Hypertonic saline- and detergent-accelerated EDTA-based decalcification better preserves mRNA of bones.

Ethylenediaminetetraacetic acid (EDTA), a classically used chelating agent of decalcification, maintains good morphological details, but its slow decalcification limits its wider applications. Many procedures have been reported to accelerate EDTA-based decalcification, involving temperature, concentration, sonication, agitation, vacuum, microwave, or combination. However, these procedures, concentrating on purely tissue-outside physical factors to increase the chemical diffusion, do not enable EDTA to exert its full capacity due to tissue intrinsic chemical resistances around the diffusion passage. The resistances, such as tissue inner lipids and electric charges, impede the penetration of EDTA. We hypothesized that delipidation and shielding electric charges would accelerate EDTA-based penetration and the subsequent decalcification. The hypothesis was verified by the observation of speedy penetration of EDTA with additives of detergents and hypertonic saline, testing on tissue-mimicking gels of collagen and adult mouse bones. Using a 26% EDTA mixture with the additives at 45°C, a conventional 7-day decalcification of adult mouse ankle joints could be completed within 24 h while the tissue morphological structure, antigenicity, enzymes, and DNA were well preserved, and mRNA better retained compared to using 15% EDTA at room temperature. The addition of hypertonic saline and detergents to EDTA decalcification is a simple, rapid, and inexpensive method that doesn't disrupt the current histological workflow. This method is equally or even more effective than the currently most used decalcification methods in preserving the morphological details of tissues. It can be highly beneficial for the related community.

Cryo-storage of porcine hides at the industrial scale for tissue engineering and regenerative medicine application.

BACKGROUND: The industrial scale cryo-storage of raw tissue materials requires a robust, low-cost and easy-to-operate method that can facilitate the down-stream process. OBJECTIVE: The study was aimed to develop the multifunctional protective solutions (MPS) for transportation at ambient conditions and also subsequent cryo-storage below -20 degree C of raw porcine hides for tissue engineering and regenerative medicine. MATERIALS AND METHODS: Protective solutions with antimicrobial activity and proteinase-inhibiting activity were developed and tested for its efficacy in preserving the extracellular matrix of porcine dermis from microbial spoilage, proteolytic degradation, freeze damage and excessive dehydration during shipping and cryo-storage. The MPSs contained phosphate-buffered saline with ethylene diamine tetra acetic acid (EDTA) added as chelator and proteinase inhibitor, as well as glycerol or maltodextrin (M180) as cryoprotectants. RESULTS: MPSs prepared with EDTA and glycerol or M180 had significant antimicrobial activity and proteinase-inhibiting activity during the period of shipping and handling. Glycerol and M180 prevented eutectic salt precipitation and excessive freeze dehydration upon cryo-storage of porcine hides. Without glycerol or M180, hides could be freeze-dehydrated to the low hydration at ~0.4 g/g dw, and formed irreversible plications after freezing. A critical hydration (0.8~0.9 g/g dw) was observed for the extracellular matrix of porcine dermis, and dehydration to a lower level could impose enormous stress and potential damage. The soaking of porcine hides in MPSs decreased water content as glycerol and M180 entered into dermis. Upon equilibration, the glycerol content in the tissue was about 94% of the incubating glycerol solution, but the M180 content in the tissue was only about 50% of the incubating M180 solution, indicating that M180 did not get into the entire aqueous domain within dermis. MPSs reduced ice formation and increased the unfrozen water content of porcine raw hides upon cryo-storage. CONCLUSION: MPSs prepared with EDTA and glycerol or M180 have antimicrobial activity and proteinase-inhibiting activity, which can be used for transportation and cryo-storage of raw hides at the industrial scale. Glycerol at 7.5% w/v and M180 at 20% w/v were sufficient to prevent freeze damage and excessive freeze dehydration. Doi.org/10.54680/fr24310110312.

Comprehensive chromatographic assessment of forced degraded in vitro transcribed mRNA.

