Multianticoagulant Pseudothrombocytopenia in a Patient with Primary Carcinoma of the Liver with Hypersplenism.

BACKGROUND: Pseudothrombocytopenia (PTCP) is a relatively rare phenomenon in vitro, the mechanism is not completely clear, and there is no unified solution for it. How to identify and solve PTCP accurately is a challenge for laboratory personnel. METHODS: According to the patient's clinical manifestations, thrombocytopenia caused by hypersplenism was excluded. PTCP was confirmed by platelet volume histograms, scattergrams and platelet clumps on the blood smears. Commonly used alternative anticoagulants such as sodium citrate or heparin were used for platelet counting. The corrective effect of the platelet count was not good, so non-anticoagulant blood was collected and tested immediately, and blood smears were used to count platelets manually. RESULTS: The PTCP of the patient could not be solved using sodium citrate and heparin anticoagulation. By collecting non-anticoagulant blood and testing immediately, the platelet count returned to normal (180 x 109/L), which is consistent with the results of manual counting on the patient's blood smears (175 x 109/L). CONCLUSIONS: When PTCP is confirmed, commonly used alternative anticoagulants can be used. If these do not work, non-anticoagulant blood can be collected and tested immediately, and blood smears can be used to count platelets manually.

The effect of the anti-coagulant EDTA on the deposition and adhesion of whole blood deposits on non-porous substrates.

An investigation into whether the addition of a commonly used anti-coagulant agent like ethylenediaminetetraacetic acid (EDTA) has an impact on the adhesion potential of blood to non-porous substrates was conducted. Two non-porous substrates (aluminum and polypropylene) exhibiting six different surface roughness categories (R1-R6) were used as test substrates upon which either whole blood or blood treated with EDTA was deposited. Samples were exposed to different drying periods (24 hours, 48 hours, and 1 week) before undergoing a tapping agitation experiment in order to evaluate the adhesion to the surface. Clear differences in adhesion potential were observed between whole blood and blood treated with EDTA. Blood treated with EDTA displayed a stronger adhesion strength to aluminum after a drying time of 24 h pre-agitation, while whole blood presented with a stronger adhesion strength at the drying time of 48 h and 1 week. Both EDTA-treated and EDTA-untreated blood was shown to dislodge less easily on polypropylene with the only difference observed on smooth surfaces (0.51-1.50 µm surface roughness). Thus, when conducting transfer studies using smooth hydrophobic substrates like polypropylene or considering the likelihood of transfer given specific case scenarios, differences in adhesion strength of blood due to hydrophobic substrate characteristics and a decreased surface area need to be considered. Overall, whole blood displayed a better adhesion strength to aluminum, emphasizing that indirect transfer probability experiments using EDTA blood on substrates like aluminum should take an increased dislodgment tendency into account in their transfer estimations.

Eco-friendly approach to decrease the harmful effects of untreated wastewater on growth, yield, biochemical constituents, and heavy metal contents of carrot (Daucus carota L.).

Here, the impact of irrigation using untreated wastewater (WW) on carrots (Daucus carota L.) was examined. We hypothesized that the addition of ethylenediaminetetraacetic acid (EDTA), dry algal powder (Spirulina platensis or Chlorella vulgaris), and Salix alba leaves powder would function as chelators for harmful contaminants in wastewater. The findings showed that irrigation of carrot plants with the sampled untreated wastewater led to significant decreases in the shoot lengths, fresh, dry weights of shoots and roots at stage I, the diameter of roots, pigment content, carotenoids, total soluble carbohydrate content, and soluble protein content. Furthermore, a significantly increased level of proline, total phenols, and the activities of polyphenol oxidase (PPO), peroxidase (POX), superoxide dismutase (SOD), and catalase (CAT) was identified in stage I samples. In contrast to the stage I, the length of the roots, the number of leaves on each plant, wet and dry weights of the stage II roots were all greatly enhanced. In spite of the increased yield due to the wastewater irrigation, carrot roots irrigated with wastewater had significantly more cadmium (Cd), nickel (Ni), cobalt (Co), and lead (Pb) than is considered safe. Our data clearly show that the application of Spirulina platensis, Chlorella vulgaris, EDTA, and leaves powder of salix was able to alleviate the toxicity of wastewater on carrot plants. For example, we recorded a significant decrease in the accumulation of carrot's Cd, Ni, Co, and Pb contents. We conclude that the treatments with Spirulina platensis and Chlorella vulgaris can be utilized as eco-friendly tools to lessen the damaging effects of wastewater irrigation on carrot plants.

