Either Ethylenediaminetetraacetic Acid Research Articles

Effects of Various Anticoagulants on Biochemistry Analytes From Domestic Pigeons (Columba livia domestica).

The objective of this study was to compare the effects of anticoagulant and no anticoagulant on routine biochemical analytes in domestic pigeons (Columba livia domestica). Blood samples were obtained from 8 clinically healthy pigeons. The sample obtained from each bird was divided into 4 blood collection tubes containing either ethylenediaminetetraacetic acid (EDTA), lithium heparin, sodium citrate, or no anticoagulant. The concentrations of creatinine, uric acid, triglyceride, total cholesterol, glucose, phosphorus, calcium, magnesium, total protein, albumin, and iron, and the activities of alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate aminotransferase (AST), were measured in blood from each of the blood collection tubes. The values of the measured parameters, with the exception of iron, were significantly lower in the citrated plasma samples compared with the serum samples, even after correcting for dilution. In the lithium heparin plasma samples, significant decreases in albumin, triglyceride, calcium, total cholesterol, and ALP, and a significant increase in iron, were observed compared with the values in the serum samples. The concentrations of total protein, creatinine, glucose, total cholesterol, triglyceride, magnesium, calcium, and phosphorus, as well as the activities for AST and ALP, were significantly lower in the EDTA plasma samples compared with the serum samples. In conclusion, the anticoagulants had significant effects on most of the measured parameters compared with serum. The findings of the present study suggest that a lithium heparin sample is the most appropriate plasma sample for the measurement of blood biochemical parameters in the domestic pigeon.

Acidification does not alter the stable isotope composition of bone collagen.

In this study, we compared the elemental and isotopic composition of modern and ancient bone samples pre-treated using different demineralization agents with acidic and neutral pH. The purpose of our research was to examine if demineralization using a mineral acid such as hydrochloric acid (HCl) significantly alters the δ15N and δ13C values of bone collagen. Evidence from the elemental and amino acid composition of the samples were incorporated alongside isotopic compositions to provide a holistic view of the effect of demineralization agents on the composition of bone collagen. The stable isotope compositions of collagen extracts were also compared against equivalent whole bone samples to assess whether whole bone has a stable isotope composition that is comparable to collagen demineralized with a neutral agent. Our results demonstrate that bone demineralization using either ethylenediaminetetraacetic acid (EDTA) or HCl yields collagen extracts with stable isotope compositions that are not significantly different, indicating that mineral acid does not alter δ15N and δ13C values of bone collagen. The results comparing whole bone and extracted collagen stable isotope compositions indicate that whole bone cannot be used as an effective replacement for bone collagen due to the significantly different stable isotope compositions between these sample materials. In ecological and archaeological studies performing stable isotope analysis on bone, sample pre-treatment to isolate collagen is a necessity to obtain the most reliable and reproducible isotopic measurements.

Optimization of EDTA–Ammonia Ratio for Chemically Deposited Layers of ZnO Nanoparticles

Layers of zinc oxide (ZnO) on glass substrates were grown by chemical bath deposition method. This method refers to solvo-thermal methods of crystal growth. When either ethylene-diamine-tetraacetic acid (EDTA) or ammonia was used as a complex agent, very weak adhesion of ZnO nanoparticles to the surface of glass substrates was noted, and when washing, the samples on the surfaces were removed by a water flow under pressure. When EDTA and ammonia were used together, the adhesion of ZnO nanoparticles to glass substrates increased dramatically. According to X-ray diffraction data, all sediments belong to the hexagonal syngony and correspond to the wurtzite-type ZnO structure. Optical absorption measurements showed that the band gap varied from 3.58 to 3.97 eV. The density of beams from ZnO nanowires on the substrate surface varied depending on the ratio between EDTA and ammonia.

