EDTA improves stability of whole blood C-peptide and insulin to over 24 hours at room temperature.

PubMed

McDonald, Timothy J; Perry, Mandy H; Peake, Roy W A; Pullan, Nicola J; O'Connor, John; Shields, Beverley M; Knight, Beatrice A; Hattersley, Andrew T

2012-01-01

C-peptide and insulin measurements in blood provide useful information regarding endogenous insulin secretion. Conflicting evidence on sample stability and handling procedures continue to limit the widespread clinical use of these tests. We assessed the factors that altered the stability of insulin and C-peptide in blood. We investigated the impact of preservative type, time to centrifugation, storage conditions and duration of storage on the stability of C-peptide and insulin on three different analytical platforms. C-peptide was stable for at least 24 hours at room temperature in both centrifuged and whole blood collected in K(+)-EDTA and serum gel tubes, with the exception of whole blood serum gel, which decreased to 78% of baseline at 24 hours, (p = 0.008). Insulin was stable at room temperature for 24 hours in both centrifuged and whole blood collected in K(+)-EDTA tubes. In contrast insulin levels decreased in serum gel tubes both centrifuged and whole blood (66% of baseline, p = 0.01 and 76% of baseline p = 0.01, by 24 hours respectively). C-peptide and insulin remained stable after 6 freeze-thaw cycles. The stability of C-peptide and insulin in whole blood K(+)-EDTA tubes negates the need to conform to strict sample handling procedures for these assays, greatly increasing their clinical utility.

Contamination of lithium heparin blood by K2-ethylenediaminetetraacetic acid (EDTA): an experimental evaluation

PubMed Central

Lima-Oliveira, Gabriel; Salvagno, Gian Luca; Danese, Elisa; Brocco, Giorgio; Guidi, Gian Cesare; Lippi, Giuseppe

2014-01-01

Introduction: The contamination of serum or lithium heparin blood with ethylenediaminetetraacetic acid (EDTA) salts may affect accuracy of some critical analytes and jeopardize patient safety. The aim of this study was to evaluate the effect of lithium heparin sample contamination with different amounts of K2EDTA. Materials and methods: Fifteen volunteers were enrolled among the laboratory staff. Two lithium heparin tubes and one K2EDTA tube were collected from each subject. The lithium-heparin tubes of each subject were pooled and divided in 5 aliquots. The whole blood of K2EDTA tube was then added in scalar amount to autologous heparinised aliquots, to obtained different degrees of K2EDTA blood volume contamination (0%; 5%; 13%; 29%; 43%). The following clinical chemistry parameters were then measured in centrifuged aliquots: alanine aminotranspherase (ALT), bilirubin (total), calcium, chloride, creatinine, iron, lactate dehydrogenase (LD), lipase, magnesium, phosphate, potassium, sodium. Results: A significant variation starting from 5% K2EDTA contamination was observed for calcium, chloride, iron, LD, magnesium (all decreased) and potassium (increased). The variation of phosphate and sodium (both increased) was significant after 13% and 29% K2EDTA contamination, respectively. The values of ALT, bilirubin, creatinine and lipase remained unchanged up to 43% K2EDTA contamination. When variations were compared with desirable quality specifications, the bias was significant for calcium, chloride, LD, magnesium and potassium (from 5% K2EDTA contamination), sodium, phosphate and iron (from 29% K2EDTA contamination). Conclusions: The concentration of calcium, magnesium, potassium, chloride and LD appears to be dramatically biased by even modest K2EDTA contamination (i.e., 5%). The values of iron, phosphate, and sodium are still reliable up to 29% K2EDTA contamination, whereas ALT, bilirubin, creatinine and lipase appear overall less vulnerable towards K2EDTA contamination. PMID:25351354

Effect of the association between citric acid and EDTA on root surface etching.

PubMed

Manzolli Leite, Fabio Renato; Nascimento, Gustavo Giacomelli; Manzolli Leite, Elza Regina; Leite, Amauri Antiquera; Cezar Sampaio, Josá Eduardo

2013-09-01

This study aims to compare the clot stabilization on root surfaces conditioned with citric acid and ethylenediamine-tetraacetic acid (EDTA). Scaled root samples (n = 100) were set in fve groups: group I-control group (saline solution); group II (24% EDTA); group III (25% citric acid); group IV (EDTA + citric acid); group V (citric acid + EDTA). Fifty samples were assessed using the root surface modifcation index (RSMI). The other 50 received a blood drop after conditioning. Clot formation was assessed using blood elements adhesion index (BEAI). A blind examiner evaluated photomicrographs. Statistical analysis considered p < 0.05. Groups-III and G-V attained the best results for RSMI and BEAI in comparison to control. The worst results for clot stabilization were seen in group-II. EDTA employment before citric acid (group-IV) reduced clot formation in comparison to citric acid use alone (group-III). Root conditioning with citric acid alone and before EDTA had the best results for smear layer removal and clot stabilization. EDTA inhibited clot stabilization on root surface and must have a residual activity once it has diminished clot adhesion to root even after citric acid conditioning. Thus, EDTA can be used to neutralize citric acid effects on periodontal cells without affecting clot stabilization. Clinical signifcance: To demonstrate that citric acid use on root surfaces previously affected by periodontal disease may favor clot stabilization and may have a benefcial effect on surgical outcomes. Also, EDTA can be used to neutralize citric acid effects on periodontal cells.

Detection of Cytomegalovirus (CMV) DNA in EDTA Whole-Blood Samples: Evaluation of the Quantitative artus CMV LightCycler PCR Kit in Conjunction with Automated Sample Preparation▿

PubMed Central

Michelin, Birgit D. A.; Hadžisejdić, Ita; Bozic, Michael; Grahovac, Maja; Hess, Markus; Grahovac, Blaženka; Marth, Egon; Kessler, Harald H.

2008-01-01

Whole blood has been found to be a reliable matrix for the detection and quantitation of cytomegalovirus (CMV) DNA. In this study, the performance of the artus CMV LightCycler (LC) PCR kit in conjunction with automated sample preparation on a BioRobot EZ1 workstation was evaluated. The accuracy, linearity, analytical sensitivity, and inter- and intra-assay variations were determined. A total of 102 clinical EDTA whole-blood samples were investigated, and results were compared with those obtained with the in vitro diagnostics (IVD)/Conformité Européene (CE)-labeled CMV HHV6,7,8 R-gene quantification kit. When the accuracy of the new kit was tested, seven of eight results were found to be within ±0.5 log10 unit of the expected panel results. Determination of linearity resulted in a quasilinear curve over more than 5 log units. The lower limit of detection of the assay was determined to be 139 copies/ml in EDTA whole blood. The interassay variation ranged from 15 to 58%, and the intra-assay variation ranged from 7 to 35%. When clinical samples were tested and the results were compared with those of the routinely used IVD/CE-labeled assay, 53 samples tested positive and 13 samples tested negative by both of the assays. One sample was found to be positive with the artus CMV LC PCR kit only, and 35 samples tested positive with the routinely used assay only. The majority of discrepant results were found with low-titer samples. In conclusion, use of the artus CMV LC PCR kit in conjunction with automated sample preparation on the BioRobot EZ1 workstation may be suitable for the detection and quantitation of CMV DNA in EDTA whole blood in the routine low-throughput laboratory; however, low-positive results may be missed by this assay. PMID:18272703

Detection of cytomegalovirus (CMV) DNA in EDTA whole-blood samples: evaluation of the quantitative artus CMV LightCycler PCR kit in conjunction with automated sample preparation.

PubMed

Michelin, Birgit D A; Hadzisejdic, Ita; Bozic, Michael; Grahovac, Maja; Hess, Markus; Grahovac, Blazenka; Marth, Egon; Kessler, Harald H

2008-04-01

Whole blood has been found to be a reliable matrix for the detection and quantitation of cytomegalovirus (CMV) DNA. In this study, the performance of the artus CMV LightCycler (LC) PCR kit in conjunction with automated sample preparation on a BioRobot EZ1 workstation was evaluated. The accuracy, linearity, analytical sensitivity, and inter- and intra-assay variations were determined. A total of 102 clinical EDTA whole-blood samples were investigated, and results were compared with those obtained with the in vitro diagnostics (IVD)/Conformité Européene (CE)-labeled CMV HHV6,7,8 R-gene quantification kit. When the accuracy of the new kit was tested, seven of eight results were found to be within +/-0.5 log(10) unit of the expected panel results. Determination of linearity resulted in a quasilinear curve over more than 5 log units. The lower limit of detection of the assay was determined to be 139 copies/ml in EDTA whole blood. The interassay variation ranged from 15 to 58%, and the intra-assay variation ranged from 7 to 35%. When clinical samples were tested and the results were compared with those of the routinely used IVD/CE-labeled assay, 53 samples tested positive and 13 samples tested negative by both of the assays. One sample was found to be positive with the artus CMV LC PCR kit only, and 35 samples tested positive with the routinely used assay only. The majority of discrepant results were found with low-titer samples. In conclusion, use of the artus CMV LC PCR kit in conjunction with automated sample preparation on the BioRobot EZ1 workstation may be suitable for the detection and quantitation of CMV DNA in EDTA whole blood in the routine low-throughput laboratory; however, low-positive results may be missed by this assay.

The effectiveness of cooling conditions on temperature of canine EDTA whole blood samples

PubMed Central

Sun, Xiaocun; Flatland, Bente

2016-01-01

Background Preanalytic factors such as time and temperature can have significant effects on laboratory test results. For example, ammonium concentration will increase 31% in blood samples stored at room temperature for 30 min before centrifugation. To reduce preanalytic error, blood samples may be placed in precooled tubes and chilled on ice or in ice water baths; however, the effectiveness of these modalities in cooling blood samples has not been formally evaluated. The purpose of this study was to evaluate the effectiveness of various cooling modalities on reducing temperature of EDTA whole blood samples. Methods Pooled samples of canine EDTA whole blood were divided into two aliquots. Saline was added to one aliquot to produce a packed cell volume (PCV) of 40% and to the second aliquot to produce a PCV of 20% (simulated anemia). Thirty samples from each aliquot were warmed to 37.7 °C and cooled in 2 ml allotments under one of three conditions: in ice, in ice after transfer to a precooled tube, or in an ice water bath. Temperature of each sample was recorded at one minute intervals for 15 min. Results Within treatment conditions, sample PCV had no significant effect on cooling. Cooling in ice water was significantly faster than cooling in ice only or transferring the sample to a precooled tube and cooling it on ice. Mean temperature of samples cooled in ice water was significantly lower at 15 min than mean temperatures of those cooled in ice, whether or not the tube was precooled. By 4 min, samples cooled in an ice water bath had reached mean temperatures less than 4 °C (refrigeration temperature), while samples cooled in other conditions remained above 4.0 °C for at least 11 min. For samples with a PCV of 40%, precooling the tube had no significant effect on rate of cooling on ice. For samples with a PCV of 20%, transfer to a precooled tube resulted in a significantly faster rate of cooling than direct placement of the warmed tube onto ice. Discussion Canine EDTA whole blood samples cool most rapidly and to a greater degree when placed in an ice-water bath rather than in ice. Samples stored on ice water can rapidly drop below normal refrigeration temperatures; this should be taken into consideration when using this cooling modality. PMID:27917319

CTC-mRNA (AR-V7) Analysis from Blood Samples—Impact of Blood Collection Tube and Storage Time

PubMed Central

Luk, Alison W. S.; Ma, Yafeng; Ding, Pei N.; Young, Francis P.; Chua, Wei; Balakrishnar, Bavanthi; Dransfield, Daniel T.; de Souza, Paul; Becker, Therese M.

2017-01-01

Circulating tumour cells (CTCs) are an emerging resource for monitoring cancer biomarkers. New technologies for CTC isolation and biomarker detection are increasingly sensitive, however, the ideal blood storage conditions to preserve CTC-specific mRNA biomarkers remains undetermined. Here we tested the preservation of tumour cells and CTC-mRNA over time in common anticoagulant ethylene-diamine-tetra-acetic acid (EDTA) and acid citrate dextrose solution B (Citrate) blood tubes compared to preservative-containing blood tubes. Blood samples spiked with prostate cancer cells were processed after 0, 24, 30, and 48 h storage at room temperature. The tumour cell isolation efficiency and the mRNA levels of the prostate cancer biomarkers androgen receptor variant 7 (AR-V7) and total AR, as well as epithelial cell adhesion molecule (EpCAM) were measured. Spiked cells were recovered across all storage tube types and times. Surprisingly, tumour mRNA biomarkers were readily detectable after 48 h storage in EDTA and Citrate tubes, but not in preservative-containing tubes. Notably, AR-V7 expression was detected in prostate cancer patient blood samples after 48 h storage in EDTA tubes at room temperature. This important finding presents opportunities for measuring AR-V7 expression from clinical trial patient samples processed within 48 h—a much more feasible timeframe compared to previous recommendations. PMID:28498319

Determining the effect of different environmental conditions on Ebola virus viability in clinically relevant specimens.

PubMed

Palyi, Bernadett; Magyar, Nora; Henczko, Judit; Szalai, Balint; Farkas, Agnes; Strecker, Thomas; Takacs, Maria; Kis, Zoltan

2018-03-29

In 2013-2016, West Africa experienced the largest and longest Ebola virus disease outbreak ever documented. The wide geographic spread and magnitude of the outbreak often limited the timely and rapid testing of diagnostic samples from patients with suspected Ebola virus disease, raising questions regarding the optimal storage and shipping conditions of clinically relevant specimens, including EDTA-whole blood, plasma, capillary blood, urine and seminal fluid (associated with sexual transmission of the Ebola virus after recovery from the disease). Therefore, the aim of our study was to identify the extent to which storage temperature and clinical specimen type influence Ebola virus viability. Virus infectivity was determined using a fluorescent focus-forming assay. In our study, we show that Ebola virus was the most stable in EDTA-whole blood and plasma samples, whereas rapid decay of infectivity was observed in simulated capillary blood, urine and semen samples, especially when these samples were stored at higher temperatures. The analysis of variance results demonstrated that both temperature and clinical specimen type have significant effects on virus viability, whereas donor differences were not observed. Repeated freeze and thaw cycles of the samples also had a notable impact on virus viability in EDTA-whole blood and urine. Due to the rapid temperature- and specimen-dependent degradation of the virus observed here, our study highlights the importance of proper clinical sample storage at low temperatures during transportation and laboratory analysis.

Effects of Storage and Type of Blood Collection Tubes on Hepatitis C Virus Level in Whole Blood Samples

PubMed Central

Kessler, Harald H.; Stelzl, Evelyn; Raggam, Reinhard B.; Haas, Josef; Kirchmeir, Franz; Hegenbarth, Karin; Daghofer, Elisabeth; Santner, Brigitte I.; Marth, Egon; Stauber, Rudolf E.

2001-01-01

In this study, we compared serum hepatitis C virus (HCV) RNA concentrations with HCV RNA concentrations in whole blood collection tubes, including two different types of EDTA tubes and nucleic acid stabilization tubes (NASTs). We also investigated the impact of a processing delay on HCV RNA concentration in these tubes. In NASTs, the mean HCV RNA concentration was comparable to the mean serum HCV RNA concentration at “date zero.” In EDTA tubes, mean baseline HCV RNA concentrations were higher. Storage at room temperature up to 96 h did not result in a decline of HCV RNA concentration in any of the whole blood collection tubes. In NASTs, HCV RNA concentrations remained stable during the whole study period, whereas a significant increase of HCV RNA was observed in both types of EDTA tubes at 96 h compared to date zero. We concluded that HCV RNA remains stable in NASTs at room temperature for at least 96 h, allowing greater flexibility in sample collection and transport. PMID:11325991

Assessing basophil activation by using flow cytometry and mass cytometry in blood stored 24 hours before analysis.

PubMed

Mukai, Kaori; Gaudenzio, Nicolas; Gupta, Sheena; Vivanco, Nora; Bendall, Sean C; Maecker, Holden T; Chinthrajah, Rebecca S; Tsai, Mindy; Nadeau, Kari C; Galli, Stephen J

2017-03-01

Basophil activation tests (BATs) have promise for research and for clinical monitoring of patients with allergies. However, BAT protocols vary in blood anticoagulant used and temperature and time of storage before testing, complicating comparisons of results from various studies. We attempted to establish a BAT protocol that would permit analysis of blood within 24 hours of obtaining the sample. Blood from 46 healthy donors and 120 patients with peanut allergy was collected into EDTA or heparin tubes, and samples were stored at 4°C or room temperature for 4 or 24 hours before performing BATs. Stimulation with anti-IgE or IL-3 resulted in strong upregulation of basophil CD203c in samples collected in EDTA or heparin, stored at 4°C, and analyzed 24 hours after sample collection. However, a CD63 hi population of basophils was not observed in any conditions in EDTA-treated samples unless exogenous calcium/magnesium was added at the time of anti-IgE stimulation. By contrast, blood samples collected in heparin tubes were adequate for quantification of upregulation of basophil CD203c and identification of a population of CD63 hi basophils, irrespective of whether the specimens were analyzed by means of conventional flow cytometry or cytometry by time-of-flight mass spectrometry, and such tests could be performed after blood was stored for 24 hours at 4°C. BATs to measure upregulation of basophil CD203c and induction of a CD63 hi basophil population can be conducted with blood obtained in heparin tubes and stored at 4°C for 24 hours. Copyright © 2016 American Academy of Allergy, Asthma & Immunology. Published by Elsevier Inc. All rights reserved.

Effective estimation of correct platelet counts in pseudothrombocytopenia using an alternative anticoagulant based on magnesium salt

PubMed Central

Schuff-Werner, Peter; Steiner, Michael; Fenger, Sebastian; Gross, Hans-Jürgen; Bierlich, Alexa; Dreissiger, Katrin; Mannuß, Steffen; Siegert, Gabriele; Bachem, Maximilian; Kohlschein, Peter

2013-01-01

Pseudothrombocytopenia remains a challenge in the haematological laboratory. The pre-analytical problem that platelets tend to easily aggregate in vitro, giving rise to lower platelet counts, has been known since ethylenediamine-tetra acetic acid EDTA and automated platelet counting procedures were introduced in the haematological laboratory. Different approaches to avoid the time and temperature dependent in vitro aggregation of platelets in the presence of EDTA were tested, but none of them proved optimal for routine purposes. Patients with unexpectedly low platelet counts or flagged for suspected aggregates, were selected and smears were examined for platelet aggregates. In these cases patients were asked to consent to the drawing of an additional sample of blood anti-coagulated with a magnesium additive. Magnesium was used in the beginning of the last century as anticoagulant for microscopic platelet counts. Using this approach, we documented 44 patients with pseudothrombocytopenia. In all cases, platelet counts were markedly higher in samples anti-coagulated with the magnesium containing anticoagulant when compared to EDTA-anticoagulated blood samples. We conclude that in patients with known or suspected pseudothrombocytopenia the magnesium-anticoagulant blood samples may be recommended for platelet counting. PMID:23808903

Analytical sensitivity and specificity of a loop-mediated isothermal amplification (LAMP) kit prototype for detection of Trypanosoma cruzi DNA in human blood samples.

PubMed

Besuschio, Susana A; Llano Murcia, Mónica; Benatar, Alejandro F; Monnerat, Severine; Cruz, Israel; Picado, Albert; Curto, María de Los Ángeles; Kubota, Yutaka; Wehrendt, Diana P; Pavia, Paula; Mori, Yasuyoshi; Puerta, Concepción; Ndung'u, Joseph M; Schijman, Alejandro G

2017-07-01

This study aimed to assess analytical parameters of a prototype LAMP kit that was designed for detection of Trypanosoma cruzi DNA in human blood. The prototype is based on the amplification of the highly repetitive satellite sequence of T.cruzi in microtubes containing dried reagents on the inside of the caps. The reaction is carried out at 65°C during 40 minutes. Calcein allows direct detection of amplified products with the naked eye. Inclusivity and selectivity were tested in purified DNA from Trypanosoma cruzi stocks belonging to the six discrete typing units (DTUs), in DNA from other protozoan parasites and in human DNA. Analytical sensitivity was estimated in serial dilutions of DNA samples from Sylvio X10 (Tc I) and CL Brener (Tc VI) stocks, as well as from EDTA-treated or heparinized blood samples spiked with known amounts of cultured epimastigotes (CL Brener). LAMP sensitivity was compared after DNA extraction using commercial fiberglass columns or after "Boil & Spin" rapid preparation. Moreover, the same DNA and EDTA-blood spiked samples were subjected to standardized qPCR based on the satellite DNA sequence for comparative purposes. A panel of peripheral blood specimens belonging to Chagas disease patients, including acute, congenital, chronic and reactivated cases (N = 23), as well as seronegative controls (N = 10) were evaluated by LAMP in comparison to qPCR. LAMP was able to amplify DNAs from T. cruzi stocks representative of the six DTUs, whereas it did not amplify DNAs from Leishmania sp, T. brucei sp, T. rangeli KPN+ and KPN-, P. falciparum and non-infected human DNA. Analytical sensitivity was 1x10-2 fg/μL of both CL Brener and Sylvio X10 DNAs, whereas qPCR detected up to 1x 10-1 fg/μL of CL Brener DNA and 1 fg/μl of Sylvio X10 DNA. LAMP detected 1x10-2 parasite equivalents/mL in spiked EDTA blood and 1x10-1 par.eq/mL in spiked heparinized blood using fiberglass columns for DNA extraction, whereas qPCR detected 1x10-2 par.eq./mL in EDTA blood. Boil & Spin extraction allowed detection of 1x10-2 par.eq /mL in spiked EDTA blood and 1 par.eq/ml in heparinized blood. LAMP was able to detect T.cruzi infection in peripheral blood samples collected from well-characterised seropositive patients, including acute, congenital, chronic and reactivated Chagas disease. To our knowledge, this is the first report of a prototype LAMP kit with appropriate analytical sensitivity for diagnosis of Chagas disease patients, and potentially useful for monitoring treatment response.

Analytical sensitivity and specificity of a loop-mediated isothermal amplification (LAMP) kit prototype for detection of Trypanosoma cruzi DNA in human blood samples

PubMed Central

Besuschio, Susana A.; Llano Murcia, Mónica; Benatar, Alejandro F.; Monnerat, Severine; Cruz, Israel; Picado, Albert; Curto, María de los Ángeles; Kubota, Yutaka; Wehrendt, Diana P.; Pavia, Paula; Mori, Yasuyoshi; Puerta, Concepción; Ndung'u, Joseph M.

2017-01-01

This study aimed to assess analytical parameters of a prototype LAMP kit that was designed for detection of Trypanosoma cruzi DNA in human blood. The prototype is based on the amplification of the highly repetitive satellite sequence of T.cruzi in microtubes containing dried reagents on the inside of the caps. The reaction is carried out at 65°C during 40 minutes. Calcein allows direct detection of amplified products with the naked eye. Inclusivity and selectivity were tested in purified DNA from Trypanosoma cruzi stocks belonging to the six discrete typing units (DTUs), in DNA from other protozoan parasites and in human DNA. Analytical sensitivity was estimated in serial dilutions of DNA samples from Sylvio X10 (Tc I) and CL Brener (Tc VI) stocks, as well as from EDTA-treated or heparinized blood samples spiked with known amounts of cultured epimastigotes (CL Brener). LAMP sensitivity was compared after DNA extraction using commercial fiberglass columns or after “Boil & Spin” rapid preparation. Moreover, the same DNA and EDTA-blood spiked samples were subjected to standardized qPCR based on the satellite DNA sequence for comparative purposes. A panel of peripheral blood specimens belonging to Chagas disease patients, including acute, congenital, chronic and reactivated cases (N = 23), as well as seronegative controls (N = 10) were evaluated by LAMP in comparison to qPCR. LAMP was able to amplify DNAs from T. cruzi stocks representative of the six DTUs, whereas it did not amplify DNAs from Leishmania sp, T. brucei sp, T. rangeli KPN+ and KPN-, P. falciparum and non-infected human DNA. Analytical sensitivity was 1x10-2 fg/μL of both CL Brener and Sylvio X10 DNAs, whereas qPCR detected up to 1x 10−1 fg/μL of CL Brener DNA and 1 fg/μl of Sylvio X10 DNA. LAMP detected 1x10-2 parasite equivalents/mL in spiked EDTA blood and 1x10-1 par.eq/mL in spiked heparinized blood using fiberglass columns for DNA extraction, whereas qPCR detected 1x10-2 par.eq./mL in EDTA blood. Boil & Spin extraction allowed detection of 1x10-2 par.eq /mL in spiked EDTA blood and 1 par.eq/ml in heparinized blood. LAMP was able to detect T.cruzi infection in peripheral blood samples collected from well-characterised seropositive patients, including acute, congenital, chronic and reactivated Chagas disease. To our knowledge, this is the first report of a prototype LAMP kit with appropriate analytical sensitivity for diagnosis of Chagas disease patients, and potentially useful for monitoring treatment response. PMID:28727723

Quantitative analysis of genomic DNA degradation in whole blood under various storage conditions for molecular diagnostic testing.

PubMed

Permenter, Jessalyn; Ishwar, Arjun; Rounsavall, Angie; Smith, Maddie; Faske, Jennifer; Sailey, Charles J; Alfaro, Maria P

2015-12-01

Proper storage of whole blood is crucial for isolating nucleic acids from leukocytes and to ensure adequate performance of downstream assays in the molecular diagnostic laboratory. Short-term and long-term storage recommendations are lacking for successful isolation of genomic DNA (gDNA). Container type (EDTA or heparin), temperature (4 °C and room temperature) and time (1-130 days) were assessed as criterion for sample acceptance policies. The percentage of integrated area (%Ti) between 150 and 10,000 bp from the 2200 TapeStation electropherogram was calculated to measure gDNA degradation. Refrigerated EDTA samples yielded gDNA with low %Ti (high quality). Heparinized samples stored at room temperature yielded gDNA of worst quality. Downstream analysis demonstrated that the quality of the gDNA correlated with the quality of the data; samples with high %Ti generated significantly lower levels of high molecular weight amplicons. Recommendations from these analyses include storing blood samples intended for nucleic acid isolation in EDTA tubes at 4 °C for long term storage (>10 days). gDNA should be extracted within 3 days when blood is stored at room temperature regardless of the container. Finally, refrigerated heparinized samples should not be stored longer than 9 days if expecting high quality gDNA isolates. Laboratories should consider many factors, in addition to the results obtained herein, to update their policies for sample acceptance for gDNA extraction intended for molecular genetic testing. Copyright © 2015 Elsevier Ltd. All rights reserved.

Evaluation of NGAL TestTM on Cobas 6000.

PubMed

Hansen, Young B L; Damgaard, Anette; Poulsen, Jørgen H

2014-01-01

Neutrophil Gelatinase-Associated Lipocalin (NGAL) is a promising biomarker for acute kidney injury (AKI). Our objectives were to evaluate the NGAL Test(TM) from Bioporto for both urine NGAL and plasma NGAL on the Cobas 6000 c501 (Roche Diagnostics, Rotkreuz, Switzerland) with matched measurements run on Hitachi 917, the method's linearity on the Cobas 6000 in urine, EDTA and Lithium-Heparin (Li-Hep), the influence of using EDTA or Li-Hep tubes and, finally, the impact of freezing and thawing on the sample. Forty matched samples of Li-Hep and EDTA plasma and 40 urine samples were analyzed for method, anticoagulant, and freeze-thaw comparisons. Linearity was assessed using high NGAL samples diluted in urine, EDTA, and Li-Hep plasma. Commercial internal controls were used for the imprecision study. The Cobas 6000 measured identically with the Hitachi 917, however, not in EDTA plasma (Median Difference = 17.50 μg/L, p < 0.0001). Freeze-thaw process reduced NGAL ((EDTA: Mean Difference = = 15.13 μg/L, p = 0.0014)(Li-Hep: Median Difference = = 6.5 μg/L, p = 0.0129)). NGAL results were higher in Li-Hep plasma than in EDTA plasma ((Non-thawed: Median Difference = = 14.5 μg/L, p < 0.0001), (Thawed: Median Difference = = 21.5 μg/L, p = 0.0003)). Linearity agreements were observed in all three specimens. Imprecision (CV%) was below 3%. The NGAL Test(TM) can be applied on the Cobas 6000 with acceptable performance, although the Cobas 6000 measured higher than the Hitachi 917 in EDTA plasma. Though clinically insignificant, we found that the freeze-thaw process had a reduced effect. NGAL results were higher in Li-Hep tubes than in EDTA tubes. Thus, for blood samples we recommend use of EDTA tubes for NGAL measurements.

A fit-for-purpose LC-MS/MS method for the simultaneous quantitation of ATP and 2,3-DPG in human K2EDTA whole blood.