Do Nanoparticles of Calcium Disodium EDTA Minimize the Toxic Effects of Cadmium in Female Rats?

The present study aims to investigate the ability of CaNa2EDTA (ethylenediaminetetraacetic acid) macroparticles and nanoparticles to treat cadmium-induced toxicity in female rats and to compare their efficacies. Forty rats were divided into 4 equal groups: control, cadmium, cadmium + CaNa2EDTA macroparticles and Cd + CaNa2EDTA nanoparticles. Cadmium was added to the drinking water in a concentration of 30 ppm for 10 weeks. CaNa2EDTA macroparticles and nanoparticles (50 mg/kg) were intraperitoneally injected during the last 4 weeks of the exposure period. Every two weeks, blood and urine samples were collected for determination of urea, creatinine, metallothionein and cadmium concentrations. At the end of the experiment, the skeleton of rats was examined by X-ray and tissue samples from the kidney and femur bone were collected and subjected to histopathological examination. Exposure to cadmium increased the concentrations of urea and creatinine in the serum and the concentrations of metallothionein and cadmium in serum and urine of rats. A decrease in bone mineralization by X-ray examination in addition to various histopathological alterations in the kidney and femur bone of Cd-intoxicated rats were also observed. Treatment with both CaNa2EDTA macroparticles and nanoparticles ameliorated the toxic effects induced by cadmium on the kidney and bone. However, CaNa2EDTA nanoparticles showed a superior efficacy compared to the macroparticles and therefore can be used as an effective chelating antidote for treatment of cadmium toxicity.

Exploring the Influence of Spacers in EDTA-ß-Cyclodextrin Dendrimers: Physicochemical Properties and In Vitro Biological Behavior.

The synthesis of a new family of ethylenediaminetetraacetic acid (EDTA) core dimers and G0 dendrimers end-capped with two and four ß-cyclodextrin (ßCD) moieties was performed by click-chemistry conjugation, varying the spacers attached to the core. The structure analyses were achieved in DMSO-d6 and the self-inclusion process was studied in D2O by 1H-NMR spectroscopy for all platforms. It was demonstrated that the interaction with adamantane carboxylic acid (AdCOOH) results in a guest-induced shift of the self-inclusion effect, demonstrating the full host ability of the ßCD units in these new platforms without any influence of the spacer. The results of the quantitative size and water solubility measurements demonstrated the equivalence between the novel EDTA-ßCD platforms and the classical PAMAM-ßCD dendrimer. Finally, we determined the toxicity for all EDTA-ßCD platforms in four different cell lines: two human breast cancer cells (MCF-7 and MDA-MB-231), human cervical adenocarcinoma cancer cells (HeLa), and human lung adenocarcinoma cells (SK-LU-1). The new EDTA-ßCD carriers did not present any cytotoxicity in the tested cell lines, which showed that these new classes of platforms are promising candidates for drug delivery.

Intra Nasal Use of Ethylene Diamine Tetra Acetic Acid for Improving Olfactory Dysfunction Post COVID-19.

BACKGROUND: COVID-19 has been associated with olfactory dysfunction in many infected patients. The rise of calcium levels in the nasal secretions plays an essential role in the olfaction process with a desensitization effect on the olfactory receptor neurons and a negative impact on the olfaction transmission. Ethylene diamine tetra acetic acid (EDTA) is a chelating agent that can bind free calcium in the nasal secretions, thereby reducing the adverse effects of calcium on olfactory function. OBJECTIVES: The objective of this work is to demonstrate the effect of intranasal EDTA on improving olfactory dysfunction following COVID-19. METHODS: Fifty patients with a history of COVID-19 and olfactory dysfunction that persisted for more than 6 months were enrolled in the current prospective randomized clinical trial. Participants were randomized into 2 equal groups. Twenty-five patients were treated with olfactory training only, while the remaining 25 patients received treatment with olfactory training and a topical nasal spray of ethylene diamine tetra acetic acid. The olfactory function was assessed before treatment and 3 months later using the Sniffin' Sticks test. Additionally, the determination of calcium level in the nasal secretions was performed using an ion-selective electrode before treatment and 3 months later. RESULTS: Eighty-eight percent of the patients treated with olfactory training in addition to EDTA exhibited clinical improvement, while 60% showed improvement in patients treated with olfactory training only. Furthermore, a significant decrease in the measured calcium level in the nasal secretions was demonstrated after the use of ethylene diamine tetra compared to patients treated with olfactory training only. CONCLUSION: Ethylene diamine tetra acetic acid may be associated with an improvement of the olfactory function post-COVID-19.

