Experimental and Theoretical Investigations of the Fragmentation of Ethylenediamine Induced by Low-Energy (<10 eV) Electrons.

Ethylenediamine is industrially used as an intermediate for the fabrication of many products. The development of new methodologies for synthesis compatible with the environment and sustainability, such as cold plasma processes, implicates reactions induced by nonthermal electrons. In this contribution, we study the interaction of low-energy (<10 eV) electrons with ethylenediamine. We show that electrons induce the fragmentation of the molecule into various anion fragments and associated neutral counterparts via dissociative electron attachment. The fragmentation mechanisms and energetics are discussed in the frame of DFT calculations. The fragmentation processes are quantified by the estimation of the cross sections and the branching ratios for competitive accessible dissociation routes.

Synthesis of Polyethyleneimines from the Manganese-Catalysed Coupling of Ethylene Glycol and Ethylenediamine.

Polyethyleneimines find many applications in products such as detergents, adhesives, cosmetics, and for processes such as tissue culture, gene therapy, and CO2 capture. The current state-of-the-art technology for the production of the branched polyethyleneimines involves aziridine feedstock which is a highly toxic, volatile and mutagenic chemical and raises significant concern to human health and environment. We report here a novel method for the synthesis of branched polyethyleneimine derivative from ethylene glycol and ethylenediamine which are much safer, environmentally benign, commercially available and potentially renewable feedstock. The polymerisation reaction is catalysed by a complex of an earth-abundant metal, manganese and liberates H2 O as the only by-product. Our mechanistic studies using a combination of DFT computation and experiment suggest that the reaction proceeds by the formation and subsequent hydrogenation of imine intermediates.

Renewable Schiff-Base Ionic Liquids for Lignocellulosic Biomass Pretreatment.

Growing interest in sustainable sources of chemicals and energy from renewable and reliable sources has stimulated the design and synthesis of renewable Schiff-base (iminium) ionic liquids (ILs) to replace fossil-derived ILs. In this study, we report on the synthesis of three unique iminium-acetate ILs from lignin-derived aldehyde for a sustainable "future" lignocellulosic biorefinery. The synthesized ILs contained only imines or imines along with amines in their structure; the ILs with only imines group exhibited better pretreatment efficacy, achieving >89% sugar release. Various analytical and computational tools were employed to understand the pretreatment efficacy of these ILs. This is the first study to demonstrate the ease of synthesis of these renewable ILs, and therefore, opens the door for a new class of "Schiff-base ILs" for further investigation that could also be designed to be task specific.

Integrated LC-MS metabolomics with dual derivatization for quantification of FFAs in fecal samples of hepatocellular carcinoma patients.

FFAs display pleiotropic functions in human diseases. Short-chain FAs (SCFAs), medium-chain FAs, and long-chain FAs are derived from different origins, and precise quantification of these FFAs is critical for revealing their roles in biological processes. However, accessing stable isotope-labeled internal standards is difficult, and different chain lengths of FFAs challenge the chromatographic coverage. Here, we developed a metabolomics strategy to analyze FFAs based on isotope-free LC-MS-multiple reaction monitoring integrated with dual derivatization. Samples and dual derivatization internal standards were synthesized using 2-dimethylaminoethylamine or dansyl hydrazine as a "light" label and N,N-diethyl ethylene diamine or N,N-diethyldansulfonyl hydrazide as a "heavy" label under mild and efficient reaction conditions. General multiple reaction monitoring parameters were designed to analyze these FFAs. The limit of detection of SCFAs varied from 0.5 to 3 nM. Furthermore, we show that this approach exhibits good linearity (R2 = 0.99374-0.99929), there is no serious substrate interference, and no quench steps are required, confirming the feasibility and reliability of the method. Using this method, we successfully quantified 15 types of SCFAs in fecal samples from hepatocellular carcinoma patients and healthy individuals; among these, propionate, butyrate, isobutyrate, and 2-methylbutyrate were significantly decreased in the hepatocellular carcinoma group compared with the healthy control group. These results indicate that the integrated LC-MS metabolomics with isotope-free and dual derivatization is an efficient approach for quantifying FFAs, which may be useful for identifying lipid biomarkers of cancer.