Heightened interest in messenger RNA (mRNA) therapeutics has accelerated the need for analytical methodologies that facilitate the production of supplies for clinical trials. Forced degradation studies are routinely conducted to provide an understanding of potential weak spots in the molecule that are exploited by stresses encountered during bulk purification, production, shipment, and storage. Consequently, temperature fluctuations and excursions are often experienced during these unit operations and may accelerate mRNA degradation. Here, we present a concise panel of chromatography-based stability-indicating assays for evaluating thermally stressed in vitro transcribed (IVT) mRNA as part of a forced degradation study. We found that addition of EDTA to the mRNAs prior to heat exposure reduced the extent of degradation, suggesting that transcripts may be fragmenting via a divalent metal-ion mediated pathway. Trace divalent metal contamination that can accelerate RNA instability is likely carried over from upstream steps. We demonstrate the application of these methods to evaluate the critical quality attributes (CQAs) of mRNAs as well as to detect intrinsic process- and product-related impurities.

Improvement of performance for flexible film dosimeter by incorporating additive agents.

Objective.To evaluate the reduction in energy dependence and aging effect of the lithium salt of pentacosa-10,-12-diynoic acid (LiPCDA) films with additives including aluminum oxide (Al2O3), propyl gallate (PG), and disodium ethylenediaminetetracetate (EDTA).Approach. LiPCDA films exhibited energy dependence on kilovoltage (kV) and megavoltage (MV) photon energies and experienced deterioration over time. Evaluations were conducted with added Al2O3and antioxidants to mitigate these issues, and films were produced with and without Al2O3to assess energy dependence. The films were irradiated at doses of 0, 3, 6, and 12 cGy at photon energies of 75 kV, 105 kV, 6 MV, 10 MV, and 15 MV. For the energy range of 75 kV to 15 MV, the mean and standard deviation (std) were calculated and compared for the values normalized to the net optical density (netOD) at 6 MV, corresponding to identical dose levels. To evaluate the aging effect, PG and disodium EDTA were incorporated into the films: sample C with 1% PG, sample D with 2% PG, sample E with 0.62% disodium EDTA added to sample D, and sample F with 1.23% disodium EDTA added to sample D.Main results. Films containing Al2O3demonstrated a maximum 15.8% increase in mean normalized values and a 15.1% reduction in std, reflecting a greater netOD reduction at kV than MV energies, which indicates less energy dependence in these films. When the OD of sample 1-4 depending on the addition of PG and disodium EDTA, was observed for 20 weeks, the transmission mode decreased by 8.7%, 8.3%, 29.3%, and 27.3%, respectively, while the reflection mode was 5.4%, 3.0%, 37.0%, and 34.5%, respectively.Significance. Al2O3effectively reduced the voltage and MV energy dependence. PG was more effective than disodium EDTA in preventing the deterioration of film performance owing to the aging effect.

Effective oxidation decomplexation of Cu-EDTA and Cu2+ electrodeposition from PCB manufacturing wastewater by persulfate-based electrochemical oxidation: Performance and mechanisms.

Complex wastewater matrices such as printed circuit board (PCB) manufacturing wastewater present a major environmental concern. In this work, simultaneous decomplexation of metal complex Cu-EDTA and reduction/electrodeposition of Cu2+ was conducted in a persulfate-based electrochemical oxidation system. Oxidizing/reductive species were simultaneously produced in this system, which realized 99.8% of Cu-EDTA decomplexation, 94.5% of Cu2+ reduction/electrodeposition under the conditions of original solution pH = 3.2, electrode distance = 3 cm, [Na2S2O8]0 = 5 mM, current density = 12 mA/cm2, and reaction time = 180 min. The total treatment cost is as low as 0.80 USD/mol Cu-EDTA. Effective mineralization (74.1% total organic carbon removal) of the solution was obtained after 3 h of treatment. •OH and SO4•- drove the Cu-EDTA decomplexation, destroying the chelating sites and finally it was effectively mineralized to CO2, H2O and Cu2+. The mechanisms of copper electrodeposition on the stainless steel cathode and persulfate activation by the BDD anode were proposed based on the electrochemical measurements. The electrodes exhibited excellent reusability and low metal (total iron and Ni2+) leaching during 20 cycles of application. This study provide an effective and sustainable method for the application of the electro-persulfate process in treating complex wastewater matrices.

Effect of sodium hypochlorite, ethylenediaminetetraacetic acid, and dual-rinse irrigation on dentin adhesion using an etch-and-rinse or self-etch approach.

