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Sample records for cdta

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

    Directory of Open Access Journals (Sweden)

    Neeta Pandey

    2013-01-01

    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. Distribution of the cytolethal distending toxin A gene (cdtA) among species of Shigella and Vibrio, and cloning and sequencing of the cdt gene from Shigella dysenteriae.

    Science.gov (United States)

    Okuda, J; Kurazono, H; Takeda, Y

    1995-03-01

    We investigated the distribution of the cytolethal distending toxin A gene (cdtA) among S. dysenteriae, Vibrio cholerae 01 and Vibrio parahaemolyticus by polymerase chain reaction (PCR) using primers constructed from the nucleotide sequences of Escherichia coli cdtA gene reported independently by Scott and Kaper (Infect Immun 1994; 62: 244-51) and by Pickett et al. (Infect Immun 1994; 62: 1046-51). The cdtA gene reported by Scott and Kaper was found to occur among eight of the 35 strains of S. dysenteriae but was not found among V. cholerae O1 and V. parahaemolyticus. The cdtA gene reported by Pickett et al. was not found among S. dysenteriae, V. cholerae O1 and V. parahaemolyticus. To further investigate the distribution of the cdtA gene among a large number of Shigella spp. (S. dysenteriae, S. flexneri, S. boydii and S. sonnei), and among Vibrio spp. (Vibrio cholerae O1, V. cholerae O139 and V. parahaemolyticus) by colony hybridization test, we constructed a cdtA gene specific DNA probe by amplifying the cdtA gene by PCR with primers designed from the nucleotide sequence of the cdtA gene reported by Scott and Kaper. The cdtA gene reported by Scott and Kaper was found to occur among eight of the 35 strains of S. dysenteriae and one of the 100 strains of S. sonnei, but was not found among other species of Shigella or among the Vibrio species examined. From one cdtA gene-positive S. dysenteriae strain that showed cytolethal distending toxin (CDT) activity on Chinese hamster ovary cells, we cloned and sequenced the entire cdt gene comprising cdtA, cdtB and cdtC genes.(ABSTRACT TRUNCATED AT 250 WORDS)

  3. 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.

  4. Single CDTA-Based Current Mode All-Pass Filter and Its Applications

    Directory of Open Access Journals (Sweden)

    Neeta Pandey

    2011-01-01

    Full Text Available This paper presents a single current difference transconductance amplifier (CDTA based all-pass current mode filter. The proposed configuration makes use of a grounded capacitor which makes it suitable for IC implementation. Its input impedance is low and output impedance is high, hence suitable for cascading. The circuit does not use any matching constraint. The nonideality analysis of the circuit is also given. Two applications, namely, a quadrature oscillator and a high Q band pass filter are developed with the proposed circuit. The functionality of the circuit is verified with SPICE simulation using 0.35  m TSMC CMOS technology parameters.

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

    NARCIS (Netherlands)

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

    2001-01-01

    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 p

  6. Demineralization effect of EDTA, EGTA, CDTA and citric acid on root dentin: a comparative study Efeito do EDTA, EGTA, CDTA e ácido cítrico na desmineralização da dentina radicular: estudo comparativo

    Directory of Open Access Journals (Sweden)

    Simone Maria Galvão Sousa

    2005-09-01

    Full Text Available The purpose of this study was to biochemically compare the decalcifying effects of 1% EDTA (pH 7.4, 1% EGTA (pH 7.4, 1% CDTA (pH 7.4, 1% citric acid solutions (pH 1.0 and 7.4 and saline solution (control on root dentin. Forty-eight single-rooted teeth were used in this study. The canals were instrumented by the step-back technique and the roots were randomly divided into six equal experimental groups (n = 8 according to the irrigating agent tested. A total of 30 µL of each solution was pipetted into the root canal and allowed to set undisturbed for 5 minutes. After this time, 15 µL of the solutions were removed from each canal using a Hamilton syringe and placed in a container with 5 mL of deionised water. The µg/mL concentration of calcium ion (Ca2+ extracted from the root canal samples was determined using inductively coupled plasma-atomic emission spectrometry (ICP-AES. Data were analysed by means of the Kruskal-Wallis and Mood's median tests. Citric acid solution at pH 1.0 removed more calcium than at pH 7.4 and than the other chelating solutions tested (p 0.05. These results indicate that citric acid at pH 1.0 is a good alternative as an irrigating solution to remove the smear layer and facilitate the biomechanical procedures.Este trabalho teve como objetivo comparar o efeito desmineralizante do EDTA (pH 7,4, EGTA (pH 7,4, CDTA (pH 7,4, ácido cítrico (pH 1,0 e 7,4 e da solução salina (controle sobre a dentina radicular. Todas as soluções teste foram preparadas na concentração de 1%. Quarenta e oito dentes unirradiculares recém-extraídos foram utilizados neste experimento. Após a instrumentação dos canais radiculares pela técnica "step-back", as raízes foram aleatoriamente divididas em 6 grupos experimentais (n = 8 de acordo com a solução teste utilizada na irrigação final. Em cada grupo, 30 µL da solução teste foram pipetados no interior de cada canal radicular e mantidos estáveis por 5 minutos. Decorrido esse per

