Ethylenediaminetetraacetic Acid Research Articles

Assessment of the Phytochemical Constituents of Methanol Extract of Ereromastax Polysperma Leaves and its Effect on the Hematological Indices in Albino Rats

The study investigated the effects of chronic administration of methanol extract of Eremomastax polysperma on the hematological parameters of albino rats. The rats were administered daily and orally for 14 days, with water as a control. On day 15, they were anesthetized using chloroform and blood was withdrawn from the heart through cardiac puncture into ethylene diamine tetra acetic acid (EDTA) specimen bottles. The phytochemical screening of crude methanolic extracts revealed the presence of secondary metabolites such as alkaloids, saponin, flavonoids, and cardiac glycosides. The statistical test employed was Analysis of Variance (ANOVA), with a predetermined significance level of p<0.05 and p<0.01. The ANOVA analysis revealed significant differences in red blood cell count (RBC), hemoglobin (HGB) percentages among the control group and the treatment groups of 500mg/kg and 1500mg/kg. The hematological evaluation showed a significant difference in RBC, HGB, and HCT, and a significant decrease in mean corpuscular hemoglobin concentration (MCHC) with increasing doses of the extract. The results suggest that E. polysperma has anti-inflammatory, antimalarial, antimicrobial, cytotoxic, antispasmodic, and pharmacological effects, potentially aiding in disease treatment.

Effects of anticoagulant (EDTA and Heparin) on blood of goat.

Blood is an important tool for accurate diagnosis of disease, forensic investigation, and hematological analysis. However this phenomenon can be thwarted occasionally if appropriate anticoagulant with storage times is not maintained properly. This research aims to compare and explore the effect of anticoagulants (such as ethylene diamine tetra acetic acid and heparin) and storage time on the hematological parameters in indigenous goats. Twenty goats were enrolled and raised for this purpose. Bloods were collected into two different tubes containing EDTA and heparin and analyzed immediately to evaluate the basal value. All tubes were divided into two aliquots and stored at 4°C and 25°C and were analyzed again at the 24th and 48th hours of collection. The statistical analysis of this result showed that there was no significant difference (P>0.05) in the anticoagulant’s effect on the hematological parameters. RBC, Hb, PCV, MCV, MCH, MCHC, total WBC, and Platelets except ESR decreased gradually along with the storage time of up to 48 hours compared to the basal value. The hematological parameters were reduced more significantly (P<0.05) when stored at 250C rather than 4°C indicating that the parameters remain in better condition in refrigeration. The samples should be stored at 4°C rather than room temperature and be used within 24 hours because the storage time modify the analyzed results. These findings demonstrate that both of the anticoagulants (EDTA and Heparin) show reliable results, therefore it can be used to store blood samples for any diagnostic purpose. Res. Agric. Livest. Fish. Vol. 11, No. 1, April 2024: 1-10

An efficient method for determining the pozzolanic reaction degrees of low-calcium supplementary cementitious materials in blended cement pastes

The pozzolanic reaction degree of supplementary cementitious materials (SCMs) is a crucial parameter influencing the performance of SCM-blended concrete. Although several methods, such as partial-or-no-known-crystal-structure (PONKCS) analysis with X-ray diffraction (XRD) testing and image analysis, have been developed to estimate the pozzolanic reaction degree of SCMs, they have their own challenges in achieving reliable estimation. Ethylenediaminetetraacetic acid (EDTA) with NaOH is an efficient and reliable agent for selectively dissolving calcium-rich phases (e.g., cement clinkers and hydration products) without dissolving low-calcium SCMs. In this study, the ionic selective dissolution method (ISDM) that combines the use of an EDTA–NaOH dissolving agent and inductively coupled plasma–optical emission spectroscopy is developed to efficiently and reliably determine the pozzolanic reaction degree of low-calcium SCMs. Furthermore, the chemical compositions of aluminium-incorporated calcium–silicate–hydrate gel are determined via the ISDM coupled with the Rietveld analysis of XRD data, according to mass conservation principles.