The aim was to investigate the influence of endodontic irrigation solutions and protocols on the micro-tensile bond strength (µTBS) to dentin using an etch-and-rinse (ER) or self-etch (SE) adhesive approach. Eighty extracted human molars were ground to dentin. After pretreating for 27 min (21 min-3 min-3 min) with five different endodontic irrigation protocols (Group 1: NaOCl-EDTA-NaOCl; Group 2: NaOCl-NaOCl-EDTA; Group 3: NaOCl-NaCl-NaOCl; Group 4: Dual Rinse-Dual Rinse-Dual Rinse; Group 5: NaCl-NaCl-NaCl), an ER (Optibond FL, Kerr) or a SE (Clearfil SE Bond, Kuraray) adhesive system was applied. After light-curing, composite build-ups were made and cut into dentin-composite sticks. µTBS and failure modes were analyzed. Nonparametric statistical analyses (α = 0.05) were performed for comparison of the five groups within each type of adhesive as well as between the two adhesive systems used. The use of an ER instead of a SE adhesive system resulted in significantly higher µTBS for all irrigation protocols except for group 1 (NaOCl-EDTA-NaOCl) and 2 (NaOCl-NaOCl-EDTA). A statistical difference between the five different endodontic irrigation protocols was only found within the SE adhesive group, where group 1 (NaOCl-EDTA-NaOCl) achieved highest values. The use of an ER adhesive system cancels out the effect of the endodontic irrigation solution. The highest µTBS was achieved when using a NaOCl-EDTA-NaOCl-irrigation protocol in combination with Clearfil SE Bond, which shows that the selection of the endodontic irrigation should match the corresponding SE adhesive system.

The effect of oscillation depolymerization on ethylenediaminetetraacetic acid-dependent platelet aggregation samples: A cross-over study.

INTRODUCTION: Ethylenediaminetetraacetic acid (EDTA)-dependent platelet aggregation (PA) can cause medical errors. Currently, there is no reliable method for completely solving this problem. This study aims to solve this problem that has plagued clinical practice for many years by using oscillation method. METHODS: Sixty-one EDTA-PA samples were collected, divided, and disaggregated using the oscillation method at various times and speeds. The samples were analyzed using routine blood tests and blood smears. RESULTS: Platelet counts (PLT) were increased significantly after oscillation. PLT in the 3000 rpm for 0.5 min group was significantly higher than that in the 500 rpm for 0.5 min group (p < 0. 01). After 3000 rpm oscillation, the PLT gradually increased with time, while compared with the 10-min group, the PLT in the 13-min group showed no significant differences. The effective disaggregation rates in the EDTA-PA samples using the oscillation method and sodium citrate anticoagulant were 96.72% and 65.57%, respectively. There were no significant changes in white blood cell (WBC) or red blood cell (RBC) counts or morphology after the use of the oscillation method. CONCLUSION: The oscillation method effectively depolymerized EDTA-PA without adverse effects on WBC and RBC. The implementation of this technique promises to resolve the issue of EDTA-PA.

Molecular engineering of a new method for effective removal of cadmium from water.

Cadmium (Cd) is a widespread and highly toxic environmental pollutant, seriously threatening animal and plant growth. Therefore, monitoring and employing robust tools to enrich and remove Cd from the environment is a major challenge. In this work, by conjugating a fluorescent indicator (CCP) with a functionalized glass slide, a special composite material (CCPB) was constructed to enrich, remove, and monitor Cd2+ in water rapidly. Then Cd2+ could be effectively eluted by immersing the Cd-enriched CCPB in an ethylenediaminetetraacetic acid (EDTA) solution. With this, the CCPB was continuously reused. Its recovery of Cd2+was above and below 100 % after multiple uses by flame atomic absorption spectrometry (FAAS), which was excellent for practical use in enriching and removing Cd2+ in real aqueous samples. Therefore, CCPB is an ideal material for monitoring, enriching, and removing Cd2+ in wastewater, providing a robust tool for future practical applications of Cd enrichment and removal in the environment.

Preparation of landscape gardening soil using undersized fraction from aged MSW by EDTA or citric acid coupled with humic acid: Effect assessment, properties, and optimization.