  7. Complejos de V(III en solución acuosa con el ácido aminopolicarboxílico CDTA

    Directory of Open Access Journals (Sweden)

    Lino Hernández

    2011-12-01

    Full Text Available Electromotive forces measurements emf (H were employed to study the formation of the Vanadium(III complexes with the aminopolycarboxylic acid trans-1,2-diaminocyclohexane-N,N,N',N'-tetraethanoic (CDTA, H4L in aqueous solution at 25 °C in 3.0mol.L-1 KCl as the ionic media. The analysis of the potentiometric data by means of the least-squares computational program LETAGROP, indicates the formation of significant quantities of the complexes [VHL], [VL]- and [V(OHL]2- together with their respective stability constants of each complex. In addition to the LETAGROP analysis UV-Vis spectrophotometric measurements were done in order to make a qualitative characterization of the complexes formed in aqueous solution

  8. CDT-a entropic theory of quantum gravity

    DEFF Research Database (Denmark)

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

    2010-01-01

    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)...

  9. Carbohydrate-deficient transferrin (CDT)--a biomarker for long-term alcohol consumption.

    Science.gov (United States)

    Golka, Klaus; Wiese, Andreas

    2004-01-01

    Carbohydrate-deficient transferrin (CDT) is a biomarker for chronic alcohol intake of more than 60 g ethanol/d. It has been reported to be superior to conventional markers like gamma-glutamyltransferase (GGT) and mean corpuscular volume MCV). This review covers theoretical and analytical aspects, with data from controlled drinking experiments and from different population subgroups such as subjects with different liver diseases or different drinking patterns. CDT determinations are particularly indicated in (1) cases of chronic alcohol consumption and relapses after withdrawal, (2) license reapplication after driving under alcohol influence, (3) differentiating patients with enzyme-inducing medication from those with alcohol abuse, 4) congenital disorders of glycosylation such as carbohydrate-deficient glycoprotein syndrome Ia (CDGS Ia), and (5) patients treated for galactosemia. The main advantage of CDT is its high specificity, as evidenced in combination with increased alcohol consumption. CDT values are not markedly influenced by medication except in immunosuppressed patients, who may show low CDT values. In general, CDT values appear less elevated after alcohol intake in women. The main disadvantage is the relatively low sensitivity. Hence, this parameter is not suitable for screening for subjects with alcohol abuse in the general population. As CDT, GGT, and MCV are connected with chronic alcohol consumption by different pathophysiological mechanisms, a combination of these parameters will further improve the diagnostic value.

  10. Clostridium difficile binary toxin CDT

    OpenAIRE

    Gerding, Dale N.; Johnson, Stuart; Rupnik, Maja; Aktories, Klaus

    2013-01-01

    Binary toxin (CDT) is frequently observed in Clostridium difficile strains associated with increased severity of C. difficile infection (CDI). CDT belongs to the family of binary ADP-ribosylating toxins consisting of two separate toxin components: CDTa, the enzymatic ADP-ribosyltransferase which modifies actin, and CDTb which binds to host cells and translocates CDTa into the cytosol. CDTb is activated by serine proteases and binds to lipolysis stimulated lipoprotein receptor. ADP-ribosylatio...

  11. 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.;

    2003-01-01

    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...

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

    Energy Technology Data Exchange (ETDEWEB)

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

    1989-07-01

    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.

  13. 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.

    2008-10-07

    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

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

    International Nuclear Information System (INIS)

    Formation constants (log Ksub(MAL)sup(MA)) of mixed ligands complexes (MAL), where M = UO22+ or Th4+, 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 25oC and at I =0.2 (mol dm-3, KNO3). The solution stabilities exhibit the sequence (i) Th4+>UO22+, (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(ML2)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 (An-). (author). 17 refs., 1 tab

  15. Recombinant Actinobacillus actinomycetemcomitans Cytolethal Distending Toxin Proteins Are Required To Interact To Inhibit Human Cell Cycle Progression and To Stimulate Human Leukocyte Cytokine Synthesis

    OpenAIRE

    Akifusa, Sumio; Poole, Stephen; Lewthwaite, Jo; Henderson, Brian; Nair, Sean P

    2001-01-01

    It has recently been discovered that Actinobacillus actinomycetemcomitans, an oral bacterium causing periodontitis, produces cytolethal distending toxin (CDT), a cell cycle-modulating toxin that has three protein subunits: CdtA, CdtB, and CdtC. In this study, we have cloned and expressed each toxin gene from A. actinomycetemcomitans in Escherichia coli and purified the recombinant Cdt proteins to homogeneity. Individual Cdt proteins failed to induce cell cycle arrest of the human epithelial c...

