored. Ni(II) capture by EDTA was consistently more than an order of magnitude faster than capture by CDTA, and too fast to quantify using our capillary electrophoresis-based technique. Using NiNTA as a reactant, Ni(II) capture by CDTA is independent of CDTA concentration and greatly enhanced by a proton-catalyzed pathway at low pH. Using NiEDDA as reactant, Ni(II) capture by CDTA is first order with respect to CDTA concentration, and the contribution from the proton-catalyzed pathway diminished by CDTA protonation. While the convention is to assign either a disjunctive

  12. CDT-a entropic theory of quantum gravity

    DEFF Research Database (Denmark)

    Ambjørn, Jan; Görlich, A.; Jurkiewicz, J.


    High Energy Physics - Theory (hep-th); General Relativity and Quantum Cosmology (gr-qc); High Energy Physics - Lattice (hep-lat)......High Energy Physics - Theory (hep-th); General Relativity and Quantum Cosmology (gr-qc); High Energy Physics - Lattice (hep-lat)...

  13. Clostridium difficile Recombinant Toxin A Repeating Units as a Carrier Protein for Conjugate Vaccines: Studies of Pneumococcal Type 14, Escherichia coli K1, and Shigella flexneri Type 2a Polysaccharides in Mice (United States)

    Pavliakova, Danka; Moncrief, J. Scott; Lyerly, David M.; Schiffman, Gerald; Bryla, Dolores A.; Robbins, John B.; Schneerson, Rachel


    Unlike the native protein, a nontoxic peptide (repeating unit of the native toxin designated rARU) from Clostridium difficile toxin A (CDTA) afforded an antigen that could be bound covalently to the surface polysaccharides of pneumococcus type 14, Shigella flexneri type 2a, and Escherichia coli K1. The yields of these polysaccharide-protein conjugates were significantly increased by prior treatment of rARU with succinic anhydride. Conjugates, prepared with rARU or succinylated (rARUsucc), were administered to mice by a clinically relevant dosage and immunization scheme. All conjugates elicited high levels of serum immunoglobulin G both to the polysaccharides and to CDTA. Conjugate-induced anti-CDTA had neutralizing activity in vitro and protected mice challenged with CDTA, similar to the rARU alone. Conjugates prepared with succinylated rARU, therefore, have potential for serving both as effective carrier proteins for polysaccharides and for preventing enteric disease caused by C. difficile. PMID:10722615

  14. The effect of physical and chemical treatment on the mechanical properties of the cell wall of the alga Chara corallina. (United States)

    Toole, Geraldine A; Smith, Andrew C; Waldron, Keith W


    Single large internode cells of the charophyte (giant alga) Chara corallina were dissected to give sheets of cell wall, which were then notched and their mechanical properties in tension determined. The cells were subjected to a thermal treatment in excess water (cf. cooking), which had little effect on strength but increased the stiffness, contrasting with the behaviour of higher-plant tissues. Extraction in CDTA (cyclohexane-trans-1,2-diamine-N,N,N',N'-tetraacetate) or 4 M KOH reduced the strength from 17 MPa to 10 MPa, although sequential extraction in CDTA and 4 M KOH reduced the strength further to 4 MPa. The stiffness decreased from 500 MPa to 300 MPa on extraction in CDTA or 4 M KOH, while falling to 70 MPa after extraction in CDTA followed by 4 M KOH. Conventional sequential extraction in CDTA, Na2CO3 at 1 degrees C and 20 degrees C, and KOH at 0.5 M, 1 M, 2 M and 4 M caused a gradual decrease in stiffness and strength after the CDTA treatment to the same lower values. This result is in keeping with mechanical properties for plant tissues, but in contrast to the removal of pectic polysaccharides from model cell wall systems, which does not reduce the stiffness.

  15. Introduction of five potentially metabolizable linking groups between [sup 111]In-cyclohexyl EDTA derivatives and F(ab')[sub 2] fragments of anti-carcinoembryonic antigen antibody: Pt. 1; A new reproducible synthetic method

    Energy Technology Data Exchange (ETDEWEB)

    Gestin, J.F.; Faivre-Chauvet, A.; Sai-Maurel, C.; Thedrez, P.; Slinkin, M.; Chatal, J.F. (INSERM, 44 - Nantes (France)); Mease, R.C.; Meinken, G.E.; Srivastava, S.C. (Brookhaven National Lab., Upton, NY (United States))


    The purpose of this study was to synthesize new bifunctional linker-chelating agents for the modification of the in vivo distribution of [sup 111]In-labeled antibodies. A general simple synthetic method of preparing cyclohexyl EDTA (CDTA) derivatives containing a linker/spacer group is described. Linkers prepared included a diester, a six carbon aliphatic chain, two thioethers and a disulfide group. The CDTA-linker compounds were coupled to F(Ab')[sub 2] fragments of anti-carcinoembryonic antigen monoclonal antibody and labeled with [sup 111]In with good retention of immunoreactivity. (author).

  16. Introduction of five potentially metabolizable linking groups between 111In-cyclohexyl EDTA derivatives and F(ab')2 fragments of anti-carcinoembryonic antigen antibody--I. A new reproducible synthetic method. (United States)

    Gestin, J F; Faivre-Chauvet, A; Mease, R C; Sai-Maurel, C; Thédrez, P; Slinkin, M; Meinken, G E; Srivastava, S C; Chatal, J F


    The purpose of this study was to synthesize new bifunctional linker-chelating agents for the modification of the in vivo distribution of 111In-labeled antibodies. A general simple synthetic method of preparing cyclohexyl EDTA (CDTA) derivatives containing a linker/spacer group is described. Linkers prepared included a diester, a six carbon aliphatic chain, two thioethers and a disulfide group. The CDTA-linker compounds were coupled to F(Ab')2 fragments of anti-carcinoembryonic antigen monoclonal antibody and labeled with 111In with good retention of immunoreactivity.

  17. Demonstration of the BNL Continuous Dual Trap Analyzer to Detect Perfluorocarbon Tracers for the Tag, Track and Location Program

    Energy Technology Data Exchange (ETDEWEB)

    Heiser,J.H.; Adams, J.; Dietz, R..; Milian, L.; Watson, T.


    The Tag, Track and Location System (TTL) Program is investigating methods of tracking an asset using perfluorocarbon tracers (PFT). The success of any TTL method requires sound detection/location instrumentation. Tracer Detection Technologies Corp (TDT), through a contract with the Office of Naval Research (ONR), is investigating different detection systems. The detections systems generally fall into two categories; proximity detectors and standoff detectors. Proximity detectors, as the name implies, need to be in close proximity (e.g., meter to 10's of meters) to the PFT source. Standoff detection searches for the PFT from a greater distance away from the source (e.g., 100's of meters to kilometers). Gas Chromatographs (GC) are generally considered a proximity detection systems, but in the case of PFTs should be considered for both proximity and standoff detection with the caveat that in standoff use the GC needs to be somewhere in the PFT plume, i.e., generally downwind of the source. With a properly sized PFT source, the right GC can afford fairly large standoff (distance from the source) distances; 100's of meters to kilometers downwind. Brookhaven National Laboratory (BNL) has such a GC system and offered to demonstrate the CDTA for TTL as a no cost addition to the TDTTTL project, of which BNL was a participant. BNL is a leading authority on the sampling, collection, release and detection of PFTs. In addition, the BNL team has extensive background in atmospheric dispersion, the application of PFTs to such studies and the development of applications utilizing PFTs such as building infiltration measurements, control room integrity determination, leak location and environmental investigations. This experience and expertise is essential in developing any PFT application were dispersion, dilution and overcoming environmental conditions and interferences are integral to success. BNL has developed sophisticated gas chromatography methods and

  18. plasma-oxygen interaction during thin films deposition by laser ...

    African Journals Online (AJOL)


    Jun 30, 2012 ... reactive oxygen atmosphere is necessary to bring closer the thin films properties to that of the bulk state [2]. However, the beginning of this reactivity is observed to occur only at a certain oxygen pressure [3]. Author Correspondence, e-mail: ICID: 1020709. Journal of Fundamental and ...

  19. Telemedicine in Remote Areas of Algeria | CRDI - Centre de ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Algeria includes a number of remote and desert areas that are cut off from adequate health services. This grant will allow the Centre de développement des technologies avancées (CDTA - centre for the development of advanced technologies) to demonstrate the feasibility and benefits of telemedicine in the country.

  20. Telemedicine in Remote Areas of Algeria | IDRC - International ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    With respect to security, the research team will test a new method of encryption combining biomeasurement and the utilization of public key infrastructure (PKI) to ensure the confidentiality of patient data. To do so, they will use an internal intranet (CDTA) certification thereby avoiding the need for expensive international ...

  1. Human capital management in a knowledge economy: The case of ...

    African Journals Online (AJOL)

    This research aims to examine the existence of the human capital management through competencies and knowledge management approach in Scientific Research Centers within knowledge based economy. The study was applied to the case of Scientific Research Centers in Algeria, such as: (CREAD, CRSTRA, CDTA, ...

  2. Fermentation characteristics of polysaccharide fractions extracted from the cell walls of soya bean cotyledons

    NARCIS (Netherlands)

    Laar, van H.; Tamminga, S.; Williams, B.A.; Verstegen, M.W.A.; Schols, H.A.


    Full-fat soya beans were separated into hulls and cotyledons. After separation the cell wall fraction was extracted from the cotyledons. These purified cell walls were sequentially extracted with 0.05 M cyclohexane-trans-1,2-diamine-N,N,N ,N -tetraacetate (CDTA) 0.05 M NH4 oxalate (extract 1), 0.05

  3. The removal of strontium from the mouse by chelating agents

    Energy Technology Data Exchange (ETDEWEB)

    Ortega, A.; Gomez, M.; Domingo, J.L.; Corbella, J.


    The effects of the chelating agents monosodium glutamate, Tiron, tartaric acid, ascorbic acid, 1,4,7,10,13,16-hexaoxacyclooctadecane (18-crown-6), 2,3-dimercaptosuccinic acid (DMSA), succinic acid, malic acid, ethylendiaminetetraacetic acid (EDTA), ethylenglycol-bis-(beta-amino-ethylether)-N,N'tetraacetic acid (EGTA), cyclohexane-diaminetetraacetic acid (CDTA) and diethylentria-minepentaacetic acid (DTPA) on the distribution and excretion of intraperitoneally injected strontium were investigated in male Swiss mice. Strontium nitrate was given at a dose equal to 3.78 mmol/kg and ten minutes after, chelators were administered intraperitoneally at doses approximately equal to one-fourth of their respective LD50 values. DTPA, followed by CDTA, EDTA and tartaric acid, was consistently the most effective in increasing the urinary excretion of strontium. Only ascorbic acid increased significantly the fecal excretion of strontium. CDTA, DTPA and ascorbic acid were also the most effective chelators in reducing the concentration of strontium found in various tissues. CDTA, DTPA and tartaric acid are the most effective agents of those tested in the removal of strontium after a single administration.

  4. Clinical evaluation of two new bifunctional chelating agents for immunoscintigraphy (I.S.) with indium-111-anti-CEA monoclonal antibody

    Energy Technology Data Exchange (ETDEWEB)

    Chatal, J.F.; Gestin, J.F.; Faivre-Chauvet, A. [Brookhaven National Lab., Upton, NY (United States)] [and others


    Previous animal studies have shown that chelates of In-111 with semi-rigid cyclohexyl EDTA (CDTA) derivatives produce improved in-vivo stability compared to non-rigid analogs. Two CDTA derivatives, CDTA-EN-SCN (dilithium salt of N-(methyl (2-isothiocyanato-ethyl) carbamide) trans-1,2-diaminocyclohexane N,N{prime},N{prime}-triacetic acid) and 4-ICE (4-isothiocyanato-trans-1,2-diaminocyclohexane-N,N,N {prime},N{prime}-tetraacetic acid), were synthesized, coupled to anti-CEA F(ab{prime}){sub 2} fragments and labeled with In-111 with a specific activity of 111MBq/mg. Five patients were injected with CDTA-EN-SCN and 4 with 4-ICE. IS was performed 3 days after injection. Mean blood activity, at the time of IS, expressed as the percentage of injected dose per ml, was 0.0038% with 4-ICE and 0.0071% with CDTA-EN-SCN. For comparative purposes, the mean value previously obtained with DTPA coupled to the same antibody was 0.0035%. Liver uptake was estimated from semi-quantitative calculations on scintigraphic images. Compared with the value obtained in a patient injected with 72.3 antibody coupled to DTFPA, a 2.5-fold reduction in liver uptake was observed. However, this reduction was sufficient to allow visualization of a large liver metastasis in one patient. With CDTA-EN-SCN, colon activity was moderate in 3 patients and intense in 1. With 4-ICE it was moderate in 2 patients and intense in 1, With CDTA-EN-SCN, true-positive IS results were observed in 2 patients (pelvis and mediastinum) and a false-negative result was observed in 1 patient (pelvis). In 3 patients, IS was negative and recurrence not yet confirmed despite high serum CEA level. With 4-FICE, true-positive IS results were observed in 3 patients (2 pelvis, 1 lung). A false-negative result was found in a patient with a confirmed liver metastasis.