Undersized fraction from aged municipal solid waste (UFAMSW), as a kind of soil-like material, has been proved effective in providing a large amount of organic matter and nutrients for soil and plants. The characteristics and effectiveness of heavy metal pollution removal in UFAMSW attracted tremendous research interest from scientists recently. In this study, the heavy metal removal efficiencies and bioavailability of washing on contaminated UFAMSW were evaluated with three washing reagents including ethylene diamine tetra acetic acid (EDTA), citric acid (CA), and humic acid (HA). The effects of chelating agent concentration, pH, and washing time on metal removal were investigated and response surface methodology (RSM) was employed to optimize the washing conditions. The results indicated that the removal efficiencies of Cu, Zn, and Mn could be 53.68%, 52.12%, and 30.63% by EDTA/HA washing and 42.36%, 39.67% and 28.49% by CA/HA washing, respectively. The European Community Bureau of Reference (BCR) sequential extraction was applied to analyze the fraction change of heavy metals in UFAMSW before and after washing, and it was found that chelating agent combined with HA could contribute to the removal of the exchangeable fraction. Physical and chemical properties of UFAMSW were improved to some extent after washing with mixed HA and chelating agent and could achieve the quality standard of landscape gardening soil. Accordingly, the mixture of HA and other chelating agents could be a promising washing process for preparation of landscape gardening soil using UFAMSW.Implications: Our manuscript studies the removal of heavy metals from the contaminated undersized fraction from aged municipal solid waste (UFAMSW). UFAMSW, as a kind of soil-like material, has been proved effective in providing a large amount of organic matter and nutrients for soil and plants however often limited by heavy metal pollution. The UFAMSW used in this experiment was collected after the excavation and screening-sorting of aged refuse from Changshankou Domestic Waste Sanitary Landfill in Wuhan City, Hubei Province, Southern China. This study investigated the effects of EDTA, CA, HA, mixed EDTA/HA, and mixed CA/HA washing on heavy metal removal (Cu, Zn, and Mn), bioavailability of residual heavy metal and properties. The effects of chelating agent concentration, pH, and washing time on metal removal were investigated and then response surface methodology was employed to optimize the washing conditions. The results showed that washing by CA/HA and EDTA/HA, had a higher removal efficiency of heavy metals (Cu, Zn, and Mn) in UFAMSW compared to single HA. Meanwhile, HA has a higher removal for exchangeable fraction of heavy metals, the exchangeable concentration of Cu, Zn, and Mn in CA/HA and EDTA/HA washed UFAMSW were lower compared with UFAMSW washed by single CA and EDTA. Thus, mixing HA with EDTA or CA makes a less risk to environmental and the removal efficiency is acceptable. Additionally, CA/HA and EDTA/HA washing tend to improve soil physicochemical properties and soil fertility. Thus, mixing HA with different washing agent are potential methods for preparation of landscape gardening soil using UFAMSW.

Remediation of cadmium-contaminated soil: GLDA-assisted extraction and sequential FeCl3-CaO-based post-stabilization.

Cadmium (Cd) contamination of farmland soils is a growing concern because of its highly toxic impact on ecosystems and human health. Chelator-assisted washing and chemical immobilization are effective remediation strategies for Cd-contaminated soils. Ethylenediaminetetraacetic acid (EDTA) has traditionally been used for soil washing, but its persistence in the environment and subsequent toxicity have raised significant ecological concerns. Consequently, biodegradable chelators have gained increasing attention as eco-friendly alternatives to the persistent chelator, EDTA. Therefore, this study evaluated the performance and efficacy of three biodegradable chelators: L-glutamate-N,N'-diacetic acid (GLDA), methylglycine-diacetic acid (MGDA), and 3-hydroxy-2,2'-iminodisuccinic acid (HIDS) in comparison to EDTA for remediating a real Cd-contaminated agricultural soil. The influence of treatment parameters, including chelator variants, washing time, chelator concentration, solution pH, and liquid-to-soil ratio (L/S) on Cd extraction was studied and optimized to attain the maximum removal rate. Following chelator-assisted washing, the efficacy of a stabilization preference combining FeCl3 and CaO in reducing the leaching potential of residual Cd in chelator-washed soil residues was also investigated. GLDA demonstrated comparable Cd extraction efficiency to EDTA, and the Cd extraction efficiency was found to be positively correlated with the soil washing parameters. However, under the optimized conditions (chelator concentration: 10 mmol L-1; washing time: 3 h; solution pH: 3; L/S ratio: 10:1), GLDA exhibited a higher Cd extraction rate than EDTA or the other chelators. Furthermore, a post-treatment process incorporating FeCl3 and CaO substantially diminished the water-leachable Cd content in the resultant soil residues. The proposed remediation strategy, which combines chemically assisted washing and stabilization, could be a practical option for extracting bulk Cd from soil and reducing the leaching potential of residual Cd.

A glycol chitosan derivative with extrafibrillar demineralization potential for self-etch dentin bonding.