Impact of ethylene diamine tetraacetic acid on physiochemical parameters and yield attribute in two varieties of Brassica juncea under lead stress.

Lead (Pb) is one of the most toxic elements on earth. The harmful effects of Pb at higher concentrations were seen on plant vegetation because plants are directly exposed towards it. Indian mustard, a well-known hyperaccumulator plant is the most promising crop for the environment, engaged in a variety of scenarios for ecological cleanup. In the present study, we used ethylene diamine tetraacetic acid (EDTA), a chelating agent that is of remarkable efficiency. The pot experiments were conducted in soil pretreated with 1000 mgkg-1 Pb with different concentrations of EDTA (2-10 mmol). All the growth parameters were reduced significantly in the plants treated with Pb and EDTA, however, a non-significant effect was observed in 5 mmol EDTA compared to Pb alone treatment. Photosynthetic pigments yield, nitrate reductase activity and NPK content were affected negatively; in contrast, superoxide dismutase and catalase activity was increased in Pb and Pb+EDTA treated in both the varieties. The Pb accumulation was elevated significantly by the augmentation of 5 mmol EDTA in both varieties. Accumulation of Pb in the shoot was higher in PM 25 than in P. Vijay, whereas root Pb accumulation showed the opposite, i.e., more Pb in roots of P. Vijay than PM 25. Moreover, The Pb accumulation per plant was observed more in P. Vijay as compared to PM 25. Hence, the present study implies that the augmentation of Pb-polluted soil with EDTA works well while dealing with B. juncea assisted phytoremediation and P. Vijay to be a stronger variety than PM 25. Further, 5 mmol of EDTA was optimum for phytoremediation of the soil polluted with up to 1000 mg Pb kg-1 soil.

A Method for Eliminating Fibroblast Contamination in Mouse and Human Primary Corneal Epithelial Cell Cultures.

PURPOSE: This study was designed to determine if previous approaches to eliminate fibroblast contamination in different cells types would be successful in eliminating fibroblast contamination from human and mouse primary corneal epithelial cell cultures, with the primary goal being to describe a simple, easy, and effective method to culture fibroblast-free primary mouse and human corneal epithelial cell cultures. METHODS: Primary human and mouse corneal stromal cells and epithelial cells were isolated and cultured from human corneal rims and mouse corneas, respectively. Several approaches previously used in other tissue types were evaluated using corneal epithelial cells and mixtures of fibroblasts and epithelial cells to determine the most effective purification method. Methods evaluated included 0.25% trypsin-EDTA, low temperature, mitomycin-C, and dispase. Degree of fibroblast contamination was examined using light microscopy evaluation of cell phenotype, immunofluorescence and western blotting using cell type-specific markers. Anti-pancytokeratin (PanCK) was used as the epithelial immunofluorescence label, and anti-α smooth muscle actin (αSMA) as the fibroblast immunofluorescence label. Epithelial western blot antibodies included PanCK, keratin 12, and E-cadherin, while αSMA, collagen 1A1 and collagen 3A1 were used to identify fibroblasts. RESULTS: Fibroblast contamination of human and mouse primary cornea epithelial cell cultures was best controlled using the 0.25% trypsin-EDTA method. The other methods examined were not effective at eliminating cornea fibroblast contamination. CONCLUSIONS: Trypsin-EDTA digestion is a simple and effective method for controlling fibroblast contamination of cultured primary human and mouse corneal epithelial cells.

Pretreatment of Root Dentin Using Photon-Induced Photoacoustic Streaming, Photodynamic Therapy, and Riboflavin with EDTA as a Final Irrigant to Improve Bond Integrity of Glass Fiber Post.