  5. Structural changes in cell wall pectins during strawberry fruit development. (United States)

    Paniagua, Candelas; Santiago-Doménech, Nieves; Kirby, Andrew R; Gunning, A Patrick; Morris, Victor J; Quesada, Miguel A; Matas, Antonio J; Mercado, José A


    Strawberry (Fragaria × anannasa Duch.) is one of the most important soft fruit. Rapid loss of firmness occurs during the ripening process, resulting in a short shelf life and high economic losses. To get insight into the role of pectin matrix in the softening process, cell walls from strawberry fruit at two developmental stages, unripe-green and ripe-red, were extracted and sequentially fractionated with different solvents to obtain fractions enriched in a specific component. The yield of cell wall material as well as the per fresh weight contents of the different fractions decreased in ripe fruit. The largest reduction was observed in the pectic fractions extracted with a chelating agent (trans-1,2- diaminocyclohexane-N,N,N'N'-tetraacetic acid, CDTA fraction) and those covalently bound to the wall (extracted with Na 2 CO 3 ). Uronic acid content of these two fractions also decreased significantly during ripening, but the amount of soluble pectins extracted with phenol:acetic acid:water (PAW) and water increased in ripe fruit. Fourier transform infrared spectroscopy of the different fractions showed that the degree of esterification decreased in CDTA pectins but increased in soluble fractions at ripen stage. The chromatographic analysis of pectin fractions by gel filtration revealed that CDTA, water and, mainly PAW polyuronides were depolymerised in ripe fruit. By contrast, the size of Na 2 CO 3 pectins was not modified. The nanostructural characteristics of CDTA and Na 2 CO 3 pectins were analysed by atomic force microscopy (AFM). Isolated pectic chains present in the CDTA fractions were significantly longer and more branched in samples from green fruit than those from red fruit. No differences in contour length were observed in Na 2 CO 3 strands between samples of both stages. However, the percentage of branched chains decreased from 19.7% in unripe samples to 3.4% in ripe fruit. The number of pectin aggregates was higher in green fruit samples of both

  6. Solution stabilities of some mixed ligand complexes of UO22+ and Th4+ with complexones and salicylic acids

    International Nuclear Information System (INIS)

    Singh, R.K.; Saxena, M.C.


    Formation constants (log Ksub(MAL)sup(MA)) of mixed ligands complexes (MAL), where M = UO 2 2+ or Th 4+ , A = IMDA, NTA, HEDTA, EDTA, CDTA or DTPA, L = salicylic acid (SA) or 5-sulphosalicylic acid (SSA), have been determined by pH titrations using Irving-Rossotti approach at 25 o C and at I =0.2 (mol dm -3 , KNO 3 ). The solution stabilities exhibit the sequence (i) Th 4+ >UO 2 2+ , (ii) IMDA>NTA>HEDTA>EDTA>CDTA>DTPA, and (iii) SA>SSA with respect to metal ions, primary ligands and secondary ligands, respectively. The formation constants log Ksub(ML)sup(M) and log Ksub(ML 2 )sup(ML) have also been determined. The Δlog K values have been found to be negative-increasing numerically with the negative charge on the deprotonated primary ligand (A n- ). (author). 17 refs., 1 tab

  7. Plate-forme d'outils en logiciel libre pour l'enseignement supérieur ...

    International Development Research Centre (IDRC) Digital Library (Canada) Chargé(e) de projet. Mustapha es-Salihe. Institution. Centre de recherche informatique de Montréal Inc. Pays d' institution. Canada. Site internet. Chargé(e) de projet. Henda Ben Ghezala. Institution. Association Scientifique de l'École Nationale d'Ingénieurs de Tunis. Pays d' institution.

  8. Stability constants of mixed ligand complexes of dioxouranium(II) and thorium(IV) with complexones and isomeric alanines

    International Nuclear Information System (INIS)

    Singh, R.K.; Saxena, M.C.


    The present work reports on the stability sequence between UO 2 II and Th IV ions for their mixed ligands complexes with the two isomeric alanines, α-alanine (α-ala) and β-alanine (β-ala) containing a complexone as primary ligand. The complexones used are iminodiacetate (IMDA), nitrilotricetate (NTA), 2-hydroxyethylenediaminetriacetate (HEDTA), ethylenediaminetetraacetate (EDTA), 1,2-diaminocyclohexanetraacetate (CDTA) and diethylenetriminepentaacetate (DTPA). (author). 9 refs., 1 tab

  9. Development of a Novel Vaccine Containing Binary Toxin for the Prevention of Clostridium difficile Disease with Enhanced Efficacy against NAP1 Strains.

    Directory of Open Access Journals (Sweden)

    Susan Secore

    Full Text Available Clostridium difficile infections (CDI are a leading cause of nosocomial diarrhea in the developed world. The main virulence factors of the bacterium are the large clostridial toxins (LCTs, TcdA and TcdB, which are largely responsible for the symptoms of the disease. Recent outbreaks of CDI have been associated with the emergence of hypervirulent strains, such as NAP1/BI/027, many strains of which also produce a third toxin, binary toxin (CDTa and CDTb. These hypervirulent strains have been associated with increased morbidity and higher mortality. Here we present pre-clinical data describing a novel tetravalent vaccine composed of attenuated forms of TcdA, TcdB and binary toxin components CDTa and CDTb. We demonstrate, using the Syrian golden hamster model of CDI, that the inclusion of binary toxin components CDTa and CDTb significantly improves the efficacy of the vaccine against challenge with NAP1 strains in comparison to vaccines containing only TcdA and TcdB antigens, while providing comparable efficacy against challenge with the prototypic, non-epidemic strain VPI10463. This combination vaccine elicits high neutralizing antibody titers against TcdA, TcdB and binary toxin in both hamsters and rhesus macaques. Finally we present data that binary toxin alone can act as a virulence factor in animal models. Taken together, these data strongly support the inclusion of binary toxin in a vaccine against CDI to provide enhanced protection from epidemic strains of C. difficile.

  10. Thermodynamic studies of the complexation of plutonium(IV) by linear and macrocyclic poly-amino-carboxylate ligands

    International Nuclear Information System (INIS)

    Burgat, Romain


    In the framework of a collaboration between the CEA (Commissariat a l Energie Atomique) of Valduc and the ICMUB (Institut de Chimie Moleculaire de l Universite de Bourgogne), a study platform of the structural and physico-chemical properties of the radioelements U, Pu and Am complexes has been implemented. The plutonium(IV) complexation has been studied in a molar nitrate medium. The affinity of three linear poly-amino-carboxylates (EDTA, CDTA and DTPA) towards plutonium(IV) has then been estimated. For the three ligands, the formation constants of the monoleptic complexes Pu(EDTA), Pu(CDTA) and [Pu(DTPA)] - have been determined in a (H,K)NO 3 1 M medium and then extrapolated at a zero ionic force with the specific interactions theory (SIT). For the three complexes, mono-hydroxylated monoleptic species have been observed. With the EDTA and the CDTA, protonated dileptic complexes of a general formula [Pu(L) 2 H h ] (4-h)- have been revealed too. Nevertheless, the steric hindrance around the metallic center is too important to allow to a second molecule of DTPA to coordinate the Pu 4+ cation. The exclusive formation of the species [Pu(DTPA)] - and [Pu(DTPA)(OH)] 2- has been confirmed by capillary electrophoresis (EC-ICP-MS). On account of the preliminary results obtained during the titration of the cyclame tetraacetic product (TETRA) in presence of plutonium(IV), the adding of a competitive ligand such as EDTA has been considered for the study of the complexation of this radioelement by macrocyclic ligands. At last, the affinity of different macrocyclic ligands containing either four amide functions (TETAMMe 2 and TETAMMEt 2 ) or carboxylate groups (TETA, DOTPr and TETPr) towards lanthanides(III) has been estimated too. Although the complexation reaction be fast with the two first ligands, these complexes are less stable than those formed with the carboxylic macrocycles. (O.M.)

  11. Télémédecine dans les régions éloignées en Algérie | IDRC ...

    International Development Research Centre (IDRC) Digital Library (Canada)

    Télémédecine dans les régions éloignées en Algérie. L'Algérie comporte des régions rurales et désertiques éloignées et coupées des services de santé adéquats. Ce projet aidera le Centre de développement des technologies avancées (CDTA) à démontrer la faisabilité et les bénéfices de la télémédecine dans le pays.

  12. The mechanism of Jurkat cells apoptosis induced by Aggregatibacter actinomycetemcomitans cytolethal distending toxin. (United States)

    Chen, Hui-Ping; Li, Lu; Chen, Xu; Yang, Mi-Fang; Ye, Yu; Wang, Xiao-Qian; Xu, Yan


    Cytolethal distending toxin (CDT) which is produced by Aggregatibacter actinomycetemcomitans causes apoptosis in lymphocytes. But the specific mechanism is not clear. The aim of our research was to investigate the effect and mechanism during this process. The wild-type CdtA, CdtB, CdtC (CdtA W , CdtB W , CdtC W ) and mutant CdtB (CdtB M ) were expressed and purified respectively and the purity of each subunit was examined by BandScan software. And the type I deoxyribonuclease and PI-3,4,5-triphosphate (PI-3,4,5-P3, PIP3) phosphatase activity were detected by DNA agarose gel electrophoresis and enzyme-linked immunosorbent assay respectively. The cell apoptosis rates were analyzed by flow cytometry. The morphological changes of apoptosis cells were observed by confocal laser scanning microscopy. The protein expression of Bax and Bcl-2 was examined by western blot. Differentially expressed apoptosis-related proteins were identified based on isobaric tags for relative and absolute quantitation technology. In the present study we found that: (i) recombinant wild-type CdtA, CdtB and CdtC (CdtA W , CdtB W , CdtC W ) and mutant CdtB (CdtB M ) were correctly expressed and the purity of each protein was higher than 80%, (ii) the average apoptosis rate in wild-type CDT (CDT W ) treated groups was 50.54%, which was significantly higher than the controls (4.71%) and mutant CDT (CDT M ) treated groups (5.58%) (p apoptosis were observed in CDT W treated cells, (iv) the expression of Bax protein was significantly increased in CDT W treated cells, while Bcl-2 protein expression was significantly decreased, (v) 17 apoptosis-related proteins were expressed differentially, among which 10 proteins (SMNDC1, TNFRSF10B, UBE2I, ITM2A, CASP3, P53, EIF1, TCF3, HMGN5, CASP8) were up-regulated and 7 proteins (RRM2, TPX2, KIF11, NUCKS1, TOP2A, XRCC1, PTPLAD1, RRM2) were down-regulated, (vi) one possible apoptotic pathway [Ubc9 (UBE2I)/P53/DR5 (TNFRSF10B)/Caspase-8 (CASP8)/ Caspase-3 (CASP3

  13. Chelant extraction of heavy metals from contaminated soils using new selective EDTA derivatives. (United States)

    Zhang, Tao; Liu, Jun-Min; Huang, Xiong-Fei; Xia, Bing; Su, Cheng-Yong; Luo, Guo-Fan; Xu, Yao-Wei; Wu, Ying-Xin; Mao, Zong-Wan; Qiu, Rong-Liang


    Soil washing is one of the few permanent treatment alternatives for removing metal contaminants. Ethylenediaminetetraacetic acid (EDTA) and its salts can substantially increase heavy metal removal from contaminated soils and have been extensively studied for soil washing. However, EDTA has a poor utilization ratio due to its low selectivity resulting from the competition between soil major cations and trace metal ions for chelation. The present study evaluated the potential for soil washing using EDTA and three of its derivatives: CDTA (trans-1,2-cyclohexanediaminetetraacetic acid), BDTA (benzyldiaminetetraacetic acid), and PDTA (phenyldiaminetetraacetic acid), which contain a cylcohexane ring, a benzyl group, and a phenyl group, respectively. Titration results showed that PDTA had the highest stability constants for Cu(2+) and Ni(2+) and the highest overall selectivity for trace metals over major cations. Equilibrium batch experiments were conducted to evaluate the efficacy of the EDTA derivatives at extracting Cu(2+), Zn(2+), Ni(2+), Pb(2+), Ca(2+), and Fe(3+) from a contaminated soil. At pH 7.0, PDTA extracted 1.5 times more Cu(2+) than did EDTA, but only 75% as much Ca(2+). Although CDTA was a strong chelator of heavy metal ions, its overall selectivity was lower and comparable to that of EDTA. BDTA was the least effective extractant because its stability constants with heavy metals were low. PDTA is potentially a practical washing agent for soils contaminated with trace metals. Copyright © 2013 Elsevier B.V. All rights reserved.

