Comparative Analysis of Alternative Calcium Chelators for the Treatment of Calcific Band Keratopathy.

PURPOSE: In this study, we compared clinically relevant biochemical properties of each chelator for pH, osmolarity, and calcium chelation potential. METHODS: In total, 0.2 M K 2 EDTA and K 3 EDTA (BD vacutainer tubes by Becton, Dickinson and Company) and Na 2 EDTA (Sigma Aldrich) solutions were made. The pH of each solution was measured (Mettler Toledo pH meter), and the theoretical osmolarity was calculated. Next, we determined the calcium chelation potential of each EDTA salt by titrating it with 10 µmol of calcium hydroxyapatite or CaCl 2 containing Patton-Reeder colorimetric indicator. Statistical significance was analyzed using analysis of variance. RESULTS: The 0.2 M solutions of Na 2 EDTA, K 2 EDTA, and K 3 EDTA have pH values of 4.43, 5.71, and 9.191 and theoretical osmolarities of 600, 600, and 800 mOsm/L, respectively. Calcium chelation ability was similar among all 3 solutions: 0.94 to 0.98 mol of EDTA was needed to fully chelate 1 mol calcium ions of CaCl 2 ( P = 0.296), 0.100 to 0.108 mol of EDTA for 1 mol calcium ions of the hydroxyapatite aqueous suspension ( P = 0.296), and 0.992 to 0.996 mol for 1 mol calcium ions of hydroxyapatite in acidic solution ( P = 0.178). Compared with the clinical standard of 3% (30 mg/mL) Na 2 EDTA, approximately 3.3% (33 mg/mL) K 2 EDTA and 3.6% (36 mg/mL) K 3 EDTA are needed to chelate an equivalent amount of calcium. CONCLUSIONS: In this article, we provide clinically relevant biochemical properties of 2 alternatives to Na 2 EDTA and demonstrate comparable calcium chelation ability among all 3 solutions. In situations where sterile sources of Na 2 EDTA are unavailable, potassium EDTA may provide a convenient and equally effective method of treatment for band keratopathy.

Iron oxide-EDTA nanoparticles for chelation-enhanced chemodynamic therapy and ion interference therapy.

As an emerging anti-tumor strategy, chemodynamic therapy (CDT) utilizes a Fenton/Fenton-like reaction to generate highly toxic hydroxyl radicals to kill tumor cells. However, the efficiency of CDT is still hindered by the low Fenton/Fenton-like reaction rate. Herein, we report the combination of ion interference therapy (IIT) and chemodynamic therapy (CDT) via an amorphous iron oxide (AIO) nanomedicine with encapsulated EDTA-2Na (EDTA). Iron ions and EDTA are released from the nanomedicine in acidic tumors and chelate to form iron ion-EDTA, which improves the efficiency of CDT and promotes the generation of reactive oxygen species (ROS). In addition, EDTA can disrupt the homeostasis of Ca2+ in tumor cells by chelating with Ca2+ ions, which induces the separation of tumor cells and affects normal physiological activities. Both in vitro and in vivo experiments show that the nano chelating drugs exhibit significant improvement in Fenton reaction performance and excellent anti-tumor activity. This study based on chelation provides a new idea for designing efficient catalysts to enhance the Fenton reaction and provides more revelations on future research on CDT.

Exceptional Response of Rare Plasmacytoid Variant Prostate Cancer Post 177Lu-PSMA Therapy Seen on 68Ga-PSMA PET/CT.

ABSTRACT: Plasmacytoid is a rare variant of acinar prostatic adenocarcinoma. The aggressive type is characterized by an aggressive clinical course, lack of responsiveness to hormonal therapies, and an overall poor prognosis. Here we present pretherapy and posttherapy 68Ga-PSMA PET/CT images showing an exceptional response to 177Lu-PSMA therapy. This case demonstrates the usefulness of both 68Ga-PSMA PET/CT in assessing the tumor PSMA avidity and the potential of 177Lu-PSMA therapy in these patients.