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

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

Oral sericin ameliorates type 2 diabetes through passive intestinal and bypass transport into the systemic circulation

Ethnopharmacological relevanceBoiled silkworm cocoons have been used to treat 'Xiaoke disease' (diabetes mellitus) recorded in Chinese medicine for over 800 years. In recent years, it has been found that the active substance silk sericin (SS) has therapeutic benefits in treating type 2 diabetes mellitus (T2DM). SS promotes pancreatic islet signalling, the proliferation of pancreatic islet cells, and insulin secretion. It is inferred that SS enters the bloodstream after oral administration and plays a role in the body's circulation. As a natural protein, SS needs to resist digestion by proteases in the gastrointestinal tract and cross the gastrointestinal barrier after oral administration. It is currently unclear how SS crosses the gastrointestinal barrier and whether it exerts therapeutic effects on T2DM by entering the circulation. Aim of the studyTo study how SS crosses the gastrointestinal barrier and whether it enters the body circulation to exert a therapeutic effect on T2DM. Materials and methodsSS was extracted from silkworm cocoons using an alkaline method with sodium carbonate. The antidigestive capacity of SS was detected using SDS-PAGE gel electrophoresis experiments. The mode of uptake and translocation of orally consumed SS in vivo was analysed using the AP-side to BL-side and BL-side-AP-side translocations, apparent Permeability coefficient (Papp), and Exocytosis rates (ER). The study compared the differences between the adSS group and the adSS + EDTA group by using Ethylenediaminetetraacetic acid (EDTA) to separate the tight junctions between Caco-2 cells. The aim was to analyze whether the transport mode of oral filaggrin proteins in vivo could be absorbed by bypass transport. By administering SS through oral and intraperitoneal injection to type 2 diabetic mice, we measured its concentration in the blood, as well as blood glucose and insulin levels, to determine its effectiveness in treating diabetes and its ability to enter the body's circulation for treatment. ResultsThe molecular weight of SS decreased from 10k∼25 kDa to 10k∼15 kDa after in vitro simulated gastrointestinal fluid digestion, indicating its good antidigestive properties. The apparent Papp was greater than 1 × 10−6 cm·s-1, and the ER was between 0.5 and 1.5, indicating that adSS was well-absorbed and mainly passively transported. The Caco-2 cell model showed that the addition of EDTA promoted the transport of adSS, resulting in significantly larger Papp and ER values, indicating that adSS was absorbed by bypass transport. After oral administration of SS, the concentration of SS in the blood was lower than after intraperitoneal injection, which is 60% of intraperitoneal administration. Mice with a T2DM model who were administered SS for 5 weeks showed significant improvement in insulin resistance and glucose tolerance. Additionally, the pancreatic tissue appeared more regular. In the treatment of T2DM, injections of SS have been shown to be more effective than oral administration. Both oral and intraperitoneal injections have been partially involved in the circulation. ConclusionsSS is enzymatically cleaved by proteolytic enzymes in the gastrointestinal tract. The smaller molecules are partially absorbed into the body's circulation through passive and paracrine transport, exerting a therapeutic effect on T2DM.

Unveiling antibiofilm potential: proteins from Priestia sp. targeting Staphylococcus aureus biofilm formation.

Staphylococcus aureus is the etiologic agent of many nosocomial infections, and its biofilm is frequently isolated from medical devices. Moreover, the dissemination of multidrug-resistant (MDR) strains from this pathogen, such as methicillin-resistant S. aureus (MRSA) strains, is a worldwide public health issue. The inhibition of biofilm formation can be used as a strategy to weaken bacterial resistance. Taking that into account, we analysed the ability of marine sponge-associated bacteria to produce antibiofilm molecules, and we found that marine Priestia sp., isolated from marine sponge Scopalina sp. collected on the Brazilian coast, secretes proteins that impair biofilm development from S. aureus. Partially purified proteins (PPP) secreted after 24 hours of bacterial growth promoted a 92% biofilm mass reduction and 4.0 µg/dL was the minimum concentration to significantly inhibit biofilm formation. This reduction was visually confirmed by light microscopy and Scanning Electron Microscopy (SEM). Furthermore, biochemical assays showed that the antibiofilm activity of PPP was reduced by ethylenediaminetetraacetic acid (EDTA) and 1,10 phenanthroline (PHEN), while it was stimulated by zinc ions, suggesting an active metallopeptidase in PPP. This result agrees with mass spectrometry (MS) identification, which indicated the presence of a metallopeptidase from the M28 family. Additionally, whole-genome sequencing analysis of Priestia sp. shows that gene ywad, a metallopeptidase-encoding gene, was present. Therefore, the results presented herein indicate that PPP secreted by the marine Priestia sp. can be explored as a potential antibiofilm agent and help to treat chronic infections.