Objective: To evaluate the outcome of post space disinfection using Er,Cr:YSGG laser (ECYL), riboflavin photosensitizer (RFP), and photon-induced photoacoustic streaming (PIPS) along with ethylene diamine tetra acetic acid (EDTA) as final irrigant on the bond values of glass fiber post (GFP) to the canal wall. Materials and methods: Forty human mandibular premolars were subjected to root canal treatment after decoronation till the cementoenamel junction. Post space was prepared using a Gates-Glidden drill leaving 4 mm of gutta perch at the apical third of the root section. Random allocation of samples was performed into four groups based on the post space disinfection (n = 10 each): Group 1: 1% RFP and 17% EDTA, group 2: 5.25% sodium hypochlorite (NaOCl) and 17% EDTA (control), group 3: PIPS and 17% EDTA, and group 4: ECYL and 17% EDTA. GFP was luted using Rely X Unicem. A universal testing machine and stereomicroscope were used to assess the push-out bond strength (PBS) and failure mode, respectively. Analysis of variance determined the PBS amid different experimental groups at three root levels. Multiple group comparison using means of tested groups was identified using post hoc Tukey. Results: The coronal third of group 2 (5.25% NaOCl +17% EDTA) demonstrated the highest PBS (8.83 ± 0.11 MPa). However, the lowest bond integrity (4.15 ± 0.52 MPa) was displayed in an apical third of group 1 (RFP +17% EDTA). The intergroup comparison revealed that 5.25% NaOCl +17% EDTA at all three levels demonstrated comparable outcomes of PBS with PIPS +17% EDTA and ECYL +17% EDTA specimens (p > 0.05). However, riboflavin +17% EDTA demonstrated the lowest bond strength (p < 0.05). Conclusions: Er,Cr: YSGG and PIPS in combination with 17% EDTA demonstrated comparable outcomes of bond score with those of conventional gold standard irrigation regime 5.25% NaOCl.

Effect of iron nanoparticles and conventional sources of Fe on growth, physiology and nutrient accumulation in wheat plants grown on normal and salt-affected soils.

Salt stress is becoming a serious problem for the global environment and agricultural sector. Different sources of iron (Fe) can provide an eco-friendly solution to remediate salt-affected soils. The Fe nanoparticles (FeNPs) and conventional sources of Fe (iron-ethylene diamine tetra acetic acid; Fe-EDTA; and iron sulfate; FeSO4) were used to evaluate their effects on wheat crop grown in normal and salt-affected soils. Application of FeNPs (25 mg/kg) on normal soil increased the dry weights of wheat roots, shoots, and grains by 46%, 59%, and 77%, respectively. In salt-affected soil, FeNPs increased the dry weights of wheat roots, shoots, and grains by 65%, 78%, and 61%, respectively. The application of FeSO4 and Fe-EDTA increased the growth parameters of wheat in both normal and salt-affected soils compared to the respective controls. The photosynthetic parameters, including chlorophyll a (50%), chlorophyll b (67%), carotenoids (62%), and total chlorophyll contents (50%), were increased with the application of FeNPs under salt stress. The FeNPs increased plant-essential nutrients like iron, zinc, calcium, magnesium, and potassium in both normal and salt-affected soils. The experiment revealed that the application of Fe plays a significant role in enhancing the growth of wheat on alkaline normal and salt-affected soils. Maximum growth response was recorded with FeNPs than other Fe sources. The future must be focused on long term field experiments to economize the application of FeNPs on a large scale for commercialization.

Effects of EDTA and saline as the final irrigation in regenerative endodontic procedures on the migration, proliferation, and differentiation of human stem cells from the apical papilla.

OBJECTIVES: To evaluate the effect of EDTA and saline as the final irrigation in regenerative endodontic procedures (REPS) on the attachment, proliferation, migration, and differentiation of stem cells from the apical papilla (SCAPs). MATERIALS AND METHODS: Dentin specimens from 140 human third molars were irrigated with various protocols-group 1: normal sterile saline (NSS), group 2: EDTA, group 3: EDTA then 5 mL NSS, or group 4: EDTA then 20 mL NSS. The specimens were used in cell assays. For cell proliferation, SCAPs were seeded on dentin, and the cell viability on days 1, 3, and 7 was determined using an MTT assay. At day 3, the attached cells' morphology was observed using SEM, and cell migration was investigated using a transwell migration assay. The ALP activity and odonto/osteogenic differentiation gene expression were evaluated at days 7, 14, and 21 using an ALP activity assay and RT-qPCR. RESULTS: On days 3 and 7, group 4 demonstrated more viable cells than group 1 (p < 0.01). The amount of migrated cells in groups 2, 3, and 4 was greater compared with group 1 (p < 0.05). Moreover, SCAP differentiation was similar between groups. CONCLUSIONS: Irrigating dentin with EDTA alone or with EDTA then NSS promoted SCAP migration. However, a final irrigation with 20 mL NSS after EDTA promoted SCAP proliferation without affecting their differentiation. CLINICAL RELEVANCE: When using a blood clot as a scaffold, a final flushing with 20 mL NSS after EDTA could be beneficial for clinical REP protocols.