PubMed

Kim, Hyeryun; Kosinski, Penelope; Kung, Charles; Dang, Lenny; Chen, Yue; Yang, Hua; Chen, Yuan-Shek; Kramer, Jordyn; Liu, Guowen

2017-09-01

Many hemolytic anemias results in major metabolic abnormalities: two common metabolite abnormalities include increased levels of 2,3-diphosphoglycerate (2,3-DPG) and decreased levels of adenosine triphosphate (ATP). To better monitor the concentration changes of these metabolites, the development of a reliable LC-MS/MS method to quantitatively profile the concentrations of 2, 3-DPG and ATP in whole blood is essential to understand the effects of investigational therapeutics. Accurate quantification of both compounds imposes great challenges to bioanalytical scientists due to their polar, ionic and endogenous nature. Here we present an LC-MS/MS method for the reliable quantification of 2,3-DPG and ATP from K 2 EDTA human whole blood (WB) simultaneously. Whole blood samples were spiked with stable isotope labeled internal standards, processed by protein precipitation extraction, and analyzed using zwitterionic ion chromatography-hydrophilic interaction chromatography (ZIC-HILIC) coupled with tandem mass spectrometry. The linear analytical range of the assay was 50-3000μg/mL. The fit-for-purpose method demonstrated excellent accuracy and precision. The overall accuracy was within ±10.5% (%RE) for both analytes and the intra- and inter-assay precision (%CV) were less than 6.7% and 6.2% for both analytes, respectively. ATP and 2,3-DPG were found to be stable in human K 2 EDTA blood for at least 8h at 4°C, 96days when stored at -70°C and after three freeze/thaw cycles. The assay has been successfully applied to K 2 EDTA human whole blood samples to support clinical studies. Copyright © 2017 Elsevier B.V. All rights reserved.

Appropriate Handling, Processing and Analysis of Blood Samples Is Essential to Avoid Oxidation of Vitamin C to Dehydroascorbic Acid

PubMed Central

Pullar, Juliet M.; Carr, Anitra C.

2018-01-01

Vitamin C (ascorbate) is the major water-soluble antioxidant in plasma and its oxidation to dehydroascorbic acid (DHA) has been proposed as a marker of oxidative stress in vivo. However, controversy exists in the literature around the amount of DHA detected in blood samples collected from various patient cohorts. In this study, we report on DHA concentrations in a selection of different clinical cohorts (diabetes, pneumonia, cancer, and critically ill). All clinical samples were collected into EDTA anticoagulant tubes and processed at 4 °C prior to storage at −80 °C for subsequent analysis by HPLC with electrochemical detection. We also investigated the effects of different handling and processing conditions on short-term and long-term ascorbate and DHA stability in vitro and in whole blood and plasma samples. These conditions included metal chelation, anticoagulants (EDTA and heparin), and processing temperatures (ice, 4 °C and room temperature). Analysis of our clinical cohorts indicated very low to negligible DHA concentrations. Samples exhibiting haemolysis contained significantly higher concentrations of DHA. Metal chelation inhibited oxidation of vitamin C in vitro, confirming the involvement of contaminating metal ions. Although EDTA is an effective metal chelator, complexes with transition metal ions are still redox active, thus its use as an anticoagulant can facilitate metal ion-dependent oxidation of vitamin C in whole blood and plasma. Handling and processing blood samples on ice (or at 4 °C) delayed oxidation of vitamin C by a number of hours. A review of the literature regarding DHA concentrations in clinical cohorts highlighted the fact that studies using colourimetric or fluorometric assays reported significantly higher concentrations of DHA compared to those using HPLC with electrochemical detection. In conclusion, careful handling and processing of samples, combined with appropriate analysis, is crucial for accurate determination of ascorbate and DHA in clinical samples. PMID:29439480

Proinsulin is stable at room temperature for 24 hours in EDTA: A clinical laboratory analysis (adAPT 3).

PubMed

Davidson, Jane; McDonald, Timothy; Sutherland, Calum; Mostazir, Mohammod; VanAalten, Lidy; Wilkin, Terence

2017-01-01

Reference laboratories advise immediate separation and freezing of samples for the assay of proinsulin, which limit its practicability for smaller centres. Following the demonstration that insulin and C-peptide are stable in EDTA at room temperature for at least 24hours, we undertook simple stability studies to establish whether the same might apply to proinsulin. Venous blood samples were drawn from six adult women, some fasting, some not, aliquoted and assayed immediately and after storage at either 4°C or ambient temperature for periods from 2h to 24h. There was no significant variation or difference with storage time or storage condition in either individual or group analysis. Proinsulin appears to be stable at room temperature in EDTA for at least 24h. Immediate separation and storage on ice of samples for proinsulin assay is not necessary, which will simplify sample transport, particularly for multicentre trials.

Conditions in blood sampling procedures that extend the ex vivo stability of eosinophil activity markers in peripheral blood from allergic patients and healthy controls.

PubMed

Halldén, G; Nopp, A; Ihre, E; Peterson, C; Lundahl, J

1999-11-01

Serum-ECP, EG2-epitope on intracellular ECP and surface expression of CD9 and CD11b in peripheral blood eosinophils (PBE) are considered to be markers that mirror clinical parameters in allergic inflammation. The aim was to investigate the impact of the blood sampling procedure on PBE markers and to identify optimal conditions for extended pre-analysis storage. Blood, from healthy individuals and patients with allergic rhinitis/asthma, was collected in tubes with EDTA, citrate, or without anti-coagulant. The expression of EG2-epitope, CD9, and CD11b were analyzed in eosinophils and neutrophils after 1, 5, and 24 hours of storage at +4 degrees C, according to the FOG-method and flow cytometry. In vitro stimulation with fMLP/PMA was used for metabolic activity analysis and CD11b mobilization. Following a 1-hour clotting period at +20 to 22 degrees C, samples were stored at +4 degrees C and serum-ECP levels were measured. The EG2-epitope, serum-ECP, and CD9 were stable in samples from both healthy controls and allergic patients at all storage conditions. The EG2-epitope, serum-ECP and PBE count were significantly increased in the patient group, whereas no differences were observed in the expression of CD9 or CD11b. Both granulocytes and monocytes retained their metabolic activity for 24 hours. Neutrophils in citrate-blood increased their ability to respond to fMLP, as compared with EDTA-blood. In vitro analysis of selected activity markers and functional tests could be performed on granulocytes from both healthy individuals and allergic patients after 24 hours storage at +4 degrees C. The anticoagulant citrate seems to be preferable to EDTA when monocytes or CD11b expression are analyzed.

Effect of glutathione on the cadmium chelation of EDTA in a patient with cadmium intoxication.

PubMed

Gil, Hyo-wook; Kang, Eun-jung; Lee, Kwon-hyun; Yang, Jong-oh; Lee, Eun-young; Hong, Sae-yong

2011-01-01

In order to evaluate the efficiency and renal protective effects of glutathione during Ca(++)-EDTA chelation therapy for chronic cadmium intoxication, we measured the renal excretion of cadmium, β(2)-microglobulin, proteinuria, and hematuria during intravenous administration of glutathione with Ca(++)-EDTA in a 54-year-old patient with chronic cadmium intoxication. We administered 500 mg of Ca(++)-EDTA and 50 mg/kg of glutathione alone or in 1 L of normal saline over the next 24 hours and repeated this over 12 consecutive days. During the first 3 days, the basal levels (only saline administration) were determined; during the second 3 days, Ca(++)-EDTA only was administered, for the third sequence of 3 days, Ca(++)-EDTA with glutathione was provided, and for the last 3 days, glutathione alone was given. One month later, the same protocol was repeated. There were six blood and urine samples to analyze in each group. The blood cadmium level was higher when the EDTA was infused together with glutathione (7.44 ± 0.73 µg/L, p < 0.01) compared to the basal level of 4.6 ± 0.44 µg/L. Also, the renal cadmium excretion was significantly higher in the EDTA with glutathione group than in the basal group (23.4 ± 15.81 µg/g creatinine vs 89.23 ± 58.52 µg/g creatinine, p < 0.01). There was no difference in the protein/creatinine and β(2)-microglobulin/creatinine ratio in the urine (p > 0.05) among the groups. Furthermore, microhematuria and proteinuria did not develop over the observation period of 6 months. These results suggest that glutathione administration with EDTA might be an effective treatment modality for patients with cadmium intoxication.

Blood Substrate Collection and Handling Procedures under Pseudo-Field Conditions: Evaluation of Suitability for Inflammatory Biomarker Measurement.

PubMed

Sugden, Karen; Danese, Andrea; Shalev, Idan; Williams, Benjamin S; Caspi, Avshalom

2015-01-01

Routine incorporation of blood-based biomarker measurements in population studies has been hampered by challenges in obtaining samples suitable for biomarker assessment outside of laboratory settings. Here, we assessed the suitability of venous blood left unprocessed for 4, 24, or 48 hours post-collection at either room temperature or 4°C for quantification of two biomarkers, Interleukin-6 (IL-6) and C-reactive protein (CRP). Blood samples were collected in both K2EDTA tubes and a dedicated plasma-preservation tube, P100. Dried blood spot (DBS) samples from the same subjects were also collected in order to compare delayed-processing plasma performance against a popular alternative collection method. We found that K2EDTA mean plasma concentrations of both IL-6 and CRP were not significantly different from concentrations in plasma processed immediately; this was observed for tubes stored up to 48 hours pre-processing at either temperature. Concentrations of IL-6 measured in P100 tubes showed significant time-dependent increases when stored at room temperature; otherwise, levels of IL-6 and CRP were similar to those found in samples processed immediately. Levels of CRP in DBS were correlated with plasma CRP levels, even when pre-processed blood was stored for up to 48 hours. These data indicate that plasma is suitable for IL-6 and CRP estimation under data collection conditions that involve processing delays.

K(3)EDTA Vacuum Tubes Validation for Routine Hematological Testing.

PubMed

Lima-Oliveira, Gabriel; Lippi, Giuseppe; Salvagno, Gian Luca; Montagnana, Martina; Poli, Giovanni; Solero, Giovanni Pietro; Picheth, Geraldo; Guidi, Gian Cesare

2012-01-01

Background and Objective. Some in vitro diagnostic devices (e.g, blood collection vacuum tubes and syringes for blood analyses) are not validated before the quality laboratory managers decide to start using or to change the brand. Frequently, the laboratory or hospital managers select the vacuum tubes for blood collection based on cost considerations or on relevance of a brand. The aim of this study was to validate two dry K(3)EDTA vacuum tubes of different brands for routine hematological testing. Methods. Blood specimens from 100 volunteers in two different K(3)EDTA vacuum tubes were collected by a single, expert phlebotomist. The routine hematological testing was done on Advia 2120i hematology system. The significance of the differences between samples was assessed by paired Student's t-test after checking for normality. The level of statistical significance was set at P < 0.05. Results and Conclusions. Different brand's tubes evaluated can represent a clinically relevant source of variations only on mean platelet volume (MPV) and platelet distribution width (PDW). Basically, our validation will permit the laboratory or hospital managers to select the brand's vacuum tubes validated according to him/her technical or economical reasons for routine hematological tests.

K3EDTA Vacuum Tubes Validation for Routine Hematological Testing

PubMed Central

Lima-Oliveira, Gabriel; Lippi, Giuseppe; Salvagno, Gian Luca; Montagnana, Martina; Poli, Giovanni; Solero, Giovanni Pietro; Picheth, Geraldo; Guidi, Gian Cesare

2012-01-01

Background and Objective. Some in vitro diagnostic devices (e.g, blood collection vacuum tubes and syringes for blood analyses) are not validated before the quality laboratory managers decide to start using or to change the brand. Frequently, the laboratory or hospital managers select the vacuum tubes for blood collection based on cost considerations or on relevance of a brand. The aim of this study was to validate two dry K3EDTA vacuum tubes of different brands for routine hematological testing. Methods. Blood specimens from 100 volunteers in two different K3EDTA vacuum tubes were collected by a single, expert phlebotomist. The routine hematological testing was done on Advia 2120i hematology system. The significance of the differences between samples was assessed by paired Student's t-test after checking for normality. The level of statistical significance was set at P < 0.05. Results and Conclusions. Different brand's tubes evaluated can represent a clinically relevant source of variations only on mean platelet volume (MPV) and platelet distribution width (PDW). Basically, our validation will permit the laboratory or hospital managers to select the brand's vacuum tubes validated according to him/her technical or economical reasons for routine hematological tests. PMID:22888448

Proinsulin is stable at room temperature for 24 hours in EDTA: A clinical laboratory analysis (adAPT 3)

PubMed Central

Davidson, Jane; McDonald, Timothy; Sutherland, Calum; Mostazir, Mohammod; VanAalten, Lidy

2017-01-01

Aims Reference laboratories advise immediate separation and freezing of samples for the assay of proinsulin, which limit its practicability for smaller centres. Following the demonstration that insulin and C-peptide are stable in EDTA at room temperature for at least 24hours, we undertook simple stability studies to establish whether the same might apply to proinsulin. Methods Venous blood samples were drawn from six adult women, some fasting, some not, aliquoted and assayed immediately and after storage at either 4°C or ambient temperature for periods from 2h to 24h. Results There was no significant variation or difference with storage time or storage condition in either individual or group analysis. Conclusion Proinsulin appears to be stable at room temperature in EDTA for at least 24h. Immediate separation and storage on ice of samples for proinsulin assay is not necessary, which will simplify sample transport, particularly for multicentre trials. PMID:28426711

A novel comprehensive set of fungal Real time PCR assays (fuPCR) for the detection of fungi in immunocompromised haematological patients-A pilot study.

PubMed

Rahn, Sebastian; Schuck, Anna; Kondakci, Mustafa; Haas, Rainer; Neuhausen, Nicole; Pfeffer, Klaus; Henrich, Birgit

2016-12-01

Fungal infections are recognized in an increasing number of patients with immunological deficits and are associated with high rates of mortality (Brown et al., 2012a). In this pilot-study, a rapid Real time PCR (fuPCR) was designed for the detection and differentiation of fungal pathogens in clinical specimens of haematological patients. The fuPCR, targeting the internal transcribed spacer region 2 (ITS2) of rDNA region, is comprised of seven multiplex reactions, which were shown to be specific and sensitive for a comprehensive spectrum of clinically relevant fungal species. This was validated by testing respective fungal DNAs in each fuPCR reaction and 28 respiratory samples of fungal pneumonia-proven patients. Clinical sample sets of throat swab, EDTA-blood and blood sera from 50 patients with severe haematological malignancies, including haematopoietic stem cell transfer (HSCT), and samples from 30 healthy individuals were then analysed. In a first step, 198 samples of immunosuppressed patients were solely examined by fuPCR; and 50.8% (33/65) respiratory swabs, 4.8% (3/63) EDTA blood samples and 1.4% (1/70) blood serum samples were tested positive. In a second step, 56 respiratory samples of immunosuppressed patients and 30 of healthy individuals were simultaneously analysed by fuPCR and standard cultivation techniques. By both methods 30.4% (17/56) swabs of the immunocompromised patients were tested positive, 37.5% (21/56) were tested negative and 32.1% (18/56) were tested fuPCR positive and culture negative. In analysing the blood samples of the immunocompromised patients 5.4% (3/56) EDTA blood samples and 16.1% (9/56) sera samples were tested fuPCR-positive, whereas all samples of 30 healthy individuals with no signs of immunological deficits were tested negative by fuPCR. 38.9% (14/36) of the fungi detected in respiratory samples of the immunosuppressed patients, belonged to Candida spp., 47.2% (17/36) to Saccharomyces spp., 5.6% (2/36) to Cladosporium spp. and 8.3% (3/36) to Alternaria spp., whereas cultivation only identified Candida spp. (10/17) and Saccharomyces spp. (7/17). In this pilot study a novel fuPCR assay was developed and validated for the simultaneous and comprehensive detection of fungal pathogens in clinical respiratory specimens of haematological patients. Future work will focus on the validation of the blood-stream detected fungi in pathogenicity of these patients. Copyright © 2016 Elsevier GmbH. All rights reserved.

[Characteristic of sample banks isolated from EDTA-blood by sedimentation method].

PubMed

Chen, Zhi-bin; Lin, Qin; Ma, Chang-hua; Liu, Kai-ning; Meng, Huan-xin

2014-02-18

To assess the characteristics of establishing the different sample banks of plasma, leukocytes and DNA by sedimentation method of isolating from ethylene diamine tetraacetic acid(EDTA)-blood and to clarify the sedimentation method of leukocyte isolation and plasma volume by comparative data and recommended procedures for applicability. In the study, 29 EDTA-bloods were obtained, the total amounts of leukocytes and the percentage of neutrophile granulocytes, and lymphocytes in the EDTA-blood detected as a control group and then assigned equally into 4 EP tubes with 1 mL EDTA-blood per tube as 4 test groups, then the 4 tubes were placed with the EDTA-blood at room temperature and the plasma layers were isolated at 0.5, 1, 2 and 3 h, receptively. The total amount of leukocytes and the percentage of neutrophile granulocytes, and lymphocytes were detected by automated hematology analyzer at the clinical laboratory. The volume of the plasma was also measured at the same time. The plasma volume at 0.5 h [(241.72 ± 101.52)μL] was substantially lower than those at 1 h[(317.24 ± 97.50)μL], at 2 h[(371.03 ± 91.66)μL], and at 3 h [(408.97 ± 97.43)μL] , P < 0.05. The plasma volume at 1 h was substantially lower than those at 2 h and 3 h (P < 0.05). The total amount of leukocytes in the plasma layer at 0.5 h (2.50 × 10(6) ± 1.48 × 10(6)) group was substantially higher than the amount of 2 or 3 h groups respectively (1.47 × 10(6) ± 7.19 × 105,1.21 × 10(6) ± 7.41 × 105), P < 0.05. Significant difference was not found between 0.5 h group and 1 h group (2.29 × 10(6)± 1.17 × 10(6)), P > 0.05. The total amount of leukocytes in the plasma layer in 1 h group was substantially higher than that in 2 h and 3 h groups (P < 0.05). There was no significant difference between 3 h group and 2 h group (P > 0.05). The total amount of leukocytes in the plasma layer of the 4 test groups was substantially lower than that in the control group (P < 0.05). The percentage of neutrophile granulocytes (54.14% ± 11.65%) in the plasma layer in 0.5 h group was substantially higher than those in 1 h, 2 h and 3 h groups (46.66% ± 12.70%,39.17% ± 12.33%,43.25% ± 14.54%), P < 0.05, respectively, which was the substantially lower than that in the control group (60.53% ± 8.46%), P < 0.05. The average value of the percentage of neutrophile granulocytes in the plasma layer in 1 h group was substantially higher than that in 2 h group (P < 0.05). There was no significant different between 3 h group and both 1 h, 2 h groups (P > 0.05). The mean percentage of lymphocytes in the plasma layer in 0.5 h group (35.09% ± 10.84%) was substantially lower than those in the plasma layer in 1 h, 2 h and 3 h groups, respectively ( 41.48% ± 12.20%, 47.96% ± 12.27%, 45.50% ± 13.71%), which was significant higher than that in the control group(30.98% ± 7.33%), P < 0.05. The average value of the percentage of lymphocytes in the plasma layer in 1 h group was substantially higher than those in the control group and 0.5 h group, but was substantially lower than those in 2 h and 3 h groups (P < 0.05). The average value of percentage of lymphocytes in the plasma layer in 2 h group was substantially higher than those in the control group, 0.5 h and 1 h groups (P < 0.05). There was no significant difference between 2 h and 3 h groups (P > 0.05). The best period of time in obtaining leukocytes is 0.5-1 h sedimentation of EDTA-blood. Both the plasma layer and leukocytes can be separated and obtained at the same time from the same sample by the sedimentation method of EDTA-blood. The sedimentation of EDTA-blood has the least interference of both chemical and physical factors, as well as a ready operation, which can establish the plasma, leukocytes and DNA sample banks for various aspects of research.

Efficacy of Minocycline and EDTA Lock Solution in Preventing Catheter-Related Bacteremia, Septic Phlebitis, and Endocarditis in Rabbits

PubMed Central

Raad, Issam; Hachem, Ray; Tcholakian, Robert K.; Sherertz, Robert

2002-01-01

To determine the efficacy of antibiotic catheter lock solution in preventing catheter-related infections, silicone catheters were tunneled and inserted into the jugular veins of 18 rabbits. The catheters were challenged with an intraluminal injection of 105 CFU of slime-producing Staphylococcus epidermidis in 0.1 ml of water. The catheters were maintained on heparin (100 IU/ml) flush for the first 3 days. On day 3, quantitative blood samples for culture were obtained from the catheters and ear veins, which documented catheter-related bacteremia, and the rabbits were randomized to have their catheters flushed as follows: five animals were continued on heparin (100 IU/ml), five animals received vancomycin (3 mg/ml) with heparin (100 IU/ml), and eight animals received 3 mg of minocycline per ml with 30 mg of EDTA per ml (M-EDTA). All animals were killed at day 7. Blood, catheters, jugular veins, and heart valves were cultured quantitatively. Animals maintained on heparin developed catheter-related colonization, bacteremia, septic phlebitis, and endocarditis. Vancomycin-heparin partially prevented catheter colonization, bacteremia, and phlebitis (P = 0.2). M-EDTA completely prevented catheter colonization, catheter-related bacteremia, and phlebitis in all of the animals (P < 0.01). Tricuspid endocarditis was equally prevented by vancomycin-heparin and M-EDTA (P ≤ 0.06). In conclusion, the M-EDTA catheter flush solution was highly efficacious in preventing catheter-related colonization, bacteremia, septic phlebitis, and endocarditis in rabbits. PMID:11796338

Behavior of lead and zinc in plasma, erythrocytes, and urine and ALAD in erythrocytes following intravenous infusion of CaEDTA in lead workers.

PubMed

Araki, S; Aono, H; Fukahori, M; Tabuki, K

1984-01-01

To evaluate the effect of calcium disodium ethylenediamine tetraacetate (CaEDTA) on concentrations of lead and zinc in plasma, erythrocytes, whole blood, and urine, we administered CaEDTA by intravenous infusion for 1 hr to seven lead workers with blood lead concentrations of 46-67 micrograms/100 g (mean 54 micrograms/100 g). The plasma lead concentration (PPb) and the mobilization yield of lead in urine by CaEDTA were highest during the period between 1 and 2 hr after the infusion was started. In contrast, the lead concentration in erythrocytes (EPb) and in whole blood (BPb) remained unchanged during the 24 hr following infusion. Plasma zinc concentration (PZn) also fell rapidly following CaEDTA infusion; the decline was followed by a gradual rise in the zinc concentration in erythrocytes (EZn) without alteration in the zinc in whole blood. The mobilization yield of zinc in urine by CaEDTA (MZn) reached its highest level within 1 hr after the start of the infusion. Delta-aminolevulinic acid dehydratase (ALAD) activity in erythrocytes gradually increased for 5 hr following CaEDTA infusion. These observations suggest that (1) PPb concentration is a more sensitive indicator of the body burden of chelatable lead than is either BPb or EPb; (2) MZn is mobilized mostly from plasma during the first several hours following the start of CaEDTA infusion, and the fall in PZn concentration following infusion is compensated first by a rise in EZn concentration and then by an immediate redistribution of zinc in other organs to the blood; and (3) Pb-inhibited ALAD activity is reactivated by the increased EZn during and shortly after CaEDTA infusion.

Stability of hematologic analytes in monkey, rabbit, rat, and mouse blood stored at 4°C in EDTA using the ADVIA 120 hematology analyzer.

PubMed

Ameri, Mehrdad; Schnaars, Henry A; Sibley, John R; Honor, David J

2011-06-01

The time from sampling to analysis can be delayed when blood samples are shipped to distant reference laboratories or when analysis cannot be readily performed. The objective of this study was to evaluate the stability of hematologic analytes in blood samples from monkeys, rabbits, rats, and mice when samples were stored for up to 72 hours at 4°C. Blood samples from 30 monkeys, 15 rabbits, 20 rats, and 30 mice were collected into EDTA-containing tubes and were initially analyzed within 1 hour of collection using the ADVIA 120 analyzer. The samples were then stored at 4°C and reanalyzed at 24, 48, and 72 hours after collection. Significant (P<.0003) changes in hematologic analytes and calculations included increased HCT and MCV and decreased MCHC and cell hemoglobin concentration mean (CHCM) at 72 hours and increased MPV at 24 hours in monkeys; increased MCV at 72 hours and MPV at 48 hours and decreased monocyte count at 24 hours in rabbits; increased MCV and decreased MCHC, CHCM, and monocyte count at 24 hours in rats; increased MCV, red cell distribution width, and MPV and decreased MCHC, CHCM, and monocyte count at 24 hours in mice. Although most of the changes in the hematologic analytes in blood from monkeys, rabbits, rats, and mice when samples were stored at 4°C were analytically acceptable and clinically negligible, the best practice in measuring hematologic analytes in these animals is timely processing of blood samples, preferably within 1 hour after collection. ©2011 American Society for Veterinary Clinical Pathology.

Blood Substrate Collection and Handling Procedures under Pseudo-Field Conditions: Evaluation of Suitability for Inflammatory Biomarker Measurement

PubMed Central

Danese, Andrea; Shalev, Idan; Williams, Benjamin S.; Caspi, Avshalom

2015-01-01

Routine incorporation of blood-based biomarker measurements in population studies has been hampered by challenges in obtaining samples suitable for biomarker assessment outside of laboratory settings. Here, we assessed the suitability of venous blood left unprocessed for four, 24 or 48 hours post-collection at either room temperature or 4°C for quantification of two biomarkers, Interleukin-6 (IL-6) and C-Reactive Protein (CRP). Blood samples were collected in both K2EDTA tubes and a dedicated plasma-preservation tube, P100. Dried Blood Spot (DBS) samples from the same subjects were also collected in order to compare delayed-processing plasma performance against a popular alternative collection method. K2EDTA mean plasma concentrations of both IL-6 and CRP were not significantly different from concentrations in plasma processed immediately; this was observed for tubes stored up to 48 hours pre-processing at either temperature. Concentrations of IL-6 measured in P100 tubes showed significant time-dependent increases when stored at room temperature; otherwise, levels of IL-6 and CRP were similar to those processed immediately. Levels of CRP in DBS were correlated with plasma CRP levels, even when pre-processed blood was stored for up to 48 hours. These data indicate that plasma is suitable for IL-6 and CRP estimation under data-collection conditions that involve processing delays. PMID:26652682

Comparison of human whole blood, plasma, and serum matrices for the determination of perfluorooctanesulfonate (PFOS), perfluorooctanoate (PFOA), and other fluorochemicals

DOE Office of Scientific and Technical Information (OSTI.GOV)

Ehresman, David J.; Froehlich, John W.; Olsen, Geary W.

2007-02-15

Interest in human exposure to perfluorinated acids, including perfluorobutanesulfonate (PFBS), perfluorohexanesulfonate (PFHS), perfluorooctanesulfonate (PFOS), and perfluorooctanoate (PFOA) has led to their measurement in whole blood, plasma and serum. Comparison of measurements in these different blood-based matrices, however, has not been rigorously investigated to allow for across-matrix comparisons. This research evaluated concentrations of PFBS, PFHS, PFOS, and PFOA in whole blood collected in heparin (lithium) and ethylenediamine tetraacetic acid (EDTA), plasma samples collected in heparin and EDTA, and serum (from whole blood allowed to clot). Blood samples were collected from 18 voluntary participants employed at 3M Company. Solid phase extraction methodsmore » were used for all analytical sample preparations, and analyses were completed using high-pressure liquid chromatography/tandem mass spectrometry methods. Serum concentrations ranged from: limit of quantitation (LOQ, 5 ng/mL) to 25 ng/mL for PFBS; LOQ (5 ng/mL) to 75 ng/mL for PFHS; LOQ (5 ng/mL) to 880 ng/mL for PFOS; and LOQ (5 or 10 ng/mL) to 7320 ng/mL for PFOA. Values less than the LOQ were not included in the statistical analyses of the mean of the ratios of individual values for the matrices. PFBS was not quantifiable in most samples. Serum to plasma ratios for PFHS, PFOS, and PFOA were 1:1 and this ratio was independent of the level of concentrations measured. Serum or plasma to whole blood ratios, regardless of the anticoagulant used, approximated 2:1. The difference between plasma and serum and whole blood corresponded to volume displacement by red blood cells, suggesting that the fluorochemicals are not found intracellularly or attached to the red blood cells.« less

Comparison of 2 electronic cowside tests to detect subclinical ketosis in dairy cows and the influence of the temperature and type of blood sample on the test results.