Flavin-Conjugated Iron Oxide Nanoparticles as Enzyme-Inspired Photocatalysts for Azo Dye Degradation

In this work, a new photocatalytic system consisting of iron oxide nanoparticles (IONPs), coated with a catechol-flavin conjugate (DAFL), is synthesized and explored for use in water remediation. In order to test the efficiency of the catalyst, the photodegradation of amaranth (AMT), an azo dye water pollutant, was performed under aerobic and anaerobic conditions, using either ethylenediaminetetraacetic acid (EDTA) or 2-(N-morpholino)ethanesulfonic acid (MES) as electron donors. Depending on the conditions, either dye photoreduction or photooxidation were observed, indicating that flavin-coated iron-oxide nanoparticles can be used as a versatile enzyme-inspired photocatalysts.

Correlating Conformational Dynamics with the Von Willebrand Factor Reductase Activity of Factor H Using Single Molecule Force Measurements.

Activation of proteins often involves conformational transitions, and these switches are often difficult to characterize in multidomain proteins. Full-length factor H (FH), consisting of 20 small consensus repeat domains (150 kD), is a complement control protein that regulates the activity of the alternative complement pathway. Different preparations of FH can also reduce the disulfide bonds linking large Von Willebrand factor (VWF) multimers into smaller, less adhesive forms. In contrast, commercially available purified FH (pFH) has little or no VWF reductase activity unless the pFH is chemically modified by either ethylenediaminetetraacetic acid (EDTA) or urea. We used atomic force microscopy single molecule force measurements to investigate different forms of FH, including recombinant FH and pFH, in the presence or absence of EDTA and urea, and to correlate the conformational changes to its activities. We found that the FH conformation depends on the method used for sample preparation, which affects the VWF reductase activity of FH.

Effects of Adding a Chelator After Ozone Microbubble Generation on Nutrient Composition, Medium Sterility, and Plant Growth

The generation of ozone microbubbles (O3MB) is an effective means of sterilizing plant culture media against bacteria and pathogens; however, the use of O3MB can induce precipitation of metal ions from the medium, such as iron (Fe) and manganese (Mn), that are important to plant growth. Here, we evaluated whether addition of a chelator, either ethylenediaminetetraacetic acid (EDTA) or diethylenetriaminepentaacetic acid (DTPA), could prevent this side effect of O3MB sterilization. We compared nutrient composition, medium sterility, and plant growth after various treatments (0, 0.1, 0.01, 0.001, or 0.0001% (w/v) of EDTA and DTPA addition after O3MB generation). Metal chelators (EDTA and DTPA) are widely used in algal and plant growth solutions to maintain iron (Fe) solubility in hydroponic solutions, EDTA is used in agriculture to remove heavy metals from heavily contaminated soils and also as a plant fertilizer, and DTPA has a wide range of applications including removal of heavy metals from agricultural soils. Thus, the available evidence indicates that addition of a chelator after O3MB generation may prevent precipitation of oxidized Fe or Mn in a culture medium. The addition of a chelator resulted in a concentration-dependent reduction in precipitates and the maintenance of Fe and Mn concentrations in the medium. Although O3MB sterilization did not affect plant growth, the addition of a chelator at concentrations of 0.1, 0.01, or 0.001% (w/v) had a significant deleterious effect. However, at a concentration of 0.0001%, the chelator had no significant effect on plant growth but did result in the maintenance of a higher Fe and Mn concentration compared to the O3MB treated control. The addition of a chelator did not alter the sterilizing effects of O3MB. These results indicate that the addition of a chelator at a concentration of 0.0001% to an O3MB-sterilized culture medium enabled the retention of dissolved Fe and Mn without affecting plant growth. The use of O3MB, plus a chelator, is an effective disinfection method in hydroponic culture.

The synergistic effects of concurrent administration to rats of EDTA and sodium salicylate on the rectal absorption of sodium cefoxitin and the effects of inhibitors.