Sorption Characteristics and Chromatographic Separation of 90Y3+ from 90Sr2+ from Aqueous Media by Chelex-100 (Anion Ion Exchange) Packed Column.

There is growing demand for separation of 90Y carrier free from 90Sr coexisting to produce high purity 90Y essential for radiopharmaceutical uses. Thus, in this context the sorption profiles of Y3+ and Sr2+ from aqueous solutions containing diethylenetriaminepenta acetic acid (DTPA), ethylenediaminetetra-acetic acid (EDTA), acetic acid, citric acid, or NaCl onto Chelex-100 (anion ion exchange) solid sorbent were critically studied for developing an efficient and low-cost methodology for selective separation of Y3+ from Sr2+ ions (1.0 × 10-5 M). Batch experiments displayed relative chemical extraction percentage (98 ± 5.4%) of Y3+ from aqueous acetic acid solution onto Chelex-100 (anion ion exchanger), whereas Sr2+ species showed no sorption. Hence, a selective separation of Y3+ from its parent 90Sr2+ has been established based upon percolation of the aqueous solution of Y3+ and Sr2+ ions containing acetic acid at pH 1-2 through Chelex-100 sorbent packed column at a 2 mL min-1 flow rate. Y3+ species were retained quantitatively while Sr2+ ions were not sorbed and passed through the sorbent packed column without extraction. The sorbed Y3+ species were then recovered from the sorbent packed column with HNO3 (1.0 M) at a 1.0 mL min-1 flow rate. A dual extraction mechanism comprising absorption associated to "weak-base anion exchanger" and "solvent extraction" of Y3+ as (YCl6)3- and an extra part for "surface adsorption" of Y3+ by the sorbent is proposed. The established method was validated by measuring the radiochemical (99.2 ± 2 1%), radionuclide purity and retardation factor (Rf = 10.0 ± 0.1 cm) of 90Y3+ recovered in the eluate. Ultimately, the sorbent packed column also presented high stability for reusing 2-3 cycles without drop in its efficiency (±5%) towards Y3+ uptake and relative chemical recovery. A proposed flow sheet describing the analytical procedures for the separation of 90Y3+ from 90Sr2+ using chelating Chelex 100 (anion exchange) packed column is also included.

Effect of Chitosan Irrigant and Lubricating Gel on Bond Strength of Resin Sealer to Radicular Dentin: An In Vitro Study.

The adhesive strength of sealers to dentin is influenced by various factors, and the presence of a smear layer is among the critical variables. Chitosan, known for its dentin compatibility, has previously demonstrated a reduction in dentin change and resin sealer bond strength comparable to ethylenediaminetetraacetic acid (EDTA) when used as an irrigant and final rinse. The study investigates the impact of chitosan, used as both a lubricating gel and final rinse, on the push-out bond strength of resin sealer. Forty single-rooted premolar teeth, each with a fully formed root and a single root canal, were collected post-extraction. During canal preparation, 1 mlsodium hypochlorite (3%) was used for irrigation at every change ofinstrument, followed by applying specific chelating gel and final rinse for each experimental group. The groups included: Group 1 (17% EDTA chelating gel, final rinse with saline), Group 2 (17% EDTA chelating gel, final rinse with 17% EDTA solution), Group 3 (chitosan chelating gel, final rinse with saline solution), and Group 4 (chitosan chelating gel, final rinse with 0.2% chitosan solution), 10 specimens in each group.After obturation, specimens were sealed and incubated for a week at 37°C with 100% humidity. The universal testing machine was used for push-out tests, and specimens were examined using a scanning electron microscope (SEM) to identify various types of bond failure. Among the four groups, Group 2 exhibited the highest mean push-out bond strength (7.33 ± 0.26 MPa), followed by Group 4 (5.33 ± 0.25 MPa), Group 1 (4.61 ± 0.30 MPa), and Group 3 (2.94 ± 0.32 MPa). The variations in bond strength suggest a notable impact of the chelating agents and final rinse solutions on the resin sealer's interaction with dentin. The study concludes that the use of EDTA as both a lubricating gel and a final rinse significantly enhances push-out bond strength, outperforming chitosan in this study. Groups with saline as the final rinse (Group 1 and Group 3) exhibited the least bond strength, highlighting the importance of the final rinse in root canal therapy.