PubMed

Iwersen, M; Klein-Jöbstl, D; Pichler, M; Roland, L; Fidlschuster, B; Schwendenwein, I; Drillich, M

2013-01-01

The objective of this study was to determine the suitability of 2 electronic hand-held devices [FreeStyle Precision (FSP), Abbott GmbH & Co. KG, Wiesbaden, Germany and GlucoMen LX Plus (GLX), A. Menarini GmbH, Vienna, Austria] for measuring β-hydroxybutyrate (BHBA) in dairy cows. Three experiments were conducted to evaluate (1) the diagnostic performance of the devices, (2) the effect of the type of blood sample, and (3) the influence of the ambient temperature on the determined results. A total of 415 blood samples from lactating Holstein and Simmental cows were collected and analyzed with both devices (whole blood) and in a laboratory (serum). Correlation coefficients between whole-blood and serum BHBA concentrations were highly significant, with 94% for the FSP and 80% for the GLX device. Based on thresholds for subclinical ketosis of 1.2 and 1.4 mmol of BHBA/L, results obtained with the hand-held devices were evaluated by receiver operating characteristics analyses. This resulted in adjusted thresholds of 1.2 and 1.4 mmol/L for the FSP and 1.1 and 1.3 mmol/L for the GLX device. Applying these thresholds, sensitivities were 98 and 100% for the FSP and 80 and 86% for the GLX device, respectively. Corresponding specificities were 90 and 97% for the FSP and 87 and 96% for the GLX device, respectively. Additionally, concentrations of BHBA were tested with both devices in whole blood, EDTA-added whole blood, and in their resulting serum and plasma, collected from 65 animals. Determined BHBA concentrations were similar within each device for whole and EDTA-added blood, and in serum and plasma, but differed between whole blood and serum and between EDTA-added blood and plasma. Blood samples with low (0.4 mmol/L), medium (1.1 mmol/L), and high (1.6 mmol/L) BHBA concentrations were stored between +5 to +32°C and analyzed repeatedly at temperature levels differing by 4°C. Additionally, devices and test strips were stored at equal conditions and used for measurement procedures. Storage temperature of the devices and test strips did not influence the differences between the results of the laboratory and the devices, whereas the temperature of the blood samples caused significant differences. Although the level of agreement between the laboratory and the GLX device was lower than for the laboratory and the FSP device, both devices are useful tools for monitoring subclinical ketosis in dairy cows. Due to their effects on the determined results, the type and temperature of the tested sample should be considered. Copyright © 2013 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Evacuated blood-collection tubes for haematological tests - a quality evaluation prior to their intended use for specimen collection.

PubMed

Gros, Nataša

2013-05-01

An inappropriate anticoagulant concentration in a blood sample can cause cell shrinkage and affect the haematocrit and mean corpuscular volume (MCV). In evacuated blood-collection tubes there are two parameters affecting the quality of the product: the anticoagulant amount introduced into the tube during its production and the internal under-pressure at the instant of the blood-specimen collection affecting the draw-volume. No testing procedures that would give an insight into the anticoagulant concentration that can be expected for blood samples after specimen collection have been available up until now. The methodology suggested here combines the draw-volume test performed with deionised water using a laboratory made measuring device, and a conductivity measurement. The corrections taking into account the air pressure and ambient temperature provide an insight into the anticoagulant concentration that can be expected for blood samples. Results presented in the form of a nomogram facilitate the routine use of the suggested methodology. Our 338-day study confirmed significant differences and variations in the quality and the anticoagulant concentrations of the K₃EDTA and K2EDTA tubes of different producers and identified different examples of non-compliance with the norms during the shelf life of the tubes. The quality evaluation of the evacuated blood-collection tubes prior to their intended use as suggested here can, in everyday laboratory practice, ensure that the tubes are used only if, and only until, their quality is adequate.

Improved detection of canine Angiostrongylus vasorum infection using real-time PCR and indirect ELISA.

PubMed

Jefferies, Ryan; Morgan, Eric R; Helm, Jenny; Robinson, Matthew; Shaw, Susan E

2011-12-01

This study reports the development of a real-time PCR assay and an indirect ELISA to improve on current detection of canine Angiostrongylus vasorum infection. A highly specific fluorescent probe-based, real-time PCR assay was developed to target the A. vasorum second internal transcribed spacer region and detected DNA in EDTA blood, lung tissue, broncho-alveolar larvage fluid, endotracheal mucus, pharyngeal swabs and faecal samples. PCR was fast (∼1 h), highly efficient when using EDTA blood samples, consistently detected a single molecule of parasite DNA and did not amplify DNA from other parasitic nematodes or definitive host species. An indirect ELISA was also developed using the soluble protein fraction from adult A. vasorum worms. Some cross-reactive antigen recognition was observed when tested against sera from dogs infected with Crenosoma vulpis (n = 8), Toxocara canis (n = 5) and Dirofilaria immitis (n = 5). This was largely overcome by setting the cut-off for a positive result at an appropriately high level. Field evaluation of the real-time PCR and ELISA was conducted by testing sera and EDTA blood from dogs with suspected A. vasorum infection (n = 148) and compared with the Baermann's larval migration test in faeces. Thirty-one dogs were positive by at least one test. Of these, 20 (65%) were detected by the Baermann method, 18 (58%) by blood PCR, 24 (77%) by ELISA and 28 (90%) by blood PCR and ELISA together. Combined testing using real-time PCR and ELISA therefore improved the detection rate of A. vasorum infection and holds promise for improved clinical diagnosis and epidemiological investigation.

A comparison of the diminution rates of lead in blood and lead mobilized by CaEDTA after termination of occupational exposure: a long-term observation in two lead workers.

PubMed

Araki, S; Murata, K; Aono, H; Yanagihara, S; Ushio, K

1983-07-01

CaEDTA 20 mg/kg was administered weekly for 3.5 years after termination of occupational exposure to two lead workers. The diminution half-lives for lead in blood and urine lead mobilized by CaEDTA were 4.8 and 3.3 years respectively for subject 1 following 28 years exposure and 3.3 and 2.0 years respectively for subject 2 following 26 years exposure. The difference in the diminution rate between lead in blood and lead mobilized by CaEDTA was significant in subject 2 (p less than 0.05).

Reliability of ⁵¹Cr-EDTA plasma and urinary clearance as a measure of residual renal function in dialysis patients.

PubMed

Kjaergaard, Krista D; Jensen, Jens D; Jespersen, Bente; Rehling, Michael

2011-12-01

In dialysis patients, longer survival is associated with a higher residual renal function. Randomized controlled trials are conducted to clarify how residual renal function can be preserved. However, existing methods for measuring residual renal function are uncertain and there is a need for establishing a standard for measurements of glomerular filtration rate (GFR) in dialysis patients. ⁵¹Cr-EDTA clearances in plasma, urine, and dialysate were evaluated in a sample of 12 hemodialysis and 12 peritoneal dialysis patients. The patients' condition was generally stable, and all patients were investigated twice within 4-10 days. Plasma clearances of ⁵¹Cr-EDTA for all patients ranged between 2.1 and 30.8 mL/min/1.73 m², whereas urinary ⁵¹Cr-EDTA clearances ranged from 0.7-20.0 mL/min/1.73 m². This difference was statistically significant (p < 0.001). Week-to-week reproducibility expressed as coefficients of variation were below or equal to 10% for plasma clearances and 13% for urinary clearances in hemodialysis patients and 14% in peritoneal dialysis patients. This study demonstrated a difference between ⁵¹Cr-EDTA plasma and urinary clearances in dialysis patients. Plasma clearance of ⁵¹Cr-EDTA had the best reproducibility. For repeated measurements as in clinical prospective trials, we recommend ⁵¹Cr-EDTA plasma clearance based on blood sampling at 5 + 24 hours with subtraction of ⁵¹Cr-EDTA dialysate clearance in peritoneal dialysis patients. Further studies are needed to corroborate our findings.

IL8 and IL16 levels indicate serum and plasma quality.

PubMed

Kofanova, Olga; Henry, Estelle; Quesada, Rocio Aguilar; Bulla, Alexandre; Linares, Hector Navarro; Lescuyer, Pierre; Shea, Kathi; Stone, Mars; Tybring, Gunnel; Bellora, Camille; Betsou, Fay

2018-02-09

Longer pre-centrifugation times alter the quality of serum and plasma samples. Markers for such delays in sample processing and hence for the sample quality, have been identified. Twenty cytokines in serum, EDTA plasma and citrate plasma samples were screened for changes in concentration induced by extended blood pre-centrifugation delays at room temperature. The two cytokines that showed the largest changes were further validated for their "diagnostic performance" in identifying serum or plasma samples with extended pre-centrifugation times. In this study, using R&D Systems ELISA kits, EDTA plasma samples and serum samples with a pre-centrifugation delay longer than 24 h had an IL16 concentration higher than 313 pg/mL, and an IL8 concentration higher than 125 pg/mL, respectively. EDTA plasma samples with a pre-centrifugation delay longer than 48 h had an IL16 concentration higher than 897 pg/mL, citrate plasma samples had an IL8 concentration higher than 21.5 pg/mL and serum samples had an IL8 concentration higher than 528 pg/mL. These robust and accurate tools, based on simple and commercially available ELISA assays can greatly facilitate qualification of serum and plasma legacy collections with undocumented pre-analytics.

Evaluation of Different Blood Collection Tubes and Blood Storage Conditions for the Preservation and Stability of Cell-Free Circulating DNA for the Analysis of the Methylated mSEPT9 Colorectal Cancer Screening Marker.

PubMed

Distler, Jurgen; Tetzner, Reimo; Weiss, Gunter; König, Thomas; Schlegel, Anne; Bagrowski, Michal

2016-01-01

For the subsequent analysis of the methylated mSEPT9 colorectal cancer screening marker in plasma, different blood collection tubes and blood storage conditions were investigated. The study demonstrated that methylated Septin 9 ( m SEPT9) can be consistently detected in plasma samples derived from whole blood samples collected with S-Monovette® K3E and BD Vacutainer ® K2EDTA tubes stored at 2-8 °C for a maximum of 24 h and for samples collected in S-Monovette CPDA tubes stored at 18-25 °C for up to 48 h.

Stability of selected hematology variables in canine blood kept at room temperature in EDTA for 24 and 48 hours.

PubMed

Médaille, C; Briend-Marchal, A; Braun, J P

2006-03-01

Most hematologic analyses are performed within a short time of blood sampling, but samples collected at the end of a week may have to be stored for up to 2 days. The stability of hematologic constituents is poorly documented. The objective of this study was to compare the results of RBC, WBC and platelet counts, hemoglobin (Hgb) concentration, and MCV before and after storage of canine blood at room temperature for 24 and 48 hours. One hundred fifty-two K3-EDTA canine blood specimens from 2 veterinary hospitals were analyzed within 4 hours of collection, then 24 and 48 hours later with a Coulter T540 hematology analyzer. Results were compared by Passing-Bablock agreement, difference plots, and according to their classification as normal or abnormal based on reference intervals. RBC count and Hgb concentration were stable for the duration of the study. Differences in WBC and platelet counts varied with the specimen, independently of the initial value. MCV increased consistently over the 2 days. However, only a few results were misclassified. Whole blood specimens stored for up to 2 days at room temperature are suitable for cell counts and Hgb measurement. However, potential variations have to be known to avoid misinterpretations, especially near the decision limits.

Metabolomic Quality Assessment of EDTA Plasma and Serum Samples.

PubMed

Malm, Linus; Tybring, Gunnel; Moritz, Thomas; Landin, Britta; Galli, Joakim

2016-10-01

Handling and processing of blood can significantly alter the molecular composition and consistency of biobank samples and can have a major impact on the identification of biomarkers. It is thus crucial to identify tools to determine the quality of samples to be used in biomarker discovery studies. In this study, a non-targeted gas chromatography/time-of-flight mass spectrometry (GC-TOFMS) metabolomic strategy was used with the aim of identifying quality markers for serum and plasma biobank collections lacking proper documentation of preanalytical handling. The effect of postcentrifugation delay was examined in serum stored in tubes with gel separation plugs and ethylenediaminetetraacetic acid (EDTA) plasma in tubes with or without gel separation plugs. The change in metabolic pattern was negligible in all sample types processed within 3 hours after centrifugation regardless of whether the samples were kept at 4°C or 22°C. After 8 and 24 hours postcentrifugation delay before aliquoting, there was a pronounced increase in the number of affected metabolites, as well as in the magnitude of the observed changes. No protective effect on the metabolites was observed in gel-separated EDTA plasma samples. In a separate series of experiments, lactate and glucose levels were determined in plasma to estimate the effect of precentrifugation delay. This separate experiment indicates that the lactate to glucose ratio may serve as a marker to identify samples with delayed time to centrifugation. Although our data from the untargeted GC-TOFMS analysis did not identify any specific markers, we conclude that plasma and serum metabolic profiles remain quite stable when plasma and serum are centrifuged and separated from the blood cells within 3 hours.

68Ga-EDTA PET/CT imaging and plasma clearance for glomerular filtration rate quantification: comparison to conventional 51Cr-EDTA.

PubMed

Hofman, Michael; Binns, David; Johnston, Val; Siva, Shankar; Thompson, Mick; Eu, Peter; Collins, Marnie; Hicks, Rodney J

2015-03-01

Glomerular filtration rate (GFR) can accurately be determined using (51)Cr-ethylenediaminetetraacetic acid (EDTA) plasma clearance counting but is time-consuming and requires technical skills and equipment not always available in imaging departments. (68)Ga-EDTA can be readily available using an onsite generator, and PET/CT enables both imaging of renal function and accurate camera-based quantitation of clearance of activity from blood and its appearance in the urine. This study aimed to assess agreement between (68)Ga-EDTA GFR ((68)Ga-GFR) and (51)Cr-EDTA GFR ((51)Cr-GFR), using serial plasma sampling and PET imaging. (68)Ga-EDTA and (51)Cr-EDTA were injected concurrently in 31 patients. Dynamic PET/CT encompassing the kidneys was acquired for 10 min followed by 3 sequential 3-min multibed step acquisitions from kidneys to bladder. PET quantification was performed using renal activity at 1-2 min (PETinitial), renal excretion at 2-10 min (PETearly), and, subsequently, urinary excretion into the collecting system and bladder (PETlate). Plasma sampling at 2, 3, and 4 h was performed, with (68)Ga followed by (51)Cr counting after positron decay. The level of agreement for GFR determination was calculated using a Bland-Altman plot and Pearson correlation coefficient (PCC). (51)Cr-GFR ranged from 10 to 220 mL/min (mean, 85 mL/min). There was good agreement between (68)Ga-GFR and (51)Cr-GFR using serial plasma sampling, with a Bland-Altman bias of -14 ± 20 mL/min and a PCC of 0.94 (95% confidence interval, 0.88-0.97). Of the 3 methods used for camera-based quantification, the strongest correlation was for plasma sampling-derived GFR with PETlate (PCC of 0.90; 95% confidence interval, 0.80-0.95). (68)Ga-GFR agreed well with (51)Cr-GFR for estimation of GFR using serial plasma counting. PET dynamic imaging provides a method to estimate GFR without plasma sampling, with the additional advantage of enabling renal imaging in a single study. Additional validation in a larger cohort is warranted to further assess utility. © 2015 by the Society of Nuclear Medicine and Molecular Imaging, Inc.

Assessment of free fetal DNA concentration in maternal plasma during the first trimester of pregnancy: comparative study between EDTA and PPT tubes - pilot study.

PubMed

Chadud, Carolina Schneider; Araujo Júnior, Edward; Martinhago, Ciro Dresh; Andari, Viviane Cristina Mello; Tedesco, Giselle Darahem; Bussamra, Luiz Claudio Silva; Aoki, Tsutomu

2015-01-01

To compare ethylenediamine tetraacetic acid (EDTA) tubes and plasma preparation tubes (PPT) for evaluating maternal plasma during the first trimester of pregnancy. A cross-sectional study was conducted on 24 male fetuses in women between 6 and 14 weeks of pregnancy. Blood samples (10 mL) were collected and stored in EDTA and PPT tubes. Subsequently, the samples were centrifuged and sent for free fetal DNA extraction by means of the polymerase chain reaction (PCR) technique. The reactions were performed in a real time PCR machine for detecting the amplification products. The genome region chosen for performing the PCR reactions was a target specific for the Y chromosome, in which the DYS-14 marker was amplified only when the DNA was of male sex. The free fetal DNA concentration was given by the threshold cycle (TC). To compare the tubes, the paired Student t-test was used. The mean gestational age was 11.08 ± 2.30 weeks (range: 6-14). The mean TC for PPT was 30.08 ± 1.05 (range: 27.08-32.61) and for EDTA, 30.23 ± 0.96 (range: 28.01-32.09), but without statistical significance (p=0.357). We did not observe any statistically significant difference in free fetal DNA concentration between the EDTA and PPT tubes.

Effects of anticoagulant, processing delay, and assay method (branched DNA versus reverse transcriptase PCR) on measurement of human immunodeficiency virus type 1 RNA levels in plasma.

PubMed

Kirstein, L M; Mellors, J W; Rinaldo, C R; Margolick, J B; Giorgi, J V; Phair, J P; Dietz, E; Gupta, P; Sherlock, C H; Hogg, R; Montaner, J S; Muñoz, A

1999-08-01

We conducted two studies to determine the potential influence of delays in blood processing, type of anticoagulant, and assay method on human immunodeficiency virus type 1 (HIV-1) RNA levels in plasma. The first was an experimental study in which heparin- and EDTA-anticoagulated blood samples were collected from 101 HIV-positive individuals and processed to plasma after delays of 2, 6, and 18 h. HIV-1 RNA levels in each sample were then measured by both branched-DNA (bDNA) and reverse transcriptase PCR (RT-PCR) assays. Compared to samples processed within 2 h, the loss (decay) of HIV-1 RNA in heparinized blood was significant (P < 0.05) but small after 6 h (bDNA assay, -0.12 log(10) copies/ml; RT-PCR, -0.05 log(10) copies/ml) and after 18 h (bDNA assay, -0.27 log(10) copies/ml; RT-PCR, -0.15 log(10) copies/ml). Decay in EDTA-anticoagulated blood was not significant after 6 h (bDNA assay, -0.002 log(10) copies/ml; RT-PCR, -0.02 log(10) copies/ml), but it was after 18 h (bDNA assay, -0.09 log(10) copies/ml; RT-PCR, -0.09 log(10) copies/ml). Only 4% of samples processed after 6 h lost more than 50% (>/=0.3 log(10) copies/ml) of the HIV-1 RNA, regardless of the anticoagulant or the assay that was used. The second study compared HIV-1 RNA levels in samples from the Multicenter AIDS Cohort Study (MACS; samples were collected in heparin-containing tubes in 1985, had a 6-h average processing delay, and were assayed by bDNA assay) and the British Columbia Drug Treatment Program (BCDTP) (collected in EDTA- or acid citrate dextrose-containing tubes in 1996 and 1997, had a 2-h maximum processing delay, and were assayed by RT-PCR). HIV-1 RNA levels in samples from the two cohorts were not significantly different after adjusting for CD4(+)-cell count and converting bDNA assay values to those corresponding to the RT-PCR results. In summary, the decay of HIV-1 RNA measured in heparinized blood after 6 h was small (-0.05 to -0.12 log(10) copies/ml), and the minor impact of this decay on HIV-1 RNA concentrations in archived plasma samples of the MACS was confirmed by the similarity of CD4(+)-cell counts and assay-adjusted HIV-1 RNA concentrations in the MACS and BCDTP.

Red Cell Properties after Different Modes of Blood Transportation

PubMed Central

Makhro, Asya; Huisjes, Rick; Verhagen, Liesbeth P.; Mañú-Pereira, María del Mar; Llaudet-Planas, Esther; Petkova-Kirova, Polina; Wang, Jue; Eichler, Hermann; Bogdanova, Anna; van Wijk, Richard; Vives-Corrons, Joan-Lluís; Kaestner, Lars

2016-01-01

Transportation of blood samples is unavoidable for assessment of specific parameters in blood of patients with rare anemias, blood doping testing, or for research purposes. Despite the awareness that shipment may substantially alter multiple parameters, no study of that extent has been performed to assess these changes and optimize shipment conditions to reduce transportation-related artifacts. Here we investigate the changes in multiple parameters in blood of healthy donors over 72 h of simulated shipment conditions. Three different anticoagulants (K3EDTA, Sodium Heparin, and citrate-based CPDA) for two temperatures (4°C and room temperature) were tested to define the optimal transportation conditions. Parameters measured cover common cytology and biochemistry parameters (complete blood count, hematocrit, morphological examination), red blood cell (RBC) volume, ion content and density, membrane properties and stability (hemolysis, osmotic fragility, membrane heat stability, patch-clamp investigations, and formation of micro vesicles), Ca2+ handling, RBC metabolism, activity of numerous enzymes, and O2 transport capacity. Our findings indicate that individual sets of parameters may require different shipment settings (anticoagulants, temperature). Most of the parameters except for ion (Na+, K+, Ca2+) handling and, possibly, reticulocytes counts, tend to favor transportation at 4°C. Whereas plasma and intraerythrocytic Ca2+ cannot be accurately measured in the presence of chelators such as citrate and EDTA, the majority of Ca2+-dependent parameters are stabilized in CPDA samples. Even in blood samples from healthy donors transported using an optimized shipment protocol, the majority of parameters were stable within 24 h, a condition that may not hold for the samples of patients with rare anemias. This implies for as short as possible shipping using fast courier services to the closest expert laboratory at reach. Mobile laboratories or the travel of the patients to the specialized laboratories may be the only option for some groups of patients with highly unstable RBCs. PMID:27471472

Time dependent reduction in platelet aggregation using the multiplate analyser and hirudin blood due to platelet clumping.

PubMed

Chapman, Kent; Favaloro, Emmanuel J

2018-05-01

The Multiplate is a popular instrument that measures platelet function using whole blood. Potentially considered a point of care instrument, it is also used by hemostasis laboratories. The instrument is usually utilized to assess antiplatelet medication or as a screen of platelet function. According to the manufacturer, testing should be performed within 0.5-3 hours of blood collection, and preferably using manufacturer provided hirudin tubes. We report time-associated reduction in platelet aggregation using the Multiplate and hirudin blood collection tubes, for all the major employed agonists. Blood for Multiplate analysis was collected into manufacturer supplied hirudin tubes, and 21 consecutive samples assessed using manufacturer supplied agonists (ADP, arachidonic acid, TRAP, collagen and ristocetin), at several time-points post-sample collection within the recommended test time period. Blood was also collected into EDTA as a reference method for platelet counts, with samples collected into sodium citrate and hirudin used for comparative counts. All platelet agonists showed a diminution of response with time. Depending on the agonist, the reduction caused 5-20% and 22-47% of responses initially in the normal reference range to fall below the reference range at 120min and 180min, respectively. Considering any agonist, 35% and 67% of initially "normal" responses became 'abnormal' at 120 min and 180 min, respectively. Platelet counts showed generally minimal changes in EDTA blood, but were markedly reduced over time in both citrate and hirudin blood, with up to 40% and 60% reduction, respectively, at 240 min. The presence of platelet clumping (micro-aggregate formation) was also observed in a time dependent manner, especially for hirudin. In conclusion, considering any platelet agonist, around two-thirds of samples can, within the recommended 0.5-3 hour testing window post-blood collection, yield a reduction in platelet aggregation that may lead to a change in interpretation (i.e., normal to reduced). Thus, the stability of Multiplate testing can more realistically be considered as being between 30-120 min of blood collection for samples collected into hirudin.

Blood antimicrobial activity varies against different Mycobacterium spp.

PubMed

Rivero-Lezcano, Octavio Miguel; Blanco-Conde, Sara; López-Medrano, Ramiro; López-Fidalgo, Eduardo; Caño-Herrero, Manuela; Nebreda-Mayoral, Teresa

2017-12-01

In vitro analysis of mycobacterial pathogenicity or host susceptibility has traditionally relied on the infection of macrophages, the target cell of mycobacteria, despite difficulties reproducing their antimycobacterial activity. We have employed alternative models, namely whole blood and leukocytes in plasma, from QuantiFERON negative individuals, and performed infections with the pathogenic M. tuberculosis, the less pathogenic M. avium, M. kansasii and M. chelonae and the occasionally pathogenic M. gordonae and M. bovis. The anticoagulant used in blood extraction, heparin or EDTA, had a major influence in the outcome of the infection. Thus, while in the heparinized models a similar number of bacteria were enumerated in the inoculum and after seven days, in the presence of EDTA a killing effect was observed, despite the inhibitory effect of EDTA on cellular functions like the production of cytokines or reactive oxygen species (ROS). A special case was the rapidly growing mycobacteria M. chelonae, that multiplied in heparinized models but was eliminated in models with EDTA. We verified that EDTA is not responsible for the bactericidal effect, but acts as a bacteriostatic agent. Further work will determine whether blood derived models are a better alternative to the classical macrophage. Copyright © 2017 Elsevier Ltd. All rights reserved.

Solidifying agent and processing of blood used for the larval diet affect screwworm (Diptera: Calliphoridae) life-history parameters.

PubMed

Chaudhury, M F; Skoda, S R; Sagel, A

2011-06-01

Spray-dried whole bovine blood and a sodium polyacrylate polymer gel as a bulking and solidifying agent are among the constituents of the current larval diet for mass rearing screwworm, Cochliomyia hominivorax (Coquerel) (Diptera: Calliphoridae). Locally available, inexpensive dietary materials could reduce rearing cost and address an uncertain commercial supply of spray-dried blood. We compared efficacy of diet prepared from fresh bovine blood after decoagulation with sodium citrate or ethylenediaminetetraacetic acid (EDTA) or after mechanical defibrination, with the diet containing spray-dried blood using either gel or cellulose fiber as the bulking and solidifying agent. Several life-history parameters were compared among insects reared on each of the blood and bulking agent diets combination. Diets containing citrated blood yielded the lightest larval and pupal weights and fewest pupae. EDTA-treated blood with the gel also caused reductions. EDTA-treated blood with fiber yielded screwworms that were heavier and more numerous than those from the diet with citrated blood but lighter than those from the control diet using spray-dried blood. A reduction in percentage of adults emerging from pupae occurred from diets with both bulking agents using citrated blood and the diet using EDTA mixed with the gel bulking agent. As a group, the cellulose-fiber diets performed better than the gel diets. Larval diet did not affect adult longevity, weight of the eggs deposited by the females that emerged or subsequent egg hatch. Parameter measurements of insects from both defibrinated blood diets were similar to those from the spray-dried blood diets, indicating that fresh, defibrinated bovine blood can successfully replace the dry blood in the screwworm rearing medium.

Prediction of blood lead levels in children before and after remediation of soil samples in the upper Meza Valley, Slovenia.

PubMed

Jez, Erika; Lestan, Domen

2015-10-15

The Meza Valley, Slovenia, has been contaminated by Pb smelting, resulting in an epidemic of lead poisoning in childhood. The potential of remediation with EDTA soil washing to mitigate the risk from Pb poisoning was investigated by applying the Integrated Exposure Uptake Bio-kinetic (IEUBK) model. Soils from 79 locations were collected and the total and bio-accessible Pb concentrations were determined before and after extraction with 60 mmol kg(-1) EDTA. Extraction reduced the soil Pb concentration in towns of Mezica, Zerjav and Crna by 53, 67 and 62%, respectively, and the concentration of in vitro bio-accessible Pb in the simulated human gastric phase by 2.6-, 3.2- and 2.9-times, respectively. The predictions of the IEUBK model based on Pb contamination data were verified with data on blood Pb levels in children. The IEUBK model predicted that, after soil remediation, the number of locations at which the expected blood Pb level in children was higher than the stipulated 10 μg d L(-1) would decrease by 90, 38 and 91% in the towns of Mezica, Zerjav and Crna, respectively. The results confirmed the feasibility of soil washing with EDTA as an efficient remediation measure in Mezica and Crna and advice for soil capping/removal for the most polluted town of Zerjav. Copyright © 2015 Elsevier B.V. All rights reserved.

Coulometric Titration of Ethylenediaminetetraacetate (EDTA) with Spectrophotometric Endpoint Detection: An Experiment for the Instrumental Analysis Laboratory

ERIC Educational Resources Information Center

Williams, Kathryn R.; Young, Vaneica Y.; Killian, Benjamin J.

2011-01-01

Ethylenediaminetetraacetate (EDTA) is commonly used as an anticoagulant in blood-collection procedures. In this experiment for the instrumental analysis laboratory, students determine the quantity of EDTA in commercial collection tubes by coulometric titration with electrolytically generated Cu[superscript 2+]. The endpoint is detected…

Meticulous plasma isolation is essential to avoid false low-level viraemia in Roche Cobas HIV-1 viral load assays.