Effect of farmyard manure, elemental sulphur and EDTA on growth and phytoextraction of cadmium by spider plants (Chlorophytum comosum L.) under Cd stress.

Cadmium (Cd) contamination is considered as a widespread concern at global scale which is serious threats to human health. Phytoremediation is an eco-friendly approach which can remove or immobilize Cd from the soil. Different organic and inorganic amendments can potentially enhance Cd phytoremediation efficiency but the comparison of farmyard manure (FM), elemental sulphur (S) and ethylenediaminetetraacetic acid (EDTA) for Cd phytoremediation through spider plants (Chlorophytum comosum L.) remained unanswered. The present study evaluated the efficiency of S (0.1 and 0.2%), EDTA (0.1 and 0.2%, represented as EDTA-0.1 and EDTA-0.2) and FM (0.5 and 1%, represented as FM-0.5 and FM-1) for remediation of Cd contaminated soils (50 and 100 mg kg-1, represented as Cd-50 and Cd-100) through spider plants. Results depicted that the highest shoots and roots dry biomass was found in FM treated plants followed by S, EDTA and control except in EDTA-0.2 treatment in which the lowest values of these parameters were observed. Application of FM-1 significantly increased the shoot dry weight (120%), root dry weight (99%), as well as photosynthetic attributes in Cd-50 as compared to control. Application of EDTA-0.2 increased the bioavailable fraction of Cd than control and the maximum increase was observed in Cd-100. The highest Cd concentrations in shoot and roots were found in EDTA treated plants followed by S, control and FM irrespective of Cd and amendment levels. Maximum Cd in roots (109%) and shoots (156%) was recorded in plants grown in Cd-100 with EDTA-0.2 than control. The maximum bioaccumulation factor, translocation index, harvest index and root to shoot translocation were observed with EDTA than control and other treatments. EDTA along with spider plants may enhance the uptake of Cd but lower biomass production in the highest dose of EDTA may questioned the efficiency of EDTA.

Effect of chitosan irrigant solutions on the release of bioactive proteins from root dentin.

OBJECTIVE: To identify the effect of two chitosan solutions on the release of root dentin matrix proteins and to describe the chemical changes observed following conditioning with chelating agents. MATERIALS AND METHODS: The release of dentin sialoprotein (DSP), transforming growth factor-beta 1 (TGF-ß1), vascular endothelial growth factor (VEGF), and platelet-derived growth factor-BB (PDGF-BB) with different chelating agents, including ethylenediaminetetraacetic acid (EDTA), chitosan solution (CS), and nanoparticulate chitosan (CSnp), was investigated. DSP was quantified using an enzyme-linked immunosorbent assay (ELISA). TGF-ß1, VEGF, and PDGF-BB were quantified using a cytokine bead panel (CBA). Raman spectroscopy was performed to identify surface chemical changes. Statistical analysis was performed using Kruskal-Wallis test with Mann-Whitney-Wilcoxon rank-sum test (p < 0.05). RESULTS: TGF-ß1, VEGF, and DSP solubilized in all irrigants tested. CSnp showed the highest concentration of DSP. PDGF-BB did not exceed the detection limits. Raman spectroscopy revealed a decrease in the phosphate and carbonate peaks, representing the chelating effect of EDTA, CS, and CSnp. Additionally, CSnp showed the greatest preservation of the amide I and III content. CONCLUSION: Proteins can be released from dentin via EDTA, CS, and CSnp conditioning. Raman spectroscopic revealed changes in the inorganic content of the root dentin after chelation. Furthermore, use of CSnp facilitated a preservation of the organic content. CLINICAL RELEVANCE: Chelation allows the release of proteins, justifying the use of chelating agents in regenerative endodontics. The chitosan-dentin matrix interaction also promotes the protection of the organic content as an additional benefit to its protein releasing effect.

Characterization of pulp calcifications and changes in their composition after treatments with citric acid and ethylenediaminetetraacetic acid solutions.