A phosphoproteomics study reveals a defined genetic program for neural lineage commitment of neural stem cells induced by olfactory ensheathing cell-conditioned medium.

Since both Olfactory ensheathing cells (OECs) and neural stem cells (NSCs) have shown certain efficacy in the cellular therapy of nerve injury and disease, there have been a series of investigations in recent years looking at the co-culture of NSCs and OECs. Protein phosphorylation forms the basis for identifying a variety of cellular signaling pathways responsible for regulating the self-renewal and differentiation of NSCs induced by OECs. To better understand the signaling cascades in the early phases of OEC-induced NSC differentiation, changes in the NSC proteome and phosphoproteome during the first 24 h were determined using dimethyl labeling and TiO2 phosphorylation enrichment coupled with Liquid chromatography-tandem mass spectrometry (LC-MS/MS). A total of 565 proteins and 2511 phosphorylation sites were identified. According to quantitative phosphoproteomics analyses of NSC differentiation induced by OECs during the first 12 and 24 h, it was speculated that there were at least two different signal waves: one peaking within 12 h after stimulation and the second upsurge after 24 h. In addition to understanding the dynamics of the proteome and phosphoproteome in the early stages of NSC differentiation, our analyses identified a key role of the TGF-ß3 protein secreted by OECs, which may be an initiating factor that promotes differentiation of NSCs into neurons induced by OECs. These findings not only redemonstrated a OECs-based therapeutic strategy in cell therapy, but also added a node to the regulatory network for the neural lineage commitment of NSCs induced by OECs.

Amperometric determination of As(III) and Cd(II) using a platinum electrode modified with acetylcholinesterase, ruthenium(II)-tris(bipyridine) and graphene oxide.

The authors describe an amperometric biosensor for the determination As(III) and Cd(II) based on the inhibition of the enzyme acetylcholineesterase (AChE). A platinum electrode was modified with ruthenium(II)-tris(bipyridyl), graphene oxide and AChE and then showed redox peaks at 0.06 and 0.2 V vs Ag/AgCl in the presence of acetylthiocholine chloride (ATChCl). Amperometry unveiled a steady-state turnover rate with the release of thiocholine. In the presence of arsenic(III) and cadmium(II), AChE showed an inhibitive response at 0.214 and 0.233 V vs Ag/AgCl, respectively. The electrode exhibits a detection limit and linear range of 0.03 µM and 0.05-0.8 µM for As(III) and 0.07 µM and 0.02-0.7 µM for Cd(II), respectively. Type of inhibition and inhibition constants induced by As(III) and Cd(II) on the catalytic sites of AChE were determined from Dixon and Lineweaver-Burk plots. The modified electrode was applied to the determination of As3+ and Cd2+ in river, tap and waste water, and the results proved that the method is sensitive and can be an alternative to chromatographic and spectroscopic techniques. Graphical abstract Schematic presentation of Pt/Ru(II)-tris(bipy)-GO/AChE electrode in absence and presence of metal ions (As3+/Cd2+).

Magnetic nanoparticles modified with organic dendrimers containing methyl methacrylate and ethylene diamine for the microextraction of rosuvastatin.