PubMed

Mortier, Virginie; Vancoillie, Leen; Dauwe, Kenny; Staelens, Delfien; Demecheleer, Els; Schauvliege, Marlies; Dinakis, Sylvie; Van Maerken, Tom; Dessilly, Géraldine; Ruelle, Jean; Verhofstede, Chris

2017-10-24

Pre-analytical sample processing is often overlooked as a potential cause of inaccurate assay results. Here we demonstrate how plasma, extracted from standard EDTA-containing blood collection tubes, may contain traces of blood cells consequently resulting in a false low-level HIV-1 viral load when using Roche Cobas HIV-1 assays. The presence of human DNA in Roche Cobas 4800 RNA extracts and in RNA extracts from the Abbott HIV-1 RealTime assay was assessed by quantifying the human albumin gene by means of quantitative PCR. RNA was extracted from plasma samples before and after an additional centrifugation and tested for viral load and DNA contamination. The relation between total DNA content and viral load was defined. Elevated concentrations of genomic DNA were detected in 28 out of 100 Cobas 4800 extracts and were significantly more frequent in samples processed outside of the AIDS Reference Laboratory. An association between genomic DNA presence and spurious low-level viraemia results was demonstrated. Supplementary centrifugation of plasma before RNA extraction eliminated the contamination and the false viraemia. Plasma isolated from standard EDTA-containing blood collection tubes may contain traces of HIV DNA leading to false viral load results above the clinical cutoff. Supplementary centrifugation of plasma before viral load analysis may eliminate the occurrence of this spurious low-level viraemia.

Performance evaluation of Samsung LABGEO(HC10) Hematology Analyzer.

PubMed

Park, Il Joong; Ahn, Sunhyun; Kim, Young In; Kang, Seon Joo; Cho, Sung Ran

2014-08-01

The Samsung LABGEO(HC10) Hematology Analyzer (LABGEO(HC10)) is a recently developed automated hematology analyzer that uses impedance technologies. The analyzer provides 18 parameters including 3-part differential at a maximum rate of 80 samples per hour. To evaluate the performance of the LABGEO(HC10). We evaluated precision, linearity, carryover, and relationship for complete blood cell count parameters between the LABGEO(HC10) and the LH780 (Beckman Coulter Inc) in a university hospital in Korea according to the Clinical and Laboratory Standards Institute guidelines. Sample stability and differences due to the anticoagulant used (K₂EDTA versus K₃EDTA) were also evaluated. The LABGEO(HC10) showed linearity over a wide range and minimal carryover (<1%) for white blood cell, hemoglobin, red blood cell, and platelet parameters. Correlation between the LABGEO(HC10) and the LH780 was good for all complete blood cell count parameters (R > 0.92) except for mean corpuscular hemoglobin concentration. The bias estimated was acceptable for all parameters investigated except for monocyte count. Most parameters were stable until 24 hours both at room temperature and at 4°C. The difference by anticoagulant type was statistically insignificant for all parameters except for a few red cell parameters. The accurate results achievable and simplicity of operation make the unit recommendable for small to medium-sized laboratories.

Behavior of heavy metals in human urine and blood following calcium disodium ethylenediamine tetraacetate injection: observations in metal workers.

PubMed

Sata, F; Araki, S; Murata, K; Aono, H

1998-06-12

To evaluate the effects of calcium disodium ethylenediamine tetraacetate (CaEDTA) on the behavior of 8 heavy metals in human urine and blood, CaEDTA was administered for 1 h by intravenous injection to 18 male metal foundry workers, whose blood lead concentrations (PbB) were between 16 and 59 (mean 34) microg/dl. Significant increases were found in urinary excretion of manganese, chromium, lead, zinc, and copper after the start of CaEDTA injection. Urinary chromium excretion reached a maximal level within 1 h after the start of injection, while urinary manganese, lead, and zinc excretion reached their highest concentrations between 1 and 2 h. Urinary copper excretion reached the highest level between 2 and 4 h. The rapid increases in urinary excretion of five metals were different from the "circadian rhythms," which are the normal, daily variations in renal glomerular filtration, reabsorption, and excretory mechanisms. Plasma lead concentrations were highest 1.5 h after the start of the 1-h injection, while plasma zinc concentration became lowest 5 h after the start of CaEDTA injection. Data suggest that manganese and chromium absorbed in human tissues might be mobilized by CaEDTA.

Sample collections from healthy volunteers for biological variation estimates' update: a new project undertaken by the Working Group on Biological Variation established by the European Federation of Clinical Chemistry and Laboratory Medicine.

PubMed

Carobene, Anna; Strollo, Marta; Jonker, Niels; Barla, Gerhard; Bartlett, William A; Sandberg, Sverre; Sylte, Marit Sverresdotter; Røraas, Thomas; Sølvik, Una Ørvim; Fernandez-Calle, Pilar; Díaz-Garzón, Jorge; Tosato, Francesca; Plebani, Mario; Coşkun, Abdurrahman; Serteser, Mustafa; Unsal, Ibrahim; Ceriotti, Ferruccio

2016-10-01

Biological variation (BV) data have many fundamental applications in laboratory medicine. At the 1st Strategic Conference of the European Federation of Clinical Chemistry and Laboratory Medicine (EFLM) the reliability and limitations of current BV data were discussed. The EFLM Working Group on Biological Variation is working to increase the quality of BV data by developing a European project to establish a biobank of samples from healthy subjects to be used to produce high quality BV data. The project involved six European laboratories (Milan, Italy; Bergen, Norway; Madrid, Spain; Padua, Italy; Istanbul, Turkey; Assen, The Netherlands). Blood samples were collected from 97 volunteers (44 men, aged 20-60 years; 43 women, aged 20-50 years; 10 women, aged 55-69 years). Initial subject inclusion required that participants completed an enrolment questionnaire to verify their health status. The volunteers provided blood specimens once per week for 10 weeks. A short questionnaire was completed and some laboratory tests were performed at each sampling consisting of blood collected under controlled conditions to provide serum, K2EDTA-plasma and citrated-plasma samples. Samples from six out of the 97 enroled subjects were discarded as a consequence of abnormal laboratory measurements. A biobank of 18,000 aliquots was established consisting of 120 aliquots of serum, 40 of EDTA-plasma, and 40 of citrated-plasma from each subject. The samples were stored at -80 °C. A biobank of well-characterised samples collected under controlled conditions has been established delivering a European resource to enable production of contemporary BV data.

Effects of Anticoagulant, Processing Delay, and Assay Method (Branched DNA versus Reverse Transcriptase PCR) on Measurement of Human Immunodeficiency Virus Type 1 RNA Levels in Plasma

PubMed Central

Kirstein, Lynn M.; Mellors, John W.; Rinaldo, Charles R.; Margolick, Joseph B.; Giorgi, Janis V.; Phair, John P.; Dietz, Edith; Gupta, Phalguni; Sherlock, Christopher H.; Hogg, Robert; Montaner, J. S. G.; Muñoz, Alvaro

1999-01-01

We conducted two studies to determine the potential influence of delays in blood processing, type of anticoagulant, and assay method on human immunodeficiency virus type 1 (HIV-1) RNA levels in plasma. The first was an experimental study in which heparin- and EDTA-anticoagulated blood samples were collected from 101 HIV-positive individuals and processed to plasma after delays of 2, 6, and 18 h. HIV-1 RNA levels in each sample were then measured by both branched-DNA (bDNA) and reverse transcriptase PCR (RT-PCR) assays. Compared to samples processed within 2 h, the loss (decay) of HIV-1 RNA in heparinized blood was significant (P < 0.05) but small after 6 h (bDNA assay, −0.12 log10 copies/ml; RT-PCR, −0.05 log10 copies/ml) and after 18 h (bDNA assay, −0.27 log10 copies/ml; RT-PCR, −0.15 log10 copies/ml). Decay in EDTA-anticoagulated blood was not significant after 6 h (bDNA assay, −0.002 log10 copies/ml; RT-PCR, −0.02 log10 copies/ml), but it was after 18 h (bDNA assay, −0.09 log10 copies/ml; RT-PCR, −0.09 log10 copies/ml). Only 4% of samples processed after 6 h lost more than 50% (≥0.3 log10 copies/ml) of the HIV-1 RNA, regardless of the anticoagulant or the assay that was used. The second study compared HIV-1 RNA levels in samples from the Multicenter AIDS Cohort Study (MACS; samples were collected in heparin-containing tubes in 1985, had a 6-h average processing delay, and were assayed by bDNA assay) and the British Columbia Drug Treatment Program (BCDTP) (collected in EDTA- or acid citrate dextrose-containing tubes in 1996 and 1997, had a 2-h maximum processing delay, and were assayed by RT-PCR). HIV-1 RNA levels in samples from the two cohorts were not significantly different after adjusting for CD4+-cell count and converting bDNA assay values to those corresponding to the RT-PCR results. In summary, the decay of HIV-1 RNA measured in heparinized blood after 6 h was small (−0.05 to −0.12 log10 copies/ml), and the minor impact of this decay on HIV-1 RNA concentrations in archived plasma samples of the MACS was confirmed by the similarity of CD4+-cell counts and assay-adjusted HIV-1 RNA concentrations in the MACS and BCDTP. PMID:10405379

Efficacy of chelation therapy to remove aluminium intoxication.

PubMed

Fulgenzi, Alessandro; De Giuseppe, Rachele; Bamonti, Fabrizia; Vietti, Daniele; Ferrero, Maria Elena

2015-11-01

There is a distinct correlation between aluminium (Al) intoxication and neurodegenerative diseases (ND). We demonstrated how patients affected by ND showing Al intoxication benefit from short-term treatment with calcium disodium ethylene diamine tetraacetic acid (EDTA) (chelation therapy). Such therapy further improved through daily treatment with the antioxidant Cellfood. In the present study we examined the efficacy of long-term treatment, using both EDTA and Cellfood. Slow intravenous treatment with the chelating agent EDTA (2 g/10 mL diluted in 500 mL physiological saline administered in 2 h) (chelation test) removed Al, which was detected (using inductively coupled plasma mass spectrometry) in urine samples collected from patients over 12 h. Patients that revealed Al intoxication (expressed in μg per g creatinine) underwent EDTA chelation therapy once a week for ten weeks, then once every two weeks for a further six or twelve months. At the end of treatment (a total of 22 or 34 chelation therapies, respectively), associated with daily assumption of Cellfood, Al levels in the urine samples were analysed. In addition, the following blood parameters were determined: homocysteine, vitamin B12, and folate, as well as the oxidative status e.g. reactive oxygen species (ROS), total antioxidant capacity (TAC), oxidized LDL (oxLDL), and glutathione. Our results showed that Al intoxication reduced significantly following EDTA and Cellfood treatment, and clinical symptoms improved. After treatment, ROS, oxLDL, and homocysteine decreased significantly, whereas vitamin B12, folate and TAC improved significantly. In conclusion, our data show the efficacy of chelation therapy associated with Cellfood in subjects affected by Al intoxication who have developed ND.

Performance Characteristics of Plasma Amyloid β 40 and 42 Assays

PubMed Central

Okereke, Olivia I.; Xia, Weiming; Irizarry, Michael C.; Sun, Xiaoyan; Qiu, Wei Q.; Fagan, Anne M.; Mehta, Pankaj D.; Hyman, Bradley T.; Selkoe, Dennis J.; Grodstein, Francine

2009-01-01

Background Identifying biomarkers of Alzheimer disease (AD) risk will be critical to effective AD prevention. Levels of circulating amyloid β (Aβ) 40 and 42 may be candidate biomarkers. However, properties of plasma Aβ assays must be established. Methods Using five different protocols, blinded samples were used to assess: intra-assay reproducibility; impact of EDTA vs. heparin anticoagulant tubes; and effect of time-to-blood processing. In addition, percent recovery of known Aβ concentrations in spiked samples was assessed. Results Median intra-assay coefficients of variation (CVs) for the assay protocols ranged from 6–24% for Aβ-40, and 8–14% for Aβ-42. There were no systematic differences in reproducibility by collection method. Plasma concentrations of Aβ (particularly Aβ-42) appeared stable in whole blood kept in ice packs and processed as long as 24 hours after collection. Recovery of expected concentrations was modest, ranging from -24% to 44% recovery of Aβ-40, and 17% to 61% of Aβ-42. Conclusions Across five protocols, plasma Aβ-40 and Aβ-42 levels were measured with generally low error, and measurements appeared similar in blood collected in EDTA vs. heparin. While these preliminary findings suggest that measuring plasma Aβ-40 and Aβ-42 may be feasible in varied research settings, additional work in this area is necessary. PMID:19221417

A comparison of four-sample slope-intercept and single-sample 51Cr-EDTA glomerular filtration rate measurements.

PubMed

Porter, Charlotte A; Bradley, Kevin M; McGowan, Daniel R

2018-05-01

The aim of this study was to verify, with a large dataset of 1394 Cr-EDTA glomerular filtration rate (GFR) studies, the equivalence of slope-intercept and single-sample GFR. Raw data from 1394 patient studies were used to calculate four-sample slope-intercept GFR in addition to four individual single-sample GFR values (blood samples taken at 90, 150, 210 and 270 min after injection). The percentage differences between the four-sample slope-intercept and each of the single-sample GFR values were calculated, to identify the optimum single-sample time point. Having identified the optimum time point, the percentage difference between the slope-intercept and optimal single-sample GFR was calculated across a range of GFR values to investigate whether there was a GFR value below which the two methodologies cannot be considered equivalent. It was found that the lowest percentage difference between slope-intercept and single-sample GFR was for the third blood sample, taken at 210 min after injection. The median percentage difference was 2.5% and only 6.9% of patient studies had a percentage difference greater than 10%. Above a GFR value of 30 ml/min/1.73 m, the median percentage difference between the slope-intercept and optimal single-sample GFR values was below 10%, and so it was concluded that, above this value, the two techniques are sufficiently equivalent. This study supports the recommendation of performing single-sample GFR measurements for GFRs greater than 30 ml/min/1.73 m.

Order of draw practices in venous blood sampling at clinical biochemistry departments in the Danish health care system.

PubMed

Jacobsen, Katja Kemp; Brandt, Ida; Christensen, Anne Vindahl; Rimsø, Bjørk Anine; Krøier, Camilla Julie; Sørensen, Michelle; Smith, Julie; Jensen, Kathrine Overgaard Foss; Larsen, Jeppe Madura

2018-06-01

Deviation in blood collection procedures is a central source of preanalytical variation affecting overall analytical and diagnostic precision. The order of draw of venous sampling is suspected to affect analytical results, in particular for coagulation analysis. Here we compare the procedures in venous blood sampling among clinical biochemistry departments to assess the uniformity of order of blood draw and adherence to international guidelines in the Danish health care system. We collected venous order of draw procedures from 49 clinical biochemistry departments at 22 public hospitals in Denmark. Procedures were compared to the international guidelines fromthe Clinical Laboratory Standards Institute (CLSI) and World Health Organization (WHO), and assessed in relation to department ISO 15189:2012 accreditation. We observed seven different order of draw procedures related to citrate, serum, heparin, and EDTA tubes, and the use of discard tubes in relation to coagulation assays. 31 departments (63.3%) were found to adhere to CLSI and WHO guidelines. A majority of departments instructs the use of discard tubes before collection for coagulation assays in citrate tubes (44 departments; 89.8%). The citrate tube was the first sample tube to be drawn for most departments (35 departments; 75.5%); and the preferred order of non-citrate tubes was serum-heparin-EDTA (36 departments; 73.5%). Adherence to the CLSI and WHO guidelines was not associated with department ISO 15189:2012 accreditation (p = .57). Venous order of draw procedures is diverse at Danish clinical biochemistry departments and show moderate adherence to international guidelines. Copyright © 2018 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Controlling of dielectrical properties of hydroxyapatite by ethylenediamine tetraacetic acid (EDTA) for bone healing applications

NASA Astrophysics Data System (ADS)

Kaygili, Omer; Ates, Tankut; Keser, Serhat; Al-Ghamdi, Ahmed A.; Yakuphanoglu, Fahrettin

2014-08-01

The hydroxyapatite (HAp) samples in the presence of various amounts of ethylenediamine tetraacetic acid (EDTA) were prepared by sol-gel method. The effects of EDTA on the crystallinity, phase structure, chemical, micro-structural and dielectric properties of HAp samples were investigated. With the addition of EDTA, the average crystallite size of the HAp samples is gradually decreased from 30 to 22 nm and the crystallinity is in the range of 65-71%. The values of the lattice parameters (a and c) and volume of the unit cell are decreased by stages with the addition of EDTA. The dielectric parameters such as relative permittivity, dielectric loss and relaxation time are affected by the adding of EDTA. The alternating current conductivity of the as-synthesized hydroxyapatites increases with the increasing frequency and obeys the universal power law behavior. The HAp samples exhibit a non-Debye relaxation mechanism. The obtained results that the dielectrical parameters of the HAp sample can be controlled by EDTA.

Assessing Glomerular Filtration in Small Animals Using [68Ga]DTPA and [68Ga]EDTA with PET Imaging.

PubMed

Gündel, Daniel; Pohle, Ulrike; Prell, Erik; Odparlik, Andreas; Thews, Oliver

2018-06-01

Determining the glomerular filtration rate (GFR) is essential for clinical medicine but also for pre-clinical animal studies. Functional imaging using positron emission tomography (PET) allows repetitive almost non-invasive measurements. The aim of the study was the development and evaluation of easily synthesizable PET tracers for GFR measurements in small animals. Diethylenetriaminepentaacetic acid (DTPA) and ethylenediaminetetraacetic acid (EDTA) were labeled with Ga-68. The binding to blood cells and plasma proteins was tested in vitro. The distribution of the tracers in rats was analyzed by PET imaging and ex vivo measurements. From the time-activity-curve of the blood compartment (heart) and the total tracer mass excreted by the kidney, the GFR was calculated. These values were compared directly with the inulin clearance in the same animals. Both tracers did not bind to blood cells. [ 68 Ga]DPTA but not [ 68 Ga]EDTA showed strong binding to plasma proteins. For this reason, [ 68 Ga]DPTA stayed much longer in the blood and only 30 % of the injected dose was eliminated by the kidney within 60 min whereas the excretion of [ 68 Ga]EDTA was 89 ± 1 %. The calculated GFR using [ 68 Ga]EDTA was comparable to the measured inulin clearance in the same animal. Using [ 68 Ga]-DPTA, the measurements led to values which were 80 % below the normal GFR. The results also revealed that definition of the volume of interest for the blood compartment affects the calculation and may lead to a slight overestimation of the GFR. [ 68 Ga]EDTA is a suitable tracer for GFR calculation from PET imaging in small animals. It is easy to be labeled, and the results are in good accordance with the inulin clearance. [ 68 Ga]DTPA led to a marked underestimation of GFR due to its strong binding to plasma proteins and is therefore not an appropriate tracer for GFR measurements.

Randomized study of minocycline and edetic acid as a locking solution for central line (port-a-cath) in children with cancer.

PubMed

Ferreira Chacon, Julieta Maria; Hato de Almeida, Emília; de Lourdes Simões, Regina; Lazzarin C Ozório, Viviane; Alves, Benaia Cândida; Mello de Andréa, Maria Lydia; Santiago Biernat, Marcela; Biernat, João Carlos

2011-01-01

Contamination of central catheters is frequent, and biofilm perpetuates infections. Heparin does not protect against infections because it has no antibiotic action. Minocycline and edetic acid (M-EDTA), a potent calcium chelating agent that destroys bacterial and fungal cell membrane and disrupts biofilm, may be an alternative to allow the associated antibiotic to act locally at a high and safe concentration. Fifty children with cancer and a port-a-cath were followed up: 26 received heparin (group 1) and 24 M-EDTA (group 2). A total of 762 serial prospective blood cultures were obtained, 387 from group 1 and 375 from group 2. In group 1 (heparin), 19 blood cultures were positive, and infection incidence was 73.1% (19/26 ports). In group 2 (M-EDTA), 5 blood cultures were positive, and the incidence rate was 20.8% (5/24 ports). M-EDTA, compared with heparin, prevents and treats catheter infections, and is a promising alternative to decrease sepsis during chemotherapy. Copyright © 2011 S. Karger AG, Basel.

Ultrasonic activation and chemical modification of photosensitizers enhances the effects of photodynamic therapy against Enterococcus faecalis root-canal isolates.

PubMed

Tennert, C; Drews, A M; Walther, V; Altenburger, M J; Karygianni, L; Wrbas, K T; Hellwig, E; Al-Ahmad, A

2015-06-01

The aim of this study was to evaluate the effect of photodynamic therapy (PDT) on Enterococcus faecalis biofilms in artificially infected root canals using modified photosensitizers and passive ultrasonic activation. Two hundred and seventy extracted human teeth with one root canal were instrumented utilizing ProTaper files, autoclaved, infected with E. faecalis T9 for 72 h and divided into different groups: irrigation with 3% sodium hypochlorite (NaOCl), 20% ethylenediaminetetraacetic acid (EDTA), or 20% citric acid, PDT without irrigation, PDT accompanied by irrigation with NaOCl, EDTA, or citric acid, PDT using an EDTA-based photosensitizer or a citric-acid-based photosensitizer and PDT with ultrasonic activation of the photosensitizer. A 15 mg/ml toluidine blue served as the photosensitizer, activated by a 100 mW LED light source. Sterile paper points were used for sampling the root canals and dentin chips were collected to assess the remaining contamination after treatment. Samples were cultured on blood agar plates and colony forming units were quantified. PDT alone achieved a reduction in E. faecalis counts by 92.7%, NaOCl irrigation alone and combined with PDT by 99.9%. The antibacterial effects increased by the combination of irrigation using EDTA or citric acid and PDT compared to irrigation alone. More than 99% of E. faecalis were killed using PDT with the modified photosensitizers and ultrasonic activation. NaOCl based disinfection achieved the highest antimicrobial effect. Using PDT with an EDTA-based or citric-acid-based phozosensitizer or activating the photosensitizer with ultrasound resulted in a significantly higher reduction in E. faecalis counts compared to conventional PDT. Copyright © 2015 Elsevier B.V. All rights reserved.

Effect of antiaggregants on the in vitro viability, cell count and stability of abalone (Haliotis iris) haemocytes.

PubMed

Grandiosa, Roffi; Bouwman, Mai-Louise; Young, Tim; Mérien, Fabrice; Alfaro, Andrea C

2018-07-01

The ability to successfully prepare and preserve haemocyte cells for microscopy and flow cytometry is critical for the investigation of animal immune systems. In this study, we observed the total cell count, in vitro viability and stability of New Zealand black-footed abalone (Haliotis iris) haemocytes with different antiaggregants and handling protocols. Haemocyte stability was evaluated by direct observation of haemocytes under the microscope and calculating the aggregation index. Haemocyte counts and viability were measured via flow cytometry and tested for the effect of different antiaggregants (Alsever's solution at three concentrations, and specialised blood collection tubes containing lithium heparin and K 2 EDTA) at different temperatures and storage times. Results showed that Alsever's solution is an effective antiaggregant at haemolymph:antiaggregant dilution ratios of 1:1, 1:2 and 1:3. Lithium heparin was ineffective as an antiaggregant, whereas K 2 EDTA was similarly as effective as Alsever's solution. The influence of different mixing techniques (vortex, pipetting and flipping) were subsequently tested using the K 2 EDTA Microtainer ® tubes, revealing that proper mixing should be performed immediately. High cell viability can be achieved by mixing samples by either 10 s of vortexing (1000 rpm), 10 times pipetting or 20 times flipping. The in vitro storage of abalone haemocytes in AS and K 2 EDTA as antiaggregants at ambient room temperature was highly effective for up to 24 h (75-85% viability; 0.05-0.15 aggregation index) and is recommended for haemocyte studies in H. iris. Utilization of K 2 EDTA Microtainer ® tubes were advantageous since they are more cost effective compared to Alsever's solution, and samples can be prepared more efficiently. Copyright © 2018. Published by Elsevier Ltd.

Controlling of dielectrical properties of hydroxyapatite by ethylenediamine tetraacetic acid (EDTA) for bone healing applications.

PubMed

Kaygili, Omer; Ates, Tankut; Keser, Serhat; Al-Ghamdi, Ahmed A; Yakuphanoglu, Fahrettin

2014-08-14

The hydroxyapatite (HAp) samples in the presence of various amounts of ethylenediamine tetraacetic acid (EDTA) were prepared by sol-gel method. The effects of EDTA on the crystallinity, phase structure, chemical, micro-structural and dielectric properties of HAp samples were investigated. With the addition of EDTA, the average crystallite size of the HAp samples is gradually decreased from 30 to 22 nm and the crystallinity is in the range of 65-71%. The values of the lattice parameters (a and c) and volume of the unit cell are decreased by stages with the addition of EDTA. The dielectric parameters such as relative permittivity, dielectric loss and relaxation time are affected by the adding of EDTA. The alternating current conductivity of the as-synthesized hydroxyapatites increases with the increasing frequency and obeys the universal power law behavior. The HAp samples exhibit a non-Debye relaxation mechanism. The obtained results that the dielectrical parameters of the HAp sample can be controlled by EDTA. Copyright © 2014 Elsevier B.V. All rights reserved.

A case report of pseudo grey platelet syndrome with citrate-induced pseudothrombocytopenia: those artifacts may interfere in the platelet numeration and lead to critical misdiagnosis.

PubMed

Herb, Agathe; Maurer, Maxime; Alamome, Isabelle; Bihl, Pierre-Adrien; Ghiura, Cosmina; Hurstel, Rémy

2017-08-01

The pseudo grey platelet syndrome is a rare artifact due to the degranulation of platelets caused, in vitro, by EDTA. This phenomenon is likely to disturb the platelet numeration and it is essential not to mistake it for a grey syndrome platelet, which is a constitutional thrombopathy with macrothrombopenia, in order to avoid specialized tests, or even misdiagnosis. Indeed, these two entities are cytologically alike, as grey platelets are found on the blood smear of a sample collected on EDTA in both cases. We here describe the case of a patient admitted in Colmar's Hospital for a chronic thrombocytopenia, associating both a pseudothrombocytopenia and a pseudo grey platelet syndrome.

Preparation of serum and plasma samples for determination of tricyclic antidepressants: effects of blood collection tubes and storage.

PubMed

Nyberg, G; Mårtensson, E

1986-01-01

The effects were tested of eight common types of blood collection tubes and two types of "plasma separators" on the stability of the tricyclic antidepressants amitriptyline, imipramine, clomipramine, and their monodemethylated metabolites in venous blood samples. Although EDTA-containing Venoject lavender and Vacutainer lavender tubes seemed to give the most stable plasma samples, and Venoject red the most stable serum samples, the differences were too small to have practical consequences. Vacutainer royal blue collection tubes gave significant losses of greater than 20% of some of the substances. The tubes with serum separator gel or filter proved unsuitable, since they were responsible for losses of greater than 40%. The losses were not caused by redistribution between blood cells and plasma but occurred mainly as a result of contact between the contents and the caps of the tubes. Experiments with freezing, thawing, and storage of samples showed that freshly sampled blood could be stored at room temperature for 24 h in Venoject green tubes without significant losses. Serum samples could be stored at refrigerator temperature for 4 weeks without important losses. Freezing, thawing, and storage at -20 degrees C did not influence the serum or plasma concentrations.

Assessment of the stability of mephedrone in ante-mortem and post-mortem blood specimens.

PubMed

Busardò, Francesco Paolo; Kyriakou, Chrystalla; Tittarelli, Roberta; Mannocchi, Giulio; Pantano, Flaminia; Santurro, Alessandro; Zaami, Simona; Baglìo, Giovanni

2015-11-01

The aim of this work is to test the stability of mephedrone added to whole blood collected from alive and dead mephedrone free-users and stored at three different temperatures (-20, +4 and +20°C) with and without preservatives up to 6 months, trying to establish the best storage condition in order to reduce possible analyte loss/degradation during the storage period. Different sources of blood were obtained as follow: 10 samples of blood came from 10 alive mephedrone free-users (mean age 34±15.8 years old) (Group 1), whereas 10 post mortem blood samples were obtained from 10 cadavers, in which the post mortem interval was between 24 and 36h (Group 2). The cause of death in post mortem cases (mean age 45±14.2 years old) was not drug related. Pools of blood were spiked with mephedrone at the concentration of 1mg/L and 1mL aliquots were transferred in 2mL Eppendorf capped tubes with and without preservatives as follow: with ethylenediaminetetraacetic acid (EDTA) 3%; with sodium fluoride/potassium oxalate (NaF/KOx) 1.67%/0.2%, respectively; without preservatives. All samples were stored at three different temperatures: -20°C, 4°C and 20°C and extracted and analyzed in duplicate by GC-MS according to a previously published method by Dickson et al., every other day during the first month and then weekly up to 6 months. our study allow us to affirm that -20°C is the best storage temperature for mephedrone stability in ante-mortem and post-mortem blood samples in comparison to the other two tested temperatures (+4 and +20°C), showing higher values in both groups in samples stored with and without preservatives (p<0.0001). The comparison of Group 1 (samples coming from alive subjects) and Group 2 (post-mortem samples) highlights a better stability of mephedrone in Group 1 (p<0.001) at all tested storage conditions. Finally, the analysis of blood specimens stored with and without preservatives in both groups suggests that specimens stored with NaF/KOx maintain mephedrone stability better than those stored with EDTA (p<0.001) and those stored without preservatives (p<0.0001), therefore, we strongly recommend in order to maintain the highest mephedrone stability in blood, to store specimens at -20°C adding NaF/KOx as preservative. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Current laboratory requirements for adrenocorticotropic hormone and renin/aldosterone sample handling are unnecessarily restrictive.