  16. Modification of Pectin and Hemicellulose Polysaccharides in Relation to Aril Breakdown of Harvested Longan Fruit

    OpenAIRE

    Duoduo Wang; Haiyan Zhang; Fuwang Wu; Taotao Li; Yuxiang Liang; Xuewu Duan

    2013-01-01

    To investigate the modification of cell wall polysaccharides in relation to aril breakdown in harvested longan fruit, three pectin fractions (WSP, water soluble pectin; CSP, CDTA-soluble pectin; ASP, alkali soluble pectin) and one hemicellulose fraction (4 M KOH-SHC, 4 M KOH-soluble hemicellulose) were extracted, and their contents, monosaccharide compositions and molecular weights were evaluated. As aril breakdown intensified, CSP content increased while ASP and 4 M KOH-SHC contents decreas...

  17. Haemophilus parasuis Encodes Two Functional Cytolethal Distending Toxins: CdtC Contains an Atypical Cholesterol Recognition/Interaction Region

    OpenAIRE

    Mingguang Zhou; Qiang Zhang; Jianping Zhao; Meilin Jin

    2012-01-01

    Haemophilus parasuis is the causative agent of Glässer's disease of pigs, a disease associated with fibrinous polyserositis, polyarthritis and meningitis. We report here H. parasuis encodes two copies of cytolethal distending toxins (Cdts), which these two Cdts showed the uniform toxin activity in vitro. We demonstrate that three Cdt peptides can form an active tripartite holotoxin that exhibits maximum cellular toxicity, and CdtA and CdtB form a more active toxin than CdtB and CdtC. Moreover...

  18. Effect of silencing the two major tomato fruit pectin methylesterase isoforms on cell wall pectin metabolism.

    Science.gov (United States)

    Wen, B; Ström, A; Tasker, A; West, G; Tucker, G A

    2013-11-01

    Post-harvest storage is largely limited by fruit softening, a result of cell wall degradation. Pectin methylesterase (PE) (EC 3.1.1.11) is a major hydrolase responsible for pectin de-esterification in the cell wall, a response to fruit ripening. Two major PE isoforms, PE1 and PE2, have been isolated from tomato (Solanum lycopersicon) pericarp tissue and both have previously been down-regulated using antisense suppression. In this paper, PE1 and PE2 double antisense tomato plants were successfully generated through crossing the two single antisense lines. In the double antisense fruit, approximately 10% of normal PE activity remained and ripening associated pectin de-esterification was almost completely blocked. However, double antisense fruit softened normally during ripening. In tomato fruit, the PE1 isoform was found to contribute little to total PE activity and have little effect on the degree of esterification of pectin. In contrast, the other dominant fruit isoform, PE2, has a major impact on de-esterification of total pectin. PE2 appears to act on non-CDTA-soluble pectin during ripening and on CDTA-soluble pectin before the start of ripening in a potentially block-wise fashion.

  19. Outer membrane vesicle-mediated release of cytolethal distending toxin (CDT from Campylobacter jejuni

    Directory of Open Access Journals (Sweden)

    Uhlin Bernt

    2009-10-01

    Full Text Available Abstract Background Background: Cytolethal distending toxin (CDT is one of the well-characterized virulence factors of Campylobacter jejuni, but it is unknown how CDT becomes surface-exposed or is released from the bacterium to the surrounding environment. Results Our data suggest that CDT is secreted to the bacterial culture supernatant via outer membrane vesicles (OMVs released from the bacteria. All three subunits (the CdtA, CdtB, and CdtC proteins were detected by immunogold labeling and electron microscopy of OMVs. Subcellular fractionation of the bacteria indicated that, apart from the majority of CDT detected in the cytoplasmic compartment, appreciable amounts (20-50% of the cellular pool of CDT proteins were present in the periplasmic compartment. In the bacterial culture supernatant, we found that a majority of the extracellular CDT was tightly associated with the OMVs. Isolated OMVs could exert the cell distending effects typical of CDT on a human intestinal cell line, indicating that CDT is present there in a biologically active form. Conclusion Our results strongly suggest that the release of outer membrane vesicles is functioning as a route of C. jejuni to deliver all the subunits of CDT toxin (CdtA, CdtB, and CdtC to the surrounding environment, including infected host tissue.