Plasma levels of cefoxitin, as enhanced by rectal coadministration of sodium salicylate, were reduced by concurrent administration of less than 0.5 mg mL-1 N-ethylmaleimide (NEM) or p-chloromercuriphenylsulphonic acid, sodium salt (p-CMP). Concentrations of these inhibitors above 1 mg mL-1 resulted in enhanced peak plasma values of cefoxitin. This did not occur after coadministration with either ethylenediaminetetraacetic acid (EDTA) or polyoxyethylene-23 lauryl ether (POE). Ouabain and 2,4-dinitrophenol (DNP) suppressed plasma cefoxitin levels in the presence of salicylate and the enhancing effects of EDTA and POE when EDTA and POE were administered at low doses. At higher concentrations of EDTA and POE, DNP had little effect, while ouabain had little effect on POE and only partially suppressed the effects of EDTA. Plasma concentrations of cefoxitin after coadministration with salicylate and POE together, or with EDTA and POE together, were about the same as expected from summing the plasma levels resulting from coadministration of each adjuvant individually at the same concentrations. However, combined administration of salicylate and EDTA with cefoxitin yielded plasma cefoxitin concentrations which were much higher than expected from the sum of their individual actions.

Clozapine activates AMP-activated protein kinase (AMPK) in C2C12 myotube cells and stimulates glucose uptake

Aims Clozapine has previously been implicated in the dysregulation of energy balance and glucose metabolism in the central nervous system, but its effects in the periphery have yet to be thoroughly elucidated. The objective of this study was to characterize the effects of clozapine on AMP-activated protein kinase (AMPK) activity in the skeletal muscles. Main methods Myotube C2C12 cells were incubated under control conditions, or with clozapine. Expression levels of phosphorylation status of AMPK and its direct downstream Acetyl-CoA carboxylase (ACC) were analyzed by Western blot. Intracellular calcium concentration was measured with calcium indicator dye, fluo-3 AM. 2-deoxyglucose uptake was assessed via the scintillation count. Key findings We reported that clozapine activated AMPK in mouse C2C12 myotubes and also stimulated glucose uptake. Clozapine also increased intracellular calcium concentrations of C2C12 cells, and pretreatment with either ethylenediaminetetraacetic acid (EDTA), an extracellular calcium chelator, or 1.8-naphthoylene benzimidazole-3-carboxylic acid (STO-609), a Ca 2+/calmodulin-dependent protein kinase kinase (CaMKK) inhibitor, blocked clozapine-induced AMPK activation. Significance These results demonstrate that clozapine increases glucose uptake through CaMKK-AMPK pathway in myotube C2C12 cells.

Ferritin-dependent radical generation in rat liver homogenates

The hypothesis of this study was that mammalian ferritin (FER) has the ability of releasing Fe in the tissue to catalyze the generation of free radicals, such as ascorbyl (A ) and hydroxyl radical ( OH), that might lead to the damage of FER itself. The rat liver homogenates exhibited an electron paramagnetic resonance (EPR) signal with the spectral features ( a H = 1.88 G, g = 2.0054) of A . The addition to the reaction medium of isolated rat liver FER increased by 3-fold the EPR signal, as compared to the recorded value in its absence. Isolated microsomes from rat liver incubated during 10 min showed a signal with the spectral features ( a H = 15 G, g = 2.0062) of OH. The addition of FER in the presence of either ethylenediamine-tetraacetic acid (EDTA) or adenosine-5′-triphosphate (ATP) significantly increased the recorded spectra. The labile Fe pool (LIP) in the homogenate was assessed by EPR. The rat liver homogenates exhibited an EPR signal with the spectral features ( g = 4.3) of the Fe 2+ and was significantly increased by the addition of FER (3-fold). The oxidation profile of the isolated FER from rat liver was analyzed after incubation with 10 mM ascorbate (AH −). A drastic increase in the width of the band suggested alterations to the protein structure. The FER content of tryptophan (Trp) and thiols was significantly lower when the incubation was performed in the presence of AH − as compared to the recorded effect in its absence. The data in tissue homogenates presented here showed that radical generation is associated to FER Fe release, and moreover that the FER protein itself was affected during this process.