PubMed

Chakera, Ali J; McDonald, Timothy J; Knight, Bridget A; Vaidya, Bijay; Jones, Angus G

2017-02-01

Samples for adrenocorticotropic hormone (ACTH) and aldosterone/renin analysis usually require rapid transport to the receiving laboratory for immediate separation and freezing. In practice, this means assessment is limited to hospital settings and many samples are rejected. We examined whether these requirements are necessary by assessing the stability of ACTH, aldosterone and renin over 48 hours in whole blood collected in serum gel and EDTA plasma from 31 participants. Our results show that ACTH collected into EDTA plasma is stable at room temperature for at least 6 hours, mean change at 6 hours -2.6% (95% CI -9.7 to 4.5). Both aldosterone and renin were stable collected on serum gel at room temperature for at least 6 hours: mean change aldosterone +0.2% (95% CI -3.6 to 4.0), renin -1.9% (95% CI -7.0 to3.2). Therefore, by using appropriate preservatives, ACTH and aldosterone/renin can be measured on samples collected at room temperature and processed within 6 hours. This would facilitate outpatient and emergency room assessment of these analytes. © Royal College of Physicians 2017. All rights reserved.

Chemical induced demineralization study in cortical bone

NASA Astrophysics Data System (ADS)

Sales, E.; da Silva, C. E. R.; Letichevsky, S.; dos Santos, R.; Leitao, R.; dos Santos, C. T.; de Oliveira, L. F.; de Avillez, R.; Monteiro, M.; Costa-Felix, R.; Paciornik, S.; dos Anjos, M.

2018-05-01

In this work we present a study of demineralization in bovine cortical bone. We selected 9 fresh cortical bone samples from 2 diaphyseal femurs for analysis. Samples were demineralized for 24 h, 48 h, 72 h and 96 h using two concentrations of EDTA with different pH: EDTA 0.1 M (pH 10, alkaline) and EDTA 0.5 M (pH 7.4, neutral). We have employed μ-X-ray fluorescence (μ-XRF) and X-ray diffraction (XRD) to assess the degree of demineralization. EDTA solutions were analyzed for Calcium (Ca) and Phosphorous (P) extractions by Atomic Absorption Spectrophotometry (AAS) and Ion Chromatography (IC), respectively. Results from AAS and IC showed that EDTA 0.5 M (pH 7.4) removed two times more Ca and 3 times more P than EDTA 0.1 M (pH 10) in the first 24 hours. μ-XRF results presented that EDTA has a high capacity to bind Calcium and Phosphorus. On the other hand, despite the differences in concentration and pH, EDTA did not bind Zn and Sr. Results from XRD showed that EDTA with high concentration had a greater impact to the samples' crystallinity causing a severe damage.

Influence of tube type, storage time, and temperature on the total and free concentration of valproic acid.

PubMed

Tarasidis, C G; Garnett, W R; Kline, B J; Pellock, J M

1986-01-01

The influence of storage conditions on the total and free concentration of valproic acid (VPA) was studied in six normal male subjects who ingested 750 mg of VPA (3 X 250 mg Depakene capsules; Abbott Laboratories). Blood samples were collected in various types of Vacutainer tubes (red top, no additives; green top, sodium heparin; blue top, sodium citrate; and purple top, EDTA) 2 h post administration of VPA. Either these samples were centrifuged immediately or stored for various periods of time at room temperature or refrigerated, or the supernate was frozen prior to analysis. Free VPA samples were obtained utilizing the Amicon ultrafiltration system. All VPA samples were analyzed by gas-liquid chromatography. Total VPA concentrations obtained from plasma collected with sodium citrate were lower (p less than 0.05) than either serum or plasma collected with other anticoagulants. There were no differences (p greater than 0.05) in total or free VPA concentrations between samples collected in serum or in plasma collected with heparin or EDTA. Storing samples for 96 h at room temperature did not alter the total VPA concentrations but was found to increase the free fraction of VPA (p less than 0.05). The refrigeration or freezing of the supernate from the blood samples for 7 days did not alter (p greater than 0.05) the total or the free fraction of VPA. The results of this study demonstrate that total and/or free VPA may be collected from either serum or plasma, provided sodium citrate is not used to collect plasma.(ABSTRACT TRUNCATED AT 250 WORDS)

Quantification of mixed chimerism by real time PCR on whole blood-impregnated FTA cards.

PubMed

Pezzoli, N; Silvy, M; Woronko, A; Le Treut, T; Lévy-Mozziconacci, A; Reviron, D; Gabert, J; Picard, C

2007-09-01

This study has investigated quantification of chimerism in sex-mismatched transplantations by quantitative real time PCR (RQ-PCR) using FTA paper for blood sampling. First, we demonstrate that the quantification of DNA from EDTA-blood which has been deposit on FTA card is accurate and reproducible. Secondly, we show that fraction of recipient cells detected by RQ-PCR was concordant between the FTA and salting-out method, reference DNA extraction method. Furthermore, the sensitivity of detection of recipient cells is relatively similar with the two methods. Our results show that this innovative method can be used for MC assessment by RQ-PCR.

Examination of blood-brain barrier permeability in dementia of the Alzheimer type with (68Ga)EDTA and positron emission tomography

DOE Office of Scientific and Technical Information (OSTI.GOV)

Schlageter, N.L.; Carson, R.E.; Rapoport, S.I.

1987-02-01

Positron emission tomography with (/sup 68/Ga)ethylenediaminetetraacetic acid ((/sup 68/Ga)EDTA) was used to examine the integrity of the blood-brain barrier (BBB) in five patients with dementia of the Alzheimer type and in five healthy age-matched controls. Within a scanning time of 90 min, there was no evidence that measurable intravascular tracer entered the brain in either the dementia or the control group. An upper limit for the cerebrovascular permeability-surface area product of (68Ga)EDTA was estimated as 2 X 10(-6) s-1 in both groups. The results provide no evidence for breakdown of the BBB in patients with dementia of the Alzheimer type.

Experiment on aggregation of red cells under microgravity on STS 51-C

NASA Astrophysics Data System (ADS)

Dintenfass, L.; Osman, P.; Maguire, B.; Jedrzejczyk, H.

Kinetics and morphology of aggregation of red cells were studied using automatic slit-capillary photo-viscometers, one situated on the middeck of the space shuttle `Discovery', and the other in the ground laboratory at KSC. Experiments were run simultaneously, blood samples being adjusted to haematocrit of 0.30 using native plasma, at temp. of 25°C, and anticoagulated by EDTA. Donors included patients with myocardial infarction, insulin-dependent diabetes, hyperlipidaemia and hypertension. Macro and microphotographs were obtained during flow and statis. There was a striking difference in the morphology of aggregates formed in space and on the ground. Aggregates formed under zero gravity showed rouleaux formation, while the same blood samples showed severe clumping on the ground, in all patients blood. Normal blood showed rouleaux on the ground, but a random swarm-like pattern in space. The shape of the red cells remained normal under zero gravity.

Rapid DNA extraction from dried blood spots on filter paper: potential applications in biobanking.

PubMed

Choi, Eun-Hye; Lee, Sang Kwang; Ihm, Chunhwa; Sohn, Young-Hak

2014-12-01

Dried blood spot (DBS) technology is a microsampling alternative to traditional plasma or serum sampling for pharmaco- or toxicokinetic evaluation. DBS technology has been applied to diagnostic screening in drug discovery, nonclinical, and clinical settings. We have developed an improved elution protocol involving boiling of blood spots dried on Whatman filter paper. The purpose of this study was to compare the quality, purity, and quantity of DNA isolated from frozen blood samples and DBSs. We optimized a method for extraction and estimation of DNA from blood spots dried on filter paper (3-mm FTA card). A single DBS containing 40 μL blood was used. DNA was efficiently extracted in phosphate-buffered saline (PBS) or Tris-EDTA (TE) buffer by incubation at 37°C overnight. DNA was stable in DBSs that were stored at room temperature or frozen. The housekeeping genes GAPDH and beta-actin were used as positive standards for polymerase chain reaction (PCR) validation of general diagnostic screening. Our simple and convenient DBS storage and extraction methods are suitable for diagnostic screening by using very small volumes of blood collected on filter paper, and can be used in biobanks for blood sample storage.

Establishing diagnostic cut-off criteria for the COBAS AmpliPrep/COBAS TaqMan HIV-1 Qualitative test through validation against the Amplicor DNA test v1.5 for infant diagnosis using dried blood spots.

PubMed

Maritz, Jean; Preiser, Wolfgang; van Zyl, Gert U

2012-02-01

As antibody testing cannot confirm HIV-1 infection in children less than 18 months of age, diagnosis in these children depends on nucleic acid testing. The COBAS(®) AmpliPrep/COBAS(®) TaqMan(®) (CAP/CTM, Roche(®) Molecular Systems, Inc., Branchburg, NJ) HIV-1 Qualitative test is a total nucleic acid real-time PCR assay utilising whole EDTA blood or dried blood spots (DBS), which recently replaced the Roche(®) AMPLICOR(®) DNA test v1.5 (Amplicor) as the diagnostic HIV PCR assay in many South African laboratories. For the Amplicor assay, stringent diagnostic criteria were previously formulated for the local population, and a comparison reported the CAP/CTM's sensitivity at 99.7% and specificity at 100% for both sample types compared to these Amplicor criteria. To validate the assay prior to introduction in our laboratory and to define stringent diagnostic cut-off criteria. Whole EDTA blood samples from patients younger than 18 months sent for routine HIV-1 diagnosis were tested by Amplicor, and positive results were confirmed from DBS. CAP/CTM assays were subsequently performed from DBS. The CAP/CTM had a sensitivity of 98.8% and a specificity of 97.1%, but a positive predictive value (PPV) of only 78.7% compared to the Amplicor assay. Samples positive by CAP/CTM but negative by Amplicor displayed poor amplification curves compared to concordant positive samples. Upon re-testing those with sufficient material available by CAP/CTM, all showed negative results. The decreased PPV may either be due to false positive CAP/CTM results, or increased sensitivity compared to the Amplicor assay. Criteria were formulated for defining presumed false-positive results. Copyright © 2011 Elsevier B.V. All rights reserved.

Specimen Collection and Submission Manual

DTIC Science & Technology

2016-06-01

immunoassays Specimen: tissue or bone marrow (100 mg); Whole EDTA blood or serum (0.5 ml) Nasopharyngeal or throat swab, dry or in transport medium; Sputum... Syndrome Coronavirus (MERS-CoV) – detection in clinical samples Methodology: molecular Specimen: If possible collect 3 specimen types (lower...guidelines-clinical-specimens.html) Shipping: ship cold on wet ice or ice packs. For delays exceeding 72 hours, ship frozen on dry ice. Turnaround: 1-2

Detection of ethyl glucuronide in blood spotted on different surfaces.

PubMed

Winkler, M; Kaufmann, E; Thoma, D; Thierauf, A; Weinmann, W; Skopp, G; Alt, A

2011-07-15

This study aims to show that sensitive detection of ethyl glucuronide in dried blood spotted onto various surfaces after a period of 24h is feasible. At present, there is insufficient information how tightly ethyl glucuronide (EtG) binds to various materials and how easily it can be eluted. 4ml aliquots of blood samples obtained from seven volunteers after consumption of alcoholic beverages were applied to six different surfaces. After drying and a 24h-storage at 20±2°C the samples were re-dissolved in water, and EtG was subsequently analyzed by a LC-MS Paul-type ion trap. A comparison was made between dried and corresponding fluid samples. EtG was detectable in all subjects' samples following consumption of alcohol. EtG was also detectable after a storage time of four weeks at 4°C in whole blood that had been preserved with EDTA. EtG was detectable in all samples dried on different surfaces and its concentration remained relatively constant irrespective of the particular condition of the material. Detection of EtG in blood spots from the scene may indicate recent alcohol consumption in cases where collection of blood remained undone or could not be performed. Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.

Investigations of blood ammonia analysis: Test matrices, storage, and stability.

PubMed

Goldstein, Brittany N; Wesler, Jordan; Nowacki, Amy S; Reineks, Edmunds; Natowicz, Marvin R

2017-06-01

An assessment of blood ammonia concentration is common medical practice in the evaluation of an individual with an unexplained mental status change or coma. The determination of a blood ammonia level is most commonly done using a glutamate dehydrogenase (GLDH)-based assay, although there are many potential sources of artifact and the literature is inconsistent regarding key preanalytic issues. Using a GLDH-based assay, we first investigated matrix effects using three anticoagulants: heparin, EDTA and oxalate. Heparin-anticoagulated plasma was substantially less precise than EDTA- and oxalate-anticoagulated plasma. Oxalate-anticoagulated plasma showed a greater baseline of apparent ammonia than either heparin- or EDTA-derived plasma, presumably due to interferants. We then evaluated the stability of EDTA-anticoagulated plasma for assessment of ammonia when stored at 4°C,-14°C or -70°C. There was a linear increase of ammonia with storage at both 4°C and -14°C. Plasma kept at -70°C for up to three weeks showed no change in measured ammonia relative to the baseline determination. This work clarifies preanalytic conditions for which a precise determination of ammonia can be accomplished using a GLDH-based assay. Copyright © 2017 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Molecular detection of vector-borne pathogens in blood and splenic samples from dogs with splenic disease.

PubMed

Movilla, Rebeca; Altet, Laura; Serrano, Lorena; Tabar, María-Dolores; Roura, Xavier

2017-03-13

The spleen is a highly perfused organ involved in the immunological control and elimination of vector-borne pathogens (VBP), which could have a fundamental role in the pathogenesis of splenic disease. This study aimed to evaluate certain VBP in samples from dogs with splenic lesions. Seventy-seven EDTA-blood and 64 splenic tissue samples were collected from 78 dogs with splenic disease in a Mediterranean area. Babesia spp., Bartonella spp., Ehrlichia/Anaplasma spp., Hepatozoon canis, Leishmania infantum, hemotropic Mycoplasma spp. and Rickettsia spp. were targeted using PCR assays. Sixty EDTA-blood samples from dogs without evidence of splenic lesions were included as a control group. More than half (51.56%) of the biopsies (33/64) were consistent with benign lesions and 48.43% (31/64) with malignancy, mostly hemangiosarcoma (25/31). PCR yielded positive results in 13 dogs with spleen alterations (16.67%), for Babesia canis (n = 3), Babesia gibsoni (n = 2), hemotropic Mycoplasma spp. (n = 2), Rickettsia massiliae (n = 1) and "Babesia vulpes" (n = 1), in blood; and for B. canis, B. gibsoni, Ehrlichia canis and L. infantum (n = 1 each), in spleen. Two control dogs (3.3%) were positive for B. gibsoni and H. canis (n = 1 each). Benign lesions were detected in the 61.54% of infected dogs (8/13); the remaining 38.46% were diagnosed with malignancies (5/13). Infection was significantly associated to the presence of splenic disease (P = 0.013). There was no difference in the prevalence of infection between dogs with benign and malignant splenic lesions (P = 0.69); however B. canis was more prevalent in dogs with hemangiosarcoma (P = 0.006). VBP infection could be involved in the pathogenesis of splenic disease. The immunological role of the spleen could predispose to alterations of this organ in infected dogs. Interestingly, all dogs with B. canis infection were diagnosed with hemangiosarcoma in the present survey. As previously reported, results support that VBP diagnosis could be improved by analysis of samples from different tissues. The sample size included here warrants further investigation.

A commercial soy-based phospholipid emulsion accelerates clot formation in normal canine whole blood and induces hemolysis in whole blood from normal and dogs with inflammatory leukograms.

PubMed

Behling-Kelly, Erica L; Wakshlag, Joseph

2018-05-01

To compare lipid emulsion-induced hemolysis in blood from dogs with inflammatory leukograms to blood from healthy dogs, and determine the impact of a prototypical soy-based phospholipid emulsion on coagulation in whole blood from healthy dogs. Ex vivo study using EDTA and citrated whole blood from healthy dogs and EDTA anticoagulated whole blood from dogs with inflammatory leukograms. University research laboratory. Healthy dogs (total of 16, 9 for hemolysis assays and 6 for thromboelastography) included student- and staff-owned animals. Blood samples from dogs with inflammatory leukograms (8) were obtained from the clinical pathology laboratory after the complete blood count was performed as part of patient care. For the purposes of this study, an inflammatory leukogram was defined as a neutrophilia with a left-shift (minimum of 3% band neutrophils) and evidence of toxic change. None. Hemolysis was measured via spectrophotometric quantification of released hemoglobin and expressed as a percent of a water-lysed control. The soy emulsion caused hemolysis in blood from healthy dogs, ranging from 3.6% to 16.4% as the dose increased, and 4.1% to 25.0% in blood from dogs with inflammatory leukograms. Hemolysis between these patient groups was significantly different at the highest dose. Coagulation was assessed by native thromboelastography. Treatment of whole blood with the lipid emulsion caused a significant decrease in the time to clot formation (R) and a shorter time to reach a clot amplitude of 20 mm (K). Soy-based lipid emulsions cause hemolysis that is more severe in blood from dogs with inflammatory leukograms and accelerate clot formation in canine blood. The in vivo significance of these findings is not clear at this time, but warrants additional investigation given the use of these emulsions in clinical practice. © Veterinary Emergency and Critical Care Society 2018.

Comparison of two real-time PCR assays for the detection of malaria parasites from hemolytic blood samples - Short communication.

PubMed

Hagen, Ralf Matthias; Hinz, Rebecca; Tannich, Egbert; Frickmann, Hagen

2015-06-01

We compared the performance of an in-house and a commercial malaria polymerase chain reaction (PCR) assay using freeze-thawed hemolytic blood samples. A total of 116 freeze-thawed ethylenediamine tetraacetic acid (EDTA) blood samples of patients with suspicion of malaria were analyzed by an in-house as well as by a commercially available real-time PCR. Concordant malaria negative PCR results were reported for 39 samples and malaria-positive PCR results for 67 samples. The in-house assay further detected one case of Plasmodium falciparum infection, which was negative in the commercial assay as well as five cases of P. falciparum malaria and three cases of Plasmodium vivax malaria, which showed sample inhibition in the commercial assay. The commercial malaria assay was positive in spite of a negative in-house PCR result in one case. In all concordant results, cycle threshold values of P. falciparum-positive samples were lower in the commercial PCR than in the in-house assay. Although Ct values of the commercial PCR kit suggest higher sensitivity in case of concordant results, it is prone to inhibition if it is applied to hemolytic freeze-thawed blood samples. The number of misidentifications was, however, identical for both real-time PCR assays.

K2-EDTA and K3-EDTA Greiner Tubes for HbA1c Measurement.

PubMed

Vrtaric, Alen; Filipi, Petra; Hemar, Marina; Nikolac, Nora; Simundic, Ana-Maria

2016-02-01

To determine whether K2-ethylenediaminetetraacetic acid (EDTA) and K3-EDTA Greiner tubes could be used interchangeably for glycosylated hemoglobin, type A1C (HbA1c) measurement via the Abbott Laboratories ARCHITECT chemiluminescent microparticle HbA1c assay on the ARCHITECT i2000SR immunoanalyzer at our university hospital. We drew blood from a total of 45 outpatients into plastic Greiner Vacuette tubes, some of which were lined with K2-EDTA and others with K3-EDTA anticoagulant. Data are presented as median and interquartile range values. We used the Wilcoxon test and Passing-Bablok regression for tube comparison. For K2-EDTA tubes median HbA1c concentration was 54 mmol/mol (41 to 71 mmol/mol) and for K3-EDTA tubes 56 mmol/mol (43 to 69 mmol/mol). There was no statistically significant difference between K2-EDTA and K3-EDTA (bias= -1.29 mmol/mol; P = 0.24). Passing-Bablok regression showed that there is no constant and proportional error: y = -0.23 (95% CI[-3.52 to 0.69]) + 1.00( 95% CI[0.98 to 1.06]) x. In this study, we provide evidence for the lack of any clinically and statistically significant bias between K2-EDTA and K3-EDTA HbA1c measurements. Thus, Greiner tubes lined with K2-EDTA and those lined with K3-EDTA can safely be used interchangeably to measure HbA1c via the Abbott Laboratories ARCHITECT assay. © American Society for Clinical Pathology, 2015. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

[DNA quantification of blood samples pre-treated with pyramidon].

PubMed

Zhu, Chuan-Hong; Zheng, Dao-Li; Ni, Rao-Zhi; Wang, Hai-Sheng; Ning, Ping; Fang, Hui; Liu, Yan

2014-06-01

To study DNA quantification and STR typing of samples pre-treated with pyramidon. The blood samples of ten unrelated individuals were anticoagulated in EDTA. The blood stains were made on the filter paper. The experimental groups were divided into six groups in accordance with the storage time, 30 min, 1 h, 3 h, 6 h, 12 h and 24h after pre-treated with pyramidon. DNA was extracted by three methods: magnetic bead-based extraction, QIAcube DNA purification method and Chelex-100 method. The quantification of DNA was made by fluorescent quantitative PCR. STR typing was detected by PCR-STR fluorescent technology. In the same DNA extraction method, the sample DNA decreased gradually with times after pre-treatment with pyramidon. In the same storage time, the DNA quantification in different extraction methods had significant differences. Sixteen loci DNA typing were detected in 90.56% of samples. Pyramidon pre-treatment could cause DNA degradation, but effective STR typing can be achieved within 24 h. The magnetic bead-based extraction is the best method for STR profiling and DNA extraction.

Comparison of home fortification with two iron formulations among Kenyan children: Rationale and design of a placebo-controlled non-inferiority trial.

PubMed

Teshome, Emily M; Otieno, Walter; Terwel, Sofie R; Osoti, Victor; Demir, Ayşe Y; Andango, Pauline E A; Prentice, Andrew M; Verhoef, Hans

2017-09-01

Home fortification powders containing iron and other micronutrients have been recommended by World Health Organisation to prevent iron deficiency anaemia in areas of high prevalence. There is evidence, however, that home fortification at this iron dose may cause gastrointestinal adverse events including diarrhoea. Providing a low dose of highly absorbable iron (3 mg iron as NaFeEDTA) may be safer because the decreased amount of iron in the gut lumen can possibly reduce the burden of these adverse effects whilst resulting in similar or higher amounts of absorbed iron. To show non-inferiority of home fortification with 3 mg iron as NaFeEDTA compared with 12.5 mg iron as encapsulated ferrous fumarate, with haemoglobin response as the primary outcome. 338 Kenyan children aged 12-36 months will be randomly allocated to daily home fortification with either: a) 3 mg iron as NaFeEDTA (experimental treatment), b) 12.5 mg iron as encapsulated ferrous fumarate (reference), or c) placebo. At baseline, after 30 days of intervention and within 100 days post-intervention, blood samples will be assessed for primary outcome (haemoglobin concentration), iron status markers, Plasmodium parasitaemia and inflammation markers. Urine and stool samples will be assessed for hepcidin concentrations and inflammation, respectively. Adherence will be assessed by self-reporting, sachet counts and by an electronic monitoring device. If daily home fortification with a low dose of iron (3 mg NaFeEDTA) has similar or superior efficacy to a high dose (12.5 mg ferrous fumarate) then it would be the preferred choice for treatment of iron deficiency anaemia in children.

Investigation of on-line chelant addition to PWR steam generators. Annual report, 1981

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tvedt, T.J.; Wallace, S.L.; Griffin, F. Jr.

1982-11-01

The thermostability of both ethylenediaminetetraacetic acid (EDTA) and hydroxyethylethylenediamininetriacetic acid (HEDTA) metal chelates in all volatile treatment water chemistry (AVT) was shown to be greater than or equal to thermostability of EDTA metal chelates in phosphate-sulfite water chemistry. HEDTA metal chelates were shown to have a much greater stability than EDTA metal chelates. Using samples taken from the EDTA metal chelate thermostability studies and samples from Commonwealth Research Corporation (CRC) model steam generators (MSG), EDTA decomposition products were determined. Active metal surfaces were shown to become passivated when exposed to EDTA and HEDTA concentrations as high as 0.1% w/w inmore » AVT. Trace amounts of iron in the water were found to increase the rate of passivation. Material balance and visual inspection data from CRC model steam generators showed that metal is being transported through and cleaning from the MSG's. EDTA metal chelates were removed from chelate solutions by passing the solutions over strong anion exchange resins.« less

Molecular characterization of Hepatozoon canis in dogs from Colombia.

PubMed

Vargas-Hernandez, Giovanni; André, Marcos R; Munhoz, Thiago D; Faria, Joice M L; Machado, Rosangela Z; Tinucci-Costa, Mirela

2012-01-01

Hepatozoonosis is a tick-borne disease whose transmission to dogs occurs by ingestion of oocysts infected ticks or feeding on preys infested by infected ticks. Until now, there is no previous report of molecular characterization of Hepatozoon sp. in dogs from Colombia. EDTA blood samples were collected from 91 dogs from central-western region of Colombia (Bogotá, Bucaramanga, and Villavicencio cities) and submitted to 18S rRNA Hepatozoon sp. PCR and blood smears confection. Phylogenetic analysis was used to access the identity of Hepatozoon species found in sampled dogs. From 91 sampled dogs, 29 (31.8%) were positive to Hepatozoon sp. (25 dogs were only positive in PCR, 1 was positive only in blood smears, and 3 were positive in both blood smears and PCR). After sequencing, the found Hepatozoon sp. DNA showed 100% of identity with Hepatozoon canis DNA isolates. The phylogenetic tree supported the identity of the found Hepatozoon sp. DNA, showing that the isolates from Colombia were placed in the same clade than other H. canis isolates from Venezuela, Spain, and Taiwan. This is the first molecular detection of H. canis in dogs from Colombia.

Influence of blood sampling methods on dopamine-receptor-blocking activities as determined by a radioreceptor assay.

PubMed

Lai, A A; Fleck, R J; Patzke, J V; Glueck, B G; Shaskan, E G; Rosenberg, B J

1982-01-01

The influence of blood collection methods on dopamine-receptor-blocking activities as determined by a radioreceptor assay kit was investigated. Thirty-one patients treated with one of six neuroleptic drugs (thioridazine, trifluoperazine, haloperidol, chlorpromazine, thiothixene, or fluphenazine) participated in this study. Blood samples were drawn from each patient into five different evacuated blood collection tubes made by the same manufacturer (red-stoppered tube containing no additives, lavender-stoppered tube containing EDTA, green-stoppered tube containing heparin, dark blue-stoppered tube containing no additives, and dark blue-stoppered tube containing heparin). The results show that for five drugs (chlorpromazine, fluphenazine, haloperidol, thiothixene, and trifluoperazine), the dark blue-stoppered tubes without additives resulted in significantly higher dopamine-receptor-blocking activities than the red-, lavender-, or green-stoppered tubes. For thioridazine, the green-stoppered tubes resulted in significantly higher blocking activities than the blue- and red-stoppered tubes. The possible effect of tris(2-butoxyethyl) phosphate, a plasticizer, on dopamine-receptor-blocking activities by neuroleptic drugs is discussed.

Cytogenetic investigations of chronic lymphocytic leukemia.

PubMed

Wren, Catherine; Moriarty, Helen; Marsden, Katherine; Tegg, Elizabeth

2010-04-15

This study aimed to determine which culture method would yield the highest culture success rate, mitotic index, banding resolution, and abnormality rate in investigation of patients with chronic lymphocytic leukemia (CLL). A range of culture techniques for conventional cytogenetic (CC) analyses was compared: 24-hour unstimulated, 72 hours incubation with additional fetal calf serum, 72 hours stimulation with interleukin 4, 72 hours stimulation with lipopolysaccharide (LPS), 72 hours stimulation with TPA (12-O-tetradecanoylphorbol 13-acetate), and 72 hours stimulation with CpG-oligonucleotide DSP30 + Interleukin-2 (IL-2). CC abnormality rates were also compared to fluorescence in situ hybridization (FISH) results using probes for CLL (LSI D13S319/13q34/CEP 12: LSI ATM/p53). Forty-five samples from 24 patients (consisting of 11 newly diagnosed and 13 previously diagnosed patients) were included. For CC, a 100.0% culture success rate was achieved (n = 45) by means of an EDTA (ethylenediaminetetraacetic acid) peripheral blood sample with an associated 62.5% CC abnormality rate (n = 24). FISH detected an abnormality rate of 75.0% (n = 24). The combined CC and FISH abnormality rate was 87.5% (n = 24). This study demonstrates that CC that uses TPA and DSP30 + IL-2 on EDTA peripheral blood is effective in the investigation of CLL and may be used as a supplement to FISH studies. Copyright 2010 Elsevier Inc. All rights reserved.

