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Effects of Pb-EDTA and EDTA on oxidative stress reactions and mineral uptake in Phaseolus vulgaris.

Sequestration of Pb by synthetic chelates has been reported to increase bioavailability, uptake, and translocation of this metal in plants. In this work the potential phytotoxic effects of Pb-EDTA were investigated in Phaseolus vulgaris L. cv. Limburgse vroege plants grown on hydroponics. Addition of 50 microM Pb-EDTA to the nutrient solution caused a significant induction of syringaldazine peroxidase (SPOD; EC 1.11.1.7) in roots and primary leaves and guaiacol peroxidase (GPOD; EC 1.11.1.7) in leaves. Addition of 100 microM Pb-EDTA further exacerbated ascorbate peroxidase (APOD; EC 1.11.1.11), GPOD, dehydroascorbate reductase (DHAR; EC 1.8.5.1), glutathione reductase (GR; EC 1.6.4.2) and malic enzyme (ME; EC 1.1.1.40) in roots and APOD and ME in primary leaves. Addition of 200 microM Pb-EDTA also induced DHAR in leaves. This induction of peroxidases (SPOD, GPOD, APOD), enzymes of the ascorbate-glutathione cycle (DHAR, GR in roots) and of an NADP+ reducing enzyme in roots and primary leaves indicates that oxidative stress has been initiated. At 200 microM Pb-EDTA, chlorophyll a and b content in leaves was significantly reduced while visible effects on root morphology and shoot length were observed, while no significant morphological effects were found in the leaves, confirming the sensitive character of the measured enzymes as plant stress indicators. Elevation of the Pb-EDTA concentration in the growth medium significantly reduced the content of Ca, Fe, Mn and Zn taken up by plants, probably due to ion leakage as a result of observed toxicity. Addition of up to 200 microM EDTA increased chelation of divalent cations in nutrient solution resulting in reduced plant uptake of Zn, Cu, Fe and Mn. This did not result in phytotoxicity.

Journal Article↗

Adsorption of EDTA and Metal-EDTA Complexes onto Goethite.

The adsorption characteristics of a variety of divalent and trivalent metal-EDTA complexes onto goethite (alpha-FeOOH) were examined in aqueous solution. Uncomplexed EDTA is adsorbed as a binuclear complex at low pH and as a mononuclear complex at high pH. Adsorption is ligand-like with a high extent of adsorption at low pH. The process can be described by formation of inner-sphere complexes by the surface complexation model with constant capacitance. The EDTA complexes of the divalent metals Ca, Zn, Ni, Cu, Co(II), and Pb, which are quinquedentate in solution (free donor atoms bound to the metal ion), all showed the same ligand-like adsorption behavior. Their adsorption as a function of pH and concentration can be described by the formation of one type of ternary surface complex and can be fitted with the same equilibrium constant. Pd(II)EDTA, which is bidentate or quadridentate in solution, is adsorbed more strongly, but also in a ligand-like manner. The EDTA complexes of the trivalent metals LaEDTA and BiEDTA are adsorbed very strongly over the whole pH range. The sexidentate complex of Co(III) is weakly adsorbed at low pH outer-spherically, i.e., by electrostatic interaction only. Fe(III)EDTA is weakly adsorbed over the whole pH range with a predominant nonspecific surface complex at low pH and a specific complex at high pH.

Journal Article↗

Zinc absorption from zinc oxide, zinc sulfate, zinc oxide + EDTA, or sodium-zinc EDTA does not differ when added as fortificants to maize tortillas.