OBJECTIVES: Extrafibrillar demineralization is an etching technique that removes only minerals from around the collagen fibrils for resin infiltration. The intrafibrillar minerals are left intact to avoid their replacement by water that is hard for adhesive resin monomers to displace. The present work reported the synthesis of a water-soluble methacryloyloxy glycol chitosan-EDTA conjugate (GCE-MA) and evaluated its potential as an extrafibrillar demineralization agent for self-etch dentin bonding. METHODS: Glycol chitosan-EDTA was functionalized with a methacryloyloxy functionality. Conjugation was confirmed using Fourier transform-infrared spectroscopy. The GCE-MA was used to prepare experimental self-etch primers. Extrafibrillar demineralization of the primers was evaluated with scaning electron microscopy and transmission electron microscopy. The feasibility of this new self-etch bonding approach was evaluated using microtensile bond strength testing and inhibition of dentin gelatinolytic activity. The antibacterial activity and cytotoxicity of GCE-MA were also analyzed. RESULTS: Conjugation of EDTA and the methacryloyloxy functionality to glycol chitosan was successful. The functionalized conjugate was capable of extrafibrillar demineralization of mineralized collagen fibrils. Tensile bond strength of the experimental self-etch primer to dentin was comparable to that of phosphoric acid-etched dentin and the commercial self-etch primer Clearfil SE Bond 2. The GCE-MA also inhibited soluble rhMMP-9. In-situ zymography detected minimal fluorescence in hybrid layers conditioned with the experimental primer. The GCE-MA was noncytotoxic and possessed antibacterial activities against planktonic bacteria. SIGNIFICANCE: Synthesis of GCE-MA brought into fruition a self-etch conditioner that selectively demineralizes the extrafibrillar mineral component of dentin. A self-etch primer prepared with GCE-MA achieved bond strengths comparable to commercial reference adhesive systems.

Mass Spectrometric Detection of Formaldehyde-Crosslinked PBMC Proteins in Cell-Free DNA Blood Collection Tubes.

Streck tubes are commonly used to collect blood samples to preserve cell-free circulating DNA. They contain imidazolidinyl urea as a formaldehyde-releasing agent to stabilize cells. We investigated whether the released formaldehyde leads to crosslinking of intracellular proteins. Therefore, we employed a shotgun proteomics experiment on human peripheral blood mononuclear cells (PBMCs) that were isolated from blood collected in Streck tubes, EDTA tubes, EDTA tubes containing formaldehyde, or EDTA tubes containing allantoin. The identified crosslinks were validated in parallel reaction monitoring LC/MS experiments. In total, we identified and validated 45 formaldehyde crosslinks in PBMCs from Streck tubes, which were also found in PBMCs from formaldehyde-treated blood, but not in EDTA- or allantoin-treated samples. Most were derived from cytoskeletal proteins and histones, indicating the ability of Streck tubes to fix cells. In addition, we confirm a previous observation that formaldehyde crosslinking of proteins induces a +24 Da mass shift more frequently than a +12 Da shift. The crosslinking capacity of Streck tubes needs to be considered when selecting blood-collection tubes for mass-spectrometry-based proteomics or metabolomic experiments.

Systematic Fe(II)-EDTA Method of Dose-Dependent Hydroxyl Radical Generation for Protein Oxidative Footprinting.

Correlating the structure and dynamics of proteins with biological function is critical to understanding normal and dysfunctional cellular mechanisms. We describe a quantitative method of hydroxyl radical generation via Fe(II)-ethylenediaminetetraacetic acid (EDTA)-catalyzed Fenton chemistry that provides ready access to protein oxidative footprinting using equipment commonly found in research and process control laboratories. Robust and reproducible dose-dependent oxidation of protein samples is observed and quantitated by mass spectrometry with as fine a single residue resolution. An oxidation analysis of lysozyme provides a readily accessible benchmark for our method. The efficacy of our oxidation method is demonstrated by mapping the interface of a RAS-monobody complex, the surface of the NIST mAb, and the interface between PRC2 complex components. These studies are executed using standard laboratory tools and a few pennies of reagents; the mass spectrometry analysis can be streamlined to map the protein structure with single amino acid residue resolution.

Functional Ag-EDTA-modified MnO2 nanocoral reef for rapid removal of hazardous copper from wastewater.