Identification of mast cells in buffy coat preparations from dogs with inflammatory skin diseases.

PubMed

Cayatte, S M; McManus, P M; Miller, W H; Scott, D W

1995-02-01

In 100 dogs with 4 inflammatory dermatologic diseases, buffy coat preparations from EDTA-treated blood samples were examined cytologically. Fifty-four dogs had atopy, 26 had flea-bite hypersensitivity, 17 had sarcoptic mange, and 3 had food allergy. Twenty-eight dogs had 2 or more concurrent skin diseases; most of these had secondary pyoderma. Dogs did not have mast cell tumors. Thirteen samples contained 1 or more mast cells/4 slides reviewed. This study revealed that dogs with inflammatory skin diseases can have a few to many mast cells evident on cytologic examination of buffy coat preparations.

Fragmentation of Red Blood Cells Caused Pseudothrombocytosis in a Patient.

PubMed

Tang, Wenjun; Tang, Chunhua; Zheng, Jianfeng; He, Yongling; Lu, Fangfang

2018-06-01

Pseudothrombocytopenia, caused by platelet (PLT) clumping, is often found in clinical studies [1]. However, pseudothrombocytosis resulting from the fragmentation of red blood cells (RBCs) is a very rare phenomenon. EDTA-K2 anticoagulation was used on a sample of venous blood extracted from the patient. A Symex XN9000 automatic blood analyzer was used to conduct CBC + DIFF mode and CBC + DIFF + RET mode tests, stained smear microscopy. The Symex XN9000 automatic blood analyzer was used to conduct CBC + DIFF mode test; PLTs were measured at 570 x 109/L. Stained smear microscopy revealed the number of PLTs did not conform to the instrument measured 570 x 109/L. "RET" alarm instrument, switch to CBC + DIFF + RET mode for testing. The second test result showed PLTs at 128 x 109/L, which accords with artificial microscopy. This was a case of a very rare phenomenon: the fragmentation of RBCs caused pseudothrombocytosis.

Most blood biomarkers related to vitamin status, one-carbon metabolism, and the kynurenine pathway show adequate preanalytical stability and within-person reproducibility to allow assessment of exposure or nutritional status in healthy women and cardiovascular patients.

PubMed

Midttun, Oivind; Townsend, Mary K; Nygård, Ottar; Tworoger, Shelley S; Brennan, Paul; Johansson, Mattias; Ueland, Per Magne

2014-05-01

Knowledge of stability during sample transportation and changes in biomarker concentrations within person over time are paramount for proper design and interpretation of epidemiologic studies based on a single measurement of biomarker status. Therefore, we investigated stability and intraindividual vs. interindividual variation in blood concentrations of biomarkers related to vitamin status, one-carbon metabolism, and the kynurenine pathway. Whole blood (EDTA and heparin, n = 12) was stored with an icepack for 24 or 48 h, and plasma concentrations of 38 biomarkers were determined. Stability was calculated as change per hour, intraclass correlation coefficient (ICC), and simple Spearman correlation. Within-person reproducibility of biomarkers was expressed as ICC in samples collected 1-2 y apart from 40 postmenopausal women and in samples collected up to 3 y apart from 551 patients with stable angina pectoris. Biomarker stability was similar in EDTA and heparin blood. Most biomarkers were essentially stable, except for choline and total homocysteine (tHcy), which increased markedly. Within-person reproducibility in postmenopausal women was excellent (ICC > 0.75) for cotinine, all-trans retinol, cobalamin, riboflavin, α-tocopherol, Gly, pyridoxal, methylmalonic acid, creatinine, pyridoxal 5'-phosphate, and Ser; was good to fair (ICC of 0.74-0.40) for pyridoxic acid, kynurenine, tHcy, cholecalciferol, flavin mononucleotide, kynurenic acid, xanthurenic acid, 3-hydroxykynurenine, sarcosine, anthranilic acid, cystathionine, homoarginine, 3-hydroxyanthranilic acid, betaine, Arg, folate, total cysteine, dimethylglycine, asymmetric dimethylarginine, neopterin, symmetric dimethylarginine, and Trp; and poor (ICC of 0.39-0.15) for methionine sulfoxide, Met, choline, and trimethyllysine. Similar reproducibilities were observed in patients with coronary heart disease. Thus, most biomarkers investigated were essentially stable in cooled whole blood for up to 48 h and had a sufficient within-person reproducibility to allow one-exposure assessment of biomarker status in epidemiologic studies. The Western Norway B Vitamin Intervention Trial was registered at clinicaltrials.gov as NTC00354081.

Transcapillary Exchange and Retention of Fluoride, Strontium, EDTA, Sucrose, and Antipyrine in Bone

PubMed Central

Lemon, Gerard J.; Davies, David R.; Hughes, Sean P.F.; Bassingthwaighte, James B.; Kelly, Patrick J.

2010-01-01

Summary The extractions of 85Sr2+, 18F−, sucrose-14C, EDTA-51Cr, and antipyrine-14C in bone were determined by the multiple indicator-dilution method. Fluoride and strontium extractions were 18 to 70% during a single transcapillary passage, and those of EDTA and sucrose were from 11 to 59%, whereas extraction of antipyrine was 87%. Injections of 85Sr2+ and 18F− made when perfusion was done alternately with blood and plasma resulted in similar fractional extractions. When flow and extraction were measured simultaneously, extraction was related inversely to flow. PMID:6770980

CaEDTA vs CaEDTA plus BAL to treat children with elevated blood lead levels.

PubMed

O'Connor, M E

1992-07-01

The effectiveness of CaEDTA alone vs CaEDTA plus BAL was compared retrospectively in a group of 72 children with lead levels between 2.41 mumol/L (50 micrograms/dL) and 2.90 mumol/L (60 micrograms/dL). The children who received both drugs had higher median zinc protoporphyrin (ZnP) concentrations at the initiation of therapy than children who received CaEDTA alone (160 micrograms/dL vs 96 micrograms/dL, p less than .01). There was a significantly increased incidence of vomiting and abnormal liver-function test results in the children who received both drugs. The children who received CaEDTA alone had a greater percent mean fall in lead level at one to three weeks postchelation (30.5% vs 18.1%, p less than .05). Children who received both CaEDTA and BAL had a greater percent decrease in ZnP at four to eight months postchelation, but there was no difference in percent decrease in lead levels. Children who received both drugs also had a greater number of repeat courses of chelation by six months. The addition of BAL to CaEDTA for treatment of children with lead levels of 2.41 mumol/L (50 micrograms/dL) to 2.90 mumol/L (60 micrograms/dL) produced greater toxicity and does not seem to prevent repeat chelations within six months.

Severe Lead Toxicosis in a Lionhead Rabbit.

PubMed

Walter, Kristen M; Bischoff, Karyn; de Matos, Ricardo

2017-03-01

Lead toxicosis occurs in veterinary patients, with few reports involving rabbits, and no previous reports using oral calcium disodium EDTA. A 7-year-old male castrated Lionhead rabbit presented to the Cornell University Hospital for Animals (CUHA) for evaluation after a 2-day history of lethargy and a 2-week history of hyporexia. The patient had been observed pulling paint from the walls of the home, a house built circa 1900, in the months prior to presentation. The patient was moderately anemic with a hematocrit of 21% with red blood cell morphological changes consistent with lead toxicosis, including basophilic stippling, nucleated red blood cells, and polychromasia. Radiographic images of the abdomen revealed excessive accumulation of gas in the gastrointestinal tract in a pattern consistent with gastric stasis and numerous small mineral to metallic opacities in the cecum. The blood lead concentration was 792 μg/dL, confirming the diagnosis of lead toxicosis with secondary gastrointestinal stasis. The rabbit was hospitalized for treatment with oral and subcutaneous calcium disodium EDTA for 4 days and then discharged home to the care of the owners. Severe lead toxicosis in a rabbit can be treated successfully with oral and subcutaneous calcium disodium EDTA and aggressive supportive treatment.

Immunological effects of CaEDTA injection: observations in two lead workers.

PubMed

Sata, F; Araki, S; Sakai, T; Nakata, A; Yamashita, K; Morita, Y; Tanigawa, T; Miki, A

1997-12-01

To evaluate the effects of calcium disodium ethylenediamine tetraacetate (CaEDTA) injection on human immune system in relation to exposure to lead, we administered CaEDTA by intravenous injection for 1 hr three times (three consecutive days) a week to two male lead workers. They had been engaged in recycling lead for 31 and 22 years, aged 61 and 53 years (workers 1 and 2), respectively. Before the treatment of CaEDTA, their blood lead concentrations (PbB) were 81 and 68 micrograms/dl, respectively. The administration of CaEDTA had been carried out to worker 1 for 10 weeks and to worker 2 for 6 weeks. A significant decrease in PbB between before and after three-times CaEDTA injection was found in both workers. Significant increases in IgG, IgA, IgM, CD8+, and CD57+ cells were found in worker 1. A significant increase in IgD was found in worker 2. During the study period, IgG in worker 1 and CD4+ cells in worker 2 were gradually increasing. There was a significant negative correlation between IgG and PbB in worker 1. It is suggested that the immunological function such as antibody formation in lead workers might be improved by CaEDTA injection.

The effect of different protease inhibitors on stability of parathyroid hormone, insulin, and prolactin levels under different lag times and storage conditions until analysis.

PubMed

Baykan, Ozgur; Yaman, Ali; Gerin, Fethullah; Sirikci, Onder; Haklar, Goncagul

2017-11-01

Proteolytic cleavage through proteases affects peptide hormone levels, which is of particular significance when the time interval between sampling and analysis is prolonged. We evaluated the stability of parathyroid hormone, insulin, and prolactin molecules (i) with different protease inhibitors such as K 2 EDTA, aprotinin, and protease inhibitor cocktail (PIC), (ii) with different lag times (6-72 hours), and (iii) under different storage temperatures (4°C vs room temperature [RT]) until analysis. Blood samples were collected into 2 sets of 5 Vacutainer ® tubes (Becton Dickinson) from 10 healthy adults. Tubes 1 and 2 were plain gel separator tubes. Tubes 3, 4, and 5 contained PIC (1%), aprotinin (500 KIU/mL), and K 2 EDTA, respectively. After centrifugation at 1300 g for 10 minutes, PIC added to tube 2 of each set. Samples were analyzed and then one set was stored at 4°C, whereas the other at RT until analysis at 6, 24, 48, and 72 hours. Hormone levels were determined with electrochemiluminescence immunoassay (ModularE170; Roche Diagnostics). The results were compared with desirable bias limits (DBL) from Westgard QC database. Insulin at RT decreases exceeding the DBL starting from 24 hours and K 2 EDTA preserved insulin. PTH exceeded the DBL at RT for 48 hours or longer and PIC addition after centrifugation inhibited its degradation. Prolactin remained stable in all tested conditions. All parameters in the plain gel separator tubes remained within DBL when stored at 4°C until 72 hours. Different proteases may degrade peptide hormones and measures should be taken to counteract these effects especially if there is a delay before analysis. © 2017 Wiley Periodicals, Inc.

Quantification of Hydroxychloroquine in Blood Using Turbulent Flow Liquid Chromatography-Tandem Mass Spectrometry (TFLC-MS/MS).

PubMed

Chambliss, Allison B; Füzéry, Anna K; Clarke, William A

2016-01-01

Hydroxychloroquine (HQ) is used routinely in the treatment of autoimmune disorders such as rheumatoid arthritis and lupus erythematosus. Issues such as marked pharmacokinetic variability and patient non-compliance make therapeutic drug monitoring of HQ a useful tool for management of patients taking this drug. Quantitative measurements of HQ may aid in identifying poor efficacy as well as provide reliable information to distinguish patient non-compliance from refractory disease. We describe a rapid 7-min assay for the accurate and precise measurement of HQ concentrations in 100 μL samples of human blood using turbulent flow liquid chromatography coupled to tandem mass spectrometry. HQ is isolated from EDTA whole blood after a simple extraction with its deuterated analog, hydroxychloroquine-d4, in 0.33 M perchloric acid. Samples are then centrifuged and injected onto the TFLC-MS/MS system. Quantification is performed using a nine-point calibration curve that is linear over a wide range (15.7-4000 ng/mL) with precisions of <5 %.

[An approach to a methodology of scientific research for assistant-students].

PubMed

Novak, Ivón T C; Bejarano, Paola Antón; Rodríguez, Fernando Marcos

2007-01-01

This work is presented from a "problematic" perspective in the attempt to establish a dialogic relationship between the educator and the student-subject, mediated by the object of knowledge. It is oriented to the integral education of the helping students departing from a closer approach to the scientific research. This work was carried out by a teacher and two hired students. This project was developed in relation with the profile required for the career of medicine in the Faculty of Medicine of the National University of Cordoba which--among other aspects- addresses the importance of "adopting a positive attitude towards research based on knowledge and the application of the scientific methodology" and towards "the development of a responsible self-learning and continuous improvements" (sic). Thus, this work tries to be aligned with this perspectives. I. Characterization of the scientific methodology. Search for bibliography and discussion of scientific works. II. Optimization of the methodology for the observation of leucocytes: blood samples donated by healthy people, non-coagulating with citrate or with EDTA (Blood reservoir of the UNC (National University of Cordoba) n = 20. a) Blood smear of full blood. b) centrifugation at 200g of plasma and aspirated leucocytes after erythro sedimentation and re suspension of the cell pellet and cyto-dispersion. Cytological and cyto-chemical techniques. I. Deeper knowledge about blood field was achieved. It generated an appropriate atmosphere to produce scientific questioning and the activities involved in the process were carried out responsibly. II. Better results were achieved using EDTA for the observation and analysis of leucocytes. It was possible to attain the objectives for an approach to a scientific research as well as for a contribution towards a responsible development in the continuous learning process.

Closed vessel miniaturized microwave assisted chelating extraction for determination of trace metals in plant materials

NASA Astrophysics Data System (ADS)

Czarnecki, Sezin; Duering, Rolf-Alexander

2013-04-01

In recent years, the use of closed vessel microwave assisted extraction (MAE) for plant samples has shown increasing research interest which will probably substitute conventional procedures in the future due to their general disadvantages including consumption of time and solvents. The objective of this study was to demonstrate an innovative miniaturized closed vessel microwave assisted extraction (µMAE) method under the use of EDTA (µMAE-EDTA) to determine metal contents (Cd, Co, Cu, Mn, Ni, Pb, Zn) in plant samples (Lolio-Cynosuretum) by inductively coupled plasma-optical emission spectrometry (ICP-OES). Validation of the method was done by comparison of the results with another miniaturized closed vessel microwave HNO3 method (µMAE-H) and with two other macro scale MAE procedures (MAE-H and MAE-EDTA) which were applied by using a mixture of nitric acid (HNO3) and hydrogen peroxide (H2O2) (MAE-H) and EDTA (MAE-EDTA), respectively. The already established MAE-H method is taken into consideration as a reference validation MAE method for plant material. A conventional plant extraction (CE) method, based on dry ashing and dissolving of the plant material in HNO3, was used as a confidence comparative method. Certified plant reference materials (CRMs) were used for comparison of recovery rates from different extraction protocols. This allowed the validation of the applicability of the µMAE-EDTA procedure. For 36 real plant samples with triplicates each, µMAE-EDTA showed the same extraction yields as the MAE-H in the determination of Cd, Co, Cu, Mn, Ni, Pb, and Zn contents in plant samples. Analytical parameters in µMAE-EDTA should be further investigated and adapted for other metals of interest. By the reduction and elimination of the use of hazardous chemicals in environmental analysis and thus allowing a better understanding of metal distribution and accumulation process in plants and also the metal transfer from soil to plants and into the food chain, µMAE-EDTA is seen as a promising technique for achieving green chemistry goals.

Mobilisation of heavy metals into the urine by CaEDTA: relation to erythrocyte and plasma concentrations and exposure indicators.

PubMed

Araki, S; Aono, H; Murata, K

1986-09-01

To investigate the effects of calcium disodium ethylenediamine tetra-acetate (CaEDTA) on the urinary excretion, erythrocyte, and plasma concentrations and exposure indicators of seven heavy metals, CaEDTA was administered by intravenous infusion to 20 workers exposed to lead, zinc, and copper. The workers' blood lead concentrations ranged from 22 to 59 micrograms/dl (mean 38 micrograms/dl (1.8 mumol/l]. The 24 hour urinary excretion of metals after CaEDTA administration (mobilisation yield) was on average 13 times the background excretion for lead, 11 times for zinc, 3.8 times for manganese, 3.4 times for cadmium, 1.3 times for copper, and 1.1 times for chromium; no significant increase was found for mercury. The mobilisation yield of lead (MPb) was significantly correlated with whole blood and erythrocyte concentrations and the urinary excretion of lead but not with its plasma concentration; similarly, the mobilisation yield of cadmium was significantly correlated with its erythrocyte concentration. In addition, MPb was significantly correlated with intra-erythrocytic enzyme delta-aminolaevulinic acid dehydratase activity and urinary coproporphyrin excretion. The relation between the mobilisation yield of heavy metals and their body burden (and toxic signs) is discussed in the light of these findings.

Mobilisation of heavy metals into the urine by CaEDTA: relation to erythrocyte and plasma concentrations and exposure indicators.

PubMed Central

Araki, S; Aono, H; Murata, K

1986-01-01

To investigate the effects of calcium disodium ethylenediamine tetra-acetate (CaEDTA) on the urinary excretion, erythrocyte, and plasma concentrations and exposure indicators of seven heavy metals, CaEDTA was administered by intravenous infusion to 20 workers exposed to lead, zinc, and copper. The workers' blood lead concentrations ranged from 22 to 59 micrograms/dl (mean 38 micrograms/dl (1.8 mumol/l]. The 24 hour urinary excretion of metals after CaEDTA administration (mobilisation yield) was on average 13 times the background excretion for lead, 11 times for zinc, 3.8 times for manganese, 3.4 times for cadmium, 1.3 times for copper, and 1.1 times for chromium; no significant increase was found for mercury. The mobilisation yield of lead (MPb) was significantly correlated with whole blood and erythrocyte concentrations and the urinary excretion of lead but not with its plasma concentration; similarly, the mobilisation yield of cadmium was significantly correlated with its erythrocyte concentration. In addition, MPb was significantly correlated with intra-erythrocytic enzyme delta-aminolaevulinic acid dehydratase activity and urinary coproporphyrin excretion. The relation between the mobilisation yield of heavy metals and their body burden (and toxic signs) is discussed in the light of these findings. PMID:3092853

Validation and determination of a reference interval for canine HbA1c using an immunoturbidimetric assay.

PubMed

Goemans, Anne F; Spence, Susanna J; Ramsey, Ian K

2017-06-01

Hemoglobin A1c (HbA1c) provides a reliable measure of glycemic control over 2-3 months in human diabetes mellitus. In dogs, presence of HbA1c has been demonstrated, but there are no validated commercial assays. The purpose of the study was to validate a commercially available automated immunoturbidimetric assay for canine HbA1c and determine an RI in a hospital population. The specificity of the assay was assessed by inducing glycosylation in vitro using isolated canine hemoglobin, repeatability by measuring canine samples 5 times in succession, long term inter-assay imprecision by measuring supplied control materials, stability using samples stored at 4°C over 5 days and -20°C over 8 weeks, linearity by mixing samples of known HbA1c in differing proportions, and the effect of anticoagulants with paired samples. An RI was determined using EDTA-anticoagulated blood samples from 60 nondiabetic hospitalized animals of various ages and breeds. Hemoglobin A1c was also measured in 10 diabetic dogs. The concentration of HbA1c increased proportionally with glucose concentration in vitro. For repeat measurements, the CV was 4.08% (range 1.16-6.10%). Samples were stable for 5 days at 4°C. The assay was linear within the assessed range. Heparin- and EDTA-anticoagulated blood provided comparable results. The RI for HbA1c was 9-18.5 mmol/mol. There was no apparent effect of age or breed on HbA1c. In diabetic dogs, HbA1c ranged from 14 to 48 mmol/mol. The assay provides a reliable method for canine HbA1c measurement with good analytic performance. © 2017 American Society for Veterinary Clinical Pathology.

EDTA retention and emissions from remediated soil.

PubMed

Jez, Erika; Lestan, Domen

2016-05-01

EDTA-based remediation is reaching maturity but little information is available on the state of chelant in remediated soil. EDTA soil retention was examined after extracting 20 soil samples from Pb contaminated areas in Slovenia, Austria, Czech Republic and USA with 120 mM kg(-1) Na2H2EDTA, CaNa2EDTA and H4EDTA for 2 and 24 h. On average, 73% of Pb was removed from acidic and 71% from calcareous soils (24 h extractions). On average, 15% and up to 64% of applied EDTA was after remediation retained in acidic soils. Much less; in average 1% and up to the 22% of EDTA was retained in calcareous soils. The secondary emissions of EDTA retained in selected remediated soil increased with the acidity of the media: the TCLP (Toxicity Characteristic Leaching Procedure) solution (average pH end point 3.6) released up to 36% of EDTA applied in the soil (28.1 mmol kg(-1)). Extraction with deionised water (pH > 6.0) did not produce measurable EDTA emissions. Exposing soil to model abiotic (thawing/freezing cycles) and biotic (ingestion by earthworms Lumbricus rubellus) ageing factors did not induce additional secondary emissions of EDTA retained in remediated soil. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effects of vigorous mixing of blood vacuum tubes on laboratory test results.

PubMed

Lima-Oliveira, Gabriel; Lippi, Giuseppe; Salvagno, Gian Luca; Montagnana, Martina; Gelati, Matteo; Volanski, Waldemar; Boritiza, Katia Cristina; Picheth, Geraldo; Guidi, Gian Cesare

2013-02-01

To evaluate the effect of tubes mixing (gentle vs. vigorous) on diagnostic blood specimens collected in vacuum tube systems by venipuncture. Blood was collected for routine coagulation, immunochemistry and hematological testing from one hundred volunteers into six vacuum tubes: two 3.6 mL vacuum tubes containing 0.4 mL of buffered sodium citrate (9NC) 0.109 mol/L: 3.2 W/V%; two 3.5 mL vacuum tubes with clot activator and gel separator; and two 3.0 mL vacuum tubes containing 5.9 mg K(2)EDTA (Terumo Europe, Belgium). Immediately after the venipuncture all vacuum tubes (each of one additive type) were processed through two different procedures: i) Standard: blood specimens in K(2)EDTA- or sodium citrate-vacuum tubes were gently inverted five times whereas the specimens in tubes with clot activator and gel separator were gently inverted ten times, as recommended by the manufacturer; ii) Vigorous mix: all blood specimens were shaken up vigorously during 3-5s independently of the additive type inside the tubes. The significance of the differences between samples was assessed by Student's t-test or Wilcoxon ranked-pairs test after checking for normality. The level of statistical significance was set at P<0.05. No significant difference (P<0.05) was detected between the procedures for all tested parameters. Surprisingly only a visual alteration (presence of foam on the top) was shown by all the tubes mixed vigorously before centrifugation (Fig. 1 A, B and C). Moreover the serum tubes from vigorous mixing procedure shows a "blood ring" on the tube top after stopper removal (Fig. 1 D). Our results drop out a paradigm suggesting that the incorrect primary blood tubes mixing promotes laboratory variability. We suggest that similar evaluation should be done using other brands of vacuum tubes by each laboratory manager. Copyright © 2012 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

An improved automated method for the measurement of red cell 2,3-diphosphoglycerate.

PubMed Central

Purcell, Y; Brozović, B

1976-01-01

A modified automated colorimetric micromethod for the determination of red cell 2,3-diphosphoglycerate (2,3-DPG) adapted from that of Grisolia et al (1969) is described. In the modified method, ethylenediaminetetra-acetic acid (EDTA) is not used and consequently concentrations of several reagents are changed. During the development of the method it was found that the presence of EDTA, either in the blood or in reagents, consistently reduced the measured value of 2,3-DPG by 15%. This effect of EDTA, not previously recognized, is independent of the EDTA concentration within the range of 5 to 50 mmol/1 and is at present unexplianed. In normal subjects (41 men and 30 women) the mean red cell 2,3-DPG was 14-5 mol/g haemoglobin (range 12-1-18-1 mol/g haemoglobin). There was no significant difference in 2,3-DPG concentrations between male and female subjects. PMID:827552

Quantitative Analysis of Sulfate in Water by Indirect EDTA Titration

ERIC Educational Resources Information Center

Belle-Oudry, Deirdre

2008-01-01

The determination of sulfate concentration in water by indirect EDTA titration is an instructive experiment that is easily implemented in an analytical chemistry laboratory course. A water sample is treated with excess barium chloride to precipitate sulfate ions as BaSO[subscript 4](s). The unprecipitated barium ions are then titrated with EDTA.…

Application of EDTA decontamination on soils affected by mining activities and impact of treatment on the geochemical partition of metal contaminants.

PubMed

Xia, Wenbin; Gao, Hui; Wang, Xianhai; Zhou, Chunhua; Liu, Yunguo; Fan, Ting; Wang, Xin

2009-05-30

Two soil samples were collected at mining areas located in southern Hunan Province, China. EDTA extraction of Pb, Zn, Cu and Cd from these two tailing soils was studied using column leaching experiments. The redistributions of heavy metals (HMs) were determined using the modified BCR (Community Bureau of Reference) sequential extraction procedure, before and after EDTA extraction. The results indicated that EDTA was an effective extractant because of its strong chelating ability for various HMs. The proportions of Pb, Zn, Cu and Cd in the four fractions varied largely after EDTA extraction. The extraction efficiency of EDTA of the acid-extractable fraction (AEX) was significant in shallow soil column, while in deeper soil column, decrease of the extraction efficiency of reduced (RED), oxidizable (OX) and residual fractions (RES) was obtained, which was mainly due to the decrease of EDTA concentration.

Determination of ethylenediaminetetraacetic acid in sea water by solid-phase extraction and high-performance liquid chromatography.

PubMed

Kemmei, Tomoko; Kodama, Shuji; Fujishima, Hironori; Yamamoto, Atsushi; Inoue, Yoshinori; Hayakawa, Kazuichi

2012-01-04

The chelating agent EDTA is widely used, and as a result is showing up widely in the aquatic environment. Here we describe a preconcentration procedure for measuring EDTA concentration in sea water samples by HPLC. The procedure consists of forming an Fe(III) complex followed by solid-phase extraction using an activated carbon cartridge. After the preconcentration, EDTA was quantified by HPLC with ultraviolet detection (260 nm). The enrichment permitted the determination of EDTA at concentrations as low as 1 nM. Good recoveries were obtained for both brackish and full-strength sea water with high repeatability (RSD<6%). The method was applied to sea water samples taken from near the mouth of the Oyabe River in Japan. Copyright © 2011 Elsevier B.V. All rights reserved.

Simultaneous detection of 15 antibiotic growth promoters in bovine muscle, blood and urine by UPLC-MS/MS.

PubMed

Wang, Zhi; Shi, Zongwei; Xi, Cunxian; Wang, Guomin; Cao, Shurui; Zhang, Lei; Tang, Bobin; Mu, Zhaode

2017-12-01

An analytical method was established for the rapid detection of antibiotic growth promoters (AGPs) in bovine muscle, and bovine blood and bovine urine, using ultra high performance liquid chromatography tandem mass spectrometry (UPLC-MS/MS). After the addition of an aqueous solution of EDTA-Na 2 , the pH of bovine urine samples was directly adjusted to 5.2 by acetic acid-ammonium acetate and purified by HLB solid-phase extraction cartridge; bovine muscle and bovine blood samples processing were extracted with acetonitrile (ACN) and ACNwater (90:10; v/v) without any purification step. The samples were then centrifuged, concentrated and analysed by UPLC-MS/MS on an ACQUITY UPLC® BEH C18 column using gradient elution. The developed method was validated and mean recovery percentages at three spiked levels were 74-119%, 76-115% and 76-119%, respectively, in bovine muscle, bovine blood, and bovine urine. The relative standard deviation (RSD) ranged from 1.0% to 14.7% in spiked bovine muscle, bovine blood and bovine urine. The limits of detection (LOD) of all analytes were in the ranges 0.11-3.82 µg kg -1 , 0.10-2.49 µg kg -1 and 0.06-4.53 µg kg -1 in bovine muscle, bovine blood, and bovine urine, respectively. The method was sensitive, accurate and was applied to monitor real samples. To the best of our knowledge, this is first method available for simultaneous determination of several classes of APGs in bovine muscle, and bovine blood and bovine urine.