Multilayer Assembly of Hyaluronic Acid/Poly(allylamine): Control of the Buildup for the Production of Hollow Capsules

The objective of this work was to investigate the formation of hollow microcapsules composed of hyaluronic acid (HA) and poly(allylamine) (PAH) by layer-by-layer adsorption on CaCO 3 microparticles and subsequent core removal by addition of chelating agents for calcium ions. We found that the molecular weight of HA as well as the HA solution concentration used during deposition are crucial parameters influencing the multilayer structure. Whereas the effect of molecular weight of HA was mainly attributed to the porous structure of the template which allows penetration of polyelectrolytes when their size is below the maximum pore size of the template ( approximately 60 nm), that of the concentration of the HA solution was related to the intrinsic properties of the polysaccharide. Indeed, as shown by quartz crystal microbalance with dissipation monitoring as well as electron microscopy techniques, the latter leads to dense structures for concentrations from five to ten times the critical overlap concentration during adsorption. Such conditions were found to be favorable for the formation of hollow shells. Regarding conditions for core dissolution, we demonstrated the possibility to use either ethylenediaminetetraacetic acid (EDTA) or citric acid as chelating agents. However, in some cases, it was necessary to chemically cross-link the shell to maintain its integrity.

Extraction of arsenate and arsenite species from soils and sediments

The primary objective of this study was to develop a simple method that can be used to extract the more readily mobilizable and bioavailable arsenic species from soil and sediment while at the same time minimizing the transformation between (As III) and (As V), the two most commonly found arsenic species in the environment. Several extraction strategies were evaluated using phosphate as extractant in combination with either ethylenediaminetetraacetic acid (EDTA), hydroxylamine hydrochloride (NH 2OH·HCl), or sodium diethyldithiocarbamate trihydrate (NaDDC). The addition of EDTA in the phosphate solution did not prevent As III from oxidation. While promising results were shown when 1% NH 2OH·HCl was added, conversion of As III began to occur with extended extraction time (>12 h). Good results were achieved using 10 mM phosphate and 0.5% NaDDC where As III oxidation was clearly minimized. The combined phosphate and NaDDC solution was applied to several soil and sediment samples. As III spiked was quantitatively recovered in all soil types tested.

Field emission in-lens SEM study of enamel and dentin.

This investigation used field emission in-lens scanning electron microscopy (FEISEM) for the study of tooth surfaces, with particular reference to adhesive bonding and acid conditioning. Dentin wafers with an intact enamel periphery were treated by either ethylenediaminetetraacetic acid (EDTA) (pH 7.4) or phosphoric acid (pH 0.7). The samples were then fixed, sequentially dehydrated in alcohol, and either air- or critical point-dried. After coating, surfaces were examined by FEISEM. For enamel, intraprismatic crystals were clearly recognizable, with the crystals showing both a longitudinal and parallel orientation to the long axis of the prisms. For dentin, the surface ultrastructure (mineral crystals and collagen banding) for the both untreated and treated samples was observed. Fine structures measuring on the order of 6 nm were also observed on samples treated by EDTA. We conclude that FEISEM can routinely provide high-resolution images of enamel and dentin, and that it has the capability of revealing the defined distribution of crystals and collagen fibers in dental tissues.

Use of reverse transcriptase-polymerase chain reaction (RT-PCR) and dot-blot hybridisation for the detection and identification of African horse sickness virus nucleic acids.

A coupled reverse transcriptase-polymerase chain reaction assay (RT-PCR) for the detection of African horse sickness virus (AHSV) dsRNA, has been developed using genome segment 7 as the target template for primers. RNA from isolates of all nine AHSV serotypes were readily detected. The potential inhibitory effects of either ethylene diamine tetra acetic acid (EDTA) or heparin on the RT-PCR were eliminated by washing blood samples before lysis of the red blood cells and storage. There was a close agreement in the sensitivity and the specificity of the RT-PCR and an indirect sandwich ELISA. Confirmation of the presence of AHSV using RT-PCR and dot-blot hybridization on blood samples collected from horses experimentally infected with AHSV serotype 4 (AHSV 4) and AHSV serotype 9 (AHSV 9), was achieved within 24 hours, compared to the period of several days required for virus isolation. The RT-PCR and virus isolation methods showed similar levels of sensitivity when used for the detection of AHSV in 3 horses infected with AHSV 4, and in 2 out of 3 horses infected with a less virulent isolate of AHSV 9. Although viraemia was detected in the third horse by virus isolation, from 6 to 14 days after infection, this animal remained consistently negative by RT-PCR. Conversely, AHSV viral RNA was detected by RT-PCR in the blood of 4 donkeys and 4 mules up to 55 days after their experimental infection despite the absence of any detectable infectious virus. RT-PCR is a sensitive and rapid method for detecting AHSV nucleic acids during either the incubation period at the start of an African horse sickness (AHS) epizootic, or for epidemiological investigations in species where clinical signs may be inapparent.