Boosting the Ni(II)-EDTA decomplexation and Ni(II) immobilization in a heterogeneous Fenton-like system with L-cysteine functionalized zero-valent iron

The remarkable stability and detrimental environmental impacts of nickel(II)-ethylenediaminetetraacetic acid (Ni(II)-EDTA) complexes originated from electroplating effluents have sparked a growing curiosity about their efficient remediation. In this study, L-cysteine functionalized zero-valent iron (C-ZVIbm) was synthesized via a ball-milling method and employed as a heterogeneous Fenton-like catalyst to decomplex Ni(II)-EDTA and immobilize Ni(II) from wastewater. The results demonstrated that the pseudo-second-order kinetic rate constant (2.096 L·mol−1·min−1) for EDTA degradation in the C-ZVIbm/H2O2 system was 123.3 times that in the ZVIbm/H2O2 system (0.017 L·mol−1·min−1). The incorporation of L-cysteine onto the surface of ZVI significantly enhanced the H2O2 activation, reactive oxygen species (OH, O2−, and 1O2) generation, and the sequential Ni(II)-EDTA decomplexation and EDTA degradation. The results of capture experiments revealed that 1O2 played a crucial role in the degradation of EDTA in the C-ZVIbm/H2O2 system. The optimization of the experimental variables indicated that the degradation of EDTA and the reduction/adsorption of Ni(II) were antagonistic in the C-ZVIbm/H2O2 system, resulting in an efficient reduction/immobilization of Ni(II) and an inhibition of EDTA degradation at a higher dosage of C-ZVIbm. Our findings provide new insights into the promising heterogeneous Fenton-like process that enables the decomplexation of Ni(II)-EDTA, the degradation of EDTA, and the synchronous immobilization of Ni(II).

Influence of Carbons on Metal Stabilization and the Reduction in Soil Phytotoxicity with the Assessment of Health Risks

Despite notable achievements in the development and application of the remediation of metal-contaminated soils, the search for and study of promising immobilizing agents continues. This article presents a new approach to using brown coal and activated carbon and their application for metal stabilization. An experiment was conducted to test Medicago falcata L. on soils from industrial areas contaminated with varying levels of lead (Pb), cadmium (Cd), chromium (Cr), zinc (Zn), copper (Cu), and nickel (Ni) with the addition of carbons. This plant is a stress-tolerant leguminous species. In this study, the total content of metals in soil and the available forms to plants (single extractions with 0.02 M ethylenediaminetetraacetic acid, EDTA) were determined, along with metal contents in the plant. The use of carbons lowered the phytoavailable forms of metals for plants and thus, resulted in a reduction in the phytotoxicity of the soils. The contents of the tested metals in the roots and shoots were lower than in the combinations of soils with no carbon added. The activated carbon had a stronger effect on limiting the availability of metals than brown coal in relation to plants growing on soils without added carbon; the percentage of reduction for the shoots was Cr (18.2%) > Zn (11.5%) > Ni (10.7%) > Cu (10.3%) > Cd (8.9%) > Pb (2.4%) and Cu (13.3%) > Cr (12.5%) > Zn (10.5%) > Pb (9.0%) > Ni (5.7%) > Cd (4.6%) for roots. Metals reduced the growth of Medicago falcata L. roots from 44 to 21%, while the growth of shoots was reduced from 25 to 2%. Adding carbons to soils in all combinations resulted in a decrease in the following pollution indices: pollution index (PI), pollution load index (PLI), and non-carcinogenic (HQ) and carcinogenic risk factors (ILCR).