Magnetic nanoparticles (MNPs) modified with organic dendrimers are shown to be a viable sorbent of the microextraction of the drug rosuvastatin (RST; also known as Crestor). The MNPs were prepared from iron(II) chloride and iron(III) chloride and then coated with silicon dioxide. The coated MNPs produced by this method have diameters ranging from 10 to 60 nm according to scanning electron microscopy. The MNPs were further modified with organic dendrimers containing methyl methacrylate and ethylene diamine. The resulting MNPs were characterized by SEM, Fourier transform infra-red and thermal gravimetry analysis. Then, the efficacy of the modified MNPs with respect to the extraction of RST was studied. The adsorption of RST by MNPs can be best described by a Langmuir isotherm. Following elution with buffer, RST was quantified by HPLC. The method was applied to the determination of RST in (spiked) human blood plasma, urine, and in tablets. RST extraction efficiencies are 54.5% in plasma, 86.6% from the drug matrix, and 94.3% in urine. The highest adsorption capacity of the RST by the MNPs adsorbent was 61 mg⋅g-1. Graphical abstract Co-precipitation was used to synthesize magnetic nanoparticles (MNPs). They were coated with a layer of SiO2 and then branched by organic dendrimers containing methyl methacrylate (MMA) and ethylene diamine (EDA). Rosuvastatin (RST) drug was trapped between dendrimer branches, therefore adsorption capacity of the drug was strongly increased.

Discovery and Correction of Spurious Low Platelet Counts due to EDTA-Dependent Pseudothrombocytopenia.

BACKGROUND: Ethylene diamine tetraacetic acid dependent pseudothrombocytopenia (EDTA-PTCP) is a laboratory artifact that may lead to unnecessary evaluation and treatment of patients. The purpose of this article is to discuss how to identify EDTA-PTCP and correct spurious low platelet counts in clinical laboratories. METHODS: We use two criteria to screen for platelet aggregation: (1) an abnormal platelet count in EDTA-treated blood from a patient lacking clinical signs of a platelet disorder, and (2) an instrument flag for platelet clumps. EDTA-PTCP was confirmed by microscopic examination for platelet agglutination and by platelet counts that corrected with citrate sample. In addition, the time course of EDTA-PTCP was investigated in samples from 26 patients anticoagulated with EDTA-K2 and sodium citrate. Amikacin (5 mg/ml) was added to tubes with EDTA-K2 or sodium citrate from seven additional cases in order to confirm its dissociative effect on platelet aggregation. RESULTS: In our laboratory, the overall incidence of EDTA-PTCP was approximately 0.09%; and the duration was between 2 weeks and 6 months. EDTA-PTCP was time-dependent and occurred as early as 10 min after sample collection. Weaker agglutination could also occur in most corresponding citrate-treated samples. The dissociative effect of amikacin on platelet agglutination was case-specific and not concentration-dependent. CONCLUSIONS: The method of screening for platelet clumping with the help of XE5000 images is convenient. The decline in the platelet count is related to the length of time and the intensity of chelation. Amikacin supplement is not always effective for correcting platelet counts in vitro.

Transfer Hydrogenation Employing Ethylene Diamine Bisborane in Water and Pd- and Ru-Nanoparticles in Ionic Liquids.

Herein we demonstrate the use of ethylenediamine bisborane (EDAB) as a suitable hydrogen source for transfer hydrogenation reactions on C-C double bonds mediated by metal nanoparticles. Moreover, EDAB also acts as a reducing agent for carbonyl functionalities in water under metal-free conditions.

Cytochrome bd-I in Escherichia coli is less sensitive than cytochromes bd-II or bo'' to inhibition by the carbon monoxide-releasing molecule, CORM-3: N-acetylcysteine reduces CO-RM uptake and inhibition of respiration.