  20. Capillary electrophoretic-ultraviolet method for the separation and estimation of zineb, maneb, and ferbam in food samples.

    Science.gov (United States)

    Aulakh, Jatinder Singh; Fekete, Agnes; Malik, Ashok Kumar; Mahajan, Rakesh Kumar; Schmitt-Kopplin, Philippe

    2007-01-01

    A simple and sensitive capillary electrophoretic method with ultraviolet detection has been developed for the separation and determination of ferbam [iron(III)-dimethyldithiocarbamate], maneb [manganese(II)-ethylenebisdithiocarbamate] and zineb [zinc(II)-ethylenebisdithiocarbamate], in borate buffer, after their acidic decomposition and complexation with CDTA (trans-1,2-diaminocyclohexane-N,N,N',N'-tetraacetic acid monohydrate), as CDTA-metal complexes of Fe+3, Mn +2, and Zn+2. The determination is dependent on the pH and the nature of the buffer solutions. In this method, the detection limit (signal-to-noise ratio = 3) is 0.0013, 0.0022, and 0.0023 mM for ferbam, maneb, and zineb, respectively. The relative standard deviation for the analysis of 1 mM of each was found to be 1.5 +/- 0.2%. The method was successfully applied for the analysis of red beans and grain samples spiked with ferbam, maneb, and zineb. The applicability of capillary electrophoresis as a useful tool for the simultaneous determination and analysis of ferbam, maneb, and zineb is demonstrated. PMID:17580637

  1. Impact of Liquidity on Islamic Banks' Profitability: Evidence from Bangladesh

    Directory of Open Access Journals (Sweden)

    Limon Moinur Rasul

    2013-04-01

    Full Text Available This study examines the impact of liquidity on Islamic banks’ profitability during an 11 years period of 2001 to 2011. To explore and interpret the results the study has taken samples from five Islamic banks that have been in operation in Bangladesh on or before 2001 to till date. In order to construct the liquidity model it used four liquidity variables namely cash & due from banks to total assets (CDTA, cash & due from banks to total deposits (CDDEP, investment to total assets (INVSTA and investment to total deposits (INVSDEP. According to adjusted R squares profitability variables return on assets (ROA, return on equity (ROE and return on deposits (ROD are respectively 17.1%, 4.5% and 24.6% dependent on independent variables. The statistical results suggest that CDTA is found insignificant with all profitability variables, whereas CDDEP is individually significant with all profitability variables except ROE. On the other hand INVSTA and INVSDEP are recognized significant with all three profitability variables. However, when ROE stands for an insignificant relationship with the overall liquidity model, ROA and ROD are identified significantly correlated with the similar model at 1% significant level. Unsurprisingly the findings do strengthen the specification that the impact of liquidity reflects adequate imposition on profitability that the Islamic banks in Bangladesh must abide by.

  2. Development of a new radiolabel (lead-203) and new chelating agents for labeling monoclonal anntibodies for imaging

    Energy Technology Data Exchange (ETDEWEB)

    Srivastava, S.C.; Mease, R.C.; Meinken, G.E.; Mausner, L.F.; Steplewski, Z.

    1988-01-01

    High liver uptake and slow body clearance presently limit the usefulness of /sup 111/In labeled antibodies for tumor imaging. We have investigated /sup 203/Pb as an alternate and better antibody label. The DTPA and cyclohexyl EDTA (CDTA) conjugates of an anticolon carcinoma antibody, 17-1A were labeled (bicyclic anhydride method) with /sup 203/Pb and /sup 111/In with 60 and 90% labeling yields, respectively. The biodistribution of /sup 203/Pb-17-1A conjugates was compared with the corresponding /sup 111/In-labeled preparations and with /sup 203/Pb-DTPA, /sup 203/Pb-nitrate and nonrelevant antibody controls in normal and human tumor (SW948) xenografted nude mice at 24, and 96 hr. Lead-203-labeled CDTA and DTPA antibody conjugates gave similar in vivo distributions. Even though the lead bound to these chelate-antibody conjugates was more labile in serum and in vivo, compared to indium, it cleared much faster from the liver and the whole body. A new series of chelating agents based on the incorporation of a trans-1,2- diaminocyclohexane moiety into the carbon backbone of polyaminocarboxylates is being synthesized. These are expected to provide stronger complexing ability for lead and produce greater in vivo stability. These ligands are also expected to be superior to EDTA and DTPA for labeling antibodies with other radiometals, including indium. 32 refs., 3 tabs.

  3. Characterization of putative cholesterol recognition/interaction amino acid consensus-like motif of Campylobacter jejuni cytolethal distending toxin C.