Dual-responsive solvatochromic sensor for simultaneous detection and mechanism of Cr(III) and Co(II) cations by a new N2O4-donor tetrabenzodiaza-crown ether Macrocyclic ligand anchored to dipyridine moieties

The present study describes the chemosensing capability of an N2O4-donor tetrabenzodiaza-crown ether macrocyclic ligand derivatized by two pyridine side arms to afford the flexible macrocyclic ligand, 5,20-bis(pyridin-4-ylmethyl)-5,6,12,13,19,20,26,27octahydrotetrabenzo [e,i,o,s][1,4,11,14]tetraoxa[7,18]diazacycloicosine (PMHBOA). A selective colorimetric behavior of PMHBOA for the prompt detection of Cr(ΙΙΙ) and Co(ΙΙ) can be observed with naked eyes. The 1H, 13C{H} NMR, and infrared spectroscopy disclosed the ligand binding capability for selective metal ions, ascribed to a 1:1 complex formation with Cr(III) and Co(II) cations in isopropanol. The limit of detection (LOD) values were calculated as 580 and 850 nM for Cr(ΙΙΙ) and Co(ΙΙ), respectively, accompanied with suitable LOQs as 1.95 µM for Cr(III) and 2.85 µM for Co(II), respectively. The reusability was confirmed in the presence of ethylenediaminetetraacetic acid (EDTA). Moreover, a TLC-based fabricated paper strip successfully applied PMHBOA as a chemosensor for synchronous visible detection of Cr(III) and Co(II) in their alcoholic solution. Meanwhile, the observed color change along with the turn-off fluorescent of PMHBOA guaranteed a felicitous choice of candidate for Cr(III) in the presence of other library metal ions in isopropanol:H2O (9/1 v/v) solution. A fast separation test on the filter paper, impregnated with PMHBOA, was also employed to separate Cr(III) and Co(II) cations from aqueous solutions by recording UV–vis spectra. The 1H NMR spectroscopy supported the macrocycle ring and side arms as the parallel coordination site of PMHBOA with Cr(III) and Co(II).

Heterojunction-Based Photocatalytic Degradation of Rose Bengal Dye via Gold-Decorated α-Fe2O3-CeO2 Nanocomposites under Visible-Light Irradiation

Developing photocatalytic nanomaterials with unique physical and chemical features using low-cost and eco-friendly synthetic methods is highly desirable in wastewater treatment. In this work, the magnetically separable α-Fe2O3-CeO2 nanocomposite (NC), with its respective metal oxides of α-Fe2O3 and CeO2 nanoparticles, was synthesized using a combination of hexadecyltrimethylammonium bromide (CATB) and ascorbic acid via the hydrothermal method. To tune the band gap, the heterojunction nanocomposite of α-Fe2O3-CeO2 was decorated with plasmonic Au nanoparticles (Au NPs). The various characterization methods, such as FTIR, UV-vis DRS, XRD, XPS, TEM, EDX, SEM, and PL, were used to determine the properties of the materials, including their morphology, elemental composition, optical properties, band gap energy, and crystalline phase. The nanocomposite of α-Fe2O3-CeO2@Au was utilized to remove Rose Bengal (RB) dye from wastewater using a photocatalytic technique when exposed to visible light. A comprehensive investigation of the impact of the catalyst concentration and initial dye concentration was conducted to establish the optimal photodegradation conditions. The maximum photocatalytic efficiency of α-Fe2O3-CeO2@Au (50 mg L−1) for RB (20 ppm) dye removal was found to be 88.9% in 120 min under visible-light irradiation at a neutral pH of 7 and 30 °C. Various scavengers, such as benzoquinone (BQ; 0.5 mM), tert-butyl alcohol (TBA; 0.5 mM), and ethylenediaminetetraacetic acid (EDTA; 0.5 mM), were used to investigate the effects of different free radicals on the photocatalytic process. Furthermore, the reusability of the α-Fe2O3-CeO2@Au photocatalyst has also been explored. Furthermore, the investigation of the potential mechanism demonstrated that the heterojunction formed between α-Fe2O3 and CeO2, in combination with the presence of deposited Au NPs, led to an enhanced photocatalytic efficiency by effectively separating the photogenerated electron (e−)–hole (h+) pairs.