  14. Enhanced Production of Fatty Acids via Redirection of Carbon Flux in Marine MicroalgaTetraselmissp. (United States)

    Han, Mi-Ae; Hong, Seong-Joo; Kim, Z-Hun; Cho, Byung-Kwan; Lee, Hookeun; Choi, Hyung-Kyoon; Lee, Choul-Gyun


    Lipids in microalgae are energy-rich compounds and considered as an attractive feedstock for biodiesel production. To redirect carbon flux from competing pathways to the fatty acid synthesis pathway of Tetraselmis sp., we used three types of chemical inhibitors that can block the starch synthesis pathway or photorespiration, under nitrogen-sufficient and nitrogen-deficient conditions. The starch synthesis pathway in chloroplasts and the cytosol can be inhibited by 3-(3,4-dichlorophenyl)-1,1-dimethylurea and 1,2-cyclohexane diamine tetraacetic acid (CDTA), respectively. Degradation of glycine into ammonia during photorespiration was blocked by aminooxyacetate (AOA) to maintain biomass concentration. Inhibition of starch synthesis pathways in the cytosol by CDTA increased fatty acid productivity by 27% under nitrogen deficiency, whereas the blocking of photorespiration in mitochondria by AOA was increased by 35% under nitrogen-sufficient conditions. The results of this study indicate that blocking starch or photorespiration pathways may redirect the carbon flux to fatty acid synthesis.

  15. PCR detection of seven virulence and toxin genes of Campylobacter jejuni and Campylobacter coli isolates from Danish pigs and cattle and cytolethal distending toxin production of the isolates

    DEFF Research Database (Denmark)

    Bang, Dang Duong; Nielsen, E.M.; Scheutz, F.


    Aims: To study the prevalence of seven virulence and toxin genes, and cytolethal distending toxin (CDT) production of Campylobacter jejuni and C. coli isolates from Danish pigs and cattle. Methods and Results: The presence of the cadF, ceuE, virB11, flaA, cdtA, cdtB, cdtC and the cdt gene cluster...... among 40 C. jejuni and C. coli isolates was detected by polymerase chain reaction. The CDT production of the isolates was determined on Vero, colon 205 and chicken embryo cells. The cadF, flaA, ceuE and cdtB genes were detected from 100% of the isolates. The cdtA and cdtC genes were found in 95.0 and 90.......0% of the isolates, respectively. The cdt gene cluster was detected in 82.5% isolates. Only 7.5% of the isolates were positive for virB11. Ninety-five per cent of the isolates produced CDT in Vero and colon 205 cell assays, and 90% of the isolates produced CDT in chicken embryo cell assays. Conclusions: High...

  16. Innovative SANEX process for trivalent actinides separation from PUREX raffinate

    International Nuclear Information System (INIS)

    Sypula, Michal


    Recycling of nuclear spent fuel and reduction of its radiotoxicity by separation of long-lived radionuclides would definitely help to close the nuclear fuel cycle ensuring sustainability of the nuclear energy. Partitioning of the main radiotoxicity contributors followed by their conversion into short-lived radioisotopes is known as partitioning and transmutation strategy. To ensure efficient transmutation of the separated elements (minor actinides) the content of lanthanides in the irradiation targets has to be minimised. This objective can be attained by solvent extraction using highly selective ligands that are able to separate these two groups of elements from each other. The objective of this study was to develop a novel process allowing co-separation of minor actinides and lanthanides from a high active acidic feed solution with subsequent actinide recovery using just one cycle, so-called innovative SANEX process. The conditions of each step of the process were optimised to ensure high actinide separation efficiency. Additionally, screening tests of several novel lipophilic and hydrophilic ligands provided by University of Twente were performed. These tests were aiming in better understanding the influence of the extractant structural modifications onto An(III)/Ln(III) selectivity and complexation properties. Optimal conditions for minor actinides separation were found and a flow-sheet of a new innovative SANEX process was proposed. Tests using a single centrifugal contactor confirmed high Eu(III)/Am(III) separation factor of 15 while the lowest SF Ln/Am obtained was 6,5 (for neodymium). In addition, a new masking agent for zirconium was found as a substitution for oxalic acid. This new masking agent (CDTA) was also able to mask palladium without any negative influence on An(III)/Ln(III). Additional tests showed no influence of CDTA on plutonium present in the feed solution unlike oxalic acid which causes Pu precipitation. Therefore, CDTA was proposed as a Zr

  17. Pectic-β(1,4)-galactan, extensin and arabinogalactan–protein epitopes differentiate ripening stages in wine and table grape cell walls (United States)

    Moore, John P.; Fangel, Jonatan U.; Willats, William G. T.; Vivier, Melané A.


    Background and Aims Cell wall changes in ripening grapes (Vitis vinifera) have been shown to involve re-modelling of pectin, xyloglucan and cellulose networks. Newer experimental techniques, such as molecular probes specific for cell wall epitopes, have yet to be extensively used in grape studies. Limited general information is available on the cell wall properties that contribute to texture differences between wine and table grapes. This study evaluates whether profiling tools can detect cell wall changes in ripening grapes from commercial vineyards. Methods Standard sugar analysis and infra-red spectroscopy were used to examine the ripening stages (green, véraison and ripe) in grapes collected from Cabernet Sauvignon and Crimson Seedless vineyards. Comprehensive microarray polymer profiling (CoMPP) analysis was performed on cyclohexanediaminetetraacetic acid (CDTA) and NaOH extracts of alcohol-insoluble residue sourced from each stage using sets of cell wall probes (mAbs and CBMs), and the datasets were analysed using multivariate software. Key Results The datasets obtained confirmed previous studies on cell wall changes known to occur during grape ripening. Probes for homogalacturonan (e.g. LM19) were enriched in the CDTA fractions of Crimson Seedless relative to Cabernet Sauvignon grapes. Probes for pectic-β-(1,4)-galactan (mAb LM5), extensin (mAb LM1) and arabinogalactan proteins (AGPs, mAb LM2) were strongly correlated with ripening. From green stage to véraison, a progressive reduction in pectic-β-(1,4)-galactan epitopes, present in both pectin-rich (CDTA) and hemicellulose-rich (NaOH) polymers, was observed. Ripening changes in AGP and extensin epitope abundance also were found during and after véraison. Conclusions Combinations of cell wall probes are able to define distinct ripening phases in grapes. Pectic-β-(1,4)-galactan epitopes decreased in abundance from green stage to véraison berries. From véraison there was an increase in abundance of

  18. Detection of virulence, antibiotic resistance and toxin (VAT) genes in Campylobacter species using newly developed multiplex PCR assays. (United States)

    Laprade, Natacha; Cloutier, Michel; Lapen, David R; Topp, Edward; Wilkes, Graham; Villemur, Richard; Khan, Izhar U H


    Campylobacter species are one of the leading causes of bacterial gastroenteritis in humans worldwide. This twofold study was sought to: i) develop and optimize four single-tube multiplex PCR (mPCR) assays for the detection of six virulence (ciaB, dnaJ, flaA, flaB, pldA and racR), three toxin (cdtA, cdtB and cdtC) and one antibiotic resistance tet(O) genes in thermophilic Campylobacter spp. and ii) apply and evaluate the developed mPCR assays by testing 470 previously identified C. jejuni, C. coli and C. lari isolates from agricultural water. In each mPCR assay, a combination of two or three sets of primer pairs for virulence, antibiotic resistance and toxin (VAT) genes was used and optimized. Assay 1 was developed for the detection of dnaJ, racR and cdtC genes with expected amplification sizes of 720, 584 and 182bp. Assay 2 generated PCR amplicons for tet(O) and cdtA genes of 559 and 370bp. Assay 3 amplified cdtB ciaB, and pldA genes with PCR amplicon sizes of 620, 527 and 385bp. Assay 4 was optimized for flaA and flaB genes that generated PCR amplicons of 855 and 260bp. The primer pairs and optimized PCR protocols did not show interference and/or cross-amplification with each other and generated the expected size of amplification products for each target VAT gene for the C. jejuni ATCC 33291 reference strain. Overall, all ten target VAT genes were detected at a variable frequency in tested isolates of thermophilic Campylobacter spp. where cdtC, flaB, ciaB, cdtB, cdtA and pldA were commonly detected compared to the flaA, racR, dnaJ and tet(O) genes which were detected with less frequency. The developed mPCR assays are simple, rapid, reliable and sensitive tools for simultaneously assessing potential pathogenicity and antibiotic resistance profiling in thermophilic Campylobacter spp. The mPCR assays will be useful in diagnostic and analytical settings for routine screening of VAT characteristics of Campylobacter spp. as well as being applicable in epidemiological

  19. Innovative SANEX process for trivalent actinides separation from PUREX raffinate

    Energy Technology Data Exchange (ETDEWEB)

    Sypula, Michal


    Recycling of nuclear spent fuel and reduction of its radiotoxicity by separation of long-lived radionuclides would definitely help to close the nuclear fuel cycle ensuring sustainability of the nuclear energy. Partitioning of the main radiotoxicity contributors followed by their conversion into short-lived radioisotopes is known as partitioning and transmutation strategy. To ensure efficient transmutation of the separated elements (minor actinides) the content of lanthanides in the irradiation targets has to be minimised. This objective can be attained by solvent extraction using highly selective ligands that are able to separate these two groups of elements from each other. The objective of this study was to develop a novel process allowing co-separation of minor actinides and lanthanides from a high active acidic feed solution with subsequent actinide recovery using just one cycle, so-called innovative SANEX process. The conditions of each step of the process were optimised to ensure high actinide separation efficiency. Additionally, screening tests of several novel lipophilic and hydrophilic ligands provided by University of Twente were performed. These tests were aiming in better understanding the influence of the extractant structural modifications onto An(III)/Ln(III) selectivity and complexation properties. Optimal conditions for minor actinides separation were found and a flow-sheet of a new innovative SANEX process was proposed. Tests using a single centrifugal contactor confirmed high Eu(III)/Am(III) separation factor of 15 while the lowest SF{sub Ln/Am} obtained was 6,5 (for neodymium). In addition, a new masking agent for zirconium was found as a substitution for oxalic acid. This new masking agent (CDTA) was also able to mask palladium without any negative influence on An(III)/Ln(III). Additional tests showed no influence of CDTA on plutonium present in the feed solution unlike oxalic acid which causes Pu precipitation. Therefore, CDTA was proposed as

  20. INL DPAH STAAR 2015 Annual Report

    Energy Technology Data Exchange (ETDEWEB)

    Peterman, Dean Richard [Idaho National Lab. (INL), Idaho Falls, ID (United States)


    Research conducted at the INL has demonstrated the synergistic extraction of americium using solvents comprised of bis(o,o-(trifluoromethyl)phenyl) dithiophosphinic acid (DPAH “1”) and trioctylphosphine oxide (TOPO), butyl bis(2,4,4-trimethylpentyl) phosphinate (BuCy272), or dibutyl butylphosphonate (DBBP). One potential drawback of this separations scheme is that soft metals such as silver, cadmium, or palladium and fission products such as zirconium are well extracted by these solvents. Several potential scrubbing reagents were examined. Of the scrubbing reagents studied, cysteine and methione exhibited some ability to scrub soft metals from the loaded solvent. More conventional scrub reagents such as ammonium fluoride or oxalic acid were not effective. Reagents like Bimet and CDTA were not soluble at the acidities used in these studies. Unfortunately, these results indicate that the identification of effective scrubbing reagents for use in a flowsheet based upon the INL DPAH is going to be very difficult.

  1. Use of activable cations as tracers in groundwater hydrology. The case of DTPA-Indium

    International Nuclear Information System (INIS)

    Lumu, Badimbayi Matu.


    The possibilities of EDTA, CDTA and DTPA metallic complexes use as activable groundwater, tracers are discussed. Indium, which has good nuclear caracteristics for activation analysis and forms complexes of great stability with polyamino carboxylic acid has been for Laboratory and field studies. For corporative studies, Rhodomine B, a fluorescent tracer have been studied together with Indium complexes. In laboratory retention studies have been carried with In-EDTA, Iodine 131 and Rhodomine B, as tracers and bentonite, zeolite 13X and Dowex-1 and Dowex-50 as sorbents. As field studies, drainage evolution flow and resident time distribution of tracers substances in water, have been carried, under artificial rain conditions realized by aspersion. Results from field studies showed good characteristics of Indium Complexes especially in very absorbent medium (argilaceous limon) where their restitution balance were superior to that of Rhodomine B

  2. Collaborative practice model for management of pain in patients with cancer. (United States)

    Hammer, Kathryn J; Segal, Eve M; Alwan, Laura; Li, Shan; Patel, Amila M; Tran, Melinda; Marshall, Helen M


    The use of a collaborative drug therapy agreement (CDTA) by oncology pharmacists in a comprehensive pain clinic is described. Recognizing the complex clinical services required by patients with cancer, the Seattle Cancer Care Alliance began offering cancer pain management through a specialized pain service. Initially, the clinic was staffed by one attending physician; however, as the volume of patient referrals increased, the clinic expanded into an interprofessional team that includes physicians, advanced practice providers, nurses, and pharmacists. Through an extensive credentialing process and under the guidance of a CDTA, pharmacists in the pain clinic are able to evaluate patients, develop treatment plans, and prescribe pain medication therapies for oncology patients. By having pharmacists provide these services, the pain clinic can improve medication dosing, ensure that medications are managed consistently, improve patients' quality of care, and save providers time by allowing tasks to be completed by appropriately trained ancillary staff. For cancer-related pain, the pharmacist, in conjunction with the attending provider, develops a pain medication plan following the principles of the World Health Organization's analgesic ladder. The pain clinic has implemented the routine use of several validated tools for screening and assessment of opioid risk as well as state guidelines for managing chronic opioid therapy. The pharmacists in the pain clinic also emphasize functional goals and improvement in functional status rather than complete relief of pain. As members of an interprofessional pain clinic team, oncology pharmacists use their specialized knowledge of cancer and pharmacotherapy to help manage and treat pain in complex cancer cases. Copyright © 2016 by the American Society of Health-System Pharmacists, Inc. All rights reserved.