A novel MnO2@EDTA-Ag nanocoral reef was constructed via a simplified redox reaction followed by EDTA and Ag nanoparticles impregnation to capture hazardous copper (II) from wastewater. A comprehensive characterization of the synthesized materials was conducted. The morphology of MnO2@EDTA-Ag in the form of a nanocoral reef was constructed of two-dimensional nanoplatelets and nanorod-like nanostructures. The optimal adsorption conditions proposed by the Plackett-Burman design (PBD) that would provide a removal % of 99.95 were pH 5.5, a contact time of 32.0 min, a Cu(II) concentration of 11.2 mg L-1, an adsorbent dose of 0.05 g, and a temperature of 40.3 °C. The loading of Ag nanoparticles onto MnO2@EDTA improved the adsorption capability of MnO2@EDTA-Ag. Additionally, the recyclability of MnO2@EDTA-Ag nanocoral reef was maintained at 80% after three adsorption-desorption cycles, and there was no significant change in the XRD analysis before and after the recycling process, implying its stability. It was found that nanocoral reef-assisted EDTA formed a chelation/complexation reaction between COO- groups and C-N bonds of EDTA with Cu(II) ions. In addition, X-ray photoelectron spectroscopy (XPS) and Fourier transform infrared spectroscopy (FTIR) analysis proved the synergistic effect of the electrostatic interaction and chelation/complexation was responsible for the removal mechanism of Cu(II). Also, the results demonstrated no significant variation in MnO2@EDTA-Ag removal efficiency for all the tested real water samples, revealing its efficacy in wastewater treatment. Therefore, the current study suggests that MnO2@EDTA-Ag has substantial potential to be used as a feasible adsorbent for probable hazardous metals remediation.

Comparative evaluation of effect of nisin-incorporated ethylenediamine tetraacetic acid and MTAD on endodontic biofilm eradication, smear layer removal, and depth of sealer penetration.

OBJECTIVES: To comparatively evaluate the nisin-incorporated ethylenediamine tetraacetic acid (N-EDTA) and MTAD on cytotoxicity, endodontic biofilm eradication potential, smear layer removal ability, and sealer penetration depth. MATERIALS AND METHODS: N-EDTA was prepared and characterized using high-performance liquid chromatography (HPLC). Minimum inhibitory, minimum bactericidal, and minimum biofilm inhibitory concentration (MBC, MIC, and MBIC) were determined on Enterococcus faecalis (E. faecalis) strain. The cytocompatibility of N-EDTA and MTAD was evaluated using 3,(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)-based colorimetric assay. Dentin specimens (n = 88 for antibacterial analysis, n = 170 for sealer penetration depth) were prepared and subjected to the classical irrigating strategy and obturation, respectively. The scanning electron microscopic evaluation (SEM) was done for the evaluation of biofilm disruption and smear layer removal. Confocal laser scanning microscopy (CLSM) evaluation was done for determining percentage of bacterial viability and sealer penetration depth. Statistical analysis of one-way ANOVA and Tukey's HSD post hoc tests for bacterial viability and Kruskal-Wallis test and Mann-Whitney test for smear layer removal and depth of penetration were done with the significance level set at p < 0.05. RESULTS: MTAD and N-EDTA showed cytocompatibility without any statistical differences from each other. For N-EDTA, the MIC and MBC values were 12.5 µg/ml (1:8), and MBIC values were 36 µg/ml. Biofilm disruption and killed bacterial percentage of N-EDTA was statistically higher than MTAD, whereas both the materials showed similar efficacy in the removal of the smear layer and sealer penetration depth. CONCLUSION: N-EDTA had negligible cytotoxicity with similar smear layer removal ability, sealer penetration, and better antibiofilm potential than MTAD. CLINICAL RELEVANCE: N-EDTA can serve as a viable alternative endodontic irrigant.

Development and Comparative Evaluation of Two Highly Sensitive Immunosensor Platforms for Trace Determination of Copper Ions in Drinking Water Using a Monoclonal Antibody Specific to Copper-EDTA Complex.