    Directory of Open Access Journals (Sweden)

    Chih-Ho Lai

    Full Text Available Cytolethal distending toxin (CDT produced by Campylobacter jejuni comprises a heterotrimeric complex formed by CdtA, CdtB, and CdtC. Among these toxin subunits, CdtA and CdtC function as essential proteins that mediate toxin binding to cytoplasmic membranes followed by delivery of CdtB into the nucleus. The binding of CdtA/CdtC to the cell surface is mediated by cholesterol, a major component in lipid rafts. Although the putative cholesterol recognition/interaction amino acid consensus (CRAC domain of CDT has been reported from several bacterial pathogens, the protein regions contributing to CDT binding to cholesterol in C. jejuni remain unclear. Here, we selected a potential CRAC-like region present in the CdtC from C. jejuni for analysis. Molecular modeling showed that the predicted functional domain had the shape of a hydrophobic groove, facilitating cholesterol localization to this domain. Mutation of a tyrosine residue in the CRAC-like region decreased direct binding of CdtC to cholesterol rather than toxin intermolecular interactions and led to impaired CDT intoxication. These results provide a molecular link between C. jejuni CdtC and membrane-lipid rafts through the CRAC-like region, which contributes to toxin recognition and interaction with cholesterol.

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

    International Nuclear Information System (INIS)

    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 SFLn/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

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

    Energy Technology Data Exchange (ETDEWEB)

    Sypula, Michal

    2013-07-01

    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

  6. INL DPAH STAAR 2015 Annual Report

    Energy Technology Data Exchange (ETDEWEB)

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

    2015-09-15

    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.

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

    International Nuclear Information System (INIS)

    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

  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

    2010-10-01

    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. Determination of synthetic ferric chelates used as fertilizers by liquid chromatography-electrospray/mass spectrometry in agricultural matrices.

    Science.gov (United States)

    Alvarez-Fernández, Ana; Orera, Irene; Abadía, Javier; Abadía, Anunciación

    2007-01-01

    A high-performance liquid chromatography-electrospray ionization/mass spectrometry (time of flight) method has been developed for the simultaneous determination of synthetic Fe(III)-chelates used as fertilizers. Analytes included the seven major Fe(III)-chelates used in agriculture, Fe(III)-EDTA, Fe(III)-DTPA, Fe(III)-HEDTA, Fe(III)-CDTA, Fe(III)-o,oEDDHA, Fe(III)-o,pEDDHA, and Fe(III)-EDDHMA, and the method was validated using isotope labeled (57)Fe(III)-chelates as internal standards. Calibration curves had R values in the range 0.9962-0.9997. Limits of detection and quantification were in the ranges 3-164 and 14-945 pmol, respectively. Analyte concentrations could be determined between the limits of quantification and 25 muM (racemic and meso Fe(III)-o,oEDDHA and Fe(III)-EDDHMA) or 50 muM (Fe(III)-EDTA, Fe(III)-HEDTA, Fe(III)-DTPA, Fe(III)-CDTA and Fe(III)-o,pEDDHA). The average intraday repeatability values were approximately 0.5 and 5% for retention time and peak area, respectively, whereas the interday repeatability values were approximately 0.7 and 8% for retention time and peak area, respectively. The method was validated using four different agricultural matrices, including nutrient solution, irrigation water, soil solution, and plant xylem exudates, spiked with Fe(III)-chelate standards and their stable isotope-labeled corresponding chelates. Analyte recoveries found were in the ranges 92-101% (nutrient solution), 89-102% (irrigation water), 82-100% (soil solution), and 70-111% (plant xylem exudates). Recoveries depended on the analyte, with Fe(III)-EDTA and Fe(III)-DTPA showing the lowest recoveries (average values of 87 and 88%, respectively, for all agricultural matrices used), whereas for other analytes recoveries were between 91 and 101%. The method was also used to determine the real concentrations of Fe(III)-chelates in commercial fertilizers. Furthermore, the method is also capable of resolving two more synthetic Fe(III)-chelates, Fe

  10. First report of Clostridium difficile NAP1/027 in a Mexican hospital.

    Directory of Open Access Journals (Sweden)