BACKGROUND: CO-releasing molecules (CO-RMs) are potential therapeutic agents, able to deliver CO - a critical gasotransmitter - in biological environments. CO-RMs are also effective antimicrobial agents; although the mechanisms of action are poorly defined, haem-containing terminal oxidases are primary targets. Nevertheless, it is clear from several studies that the effects of CO-RMs on biological systems are frequently not adequately explained by the release of CO: CO-RMs are generally more potent inhibitors than is CO gas and other effects of the molecules are evident. METHODS: Because sensitivity to CO-RMs cannot be predicted by sensitivity to CO gas, we assess the differential susceptibilities of strains, each expressing only one of the three terminal oxidases of E. coli - cytochrome bd-I, cytochrome bd-II and cytochrome bo', to inhibition by CORM-3. We present the first sensitive measurement of the oxygen affinity of cytochrome bd-II (Km 0.24µM) employing globin deoxygenation. Finally, we investigate the way(s) in which thiol compounds abolish the inhibitory effects of CORM-2 and CORM-3 on respiration, growth and viability, a phenomenon that is well documented, but poorly understood. RESULTS: We show that a strain expressing cytochrome bd-I as the sole oxidase is least susceptible to inhibition by CORM-3 in its growth and respiration of both intact cells and membranes. Growth studies show that cytochrome bd-II has similar CORM-3 sensitivity to cytochrome bo'. Cytochromes bo' and bd-II also have considerably lower affinities for oxygen than bd-I. We show that the ability of N-acetylcysteine to abrogate the toxic effects of CO-RMs is not attributable to its antioxidant effects, or prevention of CO targeting to the oxidases, but may be largely due to the inhibition of CO-RM uptake by bacterial cells. CONCLUSIONS: A strain expressing cytochrome bd-I as the sole terminal oxidase is least susceptible to inhibition by CORM-3. N-acetylcysteine is a potent inhibitor of CO-RM uptake by E. coli. GENERAL SIGNIFICANCE: Rational design and exploitation of CO-RMs require a fundamental understanding of their activity. CO and CO-RMs have multifaceted effects on mammalian and microbial cells; here we show that the quinol oxidases of E. coli are differentially sensitive to CORM-3. This article is part of a Special Issue entitled: Oxygen Binding and Sensing Proteins.

Influence of smear layer treatment on resistance to root fracture in tooth restored with epoxy fiber post.

To evaluate the influence of complete smear layer removal by EDTA on the resistance of root fracture of tooth restored with fiber post. Twenty single rooted premolars with average root length 16 ± 1 mm were randomly divided into two groups of ten each. All teeth were obturated with guttapercha after root canal preparation, sectioned 4 mm above the cement-enamel junction. In group I, teeth root canals were etched with 37 % phosphoric acid, fiber post cemented with self adhesive resin cement. In group II, teeth radicular dentin was treated with 17 % EDTA, followed by fiber post cementation with dual cure resin cement. Coronal part of the teeth reestablished with posterior composite and NiCr full veneer coping along with 2 mm circumferential ferrule. Restored teeth were loaded with incremental force at 30° to the long axis of the root until fracture occurred. The data obtained was evaluated with paired sample T test to compare the fracture resistance between the test groups. Group I had the mean failure load of 1,883.70 N while group II had 1,622.80 N mean failure load. Paired sample T test showed statistically significant (p value = 0.033) failure load difference between the groups tested. Within the limits of this study, it may be concluded that: the complete removal of smear layer by EDTA found to have reduced the fracture strength of tooth roots restored with an epoxy fiber post.

Synergistic effects of EDTA and lysozyme on the properties of hydroxypropyl starch nano antibacterial films.

Hydroxypropyl starch (HPS) nano antibacterial films incorporating Ethylene Diamine Tetraacetic Acid (EDTA) and lysozyme (LY) were fabricated via solvent casting method. The synergistic effects of EDTA and LY on the microstructure, component interactions, color, optical, mechanical, barrier and antibacterial properties of HPS nano antibacterial films were evaluated. The results indicated that EDTA and LY were well dispersed in the matrix of the HPS nano antibacterial films, the film-forming substrates have good compatibility, resulting in a dense multi-layer structure of the HPS nano antibacterial films. The addition of EDTA and LY increased the color parameters (L*, a*, b* and △E*) of the HPS nano antibacterial films. The synergistic effects of EDTA and LY significantly decreased the light transmission of the HPS nano antibacterial films. The presence of EDTA and LY increased the tensile strength (TS) and the elongation at break (EAB) of the HPS nano antibacterial films. The TS and EAB of E2.5L1 reached the highest values of 6.329 MPa and 50.24 %, respectively. The incorporation of EDTA and LY had positive effects on the improvement of water vapor permeability (WVP) and oxygen permeability (OP). The WVP and OP of E2.5L1 reached the highest values of 0.9350 × 10-12 g cm/cm2•s•Pa and 0.297 × 10 -2 g m/m2 •d, respectively. In addition, EDTA and LY had significant synergistic effects on the antibacterial activity against S. aureus (Gram-positive bacteria) and E. coli (Gram-negative bacteria). E2.5L1 exhibited the highest antibacterial activity and the inhibition zone diameters of S. aureus and E. coli were 3.69 mm and 4.28 mm, respectively. The HPS nano antibacterial films incorporating EDTA and LY are potential functional packaging materials.