Effect on emergence from anesthesia following induction with diazepam and its association with CYP2C9, CYP2C19 and CYP3A4 gene polymorphisms

Introduction & Objectives: Benzodiazepines are commonly used as adjuvants to IV anesthetic agents. Diazepam dosage varies widely depending on the desired endpoints. The rationale behind such inter-individual variability in drug responses has been attributed to genetically determined alterations in the enzyme activities. We assessed the emergence time in patients after induction of anesthesia with two different doses of diazepam and evaluated if any correlation exists between the emergence time and genetic polymorphism in drug metabolizers CYP2C19, CYP2C9, and CYP3A4 genes. Methodology: The study was conducted on randomly selected adult patients scheduled for elective surgery between 40-60 y of age with predefined inclusion and exclusion criteria. The patients were distributed into 2 groups, with group 1 receiving 0.2 mg/kg and group 2 given 0.3 mg/kg diazepam. Wake-up time after surgery was determined after a standardized verbal command played by a tape-recorder. Three ml blood in Ethylenediaminetetraacetic acid (EDTA) was collected before administering anesthesia. Results: Wake-up time between 8-18 min did not vary between the two groups. Comparatively persons having 2C19 2 took longer time to wake-up but this association was not statistically significant. However, CYP2C9 2 and 2 C9 3 and CYP3A4 genotypes had no influence on wake-up time. Conclusion: The pilot study suggests the emergence time from diazepam is longer in patients having 2C19 2 which confers slow-metabolizer phenotype. However, this study needs to be replicated in larger sample size to arrive at definitive conclusions before clinical applications. Abbreviations: BDZ- benzodiazepine; EDTA- Ethylenediaminetetraacetic acid; GA-General Anesthesia Key words: Diazepam; Emergence; Gene Polymorphism; Pharmacogenetics Citation: Mittal T, Badhe VK, Badhe DSD, Ahluwalia P, Mitta Bl, Mittal R. Effect on emergence from anesthesia following induction with diazepam and its association with CYP2C9, CYP2C19 and CYP3A4 gene polymorphisms. Anaesth. pain intensive care 2024;28(1):126-138. DOI: 10.35975/apic.v28i1.2380 Received: July 20, 2023; Reviewed: December 07, 2023; Accepted: December 07, 2023

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

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

Synthesis and toxicity evaluation of a spray consisting of silver nanoparticles, ethylenediaminetetraacetic acid, methylsulfonylmethane, and xylitol on vero cell line

Abstract Introduction: Dental plaque is a biofilm or an accumulation of bacteria that grows in the internal surfaces of the mouth and can be observed as a white to pale yellow layer over or between teeth. The continuous formation and accumulation of dental plaque lead to oral diseases. As a result, it is necessary to prevent the aggregation of dental plaque and clean it daily. Recently, ethylenediaminetetraacetic acid (EDTA) has been utilized in toothpaste to prevent plaque formation as an EDTA-transporting enhancer, methylsulfonylmethane (MSM) can effectively increase its local effect. Xylitol decreases the Streptococcus mutans count by changing the metabolic pathways. Aims: In the present study, we synthesized a solution containing silver nanoparticles, EDTA, MSM, and xylitol and evaluated its toxicity on the Vero cell line through the MTT assay. Materials and Methods: To produce silver nanoparticles, we dissolved silver nitrate in sodium citrate. Then we used a solution of distilled water and polyvinylpyrrolidone, which resulted in the encapsulation and stabilization of silver nanoparticles, and the solution was made by mixing other ingredients. We evaluated the cytotoxicity of this spray using the Vero cell line. We cultured the cells in the 10% FBS-containing RPMI culture medium. For performing the cytotoxicity test, we put 10,000 cells in each well of the 96-well plate, and the next day added the synthesized solution to each well at dilutions 0.05%, 0.1%, 0.2%, 0.5%, 1%, 2%, 5%, and 10%. As the control group, we used 4 wells containing live Vero cells without adding the solution. After 24, 48, and 72 h, we added the MTT solution to each well and incubated the plates at 37°C for 4 h. Finally, we evaluated the rate of living cells by reading the absorbance with an ELISA device at 570 nm. Results and Discussion: We used the Mann–Whitney nonparametric test to evaluate the cytotoxicity of different concentrations of the synthesized spray solution and compare the cell viability rate of groups with the controls in various periods. According to the cytotoxicity results of different concentrations of the spray solution on the Vero cell line, there was no significant difference in cytotoxicity between the 0.05%, 0.1%, 0.2%, and 0.5%, and control groups at 24, 48, and 72 h (P > 0.05). No significant difference existed in cytotoxicity between 1% and 2% concentrations and the controls 24 h after exposure; this became significant after 48 and 72 h (P = 0.014). However, a significant difference existed in cytotoxicity between 5% concentration and the controls 24, 48, and 72 h after exposure (P = 0.014). The CC50 of the spray solution was calculated at 3.51%. Conclusion: The findings of this study showed that the synthesized solution is nontoxic; therefore, this spray solution can be used safely as an oral mouthwash and spray.