    Adrián Camacho-Ortiz

    Full Text Available Clostridium difficile NAP1/ribotype 027 is associated with severe disease and high mortality rates. Our aim was to determine the prevalence of NAP1/ribotype 027 among C. difficile isolates in a tertiary care hospital, and review the main clinical data.We included 106 stool samples from 106 patients. Samples were tested for A&B toxins and were cultured on CCFA agar. The genes tcdA, tcdB, tcdC, cdtA, and cdtB were amplified using PCR in clinical isolates. The tcdA 3'-end deletion analysis, PCR-ribotyping, and pulsed-field gel electrophoresis (PFGE were also performed. Stool samples that were positive for culture were tested by the GeneXpert C. difficile assay. Clinical data were collected.Thirty-six patients tested positive for A&B toxins; and 22 patients had positive culture for C. difficile, 14 of which tested positive for the A&B toxins and all 22 patients tested positive by the GeneXpert C. difficile assay. Risk factors included an average hospital stay of 16.1 days prior to toxin detection, average antibiotic use for 16.2 days, and a median of 3 antibiotics used. The 30-day crude mortality rate was 8.4%. Six of the 22 patients died, and 3 of those deaths were directly attributed to C. difficile infection. The majority of isolates, 90.9% (20/22, carried genes tcdB, tcdA, cdtA, and cdtB; and these strains carried the corresponding downregulator gene tcdC, with an 18-bp deletion. PFGE was performed on 17 isolates, and one main pattern was observed. Analysis of the ribotyping data showed similar results.The above findings represent the clonal spread of C. difficile in our institution, which mainly includes the NAP1/027 strain. This is the first report of C. difficile ribotype NAP1/027 in Mexico.

  11. 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.

  12. 空肠弯曲菌细胞致死性肿胀毒素研究进展%Research progress of Campylobacter jejuni cytolethal distending toxin

    Institute of Scientific and Technical Information of China (English)

    陆磊; 焦新安; 黄金林

    2012-01-01

    Camp ylobacter jejuni is a food borne bacterial pathogen. It can produce cytolethal distending toxin, which has cell toxicity and can block the cell cycle at the G2/M boundary. Affected cells enlarge until they finally undergo pro grammed cell death. The toxin is composed of three subunits designated CdtA, CdtB and CdtC. This text will introduce its physicochemical properties, biological functions, mechanism of action and so on.%目的 空肠弯曲菌是一种食源性病原菌,它产生的细胞致死性肿胀毒素具有细胞毒性,能将细胞周期阻断在G2/M期,并最终导致细胞程序性死亡;它由3种多肽CdtA、CdtB和CdtC构成.本文将从空肠弯曲菌细胞致死性肿胀毒素的理化性质、生物学功能和作用机制等方面的研究进展作一介绍.

  13. Dynamic Duo-The Salmonella Cytolethal Distending Toxin Combines ADP-Ribosyltransferase and Nuclease Activities in a Novel Form of the Cytolethal Distending Toxin.

    Science.gov (United States)

    Miller, Rachel; Wiedmann, Martin

    2016-01-01

    The cytolethal distending toxin (CDT) is a well characterized bacterial genotoxin encoded by several Gram-negative bacteria, including Salmonella enterica (S. enterica). The CDT produced by Salmonella (S-CDT) differs from the CDT produced by other bacteria, as it utilizes subunits with homology to the pertussis and subtilase toxins, in place of the traditional CdtA and CdtC subunits. Previously, S-CDT was thought to be a unique virulence factor of S. enterica subspecies enterica serotype Typhi, lending to its classification as the "typhoid toxin." Recently, this important virulence factor has been identified and characterized in multiple nontyphoidal Salmonella (NTS) serotypes as well. The significance of S-CDT in salmonellosis with regards to the: (i) distribution of S-CDT encoding genes among NTS serotypes, (ii) contributions to pathogenicity, (iii) regulation of S-CDT expression, and (iv) the public health implication of S-CDT as it relates to disease severity, are reviewed here. PMID:27120620

  14. The Biology of the Cytolethal Distending Toxins

    Directory of Open Access Journals (Sweden)

    Teresa Frisan

    2011-03-01

    Full Text Available 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.

  15. The cytolethal distending toxin-IV cdt coding region in an avian pathogenic Escherichia coli (APEC) strain shows instability and irregular excision pattern.

    Science.gov (United States)

    Tóth, István; Schneider, György

    2015-12-01

    Cytolethal distending toxins (CDT) represent an emerging toxin family, widely distributed among pathogenic bacteria. The cdtABC genes in E. coli are either part of the genome of prophages, plasmid or pathogenicity island. In order to investigate the stability and the transfer potential of cdt-IV genes cdtB gene was replaced by chloramphenicol (Cm) resistance encoding cat gene in the avian pathogenic E. coli (APEC) strain E250. After consecutive passages in non-selective medium at 37 °C 7.6% (219/2900) of the investigated colonies of E250::cat strain became Cm-sensitive (Cm(S)). To reveal deletion mechanism 177 Cm(S) colonies were investigated for presence of cdtA, cdtC and cdtC associated gene by PCR. One hundred and sixteen colonies of the Cm(S) colonies (65.5%) showed partial or complete deletion in the cdt-IV region. Progressive loss of the upstream genes of the cdt cluster in E250 compared to other CDT-IV producing APEC strains and the fact that all the potential deletion patterns were identified, suggests the presence of an unstable hitherto unknown genomic region. The failure of in vitro transfer of cdt genes into a porcine EPEC E. coli strain suggests that the deletion of cdt-IV flanking genes alone do not promote the spread of cdt-IV.