Reversal of heavy arterial calcification in a rat model of chronic kidney disease using targeted ethylene diamine tetraacetic acid-loaded albumin nanoparticles.

Background: Elastin degradation and severe calcification in the medial layer of the vessel wall, known as medial arterial calcification (MAC), is typical in the aging population and patients with metabolic disorders, such as diabetes and chronic kidney disease (CKD). We have previously reported that ethylene diamine tetraacetic acid (EDTA) delivery to the site of calcification can be achieved by tagging nanoparticles with an elastin antibody that recognizes explicitly damaged elastin, and such systemic therapy can remove focal calcium deposits from the calcified arteries in CKD rodent model. The current study aims to test whether heavy calcification seen throughout arterial tree and kidneys in CKD can be reversed with nanoparticle therapy. Methods: Thirty healthy male Sprague-Dawley rats weighing approximately 300 g, were placed on an adenine diet for 21 non-consecutive days to induce kidney failure, followed by daily vitamin D3 (VitD3) injections for 4 sequential days to cause severe calcification throughout the cardiovascular system and kidneys. DiR-dye loaded and elastin antibody conjugated albumin nanoparticles were used to confirm the targeting of nanoparticles to the calcification area. The rats were divided into two groups for targeted removal of calcification starting at day 7 of the last doses of VitD3. The experimental group received biweekly IV injections of anti-elastin antibody conjugated EDTA loaded human serum albumin nanoparticles (EDTA-HSA-El-Ab NPs), while the sham controls received blank nanoparticles (Blank-HSA-El-Ab NPs) (5 injections in total). Micro-computed tomography (microCT) was used to analyze the extent of calcification. Reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry studies were performed for osteogenic markers, including bone morphogenic protein 2 (BMP2), runt-related transcription factor 2 (RUNX2), and tissue non-specific alkaline phosphatase (TNAP). For comparison, aortic ring organ cultures from healthy rats were treated with high phosphate to induce calcification in vitro, and then they were treated with EDTA. Human calcified femoral arteries were also treated ex vivo with EDTA-HSA-EL-Ab NPs to test if nanoparticles remove heavy calcification. Results: EDTA-loaded nanoparticles that specifically target degraded elastin reversed existing heavy mineral deposits in arteries, as per elemental calcium analysis (124.161±34.410 µg Ca per mg of the dry aorta in Blank-HSA-El-Ab NPs vs. 100.520±19.131 µg in EDTA-HSA-El-Ab NPs group, P=0.04) and microCT (object volume, 129.001±37.785 vs. 29.815±24.169 mm3, P=0.0005). The reversal of aortic calcification was accompanied by a significant reduction of bone-associated mRNA expression of BMP2 and RUNX2 (P=0.01). Immunohistochemistry studies corroborated RT-PCR results, showing a reduction of BMP2 and RUNX2 stains in the vessel wall. The rat aortic ring culture study also showed similar results, where osteogenic genes (BMP2, RUNX2) and proteins (BMP2, RUNX2, TNAP) were suppressed upon reversal of calcification with EDTA (P=0.001). We also show ex vivo reversal of human femoral artery calcification by microCT (calcium intensity: untreated, 57.721±28.551 vs. day 6 of treatment, 5.441±3.615, P=0.01) by EDTA nanoparticle therapy. Conclusions: This is the first study showing the removal of calcium from heavily calcified arteries by using intravenous targeted EDTA therapy. Such therapy also reversed vascular smooth muscle cell osteoblastic transition and apoptosis in the arterial tissue, thereby potentially creating an environment for suitable tissue repair.