  3. Antimicrobial Susceptibility and Genotypic Characteristic of Campylobacter spp. Isolates from Free-Living Birds in Poland. (United States)

    Krawiec, Marta; Woźniak-Biel, Anna; Bednarski, Michał; Wieliczko, Alina


    Campylobacter spp. is the most commonly reported, bacterial cause of human foodborne infection worldwide. Commercial poultry and free-living birds are natural reservoirs of three particular species: Campylobacter jejuni, Campylobacter coli, and Campylobacter lari. The aim of this study was to determine the genotypic characteristics and antibiotic susceptibility of 43 Campylobacter strains, obtained from free-living birds, in Poland. In total, 700 birds were examined. The strains were isolated from 43 birds (6.14%) from the feces of 7 wild bird species: Mallard ducks Anas platyrhynchos (29 positive/121 tested), great cormorants Phalacrocorax carbo (5/77), velvet scoters Melanitta fusca (4/30), tawny owls Strix aluco (2/5), common buzzard Buteo buteo (1/3), rook Corvus frugilegus (1/6), and Eurasian tree sparrow Passer montanus (1/30). Thirty-eight (88.37%) of obtained strains belonged to C. jejuni and five (11.63%) to C. coli. Other 428 examined birds from different bird species were Campylobacter negative. The antimicrobial susceptibility to nine antimicrobials was also studied in investigated isolates of Campylobacter spp. Sixteen of the examined strains (37.21% of all positive samples) showed susceptibility to all of the nine antimicrobials. Moreover, the prevalence of selected virulence genes, such as flaA, cadF, ceuE, virB11, cdtA, cdtB, and cdtC were all analyzed. The virulence gene that was found most frequently in total number of Campylobacter strains was ceuE (72.10%) and other genes, such as flaA, cadF, cdtA, cdtB, and cdtC, were found in over 60% of all examined strains. Variable antimicrobial susceptibility and the presence of different virulence genes of examined strains, isolated from free-living birds, suggest that special attention should be given to wild birds and any potential approaches to the control of antibiotic-resistant Campylobacter should be discussed.

  4. High quality silicon-based substrates for microwave and millimeter wave passive circuits (United States)

    Belaroussi, Y.; Rack, M.; Saadi, A. A.; Scheen, G.; Belaroussi, M. T.; Trabelsi, M.; Raskin, J.-P.


    Porous silicon substrate is very promising for next generation wireless communication requiring the avoidance of high-frequency losses originating from the bulk silicon. In this work, new variants of porous silicon (PSi) substrates have been introduced. Through an experimental RF performance, the proposed PSi substrates have been compared with different silicon-based substrates, namely, standard silicon (Std), trap-rich (TR) and high resistivity (HR). All of the mentioned substrates have been fabricated where identical samples of CPW lines have been integrated on. The new PSi substrates have shown successful reduction in the substrate's effective relative permittivity to values as low as 3.7 and great increase in the substrate's effective resistivity to values higher than 7 kΩ cm. As a concept proof, a mm-wave bandpass filter (MBPF) centred at 27 GHz has been integrated on the investigated substrates. Compared with the conventional MBPF implemented on standard silicon-based substrates, the measured S-parameters of the PSi-based MBPF have shown high filtering performance, such as a reduction in insertion loss and an enhancement of the filter selectivity, with the joy of having the same filter performance by varying the temperature. Therefore, the efficiency of the proposed PSi substrates has been well highlighted. From 1994 to 1995, she was assistant of physics at (USTHB), Algiers . From 1998 to 2011, she was a Researcher at characterization laboratory in ionized media and laser division at the Advanced Technologies Development Center. She has integrated the Analog Radio Frequency Integrated Circuits team as Researcher since 2011 until now in Microelectronic and Nanotechnology Division at Advanced Technologies Development Center (CDTA), Algiers. She has been working towards her Ph.D. degree jointly at CDTA and Ecole Nationale Polytechnique, Algiers, since 2012. Her research interest includes fabrication and characterization of microwave passive devices on porous

  5. Evaluation of a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for the identification of Campylobacter strains isolated from diarrheal patients in Japan. (United States)

    Kabir, S M Lutful; Kikuchi, Ken; Asakura, Masahiro; Shiramaru, Sachi; Tsuruoka, Naoki; Goto, Aeko; Hinenoya, Atsushi; Yamasaki, Shinji


    We have developed a cytolethal distending toxin (cdt) gene-based species-specific multiplex PCR assay for the detection and identification of Campylobacter jejuni, C. coli, and C. fetus. The applicability of this assay was evaluated with 325 Campylobacter strains isolated from diarrheal patients in Japan and the results were compared with those obtained by other genetic methods, including hipO gene detection and 16S rRNA gene sequencing. Of the 325 strains analyzed, 314 and 11 were identified as C. jejuni and C. coli, respectively, by combination of hipO gene detection and 16S rRNA gene sequencing. When the multiplex PCR assay was employed, 309, 310, and 314 strains were identified as C. jejuni on the basis of cdtA, cdtB, and cdtC gene-specific primers, respectively. Similarly, 11, 11, and 10 strains were identified as C. coli on the basis of cdtA, cdtB, and cdtC gene-specific primers, respectively. Sequence analysis of the cdt gene region of 6 strains (5 C. jejuni and 1 C. coli) which did not yield specific PCR products in any of the cdt gene-based multiplex PCR assays revealed deletions or mutations of the cdt genes. Pulsed-field gel electrophoresis indicated that C. jejuni and C. coli strains were genetically diverse. Taken together, these findings suggest that the cdtC gene-based multiplex PCR seems to be a particularly simple and rapid method for differentiating between species of Campylobacter strains, such as C. jejuni and C. coli. However, combination of these multiplex PCR assays will allow more accurate identification.

  6. Prevalence of Putative Virulence Genes in Campylobacter and Arcobacter Species Isolated from Poultry and Poultry By-Products in Tunisia. (United States)

    Jribi, Hela; Sellami, Hanen; Hassena, Amal Ben; Gdoura, Radhouane


    Campylobacter and Arcobacter spp. are common causes of gastroenteritis in humans; these infections are commonly due to undercooked poultry. However, their virulence mechanism is still poorly understood. The aim of this study was to evaluate the presence of genotypic virulence markers in Campylobacter and Arcobacter species using PCR. The prevalence of virulence and cytolethal distending toxin (CDT) genes was estimated in 71 Campylobacteraceae isolates. PCR was used to detect the presence of virulence genes (iam, cadF, virB1, flaA, cdtA, cdtB, and cdtC) using specific primers for a total of 45 Campylobacter isolates, including 37 C. jejuni and 8 C. coli. All the Campylobacter isolates were positive for the cadF gene. The plasmid gene virB11 was not detected in any strain. The invasion associated marker was not detected in C. jejuni. Lower detection rates were observed for flaA, cdtA, cdtB, and cdtC. The presence of nine putative Arcobacter virulence genes (cadF, ciaB, cj1349, mviN, pldA, tlyA, irgA, hecA, and hecB) was checked in a set of 22 Arcobacter butzleri and 4 Arcobacter cryaerophilus isolates. The pldA and mviN genes were predominant (88.64%). Lower detection rates were observed for tlyA (84.76%), ciaB (84.61%), cadF and cj1349 (76.92%), IrgA and hecA (61.53%), and hecB (57.69%). The findings revealed that a majority of the Campylobacteraceae strains have these putative virulence genes that may lead to pathogenic effects in humans.

  7. Virulence and antibiotic resistance genes in Campylobacter spp. in the Czech Republic. (United States)

    Bardoň, J; Pudová, V; Koláčková, I; Karpíšková, R; Röderová, M; Kolář, M


    Thermotolerant species of the genus Campy-lobacter are the important agents causing human foodborne infections throughout the world. The aims of this study were to evaluate the presence of nine putative virulence genes in Campylobacter spp. isolated from patients and from foods (poultry meat, pork liver), to determine the resistance of Campylobacter isolates to eight antibiotic agents and to detect four resistance genes.Matherial and methods: The presence of the virulence genes cdtA, cdtB, cdtC, virB11, ciaB, wlaN, iam, dnaJ and racR was detected by polymerase chain reaction (PCR) in 94 Campylobacter spp. isolates from humans and 123 campylobacters from foods. The phenotypic resistance to selected antimicrobial agents was tested with microdilution method in 82 human isolates and 91 food isolates. The isolates with antibiograms were tested for the presence of blaOXA-61, tet(O), aph-3-1 and cmeB genes by PCR with specific primers. In both human and food C. jejuni isolates the preva-lence of the studied virulence genes, especially dnaJ, racR, ciaB genes and the toxigenic genes cdtA, cdtB, cdtC, was considerably higher than in C. coli isolates. The only exception was the iam gene identified in only C. coli. The tested isolates of both C. jejuni and C. coli were highly resistant to quinolone antibiotics. Additionally, C. coli was also more resistant to erythromycin, streptomycin and, in case of isolates from pork liver, to tetracycline. High prevalence rates of genes encoding antibiotic resistance was noted for the blaOXA-61 and tet(O) genes in both Campylobacter species. The presented study is the first to assess the presence of genes for virulence and resistance to antibiotics in thermotolerant Campylobacter spp. isolated from humans and foods in the Czech Republic. The resistance of Campylobacter isolates to eight antibiotic agents was also assessed. The prevalence of genes responsible for virulence and resistance is rather varied in thermotolerant Campylobacter spp.

  8. Detecção dos genes da toxina citoletal distensiva em estirpes de Campylobacter jejuni isoladas de carcaças de frangos Detection of cytolethal distending toxin genes in strains of Campylobacter jejuni isolated from broiler carcasses

    Directory of Open Access Journals (Sweden)

    A.F. Carvalho


    Full Text Available Foram analisadas 80 amostras de sobrecoxas de frangos de corte resfriados provenientes de feiras livres e hipermercados do município de São Paulo, SP. Treze estirpes de Campylobacter spp. foram isoladas em 10 (12,5% sobrecoxas, sendo cinco amostras originárias de feiras livres e cinco de hipermercados. Onze estirpes foram identificadas como Campylobacter jejuni e duas como Campylobacter coli. As 11 estirpes foram confirmadas como C. jejuni pela PCR do gene da hipuricase (hip, e destas, quatro (36,4% apresentaram os três genes (cdtA, cdtB e cdtC codificantes da toxina citoletal distensiva pela multiplex-PCR, sendo três estirpes provenientes de hipermercados e uma de feira livre. Observou-se a presença de estirpes virulentas de C. jejuni, portadoras do complexo de genes cdt, nas amostras de frango resfriado, não só na linha de abate, mas até o ponto final da cadeia de distribuição, nos dois principais centros de venda a varejo.Eighty samples of refrigerated broiler thighs purchased in street markets and supermarkets in the city of São Paulo, SP, were analyzed. Thirteen Campylobacter spp. strains were isolated in 10 (12.5% thighs, five of them from street market samples and other five from supermarkets. Eleven strains were identified as Campylobacter jejuni and two of them as Campylobacter coli. The 11 strains were confirmed to be C. jejuni using PCR for hippuricase (hip gene. From these, multiplex-PCR showed that four (36.4% strains presented the three genes (cdtA, cdtB, and cdtC encoding cytolethal distending toxin: three strains from supermarket and one from street market samples. These results are important, because they demonstrate the presence of virulent C. jejuni strains in refrigerated broiler thigh samples, not only in the slaughterhouse but in the final point of the distribution chain, at the two most important food retail commercer.