Effectiveness of root canal irrigation and dressing for the treatment of apical periodontitis: A systematic review and meta-analysis of clinical trials.

BACKGROUND: Sodium hypochlorite (NaOCl) and ethylenediaminetetraacetic acid (EDTA) and/or calcium hydroxide (Ca(OH)2 ) are commonly used during root canal treatment. Evaluation of their effectiveness regarding clinical and patient-related outcomes requires further understanding. OBJECTIVES: To assess the effectiveness of root canal irrigation and dressing for the treatment of teeth with apical periodontitis (AP). METHODS: A search was conducted in the PubMed-MEDLINE, Scopus, EMBASE, Google scholar databases and available repositories, followed by hand searches, until July 2021. Eligibility criteria followed the a priori formulated Population, Intervention, Comparator, Outcomes, Timing, and Study design (PICOTS) framework. Clinical studies restricted to English language were included. The revised Cochrane risk-of-bias tool for randomised trials (RoB 2) was used to assess the quality of included studies. Meta-analyses were performed using the fixed-effect model to obtain Risk Ratio (RR) and 95% Confidence Interval (CI), with sensitivity analysis. Overall quality of evidence of meta-analyses was assessed through the Grading of Recommendations Assessment, Development, and Evaluation tool. RESULTS: The search identified 1357 records of which six fulfilled the inclusion criteria, providing data for 'irrigation' from 212 teeth and for 'dressings' from 438 teeth. Two studies reported no significant difference regarding the outcome 'pain at 7 days' using 2% chlorhexidine vs. 5.25% NaOCl and EDTA or after using different concentrations of NaOCl (1% vs. 5%). No significant difference was detected between different NaOCl concentrations regarding the reduction of AP. A meta-analysis was possible for the comparison of single-visit (SV) versus multiple-visits including the use of Ca(OH)2 demonstrating a significant effect in favour of SV (RR: 1.10; 95% CI: 1.03-1.19; p = .007; I2  = 0). RoB of included studies was moderate to low. DISCUSSION: The use of Ca(OH)2 for the treatment of AP may not be beneficial. There is scarce or no evidence fulfilling the proposed PICOTS regarding irrigants and dressings. CONCLUSIONS: There is moderate certainty that SV treatment is associated with better radiographic evidence of normal periodontal ligament space (strict criteria) compared with the use of Ca(OH)2. Reduction of AP is comparable after irrigation with 1% and 5% NaOCl, whereas postoperative pain at 7 days for the irrigants assessed is similar. REGISTRATION: PROSPERO database CRD42021260271.

USAGE OF CHLORHEXIDINE AS A FINAL IRRIGANT IN ONE-VISIT ROOT CANAL TREATMENT IN COMPARISON WITH CONVENTIONAL TWO-VISIT ROOT CANAL TREATMENT IN MANDIBULAR MOLARS: A RANDOMIZED CLINICAL TRIAL.

OBJECTIVES: The effectiveness of 1-visit root canal treatment (RCT), especially for teeth with large periapical lesions, are still being debated. Although, the use of chlorhexidine (CHX) as a final irrigant have been recommended to provide an optimized approach in 1-visit RCT, there is limited knowledge about the outcome of 1-visit RCT using CHX as the final irrigant. AIM: The aim of this randomized clinical trial was to evaluate the healing of mandibular molar teeth with large periapical lesions after RCT that was completed in 1 visit using 2% CHX as a final irrigant, comparing the results with a conventional 2-visit RCT as a control group. METHODOLOGY: This study is a prospective, parallel-designed, and single-blinded randomized clinical trial. One-hundred asymptomatic mandibular molar teeth with large periapical lesions were randomly assigned to 2 groups. All teeth were instrumented with Reciproc files with 2.5% sodium hypochloride (NaOCl) and 5% ethylenediaminetetraacetic acid (EDTA) as irrigants. In the intervention group, half of the teeth were treated in 1 visit, receiving an additional final rinse with 2% CHX before obturation; the other half of the teeth, however, were treated in 2 visits with calcium hydroxide (CH) dressings, in the control group. All root canals were obturated with Reciproc gutta-percha cones and AH Plus root canal sealer. All patients were recalled and investigated clinically and radiographically for 48 months. Periapical healing was evaluated statistically using the Mann-Whitney U test, followed by the Friedman and the Wilcoxon tests (α = 0.05). RESULTS: During the 48-month follow-up process, 86 teeth (44 in 1-visit and 42 in 2-visit group) were examined. There were no significant differences between the 2 groups regarding the radiographic healing rates (1-visit group 91% and 2-visit group 88%) (P > .05). CONCLUSION: In this study, 1-visit RCT using 2% CHX as final irrigant in asymptomatic mandibular molars with large periapical lesions produced favorable healing results, similar to 2-visit root canal treatment with calcium hydroxide.