Repetitively dosed docetaxel and ¹5³samarium-EDTMP as an antitumor strategy for metastatic castration-resistant prostate cancer.

BACKGROUND: ß-emitting bone-seeking radiopharmaceuticals have historically been administered for pain palliation whereas docetaxel prolongs life in patients with metastatic castration-resistant prostate cancer (mCRPC). In combination, these agents simultaneously target the bone stroma and cancer cell to optimize antitumor effects. The toxicity and efficacy when each agent is combined at full, recommended doses, in a repetitive fashion is not well established. METHODS: Patients with progressive mCRPC and ≥ 3 bone lesions received (153) Sm-EDTMP (samarium-153 ethylene diamine tetramethylene phosphonate) at a dose of 1.0 mCi/kg every 9 weeks and docetaxel at a dose of 75 mg/m(2) every 3 weeks. In the absence of unacceptable toxicity, patients were allowed to continue additional cycles, defined by 9 weeks of treatment, until intolerance or biochemical/radiographic disease progression. RESULTS: Of the 30 patients treated, approximately 50% were considered to be taxane-naive, 36.7% were taxane-refractory, and 13.3% had previously been exposed to taxanes but were not considered refractory. Patients received on average 2.5 cycles of treatment (6.5 doses of docetaxel and 2.5 doses of (153) Sm-EDTMP). Twelve patients (40%) demonstrated a decline in their prostate-specific antigen level of ≥ 50%. The median progression-free survival (biochemical or radiographic) was 7.0 months and the overall survival was 14.3 months. Nine patients (30%) did not recover platelet counts >100 K/mm(3) after a median of 3 cycles to allow for additional treatment, with 4 patients experiencing prolonged thrombocytopenia. The most common reasons for trial discontinuation were progressive disease and hematologic toxicity. CONCLUSIONS: The results of the current study indicate that (153) Sm-EDTMP can be safely combined with docetaxel at full doses on an ongoing basis in patients with mCRPC. Although thrombocytopenia limited therapy for some patients, preliminary efficacy supports the strategy of combining a radiopharmaceutical with chemotherapy, which is an appealing strategy given the anticipated availability of α emitters that can prolong survival.

Impaired release of corticosterone from adrenals contributes to impairment of circadian rhythms of activity in hyperammonemic rats.

Patients with liver cirrhosis may present impaired sleep-wake and circadian rhythms, relative adrenal insufficiency and altered hypothalamus-pituitary-adrenal gland (HPA) axis. The underlying mechanisms remain unclear. Circadian rhythms are modulated by corticosteroids which secretion is regulated by HPA axis. Hyperammonemia alters circadian rhythms of activity and corticosterone in rats. The aims were: (1) assessing whether corticosterone alterations are responsible for altered circadian rhythm in hyperammonemia: (2) to shed light on the mechanism by which corticosterone circadian rhythm is altered in hyperammonemia. The effects of daily corticosterone injection at ZT10 on circadian rhythms of activity, plasma corticosterone, adreno-corticotropic hormone (ACTH) and hypothalamic corticotropic releasing hormone (CRH) were assessed in control and hyperammonemic rats. ACTH-induced corticosterone release was analyzed in cultured adrenal cells. Corticosterone injection restores the corticosterone peak in hyperammonemic rats and their activity and circadian rhythm. Plasma ACTH and CRH in hypothalamus are increased in hyperammonemic rats. Corticosterone injection normalizes ACTH. Chronic hyperammonemia impairs adrenal function, reduces corticosterone content and ACTH-induced corticosterone release in adrenals, leading to reduced feedback modulation of HPA axis by corticosterone which contributes to impair circadian rhythms of activity. Impaired circadian rhythms and motor activity may be corrected in hyperammonemia and hepatic encephalopathy by corticosterone treatment.