A comparison of cobalt(II) and iron(II) hydroxyl and superoxide free radical formation

We have employed the electron spin resonance spin-trapping technique to study the reaction of Co(II) with hydrogen peroxide in a chemical system and in a microsomal system. In both cases, we employed the spin trap 5,5-dimethyl-1-pyrroline N-oxide (DMPO) and were able to detect the formation of DMPO/.OH and DMPO/.OOH. DMPO/.OOH was the predominant radical adduct formed in the chemical system, while the two adducts were of similar concentrations in the microsomal system. The formation of both of these adducts in either reaction system was inhibited by the addition of superoxide dismutase or catalase, and by chelating the cobalt with either ethylenediaminetetraacetic acid (EDTA) or diethylenetriaminepentaacetic acid (DTPA). The incorporation of the hydroxyl radical scavengers ethanol, formate, benzoate, or mannitol inhibited the formation of DMPO/.OH in both systems. We also repeated the study using Fe(II) in place of Co(II). In contrast to the Co(II) results, Fe(II) reacted with hydrogen peroxide to yield only DMPO/.OH, and this adduct formation was relatively insensitive to the presence of added superoxide dismutase. In addition, Fe(II)-mediated DMPO/.OH formation increased when the iron was chelated to either EDTA or DTPA rather than being inhibited as for Co(II). Thus, we propose that Co(II) does not react with hydrogen peroxide by the classical Fenton reaction at physiological pH values.

1/T1 NMRD profiles of solutions of Mn2+ and Gd3+ protein-chelate conjugates.

Bovine immunoglobulins (IgG) and bovine serum albumin (BSA) were multiply labeled with multidentate ligands, either ethylenediaminetetraacetic acid (EDTA) or diethylenetriaminepentaacetic acid (DTPA), and metal ions were inserted to form the ternary protein-ligand-ion conjugates. The NMRD profiles (the magnetic field dependence of 1/T1) of solutions of the ternary conjugates differ greatly from those of the corresponding binary ligand-metal-ion complexes, both in magnitude and functional form, exhibiting 5- to 10-fold greater relaxivities and prominent peaks near 20 MHz. The inference is that the protein-bound chelates are relatively rigidly attached to the macromolecules. The structure and metal ion affinities of these novel conjugates, as well as the relevance to contrast enhancement in NMR imaging, is discussed.

Interaction of aluminum species with deoxyribonucleic acid.

Interactions of aluminum with deoxyribonucleic acid (DNA) have been studied by thermal denaturation, circular dichroism, and fluorescent dye binding; a pH- and concentration-dependent alteration in the interaction of aluminum with DNA was observed. Three distinguishable complexes are produced when DNA is denaturated at pH 5.0-7.5 and in aluminum to DNA mole ratios of 0-0.7. Complex I appears at neutral pH and stabilizes a portion of DNA. Complex II is observed at acidic pH, destabilizes a fraction of the DNA double-helical molecule, and produces intrastrand cross-links. Complex III occurs at all pHs, is maximal at intermediate pH values, and is characterized by a noncooperative melting profile and cross-linking at low pH (less than 6.0). The DNA in complexes II and III can be renatured by treatment with either ethylenediaminetetraacetic acid (EDTA) or a high concentration of sodium chloride. The properties of complexes I and II are consistent with what could be expected for DNA complexes of Al(OH)2+ and Al3+, respectively. Complex III has intermediate properties that are consistent with a structure in which both ions bind the DNA simultaneously. The characteristics of complex III depend on the ratio of Al3+/Al(OH)2+ in solution. Aluminum-DNA complexes differ from other metal-DNA complexes in that melting profiles under many conditions are biphasic. Apparently more than one form of DNA can exist at any time in the presence of aluminum. The different DNA-aluminum complexes, which arise from the multiple species of aluminum in aqueous solution, lead to a variety of reactions with DNA.