Use of tetrasodium EDTA acid for the treatment of intraluminal obstruction of subcutaneous ureteral bypass devices.

OBJECTIVES: The aim of this study was to evaluate the efficacy and tolerability of a 4% tetrasodium EDTA (tEDTA) infusion protocol in the subcutaneous ureteral bypass (SUB) devices of cats with intraluminal obstruction at a veterinary teaching hospital between July 2017 and April 2020. METHODS: This was a retrospective controlled study. Cats with an obstructed SUB device underwent a 4% tEDTA infusion protocol. Obstruction of the device was diagnosed based on renal pelvic dilation, dilatation of the ureter, mineralized material within the device (cystostomy or nephrostomy catheters) seen on ultrasound, the absence of visible bubbles within the renal pelvis and/or urinary bladder following ultrasound-guided flushing of the device with saline. RESULTS: A total of 16 tEDTA infusion protocols were performed in 14 cats. The infusion protocol was considered successful in 11/16 SUB devices (68.8%). Six devices (n = 6/11; 54.5%) had recurrence of obstruction with a median time of 87 days. One or more episodes of self-limiting pollakiuria and/or hematuria following infusion was seen in eight patients (n = 8/14; 57.1%). CONCLUSIONS AND RELEVANCE: Infusions of 4% tEDTA successfully relieved intraluminal obstruction in patients with occluded SUB devices; however, the recurrence of obstruction was common. Additional studies evaluating case selection and optimal protocols are warranted.

Comparison of Ferric Sodium EDTA in Combination with Vitamin C, Folic Acid, Copper Gluconate, Zinc Gluconate, and Selenomethionine as Therapeutic Option for Chronic Kidney Disease Patients with Improvement in Inflammatory Status.

Anemia is one of the most frequent and earliest complications of chronic kidney disease (CKD), which impacts a patient's quality of life and increases the risk of adverse clinical outcomes. Patients' inflammatory status is strictly related to the occurrence of functional iron deficiency anemia (IDA) because this causes an increase in hepcidin levels with the consequent inhibition of iron absorption and release from cellular stores into blood circulation. The aim of this study was to evaluate the use of the new oral formulation based on ferric sodium EDTA in combination with vitamin C, folic acid, copper gluconate, zinc gluconate, and selenomethionine (Ferachel Forte®) in patients with moderate CKD and functional IDA, analyzing the inflammatory status in addition to iron blood parameters, in comparison with oral ferrous sulfate and liposomal iron therapies. Sixty-two elderly patients were randomly allocated to one of the following oral treatments for 6 months: ferrous sulfate (Group 1; N = 20), ferric sodium EDTA in combination (Group 2; N = 22), and ferric liposomal formulation (Group 3; N = 20). The evaluated parameters included iron profile parameters of hemoglobin (Hb), sideremia, ferritin, transferrin saturation, C-reactive protein (CRP), and hepcidin. The results showed that in Group 1, there were no improvements. In Group 2, there were statistically significant (p < 0.001) improvements in all evaluated parameters. Finally, in Group 3, there were significant improvements in all evaluated parameters except for hepcidin, which was less than that of Group 2 patients. In conclusion, the findings showed the superior efficacy of the formulation based on ferric sodium EDTA over the other oral iron sources, and that this formulation can contribute to reducing the systemic inflammatory status in patients with CKD.