This study describes the development of two highly sensitive immunosensor platforms for the trace determination of copper ions, Cu(II), in drinking water. These platforms were a microwell-based enzyme-linked immunosorbent assay (ELISA) and a kinetic exclusion assay (KinExA) with a KinExATM 3200 immunosensor. Both ELISA and KinExA were developed utilizing the same antibody and coating reagent. The antibody was a mouse monoclonal antibody, designated as 8D66, that specifically recognized Cu(II)-ethylenediamine tetraacetic acid complex (Cu(II)-EDTA) but did not recognize Cu(II)-free EDTA. The 8D66 monoclonal antibody was generated by the fusion of spleen cells of an immunized BALB/c mouse with SP2/0-Ag14 myeloma cells. The immunogen was a protein conjugate of Cu(II)-EDTA with keyhole limpet hemocyanin protein. The coating reagent was Cu(II)-EDTA covalently linked to bovine serum albumin protein (Cu(II)-EDTA-BSA). Both assays involved the competitive binding reaction between Cu(II)-EDTA complexes, formed in the sample solution, and Cu(II)-EDTA-BSA conjugate which has been immobilized onto ELISA plates (in ELISA) or polymethylmethacrylate beads (in KinExA) for a limited quantity of binding sites of the 8D66 antibody. In ELISA, color signals were generated by a peroxidase-labeled secondary antibody and 3,3',5,5'-tetramethylbenzidine substrate. In KinExA, a fluorescein isothiocyanate-labeled secondary antibody was used to generate KinExAgram (trend-line fluorescence responses vs. time). The conditions of both ELISA and KinExA were investigated, and the optimum procedures were established. Both ELISA and KinExA were validated, and all validation parameters were acceptable. Many different metal ions that are commonly encountered in drinking water did not interfere with the Cu(II) analysis by both ELISA and KinExA. Both assays were applied to the determination of Cu(II) in drinking water with satisfactory accuracy and precision. Both assays were compared favorably with inductively coupled plasma atomic emission spectroscopy in terms of their abilities to accurately and precisely determine Cu(II) in drinking water samples. A comparative evaluation of ELISA and KinExA revealed that KinExA had a higher sensitivity and better precision than ELISA, whereas both assays had comparable accuracy. Both ELISA and KinExA were superior to the existing atomic spectrometric methods for Cu(II) in terms of sensitivity, convenience, and analysis throughputs. The proposed ELISA and KinExA are anticipated to effectively contribute to assessing Cu(II) concentrations and control the exposure of humans to its potential toxicities.

Evaluation of newly designed flushing techniques for on-site remediation of arsenic-contaminated excavated debris.

Excavated debris (soil and rock) contaminated with geogenic arsenic (As) is an increasing concern for regulatory organizations and construction stakeholders. Chelator-assisted soil flushing is a promising method for practical on-site remediation of As-contaminated soil, offering technical, economic, and environmental benefits. Ethylenediaminetetraacetic acid (EDTA) is the most prevalent chelator used for remediating As-contaminated soil. However, the extensive environmental persistence and potential toxicity of EDTA necessitate the exploration of eco-compliant alternatives. In this study, the feasibility of the conventional flushing method pump-and-treat and two newly designed immersion and sprinkling techniques were evaluated at the laboratory scale (small-scale laboratory experiments) for the on-site treatment of As-contaminated excavated debris. Two biodegradable chelators, L-glutamic acid-N,N'-diacetic acid (GLDA) and 3-hydroxy-2,2'-iminodisuccinic acid (HIDS), were examined as eco-friendly substitutes for EDTA. Additionally, this study highlights a useful post-treatment measure to ensure minimal mobility of residual As in the chelator-treated debris residues. The pump-and-treat method displayed rapid As-remediation (t, 3 h), but it required a substantial volume of washing solution (100 mL g-1). Conversely, the immersion technique demonstrated an excellent As-extraction rate using a relatively smaller washing solution (0.33 mL g-1) and shorter immersion time (t, 3 h). In contrast, the sprinkling technique showed an increased As-extraction rate over an extended period (t, 48 h). Among the chelators employed, the biodegradable chelator HIDS (10 mmol L-1; pH, 3) exhibited the highest As-extraction efficiency. Furthermore, the post-treatment of chelator-treated debris with FeCl3 and CaO successfully reduced the leachable As content below the permissible limit.

Effect of ultrasound-assisted EDTA and citric acid washing on heavy metal removal, residual heavy metal mobility, and sewage sludge quality.