Cytoprotective and antioxidant effects of the edible halophyte Sarcocornia perennis L. (swampfire) against lead-induced toxicity in renal cells.

Lead (Pb) exposure is considered as a risk factor responsible for renal impairment in humans. On the other hand, the halophyte Sarcocornia perennis is a fresh vegetable crop suitable for leafy vegetable production. This study was designed to evaluate the in vitro protective activity of S. perennis against lead-induced damages in HEK293 kidney cells. Morphological and biochemical indicators were used to assess cytotoxicity and oxidative damages caused by Pb treatment on the cells. Our results showed that lead induced (1) a decrease in cell viability (MTT), (2) cell distortion and cohesion loss, (3) superoxide anion production and lipid peroxidation. Conversely, addition of S. perennis extract to the lead-containing medium alleviated every above syndrome. Thus, cell survival was increased and the production of reactive oxygen species caused by Pb treatment was inhibited. Taken together, our study revealed that S. perennis has potent cytoprotective effect against Pb-induced toxicity in HEK 293 cell. Such action would proceed through the decrease in ROS levels and resulting oxidative stress, which suggests a potential interest of this halophyte in the treatment of oxidative-stress related diseases.

Comparison of platelet count results on the Sysmex XN between citrate or MgSO4 and K2 EDTA anticoagulants.

INTRODUCTION: The aim of this study performed on Sysmex XN is to compare platelet values on citrate and MgSO4 (TBX) in patients with K2EDTA-induced platelet clusters and to identify platelet biases of these matrices compared to K2EDTA. METHODS: Sixty patients with K2EDTA-induced platelet clusters were re-sampled with K2EDTA, citrate and TBX. Platelet results were then compared, and smears were analysed for clumping. Platelet results from 120 patients without K2EDTA-induced platelet clusters were compared between K2 EDTA, citrate, and MgSO4 with impedance and fluorescence modes. Biases from regressions were analysed. RESULTS: Out of the 60 patients with K2EDTA-induced platelet clusters, none showed platelet clusters with MgSO4 whereas 50% still showed clusters with citrate. Among those without platelet clusters on citrate, the mean relative difference between (citrate- MgSO4 )/MgSO4 was -12.7% in impedance and -9.8% in fluorescence. Among the 120 patients without K2EDTA-induced platelet clusters, in fluorescence the mean relative bias with respect to K2EDTA was -2.06% for MgSO4 and -10.3% for Citrate. For the MgSO4 versus K2 EDTA regressions, the maximum absolute values of the 95% CI of the relative biases at 150 × 109 /L (5.45%) and 450 × 109 /L (3.56%) were below the desirable analytical objectives of the EFLM. CONCLUSION: In patients with K2EDTA-induced platelet clusters, MgSO4 is preferable to citrate. MgSO4 provides a bias with XN in fluorescence when compared to EDTA which is within analytical tolerance.

Efficacy of Different Root Canal Irrigating Solutions in Removing Smear Layer: A Scanning Electron Microscopic Study.

Background The aim of the present investigation was to evaluate and compare the efficacy of frequently used chemical agents in terms of their capacity to eliminate the smear layer after instrumentation, as observed via scanning electron microscopy (SEM). Materials and methods Sixty extracted single-rooted mandibular premolar teeth, each with roots 15 mm in length, were used in this study. The teeth were divided into one control group and four study groups, each containing 12 teeth. In Control Group 0, teeth were irrigated with 3 ml of saline only. In Group 1, teeth were irrigated initially with 3% sodium hypochlorite (NaOCl) and then given a final rinse with 3 ml of 17% ethylenediaminetetraacetic acid (EDTA) for one minute. In Group 2, teeth were irrigated with 3% NaOCl and given a final rinse with 3 ml of a mixture of tetracycline, acid, and detergent (MTAD, BIOPURE) for one minute. In Group 3, teeth were irrigated with saline and given a final rinse with 3 ml of 17% EDTA for one minute. In Group 4, teeth were irrigated with saline and given a final rinse with 3 ml of MTAD for one minute. One-half of each tooth was chosen and prepared for scanning electron microscopic (SEM) examination at the cervical, middle, and apical thirds. These were observed at magnifications of up to 1,000 times to check for the presence or absence of a smear layer. The data were analyzed using the Kruskal-Wallis test and post-hoc Dunn's test. Results All of the root canal irrigation protocols exhibited superior efficacy compared to the control group in the elimination of the smear layer. Group 2 (3% NaOCl with MTAD) showed the lowest mean scores, compared to all the groups, followed by Group 1 (3% NaOCl with 17% EDTA). MTAD was more effective than EDTA. The smear layer was effectively removed from the apical third, followed by the middle and coronal thirds of the root. Conclusion Initial irrigation with 3% NaOCl and one-minute final irrigation with 3 ml MTAD was the most effective root irrigant, and particularly indicated in teeth with infection of the apical third.