The trial to assess chelation therapy 2 (TACT2): Rationale and design.

BACKGROUND: Intravenous edetate disodium-based infusions reduced cardiovascular events in a prior clinical trial. The Trial to Assess Chelation Therapy 2 (TACT2) will replicate the initial study design. METHODS: TACT2 is an NIH-sponsored, randomized, 2x2 factorial, double masked, placebo-controlled, multicenter clinical trial testing 40 weekly infusions of a multi-component edetate disodium (disodium ethylenediamine tetra-acetic acid, or Na2EDTA)-based chelation solution and twice daily oral, high-dose multivitamin and mineral supplements in patients with diabetes and a prior myocardial infarction (MI). TACT2 completed enrollment of 1000 subjects in December 2020, and infusions in December 2021. Subjects are followed for 2.5 to 5 years. The primary endpoint is time to first occurrence of all-cause mortality, MI, stroke, coronary revascularization, or hospitalization for unstable angina. The trial has >;85% power to detect a 30% relative reduction in the primary endpoint. TACT2 also includes a Trace Metals and Biorepository Core Lab, to test whether benefits of treatment, if present, are due to chelation of lead and cadmium from patients. Design features of TACT2 were chosen to replicate selected features of the first TACT, which demonstrated a significant reduction in cardiovascular outcomes in the EDTA chelation arm compared with placebo among patients with a prior MI, with the largest effect in patients with diabetes. RESULTS: Results are expected in 2024. CONCLUSION: TACT2 may provide definitive evidence of the benefit of edetate disodiumbased chelation on cardiovascular outcomes, as well as the clinical importance of longitudinal changes in toxic metal levels of participants.

Chelation Therapy in Patients With Cardiovascular Disease: A Systematic Review.

Background EDTA is an intravenous chelating agent with high affinity to divalent cations (lead, cadmium, and calcium) that may be beneficial in the treatment of cardiovascular disease (CVD). Although a large randomized clinical trial showed benefit, smaller studies were inconsistent. We conducted a systematic review of published studies to examine the effect of repeated EDTA on clinical outcomes in adults with CVD. Methods and Results We searched 3 databases (MEDLINE, Embase, and Cochrane) from database inception to October 2021 to identify all studies involving EDTA treatment in patients with CVD. Predetermined outcomes included mortality, disease severity, plasma biomarkers of disease chronicity, and quality of life. Twenty-four studies (4 randomized clinical trials, 15 prospective before/after studies, and 5 retrospective case series) assessed the use of repeated EDTA chelation treatment in patients with preexistent CVD. Of these, 17 studies (1 randomized clinical trial) found improvement in their respective outcomes following EDTA treatment. The largest improvements were observed in studies with high prevalence of participants with diabetes and/or severe occlusive arterial disease. A meta-analysis conducted with 4 studies reporting ankle-brachial index indicated an improvement of 0.08 (95% CI, 0.06-0.09) from baseline. Conclusions Overall, 17 studies suggested improved outcomes, 5 reported no statistically significant effect of treatment, and 2 reported no qualitative benefit. Repeated EDTA for CVD treatment may provide more benefit to patients with diabetes and severe peripheral arterial disease. Differences across infusion regimens, including dosage, solution components, and number of infusions, limit comparisons across studies. Additional research is necessary to confirm these findings and to evaluate the potential mediating role of metals. Registration URL: https://www.crd.york.ac.uk/; Unique identifier: CRD42020166505.

Attenuation of ischemia-reperfusion injury by intracoronary chelating agent administration.