We investigated the effects of ultrasound-assisted ethylenediaminetetraacetic acid (EDTA) and citric acid (CA) washing on heavy metal (HM) removal, residual HM mobility, and sewage sludge quality. EDTA and CA washing of sewage sludge successfully reduced the total concentration of HMs after one round of washing, but the mobility of residual HMs increased significantly. The eluate had a high concentration of HMs and nutrients (nitrogen, phosphorus, potassium, and total organic carbon), although the nutritional content of the sludge remained high. The three-phase ratio of the sludge after six rounds of washing by CA was closest to the ideal three-phase ratio, and the degree of influence on the physical structure of the soil after a land application was reduced, according to the fluctuation of generalized soil structure index (GSSI) and soil three-phase structure distance (STPSD) values. The results indicate that CA as an environmental-friendly washing agent can be the superior choice for sludge HM washing; single washing of sewage sludge may increase the mobility of residual HMs, so multiple washings should be considered for land application of sludge.

Electrochemical Removal of Metal-Organic Complexes in Metal Plating Wastewater: A Comparative Study of Cu-EDTA and Ni-EDTA Removal Mechanisms.

Cu and Ni complexes with ethylenediaminetetraacetic acid (Cu/Ni-EDTA), which are commonly present in metal plating industry wastewaters, pose a serious threat to both the environment and human health due to their high toxicity and low biodegradability. In this study, the treatment of solutions containing either or both Cu-EDTA and Ni-EDTA using an electrochemical process is investigated under both oxidizing and reducing electrolysis conditions. Our results indicate that Cu-EDTA is decomplexed as a result of the cathodic reduction of Cu(II) with subsequent electrodeposition of Cu(0) at the cathode when the cathode potential is more negative than the reduction potential of Cu-EDTA to Cu(0). In contrast, the very negative reduction potential of Ni-EDTA to Ni(0) renders the direct reduction of EDTA-complexed Ni(II) at the cathode unimportant. The removal of Ni during the electrolysis process mainly occurs via anodic oxidation of EDTA in Ni-EDTA, with the resulting formation of low-molecular-weight organic acids and the release of Ni2+, which is subsequently deposited as Ni0 on the cathode. A kinetic model incorporating the key reactions occurring in the electrolysis process has been developed, which satisfactorily describes EDTA, Cu, Ni, and TOC removal. Overall, this study improves our understanding of the mechanism of removal of heavy metals from solution during the electrochemical advanced oxidation of metal plating wastewaters.

Efeito de diferentes irrigantes finais na resistência união de um cimento de silicato de cálcio: estudo in vitro / Effect of different final irrigants on the bond strength of a calcium silicate cement : in vitro study

Objetivo: Este estudo teve como objetivo avaliar a resistência de união do cimento Biodentine® à dentina radicular após a utilização de diferentes irrigantes finais. Método: Vinte dentes humanos extraídos tiveram seu terço médio radicular cortado em fatias que foram submersas em hipoclorito de sódio 2,5% e posteriormente divididas aleatoriamente em 4 grupos experimentais (n=15) conforme o irrigante final utilizado (1) água destilada (controle), (2) QMixTM, (3) ácido cítrico 10%, (4) EDTA 17%. Após a imersão na substância teste as amostras foram preenchidas com o cimento Biodentine e imersas em solução salina tamponada com fosfato (PBS) por um período de 7 dias. O teste de push out foi realizado e os valores de resistência de união em Mpa foram obtidos. Os dados foram analisados pelos testes de Kruskal Wallis e Studend- Newman-Keuls. Resultados: Os piores valores de união foram obtidos após a utilização do EDTA enquanto a água destilada, o QMix e o ácido cítrico apresentaram resultados estatisticamente semelhantes entre si. Conclusão: A remoção da smear layer não resultou em melhora nos resultados de união do cimento Biodentine.(AU)

Objective: This study aimed to evaluate the bond strength of Biodentine® cement to root dentin after the use of different final irrigants. Method: Twenty extracted human teeth had their middle root third cut into slices that were submerged in 2.5% sodium hypochlorite and then randomly divided into 4 experimental groups (n=15) according to the final irrigant used (1) distilled water (control), (2) QMixTM, (3) 10% citric acid, (4) 17% EDTA. After immersion in the test substance the samples were filled with Biodentine cement and immersed in phosphate buffered saline (PBS) for a period of 7 days. The push out test was performed and the bond strength values in MPa were obtained. The data were analyzed by Kruskal Wallis and Studend- Newman-Keuls tests. Results: The worst bond values were obtained after using EDTA while distilled water, QMix and citric acid showed statistically similar results to each other. Conclusion: Removal of the smear layer did not result in improved bonding results of Biodentine cement.(AU)