The effect of EDTA solution on corneal endothelial cells in rabbits.

Corneal disease threatens vision globally. Among corneal diseases, calcific band keratopathy has severe effects on vision owing to its unique location. Currently, ethylene diamine tetraacetic acid (EDTA) chelation remains the most important treatment. However, only the safety of low-dose topical EDTA eye drops is well established in humans. Therefore, the purpose of this study was to determine the safe dose range of EDTA for calcific band keratopathy surgery and its toxic effects on rabbit eyes. Rabbits were administered different doses of EDTA solutions (0.50, 0.20, 0.10, 0.05, and 0.01 M) for twenty minutes. In day seven, the rabbits were euthanized and pathological examination was performed for cornea. We found severe corneal edema in 0.50 M group, while milder edema in lower-concentration treated groups. Followed by corneal thickness measurement, the measured values increase to the peak in post-operative three day (0.20 M group) or one day (lower-concentration groups), then decreased. Groups comparison shown significant difference between BSS control group and higher concentration groups (0.20 M and 0.10 M) (P < 0.001) in observation period, but no significance was observed between low concentration and control group in the day seven after surgery (P > 0.05). Confocal microscopy examination suggested, the number of corneal endothelial cells significantly decreased from 3428.6 ± 180.3 cells/mm2 to 2808 ± 80.6 cells/mm2 in the 0.50 M group, while the lower-concentration groups showed lesser toxic effects on corneal endothelial cells. Finally, our histological examination demonstrated inflammation in each experimental group and dose-dependent, compared with control group. Our study found 0.05 M and 0.01 M EDTA solutions had no obvious toxic effect on the corneal endothelium compared with higher concentration. However, further study of EDTA side effect by clinical trials, and therapeutic effect observation with different concentration are necessary.

Ocular safety of intravitreal ethylene diamine tetra acetic acid (EDTA): An experimental feasibility study.

Ethylene diamine tetra acetic acid (EDTA) is a chelating component that is able to diminish oxidative reactivity and can be a potential neuroprotective drug in various ocular diseases. For assessing the safety of intravitreal EDTA, 10 rabbits were allocated and divided into 5 groups. Right eyes of the animals received intravitreal EDTA (112.5, 225, 450, 900 and 1800 µg /0.1 ml). Fellow eyes were considered as controls. Clinical examinations and electroretinography (ERG) were performed at the baseline and on day 28. The enucleated eyes were subjected to hematoxylin and eosin (H&E) staining, immunohistochemistry for glial fibrillary acidic protein (GFAP) and the terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) test. Clinical examinations, H&E staining and TUNEL assay were unremarkable. The ERG test did not exhibit any significant alteration compared to the baseline values, except for a significant decrease in just one measurement of the eyes injected with 225 µg EDTA. The mean scores of GFAP immune reactivity in the eyes injected with 112.5 and 225 µg EDTA indicated a non-significant reaction. The scores in higher doses were significant. We suggest intravitreal EDTA with a dose threshold of < 450 µg should be studied for ratification of the safe dose.