The fortification of staple foods with zinc may play an important role in achieving adequate zinc intakes in countries at risk of zinc deficiency. However, little is known about the relative bioavailability of different zinc compounds that may be used in food fortification. The objective of this study was to measure and compare fractional zinc absorption from a test meal that included a maize tortilla fortified with zinc oxide, zinc sulfate, zinc oxide + EDTA, or sodium-zinc EDTA. A double isotopic tracer ratio method ((67)Zn as oral tracer and (70)Zn as intravenous tracer) was used to estimate zinc absorption in 42 Mexican women living in a periurban community of Puebla State, Mexico. The test meal consisted of maize tortillas, yellow beans, chili sauce, and milk with instant coffee; it contained 3.3 mg zinc and had a phytate:zinc molar ratio of 17. Fractional zinc absorption did not differ significantly between the test groups (ANOVA; P > 0.05). Percent absorptions were (mean +/- SD) zinc oxide, 10.8 +/- 0.9; zinc sulfate, 10.0 +/- 0.02; zinc oxide + EDTA, 12.7 +/- 1.5; and sodium-zinc EDTA, 11.1 +/- 0.7. We conclude that there was no difference in zinc absorption from ZnO and ZnSO(4) when added as fortificants to maize tortillas and consumed with beans and milk. The addition of EDTA with zinc oxide or the use of prechelated sodium-zinc EDTA as fortificants did not result in higher zinc absorption from the test meal.

Chelating Agents↗

The in vitro effects of EDTA-tris, EDTA-tris-lysozyme, and antimicrobial agents on equine genital isolants of Pseudomonas aeruginosa.

Five isolants of Pseudomonas aeruginosa collected from clinical cases of equine genital infection and one standard strain of P. aeruginosa were exposed to various concentrations of ethylene-diaminetetraacetic acid (EDTA) and tris (hydroxymethyl) aminomethane (tris buffer pH 8) and EDTA-tris lysozyme. Colony forming units of the isolants and minimal inhibitory concentrations for 11 antimicrobial agents were determined with each isolant before and after exposure to the EDTA solutions. Decreased cellular viability was found with all six isolants after exposure to the EDTA-tris solutions. Reversal of antimicrobial resistance was variable and unpredictable. These effects were not enhanced by the addition of lysozyme. The results suggest that EDTA-tris could be a useful adjunct in treating equine genital infections caused by Pseudomonas aeruginosa.

Journal Article↗

Incorporation of an EDTA-metal complex at a rationally selected site within a protein: application to EDTA-iron DNA affinity cleaving with catabolite gene activator protein (CAP) and Cro.

We have developed a simple procedure to incorporate an EDTA-metal complex at a rationally selected site within a full-length protein. Our procedure has two steps: In step 1, we use site-directed mutagenesis to introduce a unique solvent-accessible cysteine residue at the site of interest. In step 2, we derivatize the resulting protein with S-(2-pyridylthio)cysteaminyl-EDTA-metal, a novel aromatic disulfide derivative of EDTA-metal. We have used this procedure to incorporate an EDTA-iron complex at amino acid 2 of the helix-turn-helix motif of each of two helix-turn-helix motif sequence-specific DNA binding proteins, catabolite gene activator protein (CAP) and Cro, and we have analyzed EDTA-iron-mediated DNA affinity cleavage by the resulting protein derivatives. The CAP derivative cleaves DNA at base pair 2 of the DNA half-site in the protein-DNA complex, and the Cro derivative cleaves DNA at base pairs -3 to 5 of the DNA half-site in the protein-DNA complex. We infer that amino acid 2 of the helix-turn-helix motif of CAP is close to base pair 2 of the DNA half-site in the CAP-DNA complex in solution and that amino acid 2 of the helix-turn-helix motif of Cro is close to base pairs -3 to 5 of the DNA half-site in the Cro-DNA complex in solution.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Composition↗

Agglutination of an EDTA blood sample caused by an EDTA-dependent panagglutinin.

A novel example of an EDTA-dependent panagglutinin is described. A blood sample drawn into EDTA for a routine hematologic workup demonstrated strong agglutination due to EDTA-dependent panagglutinins. Previous examples have been detected because of discrepant ABO results. This is the first report of an EDTA-dependent panagglutinin that caused agglutination in the EDTA sample collection tube and a false positive direct antiglobulin test.