  16. 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;

    2012-01-01

    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...... 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 and...

  17. Modification of Pectin and Hemicellulose Polysaccharides in Relation to Aril Breakdown of Harvested Longan Fruit

    Directory of Open Access Journals (Sweden)

    Duoduo Wang

    2013-11-01

    Full Text Available To investigate the modification of cell wall polysaccharides in relation to aril breakdown in harvested longan fruit, three pectin fractions (WSP, water soluble pectin; CSP, CDTA-soluble pectin; ASP, alkali soluble pectin and one hemicellulose fraction (4 M KOH-SHC, 4 M KOH-soluble hemicellulose were extracted, and their contents, monosaccharide compositions and molecular weights were evaluated. As aril breakdown intensified, CSP content increased while ASP and 4 M KOH-SHC contents decreased, suggesting the solubilization and conversion of cell wall components. Furthermore, the molar percentage of arabinose (Ara, as the main component of the side-chains, decreased largely in CSP and ASP while that of rhamnose (Rha, as branch point for the attachment of neutral sugar side chains, increased during aril breakdown. Analysis of (Ara + Gal/Rha ratio showed that the depolymerization of CSP and ASP happened predominantly in side-chains formed of Ara residues. For 4 M KOH-SHC, more backbones were depolymerized during aril breakdown. Moreover, it was found that the molecular weights of CSP, ASP and 4 M KOH-SHC polysaccharides tended to decrease as aril breakdown intensified. These results suggest that both enhanced depolymerization and structural modifications of polysaccharides in the CSP, ASP and 4 M KOH-SHC fractions might be responsible for aril breakdown of harvested longan fruit.

  18. Catheter-directed thrombolysis in inflammatory bowel diseases: Report of a case

    Institute of Scientific and Technical Information of China (English)

    Péter Ilonczai; Judit Tóth; László Tóth; István Altorjay; Zoltán Boda; Károly Palatka

    2012-01-01

    In patients with inflammatory bowel diseases (IBD)the prevalence of thrombosis is 6.2%,the average incidence of thromboembolism (TE) is 3.6 times higher compared to normal population.The TE is a common extraintestinal complication of IBD,squarely associated with the IBD activity.The application of anticoagulant and thrombolytic therapy in severe IBD is an unresolved issue.Herein we report the first case in literature of an active IBD patient with an upper limb acute arterial occlusion and successful catheter-directed thrombolysis (CDT).A 46-year-old male patient is reported who had Crohn's disease for 10 years.His right hand suddenly became cold and painful.Angiography proved acute occlusion of the brachial and radial artery.Vascular surgery intervention was not applicable.Endoscopy showed extended,severe inflammation of the colon.Despite the severe endoscopic findings,frequent bloody stools and moderate anaemia,CDT with recombinant tissue plasminogen activator was performed.The control angiography proved improvement,the radial artery pulse appeared.No bleeding complication was observed.This case supports that CDT-after careful estimation of the bleeding risk-can be effective and safe in patients with severe or life-threatening TE and active IBD.

  19. Cytotoxicity of the Cytolethal Distending Toxin of Haemophilus parasuis%副猪嗜血杆菌细胞致死膨胀毒素的细胞毒性研究

    Institute of Scientific and Technical Information of China (English)