  9. ICU-Onset Clostridium difficile infection in a university hospital in China: a prospective cohort study.

    Directory of Open Access Journals (Sweden)

    Xiaohui Wang

    Full Text Available A prospective study was conducted to investigate the incidence, clinical profiles and outcome of ICU-onset CDI in a 50-bed medical ICU at a university hospital in China. Stools were collected from patients who developed ICU-onset diarrhea and was screened for tcdA (toxin A gene and tcdB (toxin B gene by PCR. CDI cases were compared with the ICU-onset non-CDI diarrhea cases for demographics, comorbidities, potential risk factors, major laboratory findings and outcomes. Stool samples from CDI cases were subjected to C. difficile culture and C. difficile isolates were screened for tcdA, tcdB and the binary toxin genes (cdtA and cdtB using multiplex PCR. Strain typing of toxigenic C. difficile isolates was performed using multilocus sequence typing. There were 1,277 patients in the ICU during the study period and 124 (9.7% developed ICU-onset diarrhea, of which 31 patients had CDI. The incidence of ICU-onset CDI was 25.2 cases per 10,000 ICU days. ICU-onset CDI cases had similar features with ICU-onset non-CDI diarrhea cases including the use of proton pump inhibitors and antibacterial agents. The crude mortality rate of ICU-onset CDI was 22.6%, but the attributable mortality rate of ICU-onset CDI was only 3.2% here. Toxigenic C. difficile isolates were recovered from 28 out of the 31 patients with CDI. cdtA and cdtB were found in two strains. Seventeen STs including 11 new STs were identified. All of the 11 new STs were single-locus variants of known STs and the 17 STs identified here could be clustered into 3 clades. The incidence of ICU-onset CDI here is similar to those in Europe and North America, suggesting that CDI is likely to be a common problem in China. Toxigenic C. difficile here belonged to a variety of STs, which may represent a significant clonal expansion rather than the true clonal diversity.

  10. Cytolethal distending toxin (CDT): a bacterial weapon to control host cell proliferation? (United States)

    De Rycke, J; Oswald, E


    Cytolethal distending toxins (CDT) constitute a family of genetically related bacterial protein toxins able to stop the proliferation of numerous cell lines. This effect is due to their ability to trigger in target cells a signaling pathway that normally prevents the transition between the G2 and the M phase of the cell cycle. Produced by several unrelated Gram-negative mucosa-associated bacterial species, CDTs are determined by a cluster of three adjacent genes (cdtA, cdtB, cdtC) encoding proteins whose respective role is not yet fully elucidated. The CDT-B protein presents sequence homology to several mammalian and bacterial phosphodiesterases, such as DNase I. The putative nuclease activity of CDT-B, together with the activation by CDT of a G2 cell cycle checkpoint, strongly suggests that CDT induces an as yet uncharacterized DNA alteration. However, the effective entry of CDT into cells and subsequent translocation into the nucleus have not yet been demonstrated by direct methods. The relationship between the potential DNA-damaging properties of this original family of toxins and their role as putative virulence factors is discussed.

  11. Application of pressurized fluid extraction to determine cadmium and zinc in plants. (United States)

    Maurí-Aucejo, A R; Arnandis-Chover, T; Marín-Sáez, R; Llobat-Estellés, M


    A procedure for the determination of Cd and Zn in plants is proposed. The metals are extracted by pressurized fluid extraction (PFE). Operational conditions are: pressure 1500 psi, temperature 75 degrees C, static time 5 min, flush volume 35%, purge time 60s, cycles 1 and 1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid (CDTA) 0.01M at pH 4.5 as extracting solution. Determination of Zn is carried out by flame atomic absorption spectroscopy and depending on the concentration level, Cd content is determined by flame or electrothermal atomic absorption spectroscopy. Certified samples of Virginia tobacco leaves, tea leaves, spinach leaves, poplar leaves, a commercial spinach sample (Spinacea oleracea) and genetically modified Arabidopsis thaliana were analysed by the proposed procedure and also by microwave acid digestion and extraction with HCl-Triton X-100. Confidence intervals for Cd and Zn content obtained by the proposed procedure overlap with the certified values. The other procedures, however, provide inaccurate results for Cd. Recoveries obtained for a confidence level of 95% are 96+/-6% and 95+/-5% for Zn and Cd, respectively. Reproducibility of Zn by the proposed procedure is 7% (n=8), similar to the other tests and the detection limit is 2.6 microg. For Cd reproducibility is 8.5% (n=8), better than with HCl-Triton X-100 and similar to acid digestion, the detection limit is 3.5 ng of Cd.

  12. FT-IR study of the Chara corallina cell wall under deformation. (United States)

    Toole, Geraldine A; Kacuráková, Marta; Smith, Andrew C; Waldron, Keith W; Wilson, Reginald H


    Fourier-transform infrared (FT-IR) microspectroscopy was used to investigate both the chemical composition of, and the effects of an applied strain on, the structure of the Chara corallina cell wall. The inner layers of the cell wall are known to have a transverse cellulose orientation with a gradient through the thickness to longitudinal orientation in the older layers. In both the native state and following the removal of various biopolymers by a sequential extraction infrared dichroism was used to examine the orientation of different biopolymers in cell-wall samples subjected to longitudinal strain. In the Chara system, cellulose microfibrils were found to be aligned predominantly transverse to the long axis of the cell and became orientated increasingly transversely as longitudinal strain increased. Simultaneously, the pectic polysaccharide matrix underwent molecular orientation parallel to the direction of strain. Following extraction in CDTA, microfibrils were orientated transversely to the strain direction, and again the degree of transverse orientation increased with increasing strain. However, the pectic polysaccharides of the matrix were not detected in the dichroic difference spectra. After a full sequential extraction, the cellulose microfibrils, now with greatly reduced crystallinity, were detected in a longitudinal direction and they became orientated increasingly parallel to the direction of strain as it increased.

  13. Influence of cultivar, cooking, and storage on cell-wall polysaccharide composition of winter squash (Cucurbita maxima). (United States)

    Ratnayake, R M Sunil; Melton, Laurence D; Hurst, Paul L


    Changes in the cell-wall polysaccharides (CWP) of the edible tissues of four winter squash cultivars during storage and after cooking were investigated. A procedure for isolating cell walls of tissues containing high levels of starch was used. The starch-free CWP were sequentially fractionated using CDTA, dilute Na(2)CO(3), and 4 M KOH. Cellulose made up 40-42% of the total CWP for three cultivars (Delica, CF 2, and CF 4) at harvest but was 35% in the softer Red Warren. The pectic polysaccharides of Delica, CF 2, and CF 4 cell walls are more branched than those from Red Warren squash. The higher proportion of uronic acid in the pectic polysaccharides of Red Warren squash correlates with its lower firmness. Cooking resulted in an increase in the water-soluble pectins and a decrease in the pectins associated with cellulose. The total CWP content of the squash cultivars remained unchanged for up to 2 months of storage and then markedly decreased between 2 and 3 months of storage. The galactose content of Delica and Red Warren cell walls remained relatively constant from harvest to 2 months of storage and then decreased markedly during 2-3 months of storage.

  14. Down-regulation of POLYGALACTURONASE1 alters firmness, tensile strength and water loss in apple (Malus x domestica) fruit. (United States)

    Atkinson, Ross G; Sutherland, Paul W; Johnston, Sarah L; Gunaseelan, Kularajathevan; Hallett, Ian C; Mitra, Deepali; Brummell, David A; Schröder, Roswitha; Johnston, Jason W; Schaffer, Robert J


    While there is now a significant body of research correlating apple (Malus x domestica) fruit softening with the cell wall hydrolase ENDO-POLYGALACTURONASE1 (PG1), there is currently little knowledge of its physiological effects in planta. This study examined the effect of down regulation of PG1 expression in 'Royal Gala' apples, a cultivar that typically has high levels of PG1, and softens during fruit ripening. PG1-suppressed 'Royal Gala' apples harvested from multiple seasons were firmer than controls after ripening, and intercellular adhesion was higher. Cell wall analyses indicated changes in yield and composition of pectin, and a higher molecular weight distribution of CDTA-soluble pectin. Structural analyses revealed more ruptured cells and free juice in pulled apart sections, suggesting improved integrity of intercellular connections and consequent cell rupture due to failure of the primary cell walls under stress. PG1-suppressed lines also had reduced expansion of cells in the hypodermis of ripe apples, resulting in more densely packed cells in this layer. This change in morphology appears to be linked with reduced transpirational water loss in the fruit. These findings confirm PG1's role in apple fruit softening and suggests that this is achieved in part by reducing cellular adhesion. This is consistent with previous studies carried out in strawberry but not with those performed in tomato. In apple PG1 also appears to influence other fruit texture characters such as juiciness and water loss.

  15. Room temperature ionic liquid-based microextraction for vanadium species separation and determination in water samples by electrothermal atomic absorption spectrometry

    Energy Technology Data Exchange (ETDEWEB)

    Berton, Paula; Martinis, Estefania M. [Laboratory of Environmental Research and Services of Mendoza (LISAMEN), (CCT-CONICET-Mendoza), Av. Ruiz Leal S/N Parque General San Martin, CC. 131, M 5502 IRA Mendoza (Argentina); Martinez, Luis D. [INQUISAL-CONICET, Departamento de Quimica Analitica, Facultad de Quimica, Bioquimica y Farmacia, Universidad Nacional de San Luis (Argentina); Wuilloud, Rodolfo G., E-mail: [Laboratory of Environmental Research and Services of Mendoza (LISAMEN), (CCT-CONICET-Mendoza), Av. Ruiz Leal S/N Parque General San Martin, CC. 131, M 5502 IRA Mendoza (Argentina); Instituto de Ciencias Basicas, Universidad Nacional de Cuyo, Mendoza (Argentina)


    A simple microextraction technique based on room temperature ionic liquids (RTILs) for trace V(IV) and V(V) species separation and preconcentration in water samples was developed in this work. Vanadium species microextraction was achieved with a minimal amount of the RTIL 1-butyl-3-methylimidazolium hexafluorophosphate ([C{sub 4}mim][PF{sub 6}]) as vanadium-2-(5-bromo-2-pyridylazo)-5-diethylaminophenol (V-5-Br-PADAP) complex. The speciation analysis was performed based on a modern technique defined as temperature-controlled ionic liquid dispersive liquid phase microextraction (TILDLME). The level of V(IV) species was calculated by difference of total V and V(V) levels. Selectivity among V species was obtained with the use of 1,2-cyclohexanediaminetetraacetic acid (CDTA) as masking agent. Determination of V was developed by direct injection of the RTIL phase into the electrothermal atomic absorption spectrometer (ETAAS). A preconcentration factor of 40 was achieved with only 2 mL of sample. The limit of detection (LOD) obtained under optimum conditions was 4.9 ng L{sup -1} and the relative standard deviation for 10 replicate determinations at the 0.5 {mu}g L{sup -1} V level was 4.3%, calculated at peak heights. A correlation coefficient of 0.9961 was achieved. The method was successfully applied for the speciation analysis of V in tap and river water samples.

  16. Measurement of 90Sr radioactivity in a rapid method of strontium estimation by solvent extraction with dicarbollides

    International Nuclear Information System (INIS)

    Svoboda, K.; Kyrs, M.


    The application of liquid scintillation counting to the measurement of 90 Sr radioactivity was studied, using a previously published rapid method of strontium separation, based on solvent extraction with a solution of cobalt dicarbollide and Slovafol 909 in a nitrobenzene-carbon tetrachloride mixture and subsequent stripping of strontium with a 0.15 M Chelaton IV (CDTA) solution at pH 10.2. With liquid scintillation counting, a more efficient elimination of the effect of 90 Y β-activity on 90 Sr counting is possible than when measuring the evaporated aliquot with the use of a solid scintillator. The adverse effect of traces of dicarbollide, nitrobenzene, and CCl 4 passed over in the aqueous 90 Sr solution prepared for counting, is caused by the (poorly reproducible) shift of the 90 Sr + 90 Y β-radiation spectral curve towards lower energies, the so-called quenching. The shift is independent of the aqueous phase concentration of the organic compounds mentioned. They can be removed by shaking the aqueous reextract with an equal volume of octanol or amyl acetate so that the undesirable spectral shift does not occur. No loss of strontium was found in this washing procedure. (author) 2 tabs., 6 figs., 5 refs

  17. Separation of Ra and Th from rock matrices for alpha-spectrometry

    International Nuclear Information System (INIS)

    Ditchburn, R.G.; Whitehead, N.E.