Ischemia-reperfusion (IR) injury accelerates myocardial injury sustained during the myocardial ischemic period and thus abrogates the benefit of reperfusion therapy in patients with acute myocardial infarction. We investigated the efficacy of intracoronary ethylenediaminetetraacetic acid (EDTA) administration as an adjunctive treatment to coronary intervention to reduce IR injury in a swine model. We occluded the left anterior descending artery for 1 h. From the time of reperfusion, we infused 50 mL of EDTA-based chelating agent via the coronary artery in the EDTA group and normal saline in the control group. IR injury was identified by myocardial edema on echocardiography. Tetrazolium chloride assay revealed that the infarct size was significantly lower in the EDTA group than in the control group, and the salvage percentage was higher. Electron microscopy demonstrated that the mitochondrial loss in the cardiomyocytes of the infarcted area was significantly lower in the EDTA group than in the control group. Echocardiography after 4 weeks showed that the remodeling of the left ventricle was significantly less in the EDTA group than in the control group: end-diastolic dimension 38.8 ± 3.3 mm vs. 43.9 ± 3.7 mm (n = 10, p = 0.0089). Left ventricular ejection fraction was higher in the EDTA group (45.3 ± 10.3 vs. 34.4 ± 11.8, n = 10, respectively, p = 0.031). In a swine model, intracoronary administration of an EDTA chelating agent reduced infarct size, mitochondrial damage, and post-infarct remodeling. This result warrants further clinical study evaluating the efficacy of the EDTA chelating agent in patients with ST-segment elevation myocardial infarction.

Effect of Citric Acid Versus EDTA on Radiographic Root Development in Regenerative Endodontic Treatment: An Animal Study.

INTRODUCTION: Regenerative endodontic treatment was introduced with the aim of reinforcing the root and enabling further root development. In the process of dentin mineralization, bioactive molecules are entrapped in the dentin matrix, which can be released later by rinsing the dentin with chelating agents. This study aimed to compare the effects of citric acid and EDTA on radiographic root development in regenerative endodontic treatment in an animal model. METHODS: Premolars of 3 beagle dogs were used in this study. The teeth at 1 side of the jaw were irrigated with EDTA according to the regeneration protocol of the American Association of Endodontists. The teeth in the other quadrant were irrigated with citric acid in the second treatment session. Follow-up radiographs were obtained at 6 months, and the primary and final data were analyzed in SPSS Version 23 (IBM Corp, Armonk, NY) using the analysis of covariance. RESULTS: There was no significant difference between the 2 irrigating solutions regarding the increase in root length (P = .668); however, EDTA showed significantly superior efficacy in increasing the root dentin thickness compared with citric acid (P = .022). CONCLUSIONS: Within the limitations of this study, the use of citric acid versus EDTA was not superior on radiographic root development in regenerative endodontic treatment.

Excimer laser-assisted phototherapeutic keratectomies combined to EDTA chelation for the treatment of calcific band keratopathy.

INTRODUCTION: Calcific band keratopathy (CBK) is a relatively common chronic corneal degeneration and various forms of treatment are mentioned in the literature. CASES DESCRIPTION: Two patients (89 and 37 yo, respectively) affected by diffuse long-standing CBK in one eye and complaining of ocular pain, foreign body sensation and decreased visual acuity are reported. An ethylenediaminetetraacetic acid (EDTA) application on the ocular surface was performed associated with a customized no-touch transepithelial phototherapeutic corneal remodeling in one patient and a standard phototherapeutic keratectomy (PTK) in the second patient. Corneal transparency progressively improved in both cases since the early follow-up visits and the cornea became clear 2 weeks after surgery. In both cases, a significant reduction of ocular discomfort was reported. CONCLUSIONS: Combining EDTA chelation and excimer laser-assisted PTK represents an useful treatment of band keratopathy even in challenging cases and may help regularize corneal surface and improve corneal clarity.

Effect of 24% EDTA root conditioning on the outcome of modified coronally advanced tunnel technique with subepithelial connective tissue graft for the treatment of multiple gingival recessions: a randomized clinical trial.