The ectopic calcifications of non-mineralized tissues can occur in several forms throughout life, such as pulpal calcification. The presence of pulp stones is a challenge in endodontic treatment because they partially or fully obliterate the pulp chamber hindering access to root canals and their subsequent shaping. This study aimed to determine their crystallographic properties and evaluate the capacity of citric acid (CA) and ethylenediaminetetraacetic acid (EDTA) to promote the demineralization of pulp calcifications. The samples were obtained from patients with indications of endodontic treatment, and the radiographic examination was suggestive of pulp stone in at least one permanent tooth. The samples were isolated and analyzed by scanning electron microscopy/energy-dispersive x-ray spectroscopy (SEM/EDX). The Fourier Transform by high resolution-transmission electron microscopy, Raman microscopy, and X-ray diffraction (XRD) were used to identify the mineral phase and crystallographic characteristics. To evaluate the effect of CA and EDTA on the crystallinity of calcifications, they were submerged into these two individual solutions and the changes were assessed in situ by Raman spectroscopy. The SEM images obtained from calcifications demonstrated irregular morphologies. EDX of sample surfaces shows a high presence of oxygen, carbon, calcium, and phosphorous, however, other elements such as sodium, magnesium, nitrogen, chlorine, potassium, sulfur, and zinc were identified in less quantity. According to Raman, XRD, and high-resolution transmission electron microscopy, the predominant mineral phase identified in the pulpal calcification was a poor crystallinity apatite. According to in situ analyses, the effect of CA and EDTA was observed on the signals of PO4 3- and CH2 groups corresponding to inorganic and organic components. The changes with CA were evident at 7 min while the effect of EDTA was observed until 15 min of treatment. All results indicate that pulp stones have a heterogeneous composition principally composed of apatite with low crystallinity. The solubility of these pathological minerals is adequate using solutions such as EDTA or CA; however, the effectivity depends on the mineralization grade of calcifications, time, and concentration of exposition to this chemical.

Effects of malic acid and EDTA on oxidative stress and antioxidant enzymes of okra (Abelmoschus esculentus L.) exposed to cadmium stress.

Environmental stresses, including heavy metal pollution, are increasing at a growing rate and influencing arable lands. Chelators play an essential role in several biochemical pathways in the cells of plants treated with heavy metals. This research evaluated the modifying effect of malic acid (MA) and ethylenediaminetetraacetic acid (EDTA) on the physiological and biochemical parameters of okra plants exposed to Cd stress in which the okra plants were cultivated in hydroponic conditions. At the 4-leaf stage, they were applied with the treatments of cadmium nitrate at three levels (0, 50, and 100 mg/L), EDTA and MA at two levels (0.5 and 1 mM), and Cd + EDTA + MA at different rates for one month. The harvested plants were subjected to the measurement of the physico-biochemical factors. The results revealed that the application of Cd alone reduced leaf area (up to 21.57 %), and dissolved sugars (up to 40.51 % in the shoot and 45.19 % in the root) and increased MDA (up to 66.37 % and 76.43 % in the shoot and root, respectively), H2O2 (up to 67.14 % and 53.28 % in the shoot and root, respectively), proline (up to 52.04 % and 40.93 % in the shoot and root, respectively), and dissolved proteins (up to 14.59 % and 21.90 % in the shoot and root, respectively) contents in both shoots and roots whereas the application of MA and EDTA to the Cd-treated plants increased their leaf area and dissolved sugars and reduced MDA, H2O2, proline, and dissolved proteins content. The antioxidant enzymes, e.g., superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX), were significantly higher in the plants treated with MA, EDTA, and Cd, but the application of MA and EDTA to the Cd-treated plants reduced the activity of all these antioxidant enzymes versus the plants only treated with Cd. MA and EDTA are likely to prevent the accumulation of Cd in the cytosol by binding to it and transferring it into vacuoles, thereby mitigating Cd toxicity in the okra plants.

The Impact of the Anticoagulant Type in Blood Collection Tubes on Circulating Extracellular Plasma MicroRNA Profiles Revealed by Small RNA Sequencing.