    Dating rocks using 226 Ra/ 230 Th isochrons demands radiochemical purification of radium and thorium. This paper presents an improved method. Rocks are solubilised by nitric/hydrofluoric digestion followed by dissolution of insoluble fluorides by boric/nitric acids, and Th is extracted by passing the 8M nitric solution through an anion column in nitrate form. The eluant contains Ra and Ba which are precipitated as sulphate and redissolved in alkaline EDTA. Complete separation of Ba and Ra from sulphate is on an AG 1 x 8 anion column in EDTA form, which was found better than the chloride form. The Ba+Ra is separated on an AG 50W x 8 cation column, and uses CDTA as an eluant for Ba. Careful pH control is essential. Ra elutes later with EDTA or 4M HCl and is precipitated with 125μg barium as sulphate to yield a source suitable for α-spectrometry, or further treated to electroplate the Ra. The yield tracers used are 228 Th and 224 Ra. Because the sample contains natural 224 Ra a correction must be applied, calculated from the amount of natural 232 Th in the Th spectrum. Th may be precipitated with 100 μg of ferric iron and gives a spectrometry-quality source, but further purification and electrodeposition was found to be preferable. Variations on the method for the case of analysis of calcium-rich fish otoliths are described. (author) 37 refs.; 3 figs

  18. Determination of aminopolycarboxylic acids in river water by solid-phase extraction on activated charcoal cartridges and gas chromatography with mass spectrometric detection. Method performance characteristics and estimation of the uncertainty. (United States)

    Jiménez, Juan J


    A new sample preparation procedure to determine aminopolycarboxylic acids (ethylenediaminetetraacetic acid, EDTA, nitrilotriacetic acid, NTA, diethylenetriaminepentaacetic acid, DTPA, and cyclohexanediaminetetraacetic acid, CDTA) in river water is described. The procedure consists of the solid-phase extraction of the aminopolycaroxyllic acids on activated charcoal cartridges after increasing the ionic strength and acidifying the sample. The extract was eluted with methanol and the analytes were methylated in presence of BF3/methanol to determine them by GC with mass spectrometric detection. Recoveries were higher than 90% with good repeatabilities and inter-day precision for concentrations close to quantification limits (about 10 μg L(-1)) and higher. It has been verified that the proposed method is robust according to the Youden and Steiner test and free of matrix effects arisen from the presence of organic matter and iron(III) as deduced from statistical tests. A bottom-up approach was followed to estimate the uncertainty of the measured concentration. At concentrations close to 10 μg L(-1) the most relevant step of the method is the calculus of the interpolated concentration which has a high value of relative standard uncertainty. Copyright © 2013 Elsevier B.V. All rights reserved.

  19. Ultrasound enhances calcium absorption of jujube fruit by regulating the cellular calcium distribution and metabolism of cell wall polysaccharides. (United States)

    Zhi, Huanhuan; Liu, Qiqi; Xu, Juan; Dong, Yu; Liu, Mengpei; Zong, Wei


    Ultrasound has been applied in fruit pre-washing processes. However, it is not sufficient to protect fruit from pathogenic infection throughout the entire storage period, and sometimes ultrasound causes tissue damage. The goal of this study was to investigate the effects of calcium chloride (CaCl 2 , 10 g L -1 ) and ultrasound (350 W at 40 kHz), separately and in combination, on jujube fruit quality, antioxidant status, tissue Ca 2+ content and distribution along with cell wall metabolism at 20 °C for 6 days. All three treatments significantly maintained fruit firmness and peel color, reduced respiration rate, decay incidence, superoxide anion, hydrogen peroxide and malondialdehyde and preserved higher enzymatic (superoxide dismutase, catalase and peroxidase) and non-enzymatic (ascorbic acid and glutathione) antioxidants compared with the control. Moreover, the combined treatment was more effective in increasing tissue Ca 2+ content and distribution, inhibiting the generation of water-soluble and CDTA-soluble pectin fractions, delaying the solubilization of Na 2 CO 3 -soluble pectin and having lower activities of cell wall-modifying enzymes (polygalacturonase and pectate lyase) during storage. These results demonstrated that the combination of CaCl 2 and ultrasound has potential commercial application to extend the shelf life of jujube fruit by facilitating Ca 2+ absorption and stabilizing the cell wall structure. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

  20. Cytolethal Distending Toxin Enhances Radiosensitivity in Prostate Cancer Cells by Regulating Autophagy

    Directory of Open Access Journals (Sweden)

    Hwai-Jeng Lin


    Full Text Available Cytolethal distending toxin (CDT produced by Campylobacter jejuni contains three subunits: CdtA, CdtB, and CdtC. Among these three toxin subunits, CdtB is the toxic moiety of CDT with DNase I activity, resulting in DNA double-strand breaks (DSB and, consequently, cell cycle arrest at the G2/M stage and apoptosis. Radiation therapy is an effective modality for the treatment of localized prostate cancer (PCa. However, patients often develop radioresistance. Owing to its particular biochemical properties, we previously employed CdtB as a therapeutic agent for sensitizing radioresistant PCa cells to ionizing radiation (IR. In this study, we further demonstrated that CDT suppresses the IR-induced autophagy pathway in PCa cells by attenuating c-Myc expression and therefore sensitizes PCa cells to radiation. We further showed that CDT prevents the formation of autophagosomes via decreased high-mobility group box 1 (HMGB1 expression and the inhibition of acidic vesicular organelle (AVO formation, which are associated with enhanced radiosensitivity in PCa cells. The results of this study reveal the detailed mechanism of CDT for the treatment of radioresistant PCa.

  1. The Biology of the Cytolethal Distending Toxins (United States)

    Guerra, Lina; Cortes-Bratti, Ximena; Guidi, Riccardo; Frisan, Teresa


    The cytolethal distending toxins (CDTs), produced by a variety of Gram-negative pathogenic bacteria, are the first bacterial genotoxins described, since they cause DNA damage in the target cells. CDT is an A-B2 toxin, where the CdtA and CdtC subunits are required to mediate the binding on the surface of the target cells, allowing internalization of the active CdtB subunit, which is functionally homologous to the mammalian deoxyribonuclease I. The nature of the surface receptor is still poorly characterized, however binding of CDT requires intact lipid rafts, and its internalization occurs via dynamin-dependent endocytosis. The toxin is retrograde transported through the Golgi complex and the endoplasmic reticulum, and subsequently translocated into the nuclear compartment, where it exerts the toxic activity. Cellular intoxication induces DNA damage and activation of the DNA damage responses, which results in arrest of the target cells in the G1 and/or G2 phases of the cell cycle and activation of DNA repair mechanisms. Cells that fail to repair the damage will senesce or undergo apoptosis. This review will focus on the well-characterized aspects of the CDT biology and discuss the questions that still remain unanswered. PMID:22069704

  2. Detection of CDT toxin genes in Campylobacter spp. strains isolated from broiler carcasses and vegetables in São Paulo, Brazil

    Directory of Open Access Journals (Sweden)

    Aline Feola de Carvalho


    Full Text Available Campylobacteriosis is a worldwide distributed zoonosis. One of the main virulence factors related to Campylobacter spp. in animals and humans is the cytolethal distending toxin (CDT, encoded by three adjacent genes (cdtA, cdtB, cdtC. The occurrence of Campylobacter spp. in samples of vegetables has not been reported in Brazil yet, and has seldom been described in the international literature. The detection of CDT in these strains has not been reported, either. The objectives of the present study were to determine the occurrence of Campylobacter spp. strains carrying virulence factors in samples of poultry and vegetables (lettuce and spinach from different points of sale, thus verifying if vegetables are as an important vehicle for potentially virulent Campylobacter spp. strains as poultry. Twenty four strains were identified as Campylobacter jejuni by phenotypic and genotypic methods: 22 from broiler carcasses and two from lettuce samples. Three strains were identified as Campylobacter coli: two from broiler carcasses and one from lettuce. The presence of the cdt genes were detected in 20/24 (83.3% C. jejuni strains, and 3/3 (100% C. coli strains. The isolation of Campylobacter spp. strains with the cdt gene cluster in lettuce samples points to a new possible source of contamination, which could have an impact in the vegetable production chain and risk to public health. Results show that potentially virulent C. jejuni and C. coli strains remain viable in samples of broiler carcasses and vegetables at the points of sale.

  3. Detection of CDT toxin genes in Campylobacter spp. strains isolated from broiler carcasses and vegetables in São Paulo, Brazil. (United States)

    de Carvalho, Aline Feola; da Silva, Daniela Martins; Azevedo, Sergio Santos; Piatti, Rosa Maria; Genovez, Margareth Elide; Scarcelli, Eliana


    Campylobacteriosis is a worldwide distributed zoonosis. One of the main virulence factors related to Campylobacter spp. in animals and humans is the cytolethal distending toxin (CDT), encoded by three adjacent genes (cdtA, cdtB, cdtC). The occurrence of Campylobacter spp. in samples of vegetables has not been reported in Brazil yet, and has seldom been described in the international literature. The detection of CDT in these strains has not been reported, either. The objectives of the present study were to determine the occurrence of Campylobacter spp. strains carrying virulence factors in samples of poultry and vegetables (lettuce and spinach) from different points of sale, thus verifying if vegetables are as an important vehicle for potentially virulent Campylobacter spp. strains as poultry. Twenty four strains were identified as Campylobacter jejuni by phenotypic and genotypic methods: 22 from broiler carcasses and two from lettuce samples. Three strains were identified as Campylobacter coli: two from broiler carcasses and one from lettuce. The presence of the cdt genes were detected in 20/24 (83.3%) C. jejuni strains, and 3/3 (100%) C. coli strains. The isolation of Campylobacter spp. strains with the cdt gene cluster in lettuce samples points to a new possible source of contamination, which could have an impact in the vegetable production chain and risk to public health. Results show that potentially virulent C. jejuni and C. coli strains remain viable in samples of broiler carcasses and vegetables at the points of sale.

  4. A multiplex, internally controlled real-time PCR assay for detection of toxigenic Clostridium difficile and identification of hypervirulent strain 027/ST-1

    DEFF Research Database (Denmark)

    Hoegh, A M; Nielsen, J B; Lester, A


    The purpose of this study was to validate a multiplex real-time PCR assay capable of detecting toxigenic Clostridium difficile and simultaneously identifying C. difficile ribotype 027/ST-1 by targeting the toxin genes tcdA, tcdB and cdtA in one reaction and in a separate reaction identifying the Δ...... to confirm the correct identification of the Δ117 deletion in tcdC and C. difficile ribotype 027/ST-1, respectively. The PCR assay displayed a sensitivity, specificity, PPV and NPV of 99.0%, 97.4%, 87.4% and 99.8%, respectively, compared to toxigenic culture on 665 samples evaluable both by PCR and culture....... Sequencing of tcdC, ribotyping and MLST of cultured isolates validated the genotyping assay and confirmed the ability of the assay to correctly identify C. difficile ribotype 027/ST-1 in our current epidemiological setting. We describe the use of a combination of two separate PCR assays for sensitive...

  5. Polysaccharide compositions of collenchyma cell walls from celery (Apium graveolens L.) petioles. (United States)

    Chen, Da; Harris, Philip J; Sims, Ian M; Zujovic, Zoran; Melton, Laurence D


    Collenchyma serves as a mechanical support tissue for many herbaceous plants. Previous work based on solid-state NMR and immunomicroscopy suggested collenchyma cell walls (CWs) may have similar polysaccharide compositions to those commonly found in eudicotyledon parenchyma walls, but no detailed chemical analysis was available. In this study, compositions and structures of cell wall polysaccharides of peripheral collenchyma from celery petioles were investigated. This is the first detailed investigation of the cell wall composition of collenchyma from any plant. Celery petioles were found to elongate throughout their length during early growth, but as they matured elongation was increasingly confined to the upper region, until elongation ceased. Mature, fully elongated, petioles were divided into three equal segments, upper, middle and lower, and peripheral collenchyma strands isolated from each. Cell walls (CWs) were prepared from the strands, which also yielded a HEPES buffer soluble fraction. The CWs were sequentially extracted with CDTA, Na 2 CO 3 , 1 M KOH and 4 M KOH. Monosaccharide compositions of the CWs showed that pectin was the most abundant polysaccharide [with homogalacturonan (HG) more abundant than rhamnogalacturonan I (RG-I) and rhamnogalacturonan II (RG-II)], followed by cellulose, and other polysaccharides, mainly xyloglucans, with smaller amounts of heteroxylans and heteromannans. CWs from different segments had similar compositions, but those from the upper segments had slightly more pectin than those from the lower two segments. Further, the pectin in the CWs of the upper segment had a higher degree of methyl esterification than the other segments. In addition to the anticipated water-soluble pectins, the HEPES-soluble fractions surprisingly contained large amounts of heteroxylans. The CDTA and Na 2 CO 3 fractions were rich in HG and RG-I, the 1 M KOH fraction had abundant heteroxylans, the 4 M KOH fraction was rich in xyloglucan and

  6. Molecular characteristics of Clostridium difficile isolates from human and animals in the North Eastern region of India. (United States)

    Hussain, Isfaqul; Borah, P; Sharma, R K; Rajkhowa, S; Rupnik, M; Saikia, D P; Hasin, D; Hussain, Iftikar; Deka, N K; Barkalita, L M; Nishikawa, Y; Ramamurthy, T


    A total of 1034 samples were collected from different sources and C. difficile was isolated from 18 (9.04%) of 199 human, 9 (4.89%) of 184 cattle, 29 (12.44%) of 233 pig, and from 23 (13.94%) of 165 poultry samples. Variations were observed on the rate of isolation according to age and clinical conditions (diarrhoea). None of the samples from cow, sheep, goat, local chicken, and wild animals yielded any C. difficile. Out of those isolates, 8, 2, 19 and 6 isolates from human, cattle, pig and poultry, respectively were toxigenic. The toxigenic isolates carried both tcdA, and tcdB (A + B + ) and most of the human and the pig isolates were also positive for binary toxin genes (cdtA and cdtB). The A + B + isolates belonged to three different toxinotypes (0, VI and XXXIII). Human and pig A + B + isolates belonged to three (045, 126 and ACD 019) and four (046, 087, 126 and ACD 011) different ribotypes, respectively and the ribotypes of two cattle isolates were 014 and ACD 010. Six A + B + avian isolates belonged to six different ribotypes (014, 087, SLO 134, SLO 160, ACD 012, ACD 014). The non-toxigenic isolates from human, cattle, pig and poultry were grouped into 7, 4, 4 and 7 different ribotypes, respectively. PFGE analysis could not differentiate similar ribotypes/toxinotypes of toxigenic isolates. All the toxigenic isolates showed cytopathic effect on Vero and Hela cell monolayers at 1:100 dilutions of cell-free culture supernatants within 18-20 h of inoculation. Copyright © 2016 Elsevier Ltd. All rights reserved.