    陈西; 王湘如; 徐晓娟; 郭凤娟; 陈焕春; 蔡旭旺

    2011-01-01

    The study was aimed at prokaryotic expressing cytolethal distending toxin of Haemophilus parasuis t and acted on Pig iliac endothelial cells (PIEC) to investigate toxin effects. According to the complete genome sequence of Haemophilus parasuis SH0165 (serovar 5), which finished by our laboratory, we designed a set of specific primer, the cdtA, cdtB and cdtC gene was amplified by PCR, respectively. The 681, 834 and 531 bp amplified DNA fragment was cloned into pET-28a and the expression were induced in E. Coli BL21 (DE3) by IPTG for 3 hours. Results of Sodium docecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and Western blot assay showed that the cdtA, cdtB and cdtC gene was expressed in the form of inclusion body, and the recombinant protein was about 36, 34 and 28 kDa. Recombinant toxin acted on PIEC for 3 hours in vitro, and observed the morphological change after another culture of 72 hours. These results showed that CdtABC holotoxin induced PIEC distension, vacuolization and apoptosis, while the other test groups changed unobviously. The study indicated that CDT may play an important role in the infection and pathopoiesis of bacterial.%本研究旨在原核表达副猪嗜血杆菌细胞致死膨胀毒素(Cytolethal distending toxin,CDT),并作用于猪髋动脉内皮细胞(Pig iliac endothelial cells,PIEC),以研究其细胞毒性.根据本实验室完成的副猪嗜血杆菌SH 0165株(血清5型)全基因组序列,针对cdtA、cdtB和cdtC基因序列设计引物,扩增的基因片段大小分别约为681、834和531 bp.将靶基因克隆到原核表达载体pET28a中,再转化到E.coliBL21( DE3),IPTG诱导表达3h,SDSPAGE和Western blot检测证实表达产物以包涵体形式存在,大小分别约36、34和28 ku.通过体外重构毒素与PIEC细胞作用3h,继续培养72 h观察细胞形态学变化.结果表明,CdtABC全毒素致PIEC细胞膨胀、空泡形成、细胞凋亡等,而其他试验组变化不显著.结果提示,CDT毒素可能在

  20. Clostridium difficile infection diagnosis in a paediatric population: comparison of methodologies.

    Science.gov (United States)

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

    2014-09-01

    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. PMID:24781004

  1. Clostridium difficile infection diagnosis in a paediatric population: comparison of methodologies.

    Science.gov (United States)

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

    2014-09-01

    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.

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

    Directory of Open Access Journals (Sweden)

    Atkinson Ross G

    2012-08-01

    Full Text Available Abstract Background 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. Results 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. Conclusions 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.

  3. 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: ssimona@icf.ro [“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)

    2014-02-15

    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.

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

    International Nuclear Information System (INIS)

    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)

  5. Chromium speciation in particulate matter samples (Cr(VI)/Cr(III) stability in solutions of leaching agents)

    International Nuclear Information System (INIS)

    Complete text of publication follows. In recent years extensive research was conducted to estimate the bioavailability and toxicity of metals in environmental samples. Substantial health risk could be associated with high particulate matter concentrations in ambient air and with a consumption of contaminated food-stuffs, both accompanied by an occurrence of toxic elements. One of the main causes of exceedance of ambient air quality limit values is traffic, despite emissions reductions. Among other factors, the mobility of an element is usually related to its chemical properties and the toxicity mainly to its oxidation state. Thus, chromium in the hexavalent form, Cr(VI), has long been recognized as a carcinogen and mutagen at low sub-ppm levels. Therefore, in this work, the presence and stability of Cr(VI)/Cr(III) species have been determined in particular matter of urban dust samples (the modified BCR three step sequential extraction procedure). For testing of stability and presence of Cr species a coupled technique connecting on-line HPLC with element-sensitive detector ICP-OES has been used (chromium was detected on line 205,560 nm). The anion exchange column Hamilton PRP-X100 (250 x 4,6 mm, 5 μm, PEEK, (Hamilton, USA)) was used for separation of Cr species. Optimal conditions for the separation were following: mobile phase 50 mmol x l-1 CH3COOH and 10 mmol xl-1 NaClO4 (pH 7.0; flow rate 1.5 ml x min-1), injected 200 μl of the sample, addition of 30 μg x ml-1 CDTA to the sample for a transfer of Cr(III) to anion complex. This combined technique allowed to determine 10 μg of Cr(III) and 13 μg of Cr(VI) in the sample (absolute LOD). It was found out that all three extraction agents used for a fractionation of elements negatively influenced the stability of Cr(VI) species in the solution immediately after their contact with the sample. The quantitatively smallest influence was found for the acetic acid. Probably this is a reason why only species of Cr

  6. 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

    2015-08-01

    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.

  7. Campylobacter spp.isolation, its toxin genes detection and molecular subtyping in diarrhea patients in Shanghai in 2014%上海市2014年腹泻患者弯曲菌分离、毒力基因检测及分子分型结果

    Institute of Scientific and Technical Information of China (English)

    屠丽红; 陈洪友; 陈敏

    2015-01-01

    positive for flaA and cadF, and 93.0%(40/43) of the isolates positive for cdtA and cdtB.And 88.4%(38/43) of the isolates were posi-tive for cdtC.Only 7.0%(3/43) of the isolates were positive for virB11.Using PFGE, 43 Campylobacter jejuni and Campylobacter coli strains were subtyped into 6 clusters. [ Conclusion] The genes of flaA and cadF are ubiquitous on Campylobacter spp.isolates.The distribution of cdt gene cluster in Campylobacter spp.is high, while that of virB11 is low.The PFGE types of Campylobacter spp.isolated in Shanghai are of diversified and complicated features, which causes mainly sporatic diarrhea.