Agglutinins↗

Identification and characterization of the two-enzyme system catalyzing oxidation of EDTA in the EDTA-degrading bacterial strain DSM 9103.

In a gram-negative isolate (DSM 9103) able to grow with EDTA as the sole source of carbon, nitrogen, and energy, the first two steps of the catabolic pathway for EDTA were elucidated. They consisted of the sequential oxidative removal of two acetyl groups, resulting in the formation of glyoxylate. An enzyme complex that catalyzes the removal of two acetyl groups was purified and characterized. In the reaction, ethylenediaminetriacetate (ED3A) was formed as an intermediate and N,N'-ethylenediaminediacetate was the end product. The enzyme complex consisted of two components: component A' (cA'), most likely a monooxygenase, which catalyzes the cleavage of EDTA and ED3A while consuming oxygen and reduced flavin mononucleotide (FMN)-H2, and component B' (cB'), an NADH2:FMN oxidoreductase that provides FMNH2 for cA'. cB' could be replaced by other NADH2:FMN oxidoreductases such as component B of the nitrilotriacetate monooxygenase or the NADH2:FMN oxidoreductase from Photobacterium fischeri. The EDTA-oxidizing enzyme complex accepted EDTA as a substrate only when it was complexed with Mg2+, Zn2+, Mn2+, Co2+, or Cu2+. Moreover, the enzyme complex catalyzed the removal of acetyl groups from several other aminopolycarboxylic acids that possess three or more acetyl groups.

Edetic Acid↗

Effect of three different time periods of irrigation with EDTA-T, EDTA, and citric acid on smear layer removal.

OBJECTIVE: An in vitro study was conducted to evaluate the effect of EDTA-T, 17% EDTA, and 10% citric acid on the removal of smear layer from root canal dentin after final irrigation for 3, 10, and 15 min. STUDY DESIGN: Root canals of 90 extracted human canines were divided into 9 groups of 10 teeth; each group received a final irrigation with one of the irrigants for the established time intervals. After irrigation, the teeth were split and prepared for SEM analysis to determine the number of open dentinal tubules. RESULTS: There were statistically significant better results (P <.05) when irrigation with 10% citric acid for 3 min was compared to 10 and 15 min, and when irrigation with EDTA for 3 min was compared to 15 min. In all cases, irrigation for 3 min presented the greatest number of open dentinal tubules. There were no significant differences (P>.05) for the 3 time intervals of irrigation for EDTA-T, although there were a greater number of open tubules at 3 min. CONCLUSION: These 3 irrigants were effective at the shortest time tested and did not demonstrate an improved effect with an increase in time.

Citric Acid↗

Transverse relaxation of saline and plasma using Mn(II), HSA-EDTA-Mn, and HSA-EDTA-Gd: application to erythrocyte water exchange.

Enhancements of 1H nuclear magnetic transverse relaxation rates by Mn(II)- and Gd(III)-ethylenediaminetetraacetic acid (EDTA)-human serum albumin (HSA) complexes are compared with MnCl2 in human plasma and isotonic saline. HSA-EDTA-Mn and HSA-EDTA-Gd in saline, at physiological concentrations of HSA, afforded T2's comparable to 2 mM MnCl2 in 4% HSA saline. The above reagents are new and potentially useful relaxation agents for the measurement of H2O exchange rates in erythrocyte suspensions. The exchange rate of water across the red cell membrane was determined using these new reagents over the temperature range from 20 to 37 degrees C.

Biological Transport↗

Sodium iron EDTA [NaFe(III)EDTA] as a food fortificant does not influence absorption and urinary excretion of manganese in healthy adults.