Pre-analytical factors have a significant influence on circulating microRNA (miRNA) profiling. The aim of this study was a comprehensive assessment of the impact of the anticoagulant type in blood collection tubes on circulating plasma miRNA profiles using small RNA sequencing. Blood from ten healthy participants (five males and five females from 25 to 40 years old) was taken in collection tubes with four different anticoagulants: acid citrate dextrose (ACD-B), sodium citrate, citrate-theophylline-adenosine-dipyridamole (CTAD) and dipotassium-ethylenediaminetetraacetic acid (K2 EDTA). Platelet-free plasma samples were obtained by double centrifugation. EDTA plasma samples had elevated levels of hemolysis compared to samples obtained using other anticoagulants. Small RNA was extracted from plasma samples and small RNA sequencing was performed on the Illumina NextSeq 500 system. A total of 30 samples had been successfully sequenced starting from ~1 M reads mapped to miRNAs, allowing us to analyze their diversity and isoform content. The principal component analysis showed that the EDTA samples have distinct circulating plasma miRNA profiles compared to samples obtained using other anticoagulants. We selected 50 miRNA species that were differentially expressed between the sample groups based on the type of anticoagulant. We found that the EDTA samples had elevated levels of miRNAs which are abundant in red blood cells (RBC) and associated with hemolysis, while the levels of some platelet-specific miRNAs in these samples were lowered. The ratio between RBC-derived and platelet-derived miRNAs differed between the EDTA samples and other sample groups, which was validated by quantitative PCR. This study provides full plasma miRNA profiles of 10 healthy adults, compares them with previous studies and shows that the profile of circulating miRNAs in the EDTA plasma samples is altered primarily due to an increased level of hemolysis.

Iron Source and Medium pH Affect Nutrient Uptake and Pigment Content in Petunia hybrida 'Madness Red' Cultured In Vitro.

Deficiency or excess of iron (Fe) and improper medium pH will inhibit the growth and development of plants, reduce the transfer and utilization of energy from the root to the leaf, and affect the utilization efficiency of inorganic nutrients. The most common symptom of Fe deficiency in plants is chlorosis of the young leaves. In this study, the effects of the iron source, in combination with the medium pH, on plant growth and development, plant pigment synthesis, and nutrient uptake in a model plant Petunia hybrida cultured in vitro were investigated. Iron sulfate (FeSO4·7H2O) or iron chelated with ethylenediaminetetraacetic acid (Fe-EDTA) were supplemented to the MNS (a multipurpose nutrient solution) medium at a concentration of 2.78 mg·L-1 Fe, and the treatment without any Fe was used as the control. The pH of the agar-solidified medium was adjusted to either 4.70, 5.70, or 6.70 before autoclaving. The experiment was carried out in an environmentally controlled culture room with a temperature of 24 °C with 100 µmol·m-2·s-1 photosynthetic photon flux density (PPFD) supplied by white light emitting diodes (LEDs) during a photoperiod of 16 h a day, 18 °C for 8 h a day in the dark, and 70% relative humidity. Regardless of the Fe source including the control, the greatest number of leaves was observed at pH 4.70. However, the greatest lengths of the leaf and root were observed in the treatment with Fe-EDTA combined with pH 5.70. The contents of the chlorophyll, carotenoid, and anthocyanin decreased with increasing medium pH, and contents of these plant pigments were positively correlated with the leaf color. The highest soluble protein content and activities of APX and CAT were observed in the Fe-EDTA under pH 5.70. However, the GPX activity was the highest in the control under pH 4.70. In addition, the highest contents of ammonium (NH4+) and nitrate (NO3-) were measured in the FeSO4-4.7 and EDTA-5.7, respectively. More than that, the treatment of Fe-EDTA combined with pH 5.70 (EDTA-5.7) enhanced nutrient absorption, as proven by the highest tissue contents of P, K, Ca, Mg, Fe, and Mn. The genes' ferric reduction oxidase 1 and 8 (PhFRO1 and PhFRO8), iron-regulated transporter 1 (PhIRT1), nitrate transporter 2.5 (PhNRT2.5), and deoxyhypusine synthase (PhDHS) were expressed at the highest levels in this treatment as well. In the treatment of EDTA-5.7, the reduction and transport of chelated iron in P. hybrida leaves were enhanced, which also affected the transport of nitrate and catalyzed chlorophyll level in leaves. In conclusion, when the medium pH was adjusted to 5.70, supplementation of chelated Fe-EDTA was more conducive to promoting the growth and development of, and absorption of mineral nutrients by, the plant and the expression of related genes in the leaves.

[Effect of Chelating Agents and Organic Acids on Remediation of Cadmium and Arsenic Complex Contaminated Soil Using Xanthium sibiricum].