Accuracy of an Electronic Apex Locator (EAL) in Measuring the Working Length of Root Canals in Comparison WithRadiographs: An In Vivo Study.

Traditionally, radiographs were used to assess the working length of root canals. However, the use of Electronic Apex Locators (EAL) may be a non-invasive alternative. This study aimed to evaluate the accuracy of the electronic apex locator (EAL) compared to radiographic measurements in determining the working length of root canals. The study assessed the performance of EALs in different scenarios, including the presence of blood and pulp tissue, the use of ethylenediaminetetraacetic acid (EDTA), sodium hypochlorite (NaOCl) as an irrigant, and post-instrumentation with normal saline. An electronic apex locator(DTE DPEX-V; Woodpecker, China) was used alongside traditional radiographs to measure the working length in 144 root canal treatments. Bland-Altman analysis was used to compare the measurements between EAL and radiographs using Jamovi software, v2.4.8 (https://www.jamovi.org/). The findings revealed strong correlations between EAL and radiographic measurements across all testing environments, with Pearson's correlation coefficients ranging from 0.944 to 0.998. The Bland-Altman analysis suggests bias; the average difference was negative and close to zero (Pulp: -0.354, EDTA: -0.0972, NaOCl: -0.0382, Saline: -0.0139) when EAL measurements were compared to radiographic measurements. The working length measurement of root canals using an electronic apex locator such as DTE DPEX-V is comparable to traditional radiographic measurements. The use of irrigants improves the measurement. The EAL has the potential to be an alternative to the invasive radiograph for root canal length measurement.

Effect of Different Electroconductive Root Canal Irrigations on the Accuracy of Different Apex Locators: An In Vitro Comparative Study.

An accurate working length (WL) estimation is fundamental to a successful endodontic therapy. The objective of this comparative in vitro research was to investigate the performance of iPex II and the Root ZX Mini electronic apex locators (EALs) in measuring root canal WL with different irrigant solutions and their relation to the electrical conductivity of irrigation solutions. Seventy sound permanent lower premolar teeth, each with a single root and developed apices, were used. Under an X15 stereomicroscope, the real working length was determined with the aid of a #10 file. After that, teeth were placed into an alginate model, and the iPex II and Root ZX Mini were used for the detection of electronic working length with various irrigants. Seventy teeth were randomly distributed into seven groups, 10 per each group (group I: dry canal; group II: distilled water; group III: ozonated water; group IV: 5% sodium hypochlorite (NaOCl); group V: 2% chlorhexidine, group VI: 17% ethylenediaminetetraacetic acid (EDTA) solution, and group VII: 17% EDTA gel). The difference in WL was calculated by deducting real working length from its electronic working length. The study also evaluates the electrical conductivity of the seven endodontic irrigant solutions. The two-way analysis of variance (ANOVA) test was used for statistical analysis. Statistically, neither both types of EALs (P = 0.088) nor various irrigating solutions with varying electrical conductivities (P = 0.099) significantly affect the accuracy of EL estimation. There were no significant differences between the accuracy of the Root ZX Mini and the iPex II. The accuracy of both apex locators is unaffected in the presence of various irrigation solutions with varying electrical conductivities in this research.

Enamel Matrix Derivative, 58S5 Bioactive Glass, and Fluoride Varnish for Enamel Remineralization: A Multi-analysis Approach.