Non-invasive Serial Blood Collection in Guinea Pigs (Cavia porcellus)

DTIC Science & Technology

2007-07-01

styptic powder (Kwik-Stop® with Benzocaine , Gimborn Pet Specialties, LLC., Atlanta, GA), pre-labeled blood tubes with EDTA (K3)(Monoject® Samplette...completion of blood collection, styptic powder with a local analgesic agent (Kwik- Stop® with Benzocaine , Gimborn Pet Specialties, LLC. Atlanta, GA) was...11 Figure 13. Using Kwik-Stop with benzocaine . 13

A novel approach to the use of doxycycline-loaded biodegradable membrane and EDTA root surface etching in chronic periodontitis: a randomized clinical trial.

PubMed

Gamal, Ahmed Y; Kumper, Radi M

2012-09-01

The release profile of 25% doxycycline (DOX) gel loaded on a biodegradable collagen membrane (COL) after 24% EDTA root surface etching was evaluated. Thirty systemically healthy patients, each with at least one pair of contralateral interproximal intrabony defects ≥4 mm deep, along with an interproximal probing depth ≥6 mm and clinical attachment loss ≥4 mm, were randomized into two groups. Group 1 consisted of sites treated with open-flap debridement followed by placement of DOX gel-loaded COL (DOX-COL), whereas group 2 sites were treated with flap surgery followed by the placement of DOX-COL after EDTA etching of the exposed root surfaces (DOX-COL + EDTA). Samples of gingival crevicular fluid were obtained 1, 3, 7, 14, and 21 days after surgery. Separation was performed, and quantitative measurements of DOX were taken with a high-performance liquid chromatography. Clinical evaluation and follow-up for 6 months were performed. At 21 days, DOX-COL + EDTA group showed 5.3 μg/mL value. However, no DOX was detected in samples of the DOX-COL group. DOX-COL + EDTA-treated group retained more DOX during the periods of 3, 7, 10, and 14 days than did the DOX-COL group. EDTA root surface etching could enhance DOX availability in the gingival crevicular fluid after its release from the collagen membrane.

Evaluation of the performance of a point-of-care method for total and differential white blood cell count in clozapine users.

PubMed

Bui, H N; Bogers, J P A M; Cohen, D; Njo, T; Herruer, M H

2016-12-01

We evaluated the performance of the HemoCue WBC DIFF, a point-of-care device for total and differential white cell count, primarily to test its suitability for the mandatory white blood cell monitoring in clozapine use. Leukocyte count and 5-part differentiation was performed by the point-of-care device and by routine laboratory method in venous EDTA-blood samples from 20 clozapine users, 20 neutropenic patients, and 20 healthy volunteers. From the volunteers, also a capillary sample was drawn. Intra-assay reproducibility and drop-to-drop variation were tested. The correlation between both methods in venous samples was r > 0.95 for leukocyte, neutrophil, and lymphocyte counts. The correlation between point-of-care (capillary sample) and routine (venous sample) methods for these cells was 0.772; 0.817 and 0.798, respectively. Only for leukocyte and neutrophil counts, the intra-assay reproducibility was sufficient. The point-of-care device can be used to screen for leukocyte and neutrophil counts. Because of the relatively high measurement uncertainty and poor correlation with venous samples, we recommend to repeat the measurement with a venous sample if cell counts are in the lower reference range. In case of clozapine therapy, neutropenia can probably be excluded if high neutrophil counts are found and patients can continue their therapy. © 2016 John Wiley & Sons Ltd.

Effect of gaseous ozone on Enterococcus faecalis biofilm-an in vitro study.

PubMed

Boch, Tanja; Tennert, Christian; Vach, Kirstin; Al-Ahmad, Ali; Hellwig, Elmar; Polydorou, Olga

2016-09-01

The aim of this study was to evaluate the antimicrobial effect of gaseous ozone compared to conventional methods against Enterococcus faecalis. One hundred twenty-five teeth were infected by E. faecalis and were incubated for 72 h to form biofilm. Teeth were distributed among five groups. In the first group, ozone was used; in the second group, teeth were rinsed with 20 % ethylenediaminetetraacetic acid (EDTA); in the third group, with 3 % sodium hypochlorite (NaOCl). Group 4 combined 20 % EDTA with ozone. NaOCl and ozone were combined in group 5. After treatment, the samples with paper points were taken, followed by dentin samples taken with K-file, and cultured for 24 h. Then bacterial colonies were counted. All treatments reduced significantly (p < 0.05) the bacteria. Paper points' samples showed 85.38 % reduction after ozone. The highest reduction was observed in NaOCl group (99.98 %). EDTA reduced bacteria by 80.64 %. Combination of NaOCl and ozone eradicated 99.95 % of the bacteria. Combination of EDTA and ozone reduced E. faecalis up to 91.33 %. The dentin chips showed the following: the highest CFU counts were observed in EDTA group, followed by ozone and NaOCl group. The lowest CFU counts were found in NaOCl-ozone group and EDTA-ozone group. Ozone reduced E. faecalis, even organised in a biofilm, however, lower than NaOCl. No treatment reduced totally the bacteria. Used as an adjuvant, ozone can increase the efficacy of conventional rinsing like EDTA and presents an alternative treatment when NaOCl cannot be used e.g. in teeth with a wide-open apical foramen.

Soft plasma electrolysis with complex ions for optimizing electrochemical performance

NASA Astrophysics Data System (ADS)

Kamil, Muhammad Prisla; Kaseem, Mosab; Ko, Young Gun

2017-03-01

Plasma electrolytic oxidation (PEO) was a promising surface treatment for light metals to tailor an oxide layer with excellent properties. However, porous coating structure was generally exhibited due to excessive plasma discharges, restraining its performance. The present work utilized ethylenediaminetetraacetic acid (EDTA) and Cu-EDTA complexing agents as electrolyte additives that alter the plasma discharges to improve the electrochemical properties of Al-1.1Mg alloy coated by PEO. To achieve this purpose, PEO coatings were fabricated under an alternating current in silicate electrolytes containing EDTA and Cu-EDTA. EDTA complexes were found to modify the plasma discharging behaviour during PEO that led to a lower porosity than that without additives. This was attributed to a more homogeneous electrical field throughout the PEO process while the coating growth would be maintained by an excess of dissolved Al due to the EDTA complexes. When Cu-EDTA was used, the number of discharge channels in the coating layer was lower than that with EDTA due to the incorporation of Cu2O and CuO altering the dielectric behaviour. Accordingly, the sample in the electrolyte containing Cu-EDTA constituted superior corrosion resistance to that with EDTA. The electrochemical mechanism for excellent corrosion protection was elucidated in the context of equivalent circuit model.

Soft plasma electrolysis with complex ions for optimizing electrochemical performance

PubMed Central

Kamil, Muhammad Prisla; Kaseem, Mosab; Ko, Young Gun

2017-01-01

Plasma electrolytic oxidation (PEO) was a promising surface treatment for light metals to tailor an oxide layer with excellent properties. However, porous coating structure was generally exhibited due to excessive plasma discharges, restraining its performance. The present work utilized ethylenediaminetetraacetic acid (EDTA) and Cu-EDTA complexing agents as electrolyte additives that alter the plasma discharges to improve the electrochemical properties of Al-1.1Mg alloy coated by PEO. To achieve this purpose, PEO coatings were fabricated under an alternating current in silicate electrolytes containing EDTA and Cu-EDTA. EDTA complexes were found to modify the plasma discharging behaviour during PEO that led to a lower porosity than that without additives. This was attributed to a more homogeneous electrical field throughout the PEO process while the coating growth would be maintained by an excess of dissolved Al due to the EDTA complexes. When Cu-EDTA was used, the number of discharge channels in the coating layer was lower than that with EDTA due to the incorporation of Cu2O and CuO altering the dielectric behaviour. Accordingly, the sample in the electrolyte containing Cu-EDTA constituted superior corrosion resistance to that with EDTA. The electrochemical mechanism for excellent corrosion protection was elucidated in the context of equivalent circuit model. PMID:28281672

The influence of four different anticoagulants on dynamic light scattering of platelets.

PubMed

Raczat, T; Kraemer, L; Gall, C; Weiss, D R; Eckstein, R; Ringwald, J

2014-08-01

For testing of dynamic light scattering of platelets with ThromboLUX (TLX) in platelet-rich plasma (PRP) derived from venous whole blood (vWB), anticoagulation is needed. We compared TLX score in PRPs containing citrate, ethylene-diamine-tetraacetic-acid (EDTA), citrate-phosphate-dextrose-adenine (CPDA) or citrate-theophylline-adenosine-dipyridamole. Initial and late TLX scores were measured after 30-120 min or four to six hours, respectively. Compared with citrate, mean differences in initial TLX score were only significant for CPDA. Also, mean differences between initial and late TLX scores were only significant for CPDA. TLX failed to detect EDTA-induced platelet alterations. The clinical relevance of TLX needs further studies. © 2014 International Society of Blood Transfusion.

CaNa2EDTA chelation attenuates cell damage in workers exposed to lead--a pilot study.

PubMed

Čabarkapa, A; Borozan, S; Živković, L; Stojanović, S; Milanović-Čabarkapa, M; Bajić, V; Spremo-Potparević, B

2015-12-05

Lead induced oxidative cellular damage and long-term persistence of associated adverse effects increases risk of late-onset diseases. CaNa2EDTA chelation is known to remove contaminating metals and to reduce free radical production. The objective was to investigate the impact of chelation therapy on modulation of lead induced cellular damage, restoration of altered enzyme activities and lipid homeostasis in peripheral blood of workers exposed to lead, by comparing the selected biomarkers obtained prior and after five-day CaNa2EDTA chelation intervention. The group of smelting factory workers diagnosed with lead intoxication and current lead exposure 5.8 ± 1.2 years were administered five-day CaNa2EDTA chelation. Elevated baseline activity of antioxidant enzymes Cu, Zn-SOD and CAT as well as depleted thiols and increased protein degradation products-carbonyl groups and nitrites, pointing to Pb induced oxidative damage, were restored toward normal values following the treatment. Lead showed inhibitor potency on both RBC AChE and BChE in exposed workers, and chelation re-established the activity of BChE, while RBC AChE remained unaffected. Also, genotoxic effect of lead detected in peripheral blood lymphocytes was significantly decreased after therapy, exhibiting 18.9% DNA damage reduction. Administration of chelation reversed the depressed activity of serum PON 1 and significantly decreased lipid peroxidation detected by the post-chelation reduction of MDA levels. Lactate dehydrogenase LDH1-5 isoenzymes levels showed evident but no significant trend of restoring toward normal control values following chelation. CaNa2EDTA chelation ameliorates the alterations linked with Pb mediated oxidative stress, indicating possible benefits in reducing health risks associated with increased oxidative damage in lead exposed populations. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Isolation of human genomic DNA for genetic analysis from premature neonates: a comparison between newborn dried blood spots, whole blood and umbilical cord tissue

PubMed Central

2013-01-01

Background Genotyping requires biological sample collection that must be reliable, convenient and acceptable for patients and clinicians. Finding the most optimal procedure of sample collection for premature neonates who have a very limited blood volume is a particular challenge. The aim of the current study was to evaluate the use of umbilical cord (UC) tissue and newborn dried blood spot (DBS)-extracted genomic DNA (gDNA) as an alternative to venous blood-derived gDNA from premature neonates for molecular genetic analysis. All samples were obtained from premature newborn infants between 24-32 weeks of gestation. Paired blood and UC samples were collected from 31 study participants. gDNA was extracted from ethylenediaminetetraacetic acid (EDTA) anticoagulant-treated blood samples (~500 μl) and newborn DBSs (n = 723) using QIAamp DNA Micro kit (Qiagen Ltd., Crawley, UK); and from UC using Qiagen DNAeasy Blood and Tissue kit (Qiagen Ltd., Crawley, UK). gDNA was quantified and purity confirmed by measuring the A260:A280 ratio. PCR amplification and pyrosequencing was carried out to determine suitability of the gDNA for molecular genetic analysis. Minor allele frequency of two unrelated single nucleotide polymorphisms (SNPs) was calculated using the entire cohort. Results Both whole blood samples and UC tissue provided good quality and yield of gDNA, which was considerably less from newborn DBS. The gDNA purity was also reduced after 3 years of storage of the newborn DBS. PCR amplification of three unrelated genes resulted in clear products in all whole blood and UC samples and 86%-100% of newborn DBS. Genotyping using pyrosequencing showed 100% concordance in the paired UC and whole blood samples. Minor allele frequencies of the two SNPs indicated that no maternal gDNA contamination occurred in the genotyping of the UC samples. Conclusions gDNAs from all three sources are suitable for standard PCR and pyrosequencing assays. Given that UC provide good quality and quantity gDNA with 100% concordance in the genetic analysis with whole blood, it can replace blood sampling from premature infants. This is likely to reduce the stress and potential side effects associated with invasive sample collection and thus, greatly facilitate participant recruitment for genetic studies. PMID:24168095

Infrared Spectroscopy Determination of Lead Binding to Ethylenediaminotetraacetic Acid

NASA Astrophysics Data System (ADS)

Fitch, Alanah; Dragan, Simona

1998-08-01

In an attempt to improve a thematic lab sequence based on lead analysis of community derived samples, we have considered infrared spectroscopy as a method of determining the lead bound to ethylenediaminotetraacetic acid (EDTA). Students get acquainted with infrared spectroscopy by interpreting the spectra of EDTA, disodium ethylenediaminotetraacetate (Na2EDTA) and of lead(II) ethylenediaminotetraacetate (PbEDTA). Spectral characterization of the above compounds in the 1800-1500 cm-1 region gives information about the structural changes that sodium and lead binding to EDTA, respectively, produce. The spectra show the carboxylic carbonyl absorption band shifted from 1697 cm-1 to 1633 cm-1 in Na2EDTA, and two distinctive absorption bands at 1697 cm-1 and 1558 cm-1 in PbEDTA, the former being attributed to the "free" carboxylic group, as in EDTA, and the latter to the coordinated carboxylate with the bond order of less than 1.5 between the carbon and oxygen atoms. These features led us to the conclusion that the divalent Pb is tetra-coordinated having two covalent metal-nitrogen bonds and two ionic metal-carboxylate bonds. Based on the spectral differences between PbEDTA and EDTA, we have developed a method to quantitate the amount of lead bound to EDTA by simply comparing the peak height of the most prominent peaks in the 1800-1550 cm-1 region. A potential application of this method could be determination of lead extracted by binding it to ethylenediaminotetraacetic acid, excess EDTA being added.

Single-sample 99mTc-diethylenetriamine penta-acetate plasma clearance in advanced renal failure by the mean sojourn time approach.

PubMed

Gref, Margareta C; Karp, Kjell H

2009-03-01

The single-sample Tc-diethylenetriamine penta-acetate (DTPA) clearance method by Christensen and Groth is recommended by the Radionuclides in Nephrourology Committee on Renal Clearance for use in adults with an estimated glomerular filtration rate (GFR) > or = 30 ml/min. The purpose of this study was to test a new Tc-DTPA single-sample low clearance formula for GFR lesser than 30 ml/min. Twenty-one adult patients (29 investigations) were included. Reference clearance was calculated with both Cr-EDTA and Tc-DTPA according to Brøchner-Mortensen with samples drawn between 3 and 24 h. Single-sample clearance was calculated from a 24 h sample using the low clearance formula(Equation is included in full-text article.) C(t) is the activity of the tracer in the plasma sample t minutes after the injection and Q0 is the injected amount. ECV is the extracellular volume in ml defined as the distribution volume of the tracer. ECV is estimated from the body surface area as ECV=8116.6xbody surface area-28.2. The mean difference between reference and Tc-DTPA single-sample clearance was -0.5 ml/min (SD 1.0 ml/min) for Tc-DTPA and -0.8 ml/min (SD 1.2 ml/min) for Cr-EDTA as reference clearance. In adult patients it is possible, even with GFR lesser than 30 ml/min, to get an accurate determination of Tc-DTPA plasma clearance from a single sample using the mean sojourn time approach. The blood sample should be obtained about 24 h after injection of the GFR tracer.

Effect of sample preparation techniques for carbon and nitrogen stable isotope analysis of hydroxyapatite structures in the form of elasmobranch vertebral centra.

PubMed

Christiansen, Heather M; Hussey, Nigel E; Wintner, Sabine P; Cliff, Geremy; Dudley, Sheldon F J; Fisk, Aaron T

2014-03-15

Bulk stable isotope analysis (SIA) provides an important tool for the study of animal ecology. Elasmobranch vertebral centra can be serially sampled to obtain an isotopic history of an individual over ontogeny. The measured total δ(13)C value, however, may be misinterpreted due to the inclusion of the (13)C-rich inorganic portion. Hydrochloric acid (HCl) is commonly used to remove the inorganic portion of hydroxyapatite structures before undertaking SIA, but more recently ethylenediaminetetraacetic acid (EDTA) has been recommended for elasmobranch vertebrae. These acid treatments may introduce uncertainty on measured δ(13)C and δ(15)N values above instrument precision and the effect of small sample size remains untested for elasmobranch vertebrae. Using a non-dilution program on an isotope ratio mass spectrometer the minimum sample weight of vertebrae required to obtain accurate isotopic values was determined for three shark species: white (Carcharodon carcharias), tiger (Galeocerdo cuvier), and sand tiger (Carcharias taurus). To examine if acid treatment completely removes the inorganic component of the vertebrae or whether the technique introduces its own uncertainty on measured δ(13)C and δ(15)N values, vertebrae samples were analyzed untreated and following EDTA treatment. The minimum sample weight required for accurate stable isotope values and the percentage sample yield following EDTA treatment varied within and among species. After EDTA treatment, white shark vertebrae were all enriched in (13)C and depleted in (15) N, tiger shark vertebrae showed both enrichment and depletion of (13)C and (15)N, and sand tiger shark vertebrae were all depleted in (13)C and (15)N. EDTA treatment of elasmobranch vertebrae produces unpredictable effects (i.e. non-linear and non-correctable) among species in both the percentage sample yield and the measured δ(13)C and δ(15)N values. Prior to initiating a large-scale study, we strongly recommend investigating (i) the minimum weight of vertebral material required to obtain consistent isotopic values and (ii) the effects of EDTA treatment, specific to the study species and the isotope ratio mass spectrometer employed. Copyright © 2014 John Wiley & Sons, Ltd.

Influence of transport and time on blood variables commonly measured for the athlete biological passport.

PubMed

Robinson, Neil; Giraud, Sylvain; Schumacher, Yorck Olaf; Saugy, Martial

2016-02-01

Some recent studies have characterized the stability of blood variables commonly measured for the Athlete Biological Passport. The aim of this study was to characterize the impact of different shipments conditions and the quality of the results returned by the haematological analyzer. Twenty-two healthy male subjects provided five EDTA tubes each. Four shipment conditions (24, 36, 48, 72 h) under refrigerated conditions were tested and compared to a set of samples left in the laboratory also under refrigerated conditions (group control). All measurements were conducted using two Sysmex XT-2000i analyzers. Haemoglobin concentration, reticulocytes percentage, and OFF-score numerical data were the same for samples analyzed just after collection and after a shipment under refrigerated conditions up to 72 h. Detailed information reported especially by the differential (DIFF) channel scatterplot of the Sysmex XT-2000i indicated that there were signs of blood deterioration, but were not of relevance for the variables used in the Athlete Biological Passport. As long as the cold chain is guaranteed, the time delay between the collection and the analyses of blood variables can be extended. Copyright © 2015 John Wiley & Sons, Ltd.

A PARASITOLOGIC AND MOLECULAR SURVEY OF HEPATOZOON CANIS INFECTION IN STRAY DOGS IN NORTHEAST OF IRAN.

PubMed

Barati, Ali; Razmi, Gholamreza

2018-05-15

Canine hepatozoonosis, caused by H. canis, is a tick-borne disease in domestic and wild dogs that is transmitted by ingestion of Rhipicephalus sanguineus ticks. The aim of the study was to detect H. canis in stray dogs in Iran using blood smear examination and molecular techniques. From October 2014 to September 2015, 150 EDTA blood samples were collected from stray dogs in the northeast region of Iran. Blood smears were microscopically examined for the presence of Hepatozoon gamonts; whole blood was evaluated by PCR, with subsequent sequencing and phylogenetic analysis. Hepatozoon spp. Gamonts were observed in the neutrophils of 5/150 (3.3%) blood smears, whereas Hepatozoon spp. 18S rDNA was detected in 12/150 (8.0%) blood samples from stray dogs. There was a good agreement between microscopy and PCR methods. (Kappa= 0.756). The highest rate of infection was seasonally detected in the summer (p<0.05). The difference of frequency of Hepatozoon spp infection was not significant by gender and age factors (p>0.05). The alignment analysis of the sequenced samples showed ≥99% similarity with other nucleotide sequences of Hepatozoon spp. in GenBank. The phylogenetic tree also revealed that the nucleotide sequences in this study were clustered in the H. canis clade and different from the H. felis and H. americanum clades. According to the results, it is concluded that H. canis infection is present among dogs in northeastern region of Iran.

Infection of Domestic Dogs in Peru by Zoonotic Bartonella Species: A Cross-Sectional Prevalence Study of 219 Asymptomatic Dogs

PubMed Central

Diniz, Pedro Paulo V. P.; Morton, Bridget A.; Tngrian, Maryam; Kachani, Malika; Barrón, Eduardo A.; Gavidia, Cesar M.; Gilman, Robert H.; Angulo, Noelia P.; Brenner, Elliott C.; Lerner, Richard; Chomel, Bruno B.

2013-01-01

Bartonella species are emerging infectious organisms transmitted by arthropods capable of causing long-lasting infection in mammalian hosts. Among over 30 species described from four continents to date, 15 are known to infect humans, with eight of these capable of infecting dogs as well. B. bacilliformis is the only species described infecting humans in Peru; however, several other Bartonella species were detected in small mammals, bats, ticks, and fleas in that country. The objective of this study was to determine the serological and/or molecular prevalence of Bartonella species in asymptomatic dogs in Peru in order to indirectly evaluate the potential for human exposure to zoonotic Bartonella species. A convenient sample of 219 healthy dogs was obtained from five cities and three villages in Peru. EDTA-blood samples were collected from 205 dogs, whereas serum samples were available from 108 dogs. The EDTA-blood samples were screened by PCR followed by nucleotide sequencing for species identification. Antibodies against B. vinsonii berkhoffii and B. rochalimae were detected by IFA (cut-off of 1∶64). Bartonella DNA was detected in 21 of the 205 dogs (10%). Fifteen dogs were infected with B. rochalimae, while six dogs were infected with B. v. berkhoffii genotype III. Seropositivity for B. rochalimae was detected in 67 dogs (62%), and for B. v. berkhoffii in 43 (40%) of the 108 dogs. Reciprocal titers ≥1∶256 for B. rochalimae were detected in 19% of dogs, and for B. v. berkhoffii in 6.5% of dogs. This study identifies for the first time a population of dogs exposed to or infected with zoonotic Bartonella species, suggesting that domestic dogs may be the natural reservoir of these zoonotic organisms. Since dogs are epidemiological sentinels, Peruvian humans may be exposed to infections with B. rochalimae or B. v. berkhoffii. PMID:24040427

Dried Blood Spots Combined With Ultra-High-Performance Liquid Chromatography-Mass Spectrometry for the Quantification of the Antipsychotics Risperidone, Aripiprazole, Pipamperone, and Their Major Metabolites.

PubMed

Tron, Camille; Kloosterboer, Sanne M; van der Nagel, Bart C H; Wijma, Rixt A; Dierckx, Bram; Dieleman, Gwen C; van Gelder, Teun; Koch, Birgit C P

2017-08-01

Risperidone, aripiprazole, and pipamperone are antipsychotic drugs frequently prescribed for the treatment of comorbid behavioral problems in children with autism spectrum disorders. Therapeutic drug monitoring (TDM) could be useful to decrease side effects and to improve patient outcome. Dried blood spot (DBS) sample collection seems to be an attractive technique to develop TDM of these drugs in a pediatric population. The aim of this work was to develop and validate a DBS assay suitable for TDM and home sampling. Risperidone, 9-OH risperidone, aripiprazole, dehydroaripiprazole, and pipamperone were extracted from DBS and analyzed by ultra-high-performance liquid chromatography-tandem mass spectrometry using a C18 reversed-phase column with a mobile phase consisting of ammonium acetate/formic acid in water or methanol. The suitability of DBS for TDM was assessed by studying the influence of specific parameters: extraction solution, EDTA carryover, hematocrit, punching location, spot volume, and hemolysis. The assay was validated with respect to conventional guidelines for bioanalytical methods. The method was linear, specific without any critical matrix effect, and with a mean recovery around 90%. Accuracy and imprecision were within the acceptance criteria in samples with hematocrit values from 30% to 45%. EDTA or hemolysis did not skew the results, and no punching carryover was observed. No significant influence of the spot volume or the punch location was observed. The antipsychotics were all stable in DBS stored 10 days at room temperature and 1 month at 4 or -80°C. The method was successfully applied to quantify the 3 antipsychotics and their metabolites in patient samples. A UHPLC-MS/MS method has been successfully validated for the simultaneous quantification of risperidone, 9-OH risperidone, aripiprazole, dehydroaripiprazole, and pipamperone in DBS. The assay provided good analytical performances for TDM and clinical research applications.

Ion-pairing HPLC methods to determine EDTA and DTPA in small molecule and biological pharmaceutical formulations.

PubMed

Wang, George; Tomasella, Frank P

2016-06-01

Ion-pairing high-performance liquid chromatography-ultraviolet (HPLC-UV) methods were developed to determine two commonly used chelating agents, ethylenediaminetetraacetic acid (EDTA) in Abilify® (a small molecule drug with aripiprazole as the active pharmaceutical ingredient) oral solution and diethylenetriaminepentaacetic acid (DTPA) in Yervoy® (a monoclonal antibody drug with ipilimumab as the active pharmaceutical ingredient) intravenous formulation. Since the analytes, EDTA and DTPA, do not contain chromophores, transition metal ions (Cu 2+ , Fe 3+ ) which generate highly stable metallocomplexes with the chelating agents were added into the sample preparation to enhance UV detection. The use of metallocomplexes with ion-pairing chromatography provides the ability to achieve the desired sensitivity and selectivity in the development of the method. Specifically, the sample preparation involving metallocomplex formation allowed sensitive UV detection. Copper was utilized for the determination of EDTA and iron was utilized for the determination of DTPA. In the case of EDTA, a gradient mobile phase separated the components of the formulation from the analyte. In the method for DTPA, the active drug substance, ipilimumab, was eluted in the void. In addition, the optimization of the concentration of the ion-pairing reagent was discussed as a means of enhancing the retention of the aminopolycarboxylic acids (APCAs) including EDTA and DTPA and the specificity of the method. The analytical method development was designed based on the chromatographic properties of the analytes, the nature of the sample matrix and the intended purpose of the method. Validation data were presented for the two methods. Finally, both methods were successfully utilized in determining the fate of the chelates.

Ustur whole body case 0269: demonstrating effectiveness of i.v. CA-DTPA for Pu.

PubMed

James, A C; Sasser, L B; Stuit, D B; Glover, S E; Carbaugh, E H

2007-01-01

This whole body donation case (USTUR Registrant) involved a single acute inhalation of an acidic Pu(NO3)4 solution in the form of an aerosol 'mist'. Chelation treatment with intravenously (i.v.) Ca-EDTA was initiated on the day of the intake, and continued intermittently over 6 months. After 2.5 y with no further treatment, a course of i.v. Ca-DTPA was administered. A total of 400 measurements of 239+240Pu excreted in urine were recorded; starting on the first day (both before and during the initial Ca-EDTA chelation) and continuing for 37 y. This sampling included all intervals of chelation. In addition, 91 measurements of 239+240Pu-in-feces were recorded over this whole period. The Registrant died about 38 y after the intake, at age 79 y, with extensive carcinomatosis secondary to adenocarcinoma of the prostate gland. At autopsy, all major soft tissue organs were harvested for radiochemical analyses of their 238Pu, 239+240Pu and 241Am content. Also, all types of bone (comprising about half the skeleton) were harvested for radiochemical analyses, as well as samples of skin, subcutaneous fat and muscle. This comprehensive data set has been applied to derive 'chelation-enhanced' transfer rates in the ICRP Publication 67 plutonium biokinetic model, representing the behaviour of blood-borne and tissue-incorporated plutonium during intervals of therapy. The resulting model of the separate effects of i.v. Ca-EDTA and Ca-DTPA chelation shows that the therapy administered in this case succeeded in reducing substantially the long-term burden of plutonium in all body organs, except for the lungs. The calculated reductions in organ content at the time of death are approximately 40% for the liver, 60% for other soft tissues (muscle, skin, glands, etc.), 50% for the kidneys and 50% for the skeleton. Essentially, all of the substantial reduction in skeletal burden occurred in trabecular bone. This modelling exercise demonstrated that 3-y-delayed Ca-DTPA therapy was as effective as promptly administered Ca-EDTA.