  7. Nanocrystalline Sm{sub 0.5}Sr{sub 0.5}CoO{sub 3−δ} synthesized using a chelating route for use in IT-SOFC cathodes: Microstructure, surface chemistry and electrical conductivity

    Energy Technology Data Exchange (ETDEWEB)

    Scurtu, Rares [“IlieMurgulescu” Institute of Physical Chemistry, Romanian Academy, 202 SplaiulIndependentei, 060021 Bucharest (Romania); Romania National Institute for Research and Development in Microtechnologies(IMT), 023573 Bucharest (Romania); Somacescu, Simona, E-mail: [“IlieMurgulescu” Institute of Physical Chemistry, Romanian Academy, 202 SplaiulIndependentei, 060021 Bucharest (Romania); Calderon-Moreno, Jose Maria; Culita, Daniela [“IlieMurgulescu” Institute of Physical Chemistry, Romanian Academy, 202 SplaiulIndependentei, 060021 Bucharest (Romania); Bulimestru, Ion; Popa, Nelea; Gulea, Aurelian [Faculty of Chemistry and Chemical Technology State University of Moldova 60 Mateevici, Chisinau MD 2009 Republic of Moldova (Moldova, Republic of); Osiceanu, Petre [“IlieMurgulescu” Institute of Physical Chemistry, Romanian Academy, 202 SplaiulIndependentei, 060021 Bucharest (Romania)


    Nanocrystalline Sm{sub 0.5}Sr{sub 0.5}CoO{sub 3−δ} powders were synthesized by a chelating route using different polyfunctional H{sub x}APC acids (APC=aminopolycarboxylate; x=3, 4, 5). Different homologous aminopolycarboxylic acids, namely nitrilotriacetic (H{sub 3}nta), ethylenediaminetetraacetic (H{sub 4}edta), 1,2-cyclohexanediaminetetracetic (H{sub 4}cdta) and diethylenetriaminepentaacetic (H{sub 5}dtpa) acid, were used as chelating agents to combine Sm, Sr, Co elements into a perovskite structure. The effects of the chelating agents on the crystalline structure, porosity, surface chemistry and electrical properties were investigated. The electrical properties of the perovskite-type materials emphasized that their conductivities in the temperature range of interest (600–800 °C) depend on the nature of the precursors as well as on the presence of a residual Co oxide phase as shown by XRD and XPS analysis. The surface chemistry and the surface stoichiometries were determined by XPS revealing a complex chemical behavior of Sr that exhibits a peculiar “surface phase” and “bulk phase” chemistry within the detected volume (<10 nm). - Graphical abstract: Synthesis of nanocrystalline Sm{sub 0.5}Sr{sub 0.5}CoO{sub 3−δ} powders by a chelating route and the investigation of the microstructure, surface chemistry and electrical properties. Display Omitted - Highlights: • Nanocrystalline Sm{sub 0.5}Sr{sub 0.5}CoO{sub 3−δ} obtained by a chelating synthesis route. • Cubic perovskite structures with crystallite sizes ∼23±2 nm. • The porous nature revealed by N{sub 2} adsorption/desorption (BET). • The surface chemistry and the surface stoichiometries highlighted by XPS. • A complex chemical behavior of Sr exhibits a peculiar “surface phase” and “bulk phase” chemistry.

  8. Analytical methodology for the study of decontamination factors in mixer-settlers

    International Nuclear Information System (INIS)

    Alzabet, Horacio; Crubellati, Ricardo; Di Santo, Norberto; Eppis, Maria; Gonzalez, Guillermo; Bof, Elba; Devida, Claudio


    Due to the complexity of the samples, the choice of the analytical methodology to be applied at a radiochemical laboratory, for the evaluation of different separation steps if of a great importance. It is necessary to know the performance of mixer-settlers in two extreme situations: efficiency limit determined by fluid dynamic factors and separation factor determined by physicochemical variables. The element used to determine the efficiency limit was magnesium, due to its low distribution coefficient in tri-n-butyl phosphate (TBP). Zirconium, being one of the principal fission product, is partially extracted and was used to study the influence of chemical conditions. Atomic absorption spectrometry was selected for the determination of micro quantities of magnesium, and complexometry for macro quantities of this element. It was necessary to separate uranium using extraction chromatography with Kel F-TBP columns. Concentration range was 0.1 - 1 mg/L and 7 - 70 g/L respectively. Zirconium was determined by spectrophotometry with xylenol orange. For organic samples, it was necessary to reextract the analyte with fluoride, to evaporate with sulfuric acid and to complex the excess of fluoride with aluminum. Uranium determinations, required for mass balance, were carried out by spectrophotometry and potentiometry depending upon concentration. Because of the presence of zirconium, two methods were developed for uranium determination using liquid-liquid extraction in the presence of complexing agents. In one case, TBP was used as extractant, dibenzoylmethane as chromogenic reagent and EDTA as complexing agent. For very low quantities of uranium, tri-octyl phosphine oxide (TOPO) was the extractant, 2-(2-thiazolylazo)-5-diethylaminophenol (TAAP) the chromogenic reagent, and (1-2-cyclohexylenedinitrilo) tetraacetic acid (CDTA) the complexing agent. (Author)

  9. Genetic characterisation of virulence genes associated with adherence, invasion and cytotoxicity in Campylobacter spp. isolated from commercial chickens and human clinical cases

    Directory of Open Access Journals (Sweden)

    Samantha Reddy


    Full Text Available Virulence-associated genes have been recognised and detected in Campylobacter species. The majority of them have been proven to be associated with pathogenicity. This study aimed to detect the presence of virulence genes associated with pathogenicity and responsible for invasion, expression of adherence, colonisation and production of the cytolethal distending toxin (cdt in Campylobacter jejuni and Campylobacter coli. Commercial chicken faecal samples were randomly sampled from chicken farms within the Durban metropolitan area in South Africa. Furthermore, human clinical Campylobacter spp. isolates were randomly sampled from a private pathology laboratory in South Africa. Out of a total of 100 chicken faecal samples, 78% (n = 78 were positive for Campylobacter growth on modified charcoal cefoperazone deoxycholate and from the random laboratory collection of 100 human clinical isolates, 83% (n = 83 demonstrated positive Campylobacter spp. growth following culturing methods. These samples were screened for the presence of the following virulence genes: cadF, hipO, asp, ciaB, dnaJ, pldA, cdtA, cdtB and cdtC. As expected, the cadF gene was present in 100% of poultry (n = 78 and human clinical isolates (n = 83. Campylobacter jejuni was the main species detected in both poultry and human clinical isolates, whilst C. coli were detected at a significantly lower percentage (p < 0.05. Eight per cent of the C. jejuni from human clinical isolates had all virulence genes that were investigated. Only one C. coli isolate demonstrated the presence of all the virulence genes investigated; however, the pldA virulence gene was detected in 100% of the C. coli isolates in poultry and a high percentage (71% in human clinical C. coli isolates as well. The detection of cdt genes was found at higher frequency in poultry than human clinical isolates. The high prevalence rates of virulence genes detected in poultry and human clinical isolates demonstrate their

  10. Hollow-fibre liquid phase microextraction for separation and preconcentration of vanadium species in natural waters and their determination by electrothermal vaporization-ICP-OES. (United States)

    Li, Li; Hu, Bin


    For separation and determination of vanadium(IV/V) species, a fast and sensitive method by combining hollow-fibre liquid phase microextraction (HF-LPME) with electrothermal vaporization (ETV)-ICP-OES has been developed. Two vanadium species (V(IV) and V(V)) were separated by HF-LPME with the use of ammonium pyrrolidinecarbodithioate (APDC) as chelating agent for complexing with different V species and carbon tetrachloride as the extraction solvent, and the vanadium in the post-extraction organic phase was injected into the graphite furnace for ETV-ICP-OES detection, in which APDC was acted as the chemical modifier. At pH 5.0, both V(IV)-APDC and V(V)-APDC were extracted quantitatively into CCl(4) for determination of total V. For speciation studies, 1,2-cyclohexanediaminetetraacetic acid (CDTA) was added to the sample for masking V(IV), so that only V(V)-APDC was extracted and determined. The concentration of V(IV) was calculated by subtracting the V(V) concentration from the total concentration of V. Under the optimized experimental conditions, the enrichment factor was 74 and the detection limits for V(IV) and V(V) were 86pgmL(-1) and 71pgmL(-1), respectively. The proposed method has been applied to the speciation of V in environmental water samples, and the recovery was in the range of 94%-107%. The results show that V(V) is the dominant existence form in oxygenic water and V(IV) could not been detected. In order to validate the developed procedure, a NIES No.8 vehicle exhaust particulates certified reference material was analyzed, and the results obtained for total vanadium are in good agreement with the certified values. The proposed method is simple, rapid, selective, and sensitive and no oxidation/reduction is required, it is applicable to the speciation of vanadium in environmental samples with the complicated matrix.

  11. Cell wall structures leading to cultivar differences in softening rates develop early during apple (Malus x domestica) fruit growth. (United States)

    Ng, Jovyn K T; Schröder, Roswitha; Sutherland, Paul W; Hallett, Ian C; Hall, Miriam I; Prakash, Roneel; Smith, Bronwen G; Melton, Laurence D; Johnston, Jason W


    There is a paucity of information regarding development of fruit tissue microstructure and changes in the cell walls during fruit growth, and how these developmental processes differ between cultivars with contrasting softening behaviour. In this study we compare two apple cultivars that show different softening rates during fruit development and ripening. We investigate whether these different softening behaviours manifest themselves late during ethylene-induced softening in the ripening phase, or early during fruit expansion and maturation. 'Scifresh' (slow softening) and 'Royal Gala' (rapid softening) apples show differences in cortical microstructure and cell adhesion as early as the cell expansion phase. 'Scifresh' apples showed reduced loss of firmness and greater dry matter accumulation compared with 'Royal Gala' during early fruit development, suggesting differences in resource allocation that influence tissue structural properties. Tricellular junctions in 'Scifresh' were rich in highly-esterified pectin, contributing to stronger cell adhesion and an increased resistance to the development of large airspaces during cell expansion. Consequently, mature fruit of 'Scifresh' showed larger, more angular shaped cells than 'Royal Gala', with less airspaces and denser tissue. Stronger cell adhesion in ripe 'Scifresh' resulted in tissue fracture by cell rupture rather than by cell-to-cell-separation as seen in 'Royal Gala'. CDTA-soluble pectin differed in both cultivars during development, implicating its involvement in cell adhesion. Low pectin methylesterase activity during early stages of fruit development coupled with the lack of immuno-detectable PG was associated with increased cell adhesion in 'Scifresh'. Our results indicate that cell wall structures leading to differences in softening rates of apple fruit develop early during fruit growth and well before the induction of the ripening process.