This study aimed to evaluate the enamel remineralization efficacy of enamel matrix derivative (EMD), experimental bioactive glass (BAG), and fluoride varnish in vitro. Artificial initial caries lesions were developed on fifty human enamel specimens using demineralization solution (pH 4.5, 37°C, 96 hours). Specimens were randomly assigned to five groups (n=10): I-5% NaF varnish (Enamelast), II-experimental 58S5 BAG+37% phosphoric acid (PA), III-EMD (Emdogain) + Ethylenediaminetetraacetic Acid (EDTA), IV-EMD+37% PA, V-Control (untreated). All remineralization agents were applied with pH cycling for seven days. The specimens were scanned by spectral domain optical coherence tomography (SD-OCT) at baseline, at demineralization, and after pH cycling. Lesion depths were measured using image analysis software (ImageJ). Lesions were evaluated using surface microhardness (SMH) and two fluorescence methods (FluoreCam and DIAGNOdent Pen [DDPen]). The data were statistically analyzed by Kruskal Wallis, Friedman, and Wilcoxon tests (α=0.05). According to SD-OCT results, fluoride varnish was found to be the most effective agent in reducing lesion depth (p=0.005). All agents increased the SMH values after pH cycling. No significant difference was found among fluoride varnish, BAG, and EMD+PA groups. These SMH values were significantly higher than EMD+EDTA and control groups (p<0.001). All groups showed lower DDPen scores compared with the control group (p<0.001), however, no significant difference was found among the remineralization agents. In FluoreCam assessment, size and intensity values of all treated groups showed improvement. However, there was no significant difference between the treatment groups in terms of FluoreCam size measurements (p=0.186). 58S5 BAG and EMD+PA have remineralization capacity as effective as fluoride varnish. EMD+PA showed better SMH and lesion intensity results than EMD+EDTA.

Effect of different root canal irrigants on surface roughness and microhardness of Biodentine combined with triple antibiotic paste: An in vitro study

Background: Biodentine is widely used for endodontic applications; recently, it has been incorporated with triple antibiotic paste (TAP). The effect of endodontic irrigants on the physical characteristics of this new combination needs to be studied. Aims: The aim of the study was to evaluate the surface roughness and microhardness of Biodentine incorporated with TAP subjected to various endodontic irrigants. Materials and Methods: Hundred cylindrical discs (6 mm × 3 mm) were prepared by mixing the Biodentine with TAP (3:1). The specimens were subjected to different irrigating solutions for 5 min in 5 groups (n = 20): Group 1: distilled water (control), Group 2: 2.5% sodium hypochlorite, Group 3: 17% ethylenediaminetetraacetic acid, Group 4: 2% chlorhexidine (CHX), and Group 5: 2% chitosan nanoparticles (CSNs). Half of the specimens in each group were subjected to surface roughness (n = 10) and another half to microhardness (n = 10). Surface roughness was measured using a surface roughness tester, and digital Vickers microhardness testing was performed on each specimen. Statistical Analysis: One-way ANOVA and post hoc Tukey’s tests (P ≤ 0.05) were used. Results: The highest microhardness was found with 2% CSN, whereas 2% CSN and 2% CHX had a minimal effect on the surface roughness of Biodentine incorporated with TAP (P ≤ 0.05). Conclusion: The root canal irrigant 2% CSN exhibited the highest microhardness and least surface roughness of modified Biodentine with TAP.

Effect of fetal distress on viability and yield of umbilical cord blood stem cells–a prospective comparative study

IntroductionDifferent factors affect the quality and viability of cord blood stem cells, and therefore the efficacy of umbilical cord stem cell transplantation. Fetal distress is one factor affecting the quantity of CD34+ cells in cord blood. This study was designed to compare the viability and yield of the umbilical cord blood stem cells of women who have undergone emergency lower segment caesarean section for fetal distress or for other causes. Materials and methodsThis cross-sectional analytical study was performed at a tertiary care hospital facility with a total sample size of 68: 34 participants had undergone emergency C-section for fetal distress, and 34 had undergone emergency C-section for other causes. Umbilical cord blood was collected ex-utero in a 350 mL bag with citrate-phosphate-dextrose solution with adenine. Three milliliter of blood were transferred to an ethylenediaminetetraacetic acid (EDTA) tube for cell counts and flow cytometry testing for CD34+. The chi-square test was used to compare the total mononuclear cell, CD34+, and viability between the groups. ResultsThe CD34+ count [mean 4.9 versus 1.1 (× 106 cells/unit)] and total nucleated cell count [mean 14.2 versus 7.5 (× 108/unit)] were significantly higher in cord blood units collected from women who delivered by C-section for fetal distress (p-value <0.05). However, the volume of umbilical cord stem cells and viability of stem cells did not vary significantly based on the presence or absence of fetal distress (p-value >0.05). ConclusionThe current study shows that umbilical cord blood collected during fetal distress has a significantly higher content of stem cells and total nucleated cells than the non-fetal distress group