In order to improve the phytoextraction efficiency of Xanthium sibiricum on farmland soil that had been contaminated by Cd and As, this study explored the effects of chelating agents and organic acids (EDTA, SAP, CA, and MA) on the extraction of Cd and As heavy metals using X. sibiricum. The results showed that the four different chelating agents and organic acids had little effect on the biomass of the roots, stems, and leaves of X. sibiricum. However, they had different effects on the concentrations and accumulation of Cd and As in various organs of X. sibiricum. Compared with the those in the CK treatment, EDTA, SAP, CA, and MA significantly increased the Cd concentrations in the leaves of X. sibiricum by 44.1%, 32.4%, 41.2%, and 38.2% and the As concentrations in the roots of X. sibiricum by 89.6%, 7.4%, 94.8%, and 61.5%, respectively. The four treatments (EDTA, SAP, CA, and MA) improved the total Cd accumulation of X. sibiricum, with increasing ranges, respectively, of 70.2%, 29.4%, 28.9%, and 33.1%, and the As accumulation increased by 67.0%, 19.6%, 81.9%, and 40.8%, respectively, compared with that of the CK treatment. The four chelating agents and organic acids had different effects on the Cd and As bioconcentration factor and transfer factor of various organs of X. sibiricum. Treatments with EDTA, SAP, CA, and MA resulted in a decrease of 32.7%-38.2% in soil Cd concentrations and a decrease of 14.6%-20.5% in soil As concentrations. These four chelating agents can be used for enhancing the efficiency of extraction Cd and As heavy metals by X. sibiricum.

Ethylenediaminetetraacetic acid (EDTA) enhances cAMP production in human TDAG8-expressing cells.

Ethylenediaminetetraacetic acid (EDTA) is a chelating agent that binds tightly to metal ions. We found that cAMP response element (CRE)-driven promoter activity by protons was enhanced by EDTA in human T-cell death-associated gene 8 (TDAG8)-overexpressed HEK293T cells. The enhancing action by EDTA was also detected by proton-induced cAMP production that is located upstream from the CRE-driven promoter activity even at physiological proton concentration pH7.4. The proton-induced CRE-driven promoter activity was not enhanced by other chelating agents, ethylene glycol tetraacetic acid (EGTA) and sodium citrate. The enhanced CRE-driven promoter activity by EDTA was not attenuated by increasing the extracellular calcium ion concentration. These results indicate that the EDTA-enhancing action may not be due to its chelating action but might rather be another EDTA-specific effect. Enhanced cAMP production by EDTA was also detected in a human leukemia cell line HL-60, in which TDAG8 and OGR1 (ovarian cancer G-protein-coupled receptor 1) were endogenously expressed, suggesting that the medical use of EDTA would influence the physiological and pathophysiological functions of hematopoietic cells.

Priming with EDTA, IAA and Fe alleviates Pb toxicity in Trigonella Foneum graecum L. growth: Phytochemicals and secondary metabolites.

This study evaluated the effects of the exogenous application of ethylenediaminetetraacetic acid (EDTA), indole-3-acetic acid (IAA) and iron sulfate (FeSO4) upon the phytochemical mechanisms of fenugreek grown under Pb-excess (2000 mg L-1 PbCl2). The results showed that chemical additives of EDTA and IAA as well as FeSO4 improved fenugreek germination parameters. The radicle length and the amylase activity were significantly improved under IAA treatment compared to EDTA and FeSO4. Exogenous FeSO4 was more effective to improving growth parameters. Moreover, the decrease in hydrogen peroxide (H2O2) and malondialdehyde (MDA) levels was noted under all chemical additives especially under IAA application. In addition, it was more effective than EDTA and Fe in increasing catalase, glutathione (GSH), ascorbate peroxidase (APX), flavonoids and phenols while the increment superoxide dismutase (SOD) production was more pronounced under EDTA addition to Pb than other chelators. HPLC analysis revealed that the gallic was the major phenol produced under all chelators addition especially with IAA. In addition, the syringic acid was only produced with exogenous IAA while the quercetin was only detected under EDTA addition. Our results exhibited a higher IAA efficiency than EDTA and FeSO4 in mitigating Pb stress in fenugreek through up-regulated mechanisms of the antioxidant system for reducing reactive oxygen species (ROS) activities and enhancing special phenols.