OBJECTIVES: To investigate effects of root conditioning with 24% ethylenediaminetetraacetic acid (EDTA) on the 12-month outcomes after treatment of multiple gingival recessions (GR) with modified coronally advanced tunnel (MCAT) and subepithelial connective tissue graft (SCTG). MATERIALS AND METHODS: Twenty patients with 142 GR were treated (72 test sites: SCTG + EDTA and 70 control sites: SCTG). Average and complete root coverage (ARC, CRC), gain in keratinized tissue width (KTW), gain in gingival thickness (GT), root esthetic coverage score (RES), and patient-reported outcome measures (PROMs) were evaluated at 12 months post-operatively. RESULTS: Differences between pre- and post-operative values were statistically significant only within but not between treatment modalities. At 12 months, ARC was 86.0% for SCTG + EDTA-treated and 84.6 for SCTG-treated defects (p = 0.6636). CRC was observed in 90.2% (tests) and 91.4% (controls) of all cases (p = 0.9903). Professional assessment of esthetic outcomes using RES showed highly positive results reaching the value of 8.9 in case of test sites and 8.7 for control sites (p = 0.3358). Severity of pain and swelling did not differ between sites, regardless of whether EDTA was used. CONCLUSIONS: Test and control sites presented similarly positive outcomes related to root coverage, periodontal and esthetic parameters, and patient satisfaction and self-reported morbidity with no statistical differences between them 12 months after surgery. No significant differences in evaluated variables were observed between sites treated with and without 24% EDTA. Clinical relevance Considering the limitations of the present study, the use of 24% EDTA for root conditioning did not improve 12-month outcomes after treatment of multiple RT1 and RT2 gingival recessions with MCAT and SCTG. Trial registration ClinicalTrials.gov identifier: NCT03354104.

Development, evaluation and biodistribution of stealth liposomes of 5-fluorouracil for effective treatment of breast cancer.

The current research was undertaken to design stealth liposomes of 5-Fluorouracil for reducing its cardiotoxicity and prolong the half-life by developing long-circulating liposomes. The liposomes were prepared by the NH4EDTA gradient method, where EDTA is used as a cardioprotectant. Ascorbyl-6-palmitate was also used which helped for the synergistic effect of 5-Fluorouracil to counteract the cancer cells and provide promising application in the treatment of breast cancer cells. Taguchi design was used for screening of formulation and HSPC phospholipid was selected. The drug-excipient compatibility was checked through FTIR which showed all the excipients were compatible with the drug. The formulation was optimized by using 32 factorial design. The drug to lipid ratio (1:5) and Ascorbyl-6-Palmitate concentration (15 mg) were selected. The vesicle size of the prepared liposomes was found to be 70.12 ± 0.58 nm and uniform distribution was observed. The zeta potential and entrapment efficiency of the stealth liposomes were found -16.28 mV and 92 ± 0.007% respectively. In-vitro drug release study of formulation showed drug release of 63.50 ± 0.94% in 24 hrs. The formulation was sterilized by 0.22 µm Mixed cellulose esters (MCE) membrane filter and passed sterility test. Moreover, a biodistribution study was performed by Fluorescence microscopy and by HPLC method, which showed formulation was circulated for 24 hours. Finally, a cell line study indicated that prepared formulation possess greater anti-tumour activity. The cardiotoxicity study revealed that the stealth liposomes have minimum cardiotoxicity as compare to the plain drug.

Novel colistin-EDTA combination for successful eradication of colistin-resistant Klebsiella pneumoniae catheter-related biofilm infections.