  12. Comparative glycan profiling of Ceratopteris richardii 'C-Fern' gametophytes and sporophytes links cell-wall composition to functional specialization. (United States)

    Eeckhout, Sharon; Leroux, Olivier; Willats, William G T; Popper, Zoë A; Viane, Ronald L L


    Innovations in vegetative and reproductive characters were key factors in the evolutionary history of land plants and most of these transformations, including dramatic changes in life cycle structure and strategy, necessarily involved cell-wall modifications. To provide more insight into the role of cell walls in effecting changes in plant structure and function, and in particular their role in the generation of vascularization, an antibody-based approach was implemented to compare the presence and distribution of cell-wall glycan epitopes between (free-living) gametophytes and sporophytes of Ceratopteris richardii 'C-Fern', a widely used model system for ferns. Microarrays of sequential diamino-cyclohexane-tetraacetic acid (CDTA) and NaOH extractions of gametophytes, spores and different organs of 'C-Fern' sporophytes were probed with glycan-directed monoclonal antibodies. The same probes were employed to investigate the tissue- and cell-specific distribution of glycan epitopes. While monoclonal antibodies against pectic homogalacturonan, mannan and xyloglucan widely labelled gametophytic and sporophytic tissues, xylans were only detected in secondary cell walls of the sporophyte. The LM5 pectic galactan epitope was restricted to sporophytic phloem tissue. Rhizoids and root hairs showed similarities in arabinogalactan protein (AGP) and xyloglucan epitope distribution patterns. The differences and similarities in glycan cell-wall composition between 'C-Fern' gametophytes and sporophytes indicate that the molecular design of cell walls reflects functional specialization rather than genetic origin. Glycan epitopes that were not detected in gametophytes were associated with cell walls of specialized tissues in the sporophyte. © The Author 2014. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved. For Permissions, please email:

  13. Cytolethal Distending Toxin From Campylobacter jejuni Requires the Cytoskeleton for Toxic Activity. (United States)

    Méndez-Olvera, Estela T; Bustos-Martínez, Jaime A; López-Vidal, Yolanda; Verdugo-Rodríguez, Antonio; Martínez-Gómez, Daniel


    Campylobacter jejuni is one of the major causes of infectious diarrhea worldwide. The distending cytolethal toxin (CDT) of Campylobacter spp. interferes with normal cell cycle progression. This toxic effect is considered a result of DNase activity that produces chromosomal DNA damage. To perform this event, the toxin must be endocytosed and translocated to the nucleus. The aim of this study was to evaluate the role of the cytoskeleton in the translocation of CDT to the nucleus. Campylobacter jejuni ATCC 33291 and seven isolates donated from Instituto de Biotecnologia were used in this study. The presence of CDT genes in C. jejuni strains was determined by PCR. To evaluate the effect of CDT, HeLa cells were treated with bacterial lysate, and the damage and morphological changes were analyzed by microscopy, immunofluorescence staining, and flow cytometry. To evaluate the role of the cytoskeleton, HeLa cells were treated with either latrunculin A or by nocodazole and analyzed by microscopy, flow cytometry, and immunoquantification (ELISA). The results obtained showed that the eight strains of C. jejuni , including the reference strain, had the ability to produce the toxin. Usage of latrunculin A and nocodazole, two cytoskeletal inhibitors, blocked the toxic effect in cells treated with the toxin. This phenomenon was evident in flow cytometry analysis and immunoquantification of Cdc2-phosphorylated. This work showed that the cytotoxic activity of the C. jejuni CDT is dependent on its endocytosis. The alteration in the microtubules and actin filaments caused a blockage transit of the toxin, preventing it from reaching the nucleus of the cell, as well as preventing DNA fragmentation and alteration of the cell cycle. The CDT toxin appears to be an important element for the pathogenesis of campylobacteriosis, since all clinical isolates showed the presence of cdtA , cdtB and cdtC genes.

  14. Diversity of moxifloxacin resistance during a nosocomial outbreak of a predominantly ribotype ARU 027 Clostridium difficile diarrhea. (United States)

    Carman, Robert J; Genheimer, Christopher W; Rafii, Fatemeh; Park, Miseon; Hiltonsmith, Megan F; Lyerly, David M


    To characterize the extent and diversity of moxifloxacin resistance among Clostridium difficile isolates recovered during a predominantly Anaerobe Reference Unit (ARU) ribotype 027-associated nosocomial outbreak of antibiotic associated diarrhea we measured the susceptibility of 34 field isolates and 6 laboratory strains of C. difficile to moxifloxacin. We ribotyped the isolates as well as assaying them by PCR for the metabolic gene, gdh, and the virulence genes, tcdA, tcdB, tcdC, cdtA and cdtB. All the laboratory isolates, including the historical ARU 027 isolate Cd196, were susceptible to moxifloxacin (or=16 microg/mL (high resistance). We sequenced the quinolone resistance determining regions of gyrA (position 71-460) and gyrB (position 1059-1448) from two susceptible laboratory strains, all five isolates with moderate resistance and two highly resistant isolates. Two highly resistant isolates (Pitt 40, ribotype ARU 027 and Pitt 33, ribotype ARU 001) had the same C245T (Thr(82)Delta Ile) mutation. No other changes were seen. Amplification with primer pairs specific for the C245T mutant gyrA and for the wild type gene respectively confirmed all 16 highly resistant ARU 027 isolates, as well as the highly resistant isolates from other ribotypes, had the C245T mutation and that the mutation was absent from all other isolates. Among the five isolates with moderate resistance we found combinations of mutations within gyrA (T128A, Val(43)Delta Asp and G349T, Ala(117)Delta Ser) and gyrB (G1276A, Arg(426)Delta Asn). The G1396A (Glu(466)Delta Lys) mutation was not associated with increased resistance.

  15. Clostridium difficile infection diagnosis in a paediatric population: comparison of methodologies. (United States)

    Hart, J; Putsathit, P; Knight, D R; Sammels, L; Riley, T V; Keil, A


    The increasing incidence of Clostridium difficile infection (CDI) in paediatric hospitalised populations, combined with the emergence of hypervirulent strains, community-acquired CDI and the need for prompt treatment and infection control, makes the rapid, accurate diagnosis of CDI crucial. We validated commonly used C. difficile diagnostic tests in a paediatric hospital population. From October 2011 to January 2012, 150 consecutive stools were collected from 75 patients at a tertiary paediatric hospital in Perth, Western Australia. Stools were tested using: C. Diff Quik Chek Complete, Illumigene C. difficile, GeneOhm Cdiff, cycloserine cefoxitin fructose agar (CCFA) culture, and cell culture cytotoxin neutralisation assay (CCNA). The reference standard was growth on CCFA or Cdiff Chromagar and PCR on isolates to detect tcdA, tcdB, cdtA, and cdtB. Isolates were PCR ribotyped. The prevalence of CDI was high (43 % of patients). Quik Chek Complete glutamate dehydrogenase (GDH) demonstrated a low negative predictive value (NPV) (93 %). Both CCNA and Quik Chek Complete toxin A/B had poor sensitivity (33 % and 29 % respectively). Molecular methods both had 89 % sensitivity. Algorithms using GDH + Illumigene or GeneOhm reduced the sensitivity to 85 % and 83 % respectively. Ribotype UK014/20 predominated. GDH NPV and GeneOhm and Illumigene sensitivities were reduced compared with adult studies. Quik Chek Complete and CCNA cannot reliably detect toxigenic CDI. A GDH first algorithm showed reduced sensitivity. In a high prevalence paediatric population, molecular methods alone are recommended over the use of GDH algorithm or culture and CCNA, as they demonstrate the best test performance characteristics.

  16. Detection of seven virulence and toxin genes of Campylobacter jejuni isolates from Danish turkeys by PCR and cytolethal distending toxin production of the isolates. (United States)

    Bang, Dang Duong; Borck, Birgitte; Nielsen, Eva Møller; Scheutz, Flemming; Pedersen, Karl; Madsen, Mogens


    A total of 117 Campylobacter jejuni isolates from Danish turkeys were tested for the presence of seven virulence and toxin genes by PCR. One hundred seventeen (100%) isolates were positive for flaA, cadF, and ceuE gene primers. One hundred three (88%) isolates were positive for cdt gene cluster PCR detection (cdt gene cluster-PCR), whereas 101 (86.3%), 102 (87.2%), and 110 (94%) isolates were positive for cdtA-, cdtB-, and cdtC-PCR, respectively. Only 39 (33.3%) isolates were positive for virB11. Of 117 isolates, 114 (97.4%) produced cytolethal distending toxin (CDT) in Vero cell assays, 105 (89.7%) in Colon 205 assays, and 109 (93.2%) in chicken embryo cell assays. The CDT titers were determined in Vero cell assays. Of 117 isolates, 50 (42.7%) produced a CDT titer of 1:100, 29 (24.8%) of 1:50, and 27 (23%) of 1:5 to 1:10; 8 (6.8%) produced a CDT titer at undiluted supernatants and 3 (2.6%) produced no toxin. Twenty-nine C. jejuni isolates that were PCR negative for one or more individual cdt toxin genes also produced low or no CDT toxin. The high prevalence of the seven virulence and toxin genes demonstrates that these putative pathogenic determinants are widespread among Campylobacter isolates from turkeys and calls for further investigation for the elimination of Campylobacter infection in industrial turkey production and in industrial food chains.

  17. Determination of alternative and conventional chelating agents as copper(II) complexes by capillary zone electrophoresis--the first use of didecyldimethylammonium bromide as a flow reversal reagent. (United States)

    Laamanen, Pirkko-Leena; Matilainen, Rose


    A capillary zone electrophoresis (CZE) method for analyzing 11 chelating agents [beta-alaninediacetic acid (beta-ADA), trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid (CDTA), diethylenetriaminepentaacetic acid (DTPA), ethylenediaminetetraacetic acid (EDTA), N-(2-hydroxyethyl)ethylenediamine-N,N',N'-triacetic acid (HEDTA), N-(2-hydroxyethyl)iminodiacetic acid (HEIDA), iminodiacetic acid (IDA), methylglycinediacetic acid (MGDA), nitrilotriacetic acid (NTA), 1,3-diaminopropane-N,N,N',N'-tetraacetic acid (PDTA) and triethylenetetraaminehexaacetic acid (TTHA)] as negatively charged copper(II) complexes has been established. Both conventional and alternative chelating agents were included in this study, because they are used side by side in industrial applications. In this study, didecyldimethylammonium bromide (DMDDAB) was successfully used as a flow reversal reagent for the first time in an aqueous CZE method based on phosphate BGE with UV spectrophotometric detection. In addition this new flow modifier was compared to common TTAB. Method development was done using a fused silica capillary (61 cm x 50 microm i.d.). The optimized BGE was a 105 mmol L(-1) phosphate buffer with TTAB or DMDDAB in the concentration 0.5 mmol L(-1) at pH 7.1. The measurements were done with -20 kV voltage using direct UV detection at 254 nm. In both CZE methods all 11 analyte zones were properly separated (resolutions > or =2.4), and the calibrations gave excellent correlation coefficients (> or =0.998; linear range tested 0.5-2.0 mmol L(-1)). The limits of detection were < or =34 and < or =49 micromol L(-1) with the method of DMDDAB and TTAB, respectively. A clear benefit of both methods was the short analysis time; all 11 complexes were detected in less than 6 and 5.5 min with the methods of TTAB and DMDDAB, respectively. The two methods were tested with dishwashing detergents and paper mill wastewater samples and proved to be suitable for practical use.

  18. Detecção dos genes codificantes da toxina CDT, e pesquisa de fatores que influenciam na produção de hemolisinas em amostras de Campylobacter jejuni de origem avícola

    Directory of Open Access Journals (Sweden)

    Michele M. Trindade


    Full Text Available Resumo: Membros termofílicos do gênero Campylobacter são reconhecidos como importantes enteropatógenos para o ser humano e animais. A grande diversidade ecológica destes micro-organismos em diferentes habitats tais como água, animais e alimentos predispõem ao aparecimento de novos fatores de virulência. Este trabalho teve por objetivo detectar os genes codificantes da Toxina Distensiva Citoletal (CDT por meio da técnica de PCR, pesquisar a atividade de hemolisinas e a influência de soluções quelantes e de íons nesta atividade. Foram utilizadas 45 amostras de Campylobacter jejuni de origem avícola para pesquisa de atividade hemolítica, cultivadas em Caldo Triptona de Soja (TSB. Após o crescimento bacteriano, as amostras foram semeadas em Ágar tríptico de soja (TSA contendo 5% de sangue de ovino. Para verificar a influência de agentes quelantes e solução de íons na atividade hemolítica, as amostras de C. jejuni foram cultivadas em TSB contendo separadamente os quelantes EDTA, ácido acético, soluções de íons CaCl2, MgCl2 e FeCl3, em atmosfera de microaerofilia. Quanto à atividade de hemolisina de C. jejuni em placas de TSA - sangue ovino foi possível observar que houve hemólise em 40% das amostras analisadas apenas com caldo TSB. Somente o ácido acético apresentou ação quelante sobre a atividade de hemolisinas em amostras de C. jejuni semeadas em placas de TSA - sangue ovino. Para detecção dos genes cdtA, cdtB e cdtC através da técnica da Reação em Cadeia da Polimerase (PCR foram utilizadas 119 amostras de C. jejuni de origem avícola. Foi possível observar que 37,8% possuíam o perfil de genes cdtABC. Os resultados demonstraram em amostras avícolas a presença de cepas de C. jejuni com potencial virulento, devido à presença dos genes da toxina CDT e potencial hemolítico, que apresentou ação reduzida in vitro com ácido acético.