NaFe(III)EDTA is a promising iron (Fe) compound for food fortification programs because of its high Fe bioavailability from meals containing dietary inhibitors of Fe absorption such as phytic acid. However, this Fe compound is not currently used in any large-scale fortification program because of concern over its possible negative influence on the metabolism of other essential minerals or its possible influence on the absorption of potentially toxic elements, such as manganese (Mn). In this study, Mn absorption and urinary excretion were studied in adults after intake of an Fe-fortified weaning cereal labeled with 54Mn. In a crossover design, the fortification of the weaning cereal with Fe as NaFeEDTA was compared with ferrous sulfate. Manganese absorption was measured by extrapolation from whole-body retention data 10-30 d after intake, and urinary excretion of 54Mn was measured over 7 d. No significant differences in 54Mn absorption or urinary excretion were found; 1.1 +/- 0.15 and 0.91 +/- 0.35% of the ingested dose was absorbed from the cereal fortified with NaFe(III)EDTA and FeSO4, respectively. Urinary excretion of 54Mn was very low; the total radioactivity in urine represented 1.1 +/- 0.55% of the absorbed dose with NaFe(III)EDTA and 0.72 +/- 0.53% of the absorbed dose with FeSO4. Until now, Fe-fortification programs have met with only limited success. The introduction of NaFeEDTA as a food fortificant could be a useful tool to provide bioavailable Fe to vulnerable groups in the population and thus aid in combating Fe deficiency.

Absorption↗

Apparent pore size of polyacrylamide gels: comparison of gels cast and run in Tris-acetate-EDTA and Tris-borate-EDTA buffers.

The electrophoretic mobilities of DNA molecules in three different molecular weight ladders were measured in polyacrylamide gels containing different acrylamide concentrations (%T) and cross-linker ratios (%C), cast and run in Trisacetate-EDTA (TAE) buffer. The apparent pore radius of each gel was estimated from Ferguson plots of the relative mobilities of each of the DNA molecules, using the mobility of the monomer fragment in each molecular weight ladder as the reference mobility. The effective size of each of the DNA molecules was estimated from its radius of gyration. The apparent gel pore radii calculated in this manner ranged from 21 nm in gels containing 10.5%T, 5%C to 200 nm in gels with 4.6%T, 2%C, similar to the values observed for polyacrylamide gels cast and run in Tris-borate-EDTA (TBE) buffer (Holmes and Stellwagen, Electrophoresis 1991, 12, 612-619). Hence, the effective pore size of polyacrylamide gels is essentially independent of whether the gels are cast and run in TAE or TBE buffer.

Acetates↗

Purification and characterization of a lyase from the EDTA-degrading bacterial strain DSM 9103 that catalyzes the splitting of [S,S]-ethylenediaminedisuccinate, a structural isomer of EDTA.

The bacterial strain DSM 9103, able to utilize EDTA as a sole source of carbon, nitrogen, and energy, is also capable to grow with [S,S]-ethylenediaminedisuccinate ([S,S]-EDDS), a structural isomer of EDTA. In cell-free extracts of [S,S]-EDDS-grown bacteria, [S,S]-EDDS degradation was observed in the absence of any cofactors. An enzyme was purified 41-fold that catalyzed the non-hydrolytic splitting of [S,S]-EDDS leading to the formation of fumarate and N-(2-aminoethyl) aspartic acid. These data strongly suggest that the enzyme belongs to the group of carbon-nitrogen lyases. The splitting reaction was reversible, and an equilibrium constant of approximately 43.0 10(-1) M was determined. Out of the three stereo-isomers of EDDS, [S,S]- and [R,S]-EDDS were accepted as substrates by the lyase, whereas [R,R]-EDDS remained unchanged in assays with both cell-free extracts and pure enzyme. The enzyme catalyzed the transformation of free [S,S]-EDDS and of [S,S]-EDDS-metal complexes with stability constant lower than 10, namely of MgEDDS, CaEDDS, BaEDDS and to a small extent also of MnEDDS; FeIIIEDDS, NiEDDS, CuEDDS, CoEDDS and ZnEDDS were not transformed.

Bacteria↗