Development of an effective therapy to overcome colistin resistance in Klebsiella pneumoniae, a common pathogen causing catheter-related biofilm infections in vascular catheters, has become a serious therapeutic challenge that must be addressed urgently. Although colistin and EDTA have successful roles for eradicating biofilms, no in vitro and in vivo studies have investigated their efficacy in catheter-related biofilm infections of colistin-resistant K. pneumoniae. In this study, colistin resistance was significantly reversed in both planktonic and mature biofilms of colistin-resistant K. pneumoniae by a combination of colistin (0.25-1 µg/ml) with EDTA (12 mg/ml). This novel colistin-EDTA combination was also demonstrated to have potent efficacy in eradicating colistin-resistant K. pneumoniae catheter-related biofilm infections, and eliminating the risk of recurrence in vivo. Furthermore, this study revealed significant therapeutic efficacy of colistin-EDTA combination in reducing bacterial load in internal organs, lowering serum creatinine, and protecting treated mice from mortality. Altered in vivo expression of different virulence genes indicate bacterial adaptive responses to survive in hostile environments under different treatments. According to these data discovered in this study, a novel colistin-EDTA combination provides favorable efficacy and safety for successful eradication of colistin-resistant K. pneumonia catheter-related biofilm infections.

Simplified, Readily Available Method for the Treatment of Band Keratopathy With Ethylenediaminetetraacetic Acid.

PURPOSE: To compare 3 methods for creating ethylenediaminetetraacetic acid (EDTA) solution using readily available Vacutainer tubes for the treatment of band keratopathy. METHODS: All 3 protocols used commercially available Vacutainer blood collection tubes coated with K2EDTA. An osmometer was used to measure and compare the concentration of EDTA created using 3 different protocols. The time required for preparation of the solution was measured and compared to evaluate its efficiency for everyday clinical use. In addition, volume of EDTA solution obtained was measured for method 1. The most promising protocol for clinical use was then used for treatment of a series of patients. RESULTS: Average osmolarity was 532, 285, and 422 for methods 1, 2, and 3, respectively (ANOVA P < 0.01, all Tukey honestly significant difference P < 0.01). For the respective mixtures, average concentration was 65, 35, and 52 mg/mL, and average time to create solution was 189, 38, and 83 seconds (ANOVA P < 0.01, all Tukey honestly significant difference P < 0.01). The most promising, method 3, was found to be safe and effective in removing calcium from the corneal stroma in a series of 5 patients with 6 eyes treated. It also yielded 25% more solution for clinical use than method 1. CONCLUSIONS: Method 3 using a single 10-mL Vacutainer tube with 18 mg of K2-EDTA had the best balance of effective concentration of EDTA, time to preparation, and simplicity of methodology, when compared with previously published methods 1 and 2. It also yielded a greater final volume of solution.

SARS-CoV-2 attachment to host cells is possibly mediated via RGD-integrin interaction in a calcium-dependent manner and suggests pulmonary EDTA chelation therapy as a novel treatment for COVID 19.

SARS-CoV-2 is a highly contagious virus that has caused serious health crisis world-wide resulting into a pandemic situation. As per the literature, the SARS-CoV-2 is known to exploit humanACE2 receptors (similar toprevious SARS-CoV-1) for gaining entry into the host cell for invasion, infection, multiplication and pathogenesis. However, considering the higher infectivity of SARS-CoV-2 along with the complex etiology and pathophysiological outcomes seen in COVID-19 patients, it seems that there may be an alternate receptor for SARS-CoV-2. I performed comparative protein sequence analysis, database based gene expression profiling, bioinformatics based molecular docking using authentic tools and techniques for unveiling the molecular basis of high infectivity of SARS-CoV-2 as compared to previous known coronaviruses. My study revealed that SARS-CoV-2 (previously known as 2019-nCoV) harbors a RGD motif in its receptor binding domain (RBD) and the motif is absent in all other previously known SARS-CoVs. The RGD motif is well known for its role in cell-attachment and cell-adhesion. My hypothesis is that the SARS-CoV-2 may be (via RGD) exploiting integrins, that have high expression in lungs and all other vital organs, for invading host cells. However, an experimental verification is required. The expression of ACE2, which is a known receptor for SARS-CoV-2, was found to be negligible in lungs. I assume that higher infectivity of SARS-CoV-2 could be due to this RGD-integrin mediated acquired cell-adhesive property. Gene expression profiling revealed that expression of integrins is significantly high in lung cells, in particular αvß6, α5ß1, αvß8 and an ECM protein, ICAM1. The molecular docking experiment showed the RBD of spike protein binds with integrins precisely at RGD motif in a similar manner as a synthetic RGD peptide binds to integrins as found by other researchers. SARS-CoV-2 spike protein has a number of phosphorylation sites that can induce cAMP, PKC, Tyr signaling pathways. These pathways either activate calcium ion channels or get activated by calcium. In fact, integrins have calcium & metal binding sites that were predicted around and in vicinity of RGD-integrin docking site in our analysis which suggests that RGD-integrins interaction possibly occurs in calcium-dependent manner. The higher expression of integrins in lungs along with their previously known high binding affinity (~KD = 4.0 nM) for virus RGD motif could serve as a possible explanation for high infectivity of SARS-CoV-2. On the contrary, human ACE2 has lower expression in lungs and its high binding affinity (~KD = 15 nM) for spike RBD alone could not manifest significant virus-host attachment. This suggests that besides human ACE2, an additional or alternate receptor for SARS-CoV-2 is likely to exist. A highly relevant evidence never reported earlier which corroborate in favor of RGD-integrins mediated virus-host attachment is an unleashed cytokine storm which causes due to activation of TNF-α and IL-6 activation; and integrins role in their activation is also well established. Altogether, the current study has highlighted possible role of calcium and other divalent ions in RGD-integrins interaction for virus invasion into host cells and suggested that lowering divalent ion in lungs could avert virus-host cells attachment.

Evaluation on curative effects of ethylene diamine tetra-acetic acid chelation therapy in treating with atherosclerotic cardiovascular disease: A protocol for systematic review and meta-analysis.

BACKGROUND: Ethylene diamine tetra-acetic acid (EDTA) is a chelating agent which attach to metals such as calcium and enables their elimination. In particular, some researchers suggest chelation with EDTA to treat cardiovascular disease with the hypothesis of moderating calcium to decrease atherosclerotic calcification of arteries. However, chelation with EDTA therapeutic effects in atherosclerotic cardiovascular disease is still unclear. Therefore, we propose to undertake a meta-analysis to assess the curative effects of EDTA chelation therapy in patients with atherosclerotic cardiovascular disease. METHODS: In the current study, we set to perform a systematic literature search using the electronic databases of 4 most commonly used English databases (EMBASE, MEDLINE, Cochrane Library, and ClinicalTrials.gov trials register), as well as 3 most commonly employed Chinese databases (China Nation Knowledge Infrastructure, Wan Fang, and VIP), from the date of database inception until September 30, 2020 to identify relevant randomized controlled studies of the evaluation on curative effects of EDTA chelation therapy in patients with atherosclerotic cardiovascular disease. In the study, 2 authors worked independently to screen search results, chose studies for inclusion, then they extracted pertinent data to evaluate and study quality based on Cochrane Risk of Bias Tool V.2.0. Additionally, we will address discrepancies by consultation with a third author. We also intend to use pooled risk ratio (RR) and pooled mean difference (MD) or pooled standardized mean difference (SMD) with 95% confidence intervals (CI) to approximate the relative strength of curative effects of EDTA chelation therapy in patients with atherosclerotic cardiovascular disease. RESULTS: The results of the current study will systematically assess curative effects of EDTA chelation therapy in patients with atherosclerotic cardiovascular disease. CONCLUSION: The study will infer the currently published evidence to evaluate curative effects of EDTA chelation therapy in patients with atherosclerotic cardiovascular disease, which might be beneficial to these patients. ETHICS AND DISSEMINATION: The present study is a systematic review, hence the pooled results are founded upon the published evidence. Therefore, ethical approval is not necessary for the study. OPEN SCIENCE FRAMEWORK REGISTRATION NUMBER: October 20, 2020.osf.io/tvmk8. (https://osf.io/tvmk8/).