USTUR WHOLE BODY CASE 0269: DEMONSTRATING EFFECTIVENESS OF I.V. CA-DTPA FOR PU

DOE Office of Scientific and Technical Information (OSTI.GOV)

James, Anthony C.; Sasser , Lyle B.; Stuit, Dorothy B.

2008-01-28

This whole body donation case (USTUR Registrant) involved a single acute inhalation of an acidic Pu(NO3)4 solution in the form of an aerosol ‘mist.’ Chelation treatment with i.v. Ca-EDTA was initiated on the day of the intake, and continued intermittently over 6 months. After 2½ years with no further treatment, a course of i.v. Ca-DTPA was administered. A total of 400 measurements of 239+240Pu excreted in urine were recorded; starting on the first day (both before and during the initial Ca-EDTA chelation), and continuing for 37 years. This sampling included all intervals of chelation. In addition, 91 measurements of 239+240Pu-in-fecesmore » were recorded over this whole period. The Registrant died about 38 years after the intake, at age 79 y, with extensive carcinomatosis secondary to adenocarcinoma of the prostate gland. At autopsy, all major soft tissue organs were harvested for radiochemical analyses of their 238Pu, 239+240Pu and 241Am content. Also, all types of bone (comprising about half the skeleton) were harvested for radiochemical analyses, as well as samples of skin, subcutaneous fat and muscle. This comprehensive dataset has been applied to derive ‘chelation-enhanced’ transfer rates in the ICRP Publication 67 plutonium biokinetic model, representing the behaviour of blood-borne and tissue-incorporated plutonium during intervals of therapy. The resulting model of the separate effects of i.v. Ca-EDTA and Ca-DTPA chelation shows that the therapy administered in this case succeeded in reducing substantially the long-term burden of plutonium in all body organs, except for the lungs. The calculated reductions in organ content at the time of death are approximately 40% for the liver, 60% for other soft tissues (muscle, skin, glands, etc.), 50% for the kidneys, and 50% for the skeleton. Essentially all of the substantial reduction in skeletal burden occurred in trabecular bone. This modeling exercise demonstrated that 3-y-delayed Ca-DTPA therapy was as effective as promptly administered Ca-EDTA.« less

EDTA assisted synthesis of hydroxyapatite nanoparticles for electrochemical sensing of uric acid.

PubMed

Kanchana, P; Sekar, C

2014-09-01

Hydroxyapatite nanoparticles have been synthesized using EDTA as organic modifier by a simple microwave irradiation method and its application for the selective determination of uric acid (UA) has been demonstrated. Electrochemical behavior of uric acid at HA nanoparticle modified glassy carbon electrode (E-HA/GCE) has been investigated by electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), linear sweep voltammetry (LSV) and amperometry. The E-HA modified electrode exhibits efficient electrochemical activity towards uric acid sensing without requiring enzyme or electron mediator. Amperometry studies revealed that the fabricated electrode has excellent sensitivity for uric acid with the lowest detection limit of 142 nM over a wide concentration range from 1 × 10(-7) to 3 × 10(-5)M. Moreover, the studied E-HA modified GC electrode exhibits a good reproducibility and long-term stability and an admirable selectivity towards the determination of UA even in the presence of potential interferents. The analytical performance of this sensor was evaluated for the detection of uric acid in human urine and blood serum samples. Copyright © 2014. Published by Elsevier B.V.

[Heavy metals removal and its kinetics in contaminated soil under effects of EDTA washing].

PubMed

Ke, Xin; Li, Pei-Jun; Zhang, Yun; Sun, Tie-Heng

2007-03-01

In this paper, batch experiments were made to examine the effects of different EDTA concentrations, pH, and washing duration of EDTA on the removal of heavy metals from contaminated soil. An empirical model was employed to describe the kinetics of heavy metals dissolution and desorption, and the form changes of test heavy metals were determined before and after EDTA washing. The results showed that EDTA was effective to the removal of heavy-metals from contaminated soil. At 0.1 mol x L(-1) and pH 7 of EDTA and within 24 h, the removal rate of test heavy metals was the maximum, being 89.14% for Cd, 34.78% for Pb, 14.96% for Cu, and 45.14% for Zn. The mass transfer coefficient was in the order of Cd > Zn > Pb > Cu. Sequential fractionations of treated and untreated soil samples showed that EDTA was effective in removing the exchangeable, carbonate and oxide forms of Cd, Pb, Zn and Cu, but ineffective for the organic and residual forms of test heavy metals.

Biomonitoring of 29 trace elements in whole blood from inhabitants of Cotonou (Benin) by ICP-MS.

PubMed

Yedomon, Brice; Menudier, Alain; Etangs, Florence Lecavelier Des; Anani, Ludovic; Fayomi, Benjamin; Druet-Cabanac, Michel; Moesch, Christian

2017-09-01

This study aimed to investigate the blood concentration of 29 trace elements, metals or metalloids, in a healthy population of Cotonou not directly exposed to metals in order to propose reference values. Blood samples from 70 blood donors were collected in K2 EDTA tubes for trace elements during September 2015 and a questionnaire was used to assess lifestyle exposure. Blood metal concentrations were determined by inductively coupled plasma mass spectrometry (ICP-MS) equipped with a quadrupole-based reaction cell. Among the selected blood donors 51.4% were aged from 18 to 36 years and 49.6% from 37 to 65 years. Among the 29 elements analyzed As, Pb, Mn, Pd, Sb, Co, Se, Sr showed blood concentrations higher than the reference values found in the literature for non-exposed healthy European populations and their geometric means were respectively 5.81; 47.39; 19.71; 1.91; 7.50; 0.66; 163.01; 30.53μg/L. This study provides the first reference value (5th-95th percentiles) for each element in Cotonou, which enables us to carry out further investigations on environmental and occupational exposure. Copyright © 2016 Elsevier GmbH. All rights reserved.

The comparison between two irrigation regimens on the dentine wettability for an epoxy resin based sealer by measuring its contact angle formed to the irrigated dentine.

PubMed

Mohan, Rayapudi Phani; Pai, Annappa Raghavendra Vivekananda

2015-01-01

The aim was to assess the influence of two irrigation regimens having ethylenediaminetetraacetic acid (EDTA) and ethylenediaminetetraacetic acid with cetrimide (EDTAC) as final irrigants, respectively, on the dentine wettability for AH Plus sealer by comparing its contact angle formed to the irrigated dentine. Study samples were divided into two groups (n = 10). The groups were irrigated with 3% sodium hypochlorite (NaOCl) solution followed by either 17% EDTA or 17% EDTAC solution. AH Plus was mixed, and controlled volume droplet (0.1 mL) of the sealer was placed on the dried samples. The contact angle was measured using a Dynamic Contact Angle Analyzer and results were analyzed using SPSS 21.0 and 2 sample t-test. There was a significant difference in the contact angle of AH Plus formed to the dentine irrigated with the above two regimens. AH Plus showed significantly lower contact angle with the regimen having EDTAC as a final irrigant than the one with EDTA (P < 0.05). An irrigation regimen consisting of NaOCl with either EDTA or EDTAC solution as a final irrigant influences the dentine wettability and contact angle of a sealer. EDTAC as a final irrigant facilitates better dentin wettability than EDTA for AH Plus to promote its better flow and adhesion.

Effects of delayed laboratory processing on platelet serotonin levels.

PubMed

Sanner, Jennifer E; Frazier, Lorraine; Udtha, Malini

2013-01-01

Despite the availability of established guidelines for measuring platelet serotonin, these guidelines may be difficult to follow in a hospital setting where time to processing may vary from sample to sample. The purpose of this study was to evaluate the effect of the time to processing of human blood samples on the stability of the enzyme-linked immunosorbent assay (ELISA) for the determination of platelet serotonin levels in human plasma. Human blood samples collected from a convenience sample of eight healthy volunteers were analyzed to determine platelet serotonin levels from plasma collected in ethylene diamine tetra acetic acid (EDTA) tubes and stored at 4°C for 3 hr, 5 hr, 8 hr, and 12 hr. Refrigeration storage at 4°C for 3 hr, 5 hr, 8 hr, and 12 hr altered the platelet serotonin measurement when compared to immediate processing. The bias for the samples stored at 4°C for 3 hr was 102.3 (±217.39 ng/10(9) platelets), for 5 hr was 200.1 (±132.76 ng/10(9) platelets), for 8 hr was 146.9 (±221.41 ng/10(9) platelets), and for 12 hr was -67.6 (±349.60 ng/10(9) platelets). Results from this study show that accurate measurement of platelet serotonin levels is dependent on time to processing. Researchers should therefore follow a standardized laboratory guideline for obtaining immediate platelet serotonin levels after blood sample collection.

Effect of EDTA Conditioning and Carbodiimide Pretreatment on the Bonding Performance of All-in-One Self-Etch Adhesives

PubMed Central

Singh, Shipra; Nagpal, Rajni; Tyagi, Shashi Prabha; Manuja, Naveen

2015-01-01

Objective. This study evaluated the effect of ethylenediaminetetraacetic acid (EDTA) conditioning and carbodiimide (EDC) pretreatment on the shear bond strength of two all-in-one self-etch adhesives to dentin. Methods. Flat coronal dentin surfaces were prepared on one hundred and sixty extracted human molars. Teeth were randomly divided into eight groups according to two different self-etch adhesives used [G-Bond and OptiBond-All-In-One] and four different surface pretreatments: (a) adhesive applied following manufacturer's instructions; (b) dentin conditioning with 24% EDTA gel prior to application of adhesive; (c) EDC pretreatment followed by application of adhesive; (d) application of EDC on EDTA conditioned dentin surface followed by application of adhesive. Composite restorations were placed in all the samples. Ten samples from each group were subjected to immediate and delayed (6-month storage in artificial saliva) shear bond strength evaluation. Data collected was subjected to statistical analysis using three-way ANOVA and post hoc Tukey's test at a significance level of p < 0.05.  Results and Conclusion. EDTA preconditioning as well as EDC pretreatment alone had no significant effect on the immediate and delayed bond strengths of either of the adhesives. However, EDC pretreatment on EDTA conditioned dentin surface resulted in preservation of resin-dentin bond strength of both adhesives with no significant fall over six months. PMID:26557850

Optimizing anti-coking abilities of zeolites by ethylene diamine tetraacetie acid modification on catalytic fast pyrolysis of corn stalk

NASA Astrophysics Data System (ADS)

Zhang, Bo; Zhong, Zhaoping; Song, Zuwei; Ding, Kuan; Chen, Paul; Ruan, Roger

2015-12-01

In order to minimize coke yield during biomass catalytic fast pyrolysis (CFP) process, ethylene diamine tetraacetie acid (EDTA) chemical modification method is carried out to selectively remove the external framework aluminum of HZSM-5 catalyst. X-ray diffraction (XRD), nitrogen (N2)-adsorption and ammonia-temperature programmed desorption (NH3-TPD) techniques are employed to investigate the porosity and acidity characteristics of original and modified HZSM-5 samples. Py-GC/MS and thermo-gravimetric analyzer (TGA) experiments are further conducted to explore the catalytic effect of modified HZSM-5 samples on biomass CFP and to verify the positive effect on coke reduction. Results show that EDTA treatment does not damage the crystal structure of HZSM-5 zeolites, but leads to a slight increase of pore volume and pore size. Meanwhile, the elimination of the strong acid peak indicates the dealumination of outer surface of HZSM-5 zeolites. Treatment time of 2 h (labeled EDTA-2H) is optimal for acid removal and hydrocarbon formation. Among all modified catalysts, EDTA-2H performs the best for deacidification and can obviously increase the yields of positive chemical compositions in pyrolysis products. Besides, EDTA modification can improve the anti-coking properties of HZSM-5 zeolites, and EDTA-2H gives rise to the lowest coke yield.

Sensory Acceptability of Iron-Fortified Red Lentil (Lens culinaris Medik.) Dal.

PubMed

Podder, Rajib; Khan, Shaan M; Tar'an, Bunyamin; Tyler, Robert T; Henry, Carol J; Jalal, Chowdhury; Shand, Phyllis J; Vandenberg, Albert

2018-03-01

Panelists in Saskatoon, Canada (n = 45) and Dhaka, Bangladesh (n = 98) participated in sensory evaluations of the sensory properties of both cooked and uncooked dehulled red lentil dal fortified with FeSO 4 ·7H 2 O, NaFeEDTA or FeSO 4 ·H 2 O at fortificant Fe concentrations of 800, 1,600 (both cooked and uncooked), or 2,800 ppm. Appearance, odor, and overall acceptability of cooked and uncooked samples were rated using a 9-point hedonic scale (1 = dislike extremely to 9 = like extremely). Taste and texture were rated for the cooked samples prepared as typical south Asian lentil meals. Significant differences in sensory quality were observed among all uncooked and cooked samples at both locations. Overall, scores for all sensory attributes and acceptability of uncooked lentil decreased with increasing concentration of Fe in the fortificant; however, Fe fortification (particularly with NaFeEDTA) had small effects on acceptability. Panelists from Saskatoon provided a wider range of scores than those from Bangladesh for all attributes of cooked lentil. Overall, sensory evaluation of Fe fortification using NaFeEDTA minimally affected consumer perception of color, taste, texture, odor, and overall acceptability of cooked lentil. Reliability estimates (Cronbach's alpha [CA]) indicated that consumer scores were generally consistent for all attributes of all lentil samples (mean CA > 0.80). NaFeEDTA was found to be the most suitable Fe fortificant for lentil based on consumer acceptability. Consumption of 45 to 50 g of NaFeEDTA-fortified lentil (fortificant Fe concentration of 1,600 ppm) per day meets the estimated average requirements (EARs) of Fe for humans (10.8 to 29.4 mg). Iron fortification of dehulled lentil dal may change organoleptic attributes that can influence consumer acceptability. Sensory evaluation by consumers helps to determine the effect on appearance, odor, taste, texture, and overall acceptability of fortified lentils. In this study, consumer acceptability was evaluated with panelists who consume lentil regularly. Panelists provided significantly different scores for 5 sensory attributes for 10 uncooked and 3 cooked lentil samples. Panelists reliably preferred NaFeEDTA as the most suitable Fe fortificant for dehulled lentils for 5 attributes. Overall, lentil dal fortified with NaFeEDTA can offer a simple and low-cost solution to human health problems associated with iron-related malnutrition. © 2018 Institute of Food Technologists®.

Variability in, variability out: best practice recommendations to standardize pre-analytical variables in the detection of circulating and tissue microRNAs.

PubMed

Khan, Jenna; Lieberman, Joshua A; Lockwood, Christina M

2017-05-01

microRNAs (miRNAs) hold promise as biomarkers for a variety of disease processes and for determining cell differentiation. These short RNA species are robust, survive harsh treatment and storage conditions and may be extracted from blood and tissue. Pre-analytical variables are critical confounders in the analysis of miRNAs: we elucidate these and identify best practices for minimizing sample variation in blood and tissue specimens. Pre-analytical variables addressed include patient-intrinsic variation, time and temperature from sample collection to storage or processing, processing methods, contamination by cells and blood components, RNA extraction method, normalization, and storage time/conditions. For circulating miRNAs, hemolysis and blood cell contamination significantly affect profiles; samples should be processed within 2 h of collection; ethylene diamine tetraacetic acid (EDTA) is preferred while heparin should be avoided; samples should be "double spun" or filtered; room temperature or 4 °C storage for up to 24 h is preferred; miRNAs are stable for at least 1 year at -20 °C or -80 °C. For tissue-based analysis, warm ischemic time should be <1 h; cold ischemic time (4 °C) <24 h; common fixative used for all specimens; formalin fix up to 72 h prior to processing; enrich for cells of interest; validate candidate biomarkers with in situ visualization. Most importantly, all specimen types should have standard and common workflows with careful documentation of relevant pre-analytical variables.

Synthesis of SrFe12O19 magnetic nanoparticles by EDTA complex method

NASA Astrophysics Data System (ADS)

Wang, Shifa; Li, Danming; Xiao, Yuhua; Dang, Wenqiang; Feng, Jie

2017-10-01

A modified polyacrylamide gel route was used to prepare SrFe12O19 magnetic nanoparticles; ethylenediaminetetraacetic acid (EDTA) was used as a carboxyl chelating agent. The phase purity, morphology and magnetic properties of as-prepared samples were analyzed via X-ray diffraction (XRD), Raman spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM) and vibrating sample magnetometery (VSM). XRD analysis indicates that high-purity SrFe12O19 magnetic nanoparticles can be synthesized at 700°C in air. The characteristic peaks of as-prepared sample at 210, 283, 321, 340, 381, 411, 432, 475, 532, 618, 686, and 726 cm-1 were observed in Raman spectra. SEM and TEM show that the synthesized SrFe12O19 magnetic nanoparticles are uniform with the mean particle size of 60 nm. VSM measurement shows that the maximum magnetic energy product (BH)max of sample prepared using EDTA as a chelating agent is higher than that of sample prepared using citric acid as a chelating agent.

Chlorhexidine controlled-release profile after EDTA root surface etching: an in vivo study.

PubMed

Gamal, Ahmed Y; Kumper, Radi M; Sadek, Hesham S; El Destawy, Mahmoud T

2011-05-01

The main objective of the present study was to quantify chlorhexidine (CHX) release after the use of CHX-EDTA root surface treatment as a local-delivery antimicrobial vehicle. Twenty non-smoking patients clinically diagnosed as having moderate-to-severe chronic periodontitis were selected to participate in this study. After cause-related therapy, one site in every patient received defect overfill with CHX gel 2% (20 sites). In addition, twenty contralateral sites received defect fill of CHX gel after 3 minutes of 24% EDTA gel root surface etching (20 sites). Gingival crevicular fluid samples were collected at 1, 3, 7, and 14 days post-therapy. The CHX-EDTA group showed statistically significantly higher levels of CHX than those of the control group at 1, 3, and 7 days. At 14 days, the CHX-EDTA group showed 0.8 mg/mL values. The use of CHX-EDTA root surface treatment as a local-delivery antimicrobial improves CHX substantivity.

Ethylenediaminetetraacetic acid induces antioxidant and anti-inflammatory activities in experimental liver fibrosis.

PubMed

González-Cuevas, J; Navarro-Partida, J; Marquez-Aguirre, A L; Bueno-Topete, M R; Beas-Zarate, C; Armendáriz-Borunda, J

2011-01-01

Experimental liver fibrosis induced by carbon tetrachloride (CCl(4)) is associated with oxidative stress, lipid peroxidation, and inflammation. This work was focused on elucidating the anti-inflammatory and antioxidant effects of ethylenediaminetetraacetic acid (EDTA) in this model of hepatotoxicity. Wistar male rats were treated with CCl(4) and EDTA (60, 120, or 240 mg/kg). Morphometric analyses were carried out in Masson's stained liver sections to determine fibrosis index. Coagulation tests prothrombin time (PT) and partial thromboplastin time (PTT) were also determined. Gene expression for transforming growth factor beta (TGF-beta1), alpha1(I) procollagen gene (alpha1 Col I), tumor necrosis factor alpha (TNF-alpha), interleukin-6 (IL-6), and superoxide dismutase (SOD) was monitored by real-time PCR. Antioxidant effect of EDTA was measured by its effects on lipid peroxidation; biological activity of ceruloplasmin (Cp), SOD, and catalase (Cat) were analyzed by zymography assays. Animals with CCl(4)-hepatic injury that received EDTA showed a decrement in fibrosis (20%) and lipid peroxidation (22%). The mRNA expression for TNF-alpha (55%), TGF-beta1 (50%), IL-6 (52%), and alpha1 Col I (60%) was also decreased. This group of animals showed increased Cp (62%) and SOD (25%) biological activities. Coagulation blood tests, Cat activity, and gene expression for SOD were not modified by EDTA treatment. This study demonstrates that EDTA treatment induces the activity of antioxidant enzymes, decreases lipid peroxidation, hepatic inflammation, and fibrosis in experimental liver fibrosis induced by CCl(4).

The effect of polymorphism in gene of insulin-like growth factor-I on the serum periparturient concentration in Holstein dairy cows.

PubMed

Mirzaei, A; Sharifiyazdi, H; Ahmadi, M R; Ararooti, T; Ghasrodashti, A Rowshan; Kadivar, A

2012-10-01

To investigate the relationship between polymorphism within the 5'-untranslated region (5'-UTR) of IGF-I gene and its periparturient concentration in Iranian Holstein dairy cows. Blood samples (5 mL, n = 37) were collected by caudal venipuncture from each animal into sample tubes containing the EDTA and DNA was extracted from blood. In order to measure IGF-I concentration the collection of blood samples (n = 111) was also done at 14 d before calving (prepartum), 25 and 45 d postpartum. We found evidence for a significant effect of C to T mutation in position 512 of IGF-I gene on its serum concentration in dairy cows in Iran. Cows with CC genotype had significantly higher concentration (Mean±SD) of IGF-I at 14 d prepartum (91.8±18.1) µg/L compared to those with TT genotype (73.3±14.4) µg/L (P=0.04). A significant trend (quadratic) was found for IGF-I concentration, as higher in CC cows compared to ones with TT genotype, during the 14 d before calving to 45 d postpartum (P=0.01). We concluded that C/T transition in the promoter region of IGF-I gene can influence the serum concentration of IGF-I in periparturient dairy cows.

Parathyroid Hormone, Calcitonin, and Vitamin D

NASA Technical Reports Server (NTRS)

Potts, J. T.

1972-01-01

Analyses of secretion of parathyroid hormone during tests of stimulation and suppression of hormone-secretory activity using infusions of EDTA and calcium, respectively, have established that, in contrast to previous views, secretion of the hormone is not autonomous in many patients that have adenomatous hyperparathyroidism, but is responsive to changes in blood-calcium concentration. These findings have led to a new understanding of the pathophysiology of hormone production in hyperparathy-roidism. A related application of the diagnostic use of the radioimmunoassay is the preoperative localization of parathyroid tumors and the distinction between adenomas and chief-cell hyperplasia. Work involving catheterization and radioimmunoassay of blood samples obtained from the subclavin and innominate veins and the venae cavae, led to localization in a high percentage of patients. However, this procedure has been adopted recently to detect hormone concentration in the small veins directly draining the parathyroid glands.

Dried blood spots for the enzymatic diagnosis of lysosomal storage diseases in dogs and cats.

PubMed

Sewell, Adrian C; Haskins, Mark E; Giger, Urs

2012-12-01

In people, lysosomal storage diseases (LSD) can be diagnosed by assaying enzyme activities in dried blood spots (DBS). The aim of this study was to evaluate the feasibility of using DBS samples from dogs and cats to measure lysosomal enzymatic activities and diagnose LSD. Drops of fresh whole blood collected in EDTA from dogs and cats with known or suspected LSD and from clinically healthy dogs and cats were placed on neonatal screening cards, dried, and mailed to the Metabolic Laboratory, University Children's Hospital, Frankfurt, Germany. Activities of selected lysosomal enzymes were measured using fluorescent substrates in a 2-mm diameter disk (~2.6 μL blood) punched from the DBS. Results were expressed as nmol substrate hydrolyzed per mL of blood per minute or hour. Reference values were established for several lysosomal enzyme activities in DBS from dogs and cats; for most enzymes, activities were higher than those published for human samples. Activities of β-glucuronidase, N-acetylglucosamine-4-sulfatase (arylsulfatase B), α-mannosidase, α-galactosidase, α-fucosidase, and hexosaminidase A were measureable in DBS from healthy cats and dogs; α-iduronidase activity was measureable only in cats. In samples from animals with LSD, markedly reduced activity of a specific enzyme was found. In contrast, in samples from cats affected with mucolipidosis II, activities of lysosomal enzymes were markedly increased. Measurement of lysosomal enzyme activities in DBS provides an inexpensive, simple, and convenient method to screen animals for suspected LSD and requires only a small sample volume. For diseases in which the relevant enzyme activity can be measured in DBS, a specific diagnosis can be made. © 2012 American Society for Veterinary Clinical Pathology.

Molecular characterization of Hepatozoon sp. in cats from São Luís Island, Maranhão, Northeastern Brazil.

PubMed

de Bortoli, Caroline P; André, Marcos R; Braga, Maria do Socorro C; Machado, Rosangela Zacarias

2011-10-01

Few molecular studies have been done concerning the molecular characterization of Hepatozoon species among domestic and wild felids. The present work aimed to characterize molecularly the presence of Hepatozoon sp. DNA in cat blood samples from São Luís Island, Maranhão state, Northeastern Brazil. EDTA-whole blood samples were collected from 200 domestic cats with outdoor and wood areas access from São Luís, Maranhão, Brazil. Each sample of extracted DNA was used as a template in PCR reactions aiming to amplify a partial sequence of 18S rRNA of Hepatozoon spp. We also performed sequence alignment to establish the identity of the parasite species infecting these animals using DNA sequences based on 18S rRNA. From 200 sampled cats, Hepatozoon DNA was only found in one animal (0.5%). The found Hepatozoon DNA showed 97% of identity with Hemobartonella felis isolates 1 and 2 from Spain. When analyzing the phylogenetic tree, the found Hepatozoon DNA was in the same clade than H. felis isolates. Our findings suggest that more than one species of Hepatozoon could infect felids in Brazil.

Comparison of digoxin concentration in plastic serum tubes with clot activator and heparinized plasma tubes.

PubMed

Dukić, Lora; Simundić, Ana-Maria; Malogorski, Davorin

2014-01-01

Sample type recommended by the manufacturer for the digoxin Abbott assay is either serum collected in glass tubes or plasma (sodium heparin, lithium heparin, citrate, EDTA or oxalate as anticoagulant) collected in plastic tubes. In our hospital samples are collected in plastic tubes. Our hypothesis was that the serum sample collected in plastic serum tube can be used interchangeably with plasma sample for measurement of digoxin concentration. Our aim was verification of plastic serum tubes for determination of digoxin concentration. Concentration of digoxin was determined simultaneously in 26 venous blood plasma (plastic Vacuette, LH Lithium heparin) and serum (plastic Vacuette, Z Serum Clot activator; both Greiner Bio-One GmbH, Kremsmünster, Austria) samples, on Abbott AxSYM analyzer using the original Abbott Digoxin III assay (Abbott, Wiesbaden, Germany). Tube comparability was assessed using the Passing Bablok regression and Bland-Altman plot. Serum and plasma digoxin concentrations are comparable. Passing Bablok intercept (0.08 [95% CI = -0.10 to 0.20]) and slope (0.99 [95% CI = 0.92 to 1.11]) showed there is no constant or proportional error. Blood samples drawn in plastic serum tubes and plastic plasma tubes can be interchangeably used for determination of digoxin concentration.

Comparison of digoxin concentration in plastic serum tubes with clot activator and heparinized plasma tubes

PubMed Central

Dukić, Lora; Šimundić, Ana-Maria; Malogorski, Davorin

2014-01-01

Introduction: Sample type recommended by the manufacturer for the digoxin Abbott assay is either serum collected in glass tubes or plasma (sodium heparin, lithium heparin, citrate, EDTA or oxalate as anticoagulant) collected in plastic tubes. In our hospital samples are collected in plastic tubes. Our hypothesis was that the serum sample collected in plastic serum tube can be used interchangeably with plasma sample for measurement of digoxin concentration. Our aim was verification of plastic serum tubes for determination of digoxin concentration. Materials and methods: Concentration of digoxin was determined simultaneously in 26 venous blood plasma (plastic Vacuette, LH Lithium heparin) and serum (plastic Vacuette, Z Serum Clot activator; both Greiner Bio-One GmbH, Kremsmünster, Austria) samples, on Abbott AxSYM analyzer using the original Abbott Digoxin III assay (Abbott, Wiesbaden, Germany). Tube comparability was assessed using the Passing Bablok regression and Bland-Altman plot. Results: Serum and plasma digoxin concentrations are comparable. Passing Bablok intercept (0.08 [95% CI = −0.10 to 0.20]) and slope (0.99 [95% CI = 0.92 to 1.11]) showed there is no constant or proportional error. Conclusion: Blood samples drawn in plastic serum tubes and plastic plasma tubes can be interchangeably used for determination of digoxin concentration. PMID:24627723

Tris-EDTA significantly enhances antibiotic efficacy against multidrug-resistant Pseudomonas aeruginosa in vitro.

PubMed

Buckley, Laura M; McEwan, Neil A; Nuttall, Tim

2013-10-01

Multidrug-resistant Pseudomonas aeruginosa commonly complicates chronic bacterial otitis in dogs. The aim of this in vitro study was to determine the effect of ethylenediaminetetraacetic acid-tromethamine (Tris-EDTA) on the minimal bactericidal concentrations (MBCs) and minimal inhibitory concentrations (MICs) of marbofloxacin and gentamicin for multidrug-resistant P. aeruginosa isolates from cases of canine otitis. Eleven isolates were identified as multidrug resistant on disc diffusion; 10 were resistant to marbofloxacin and two were resistant to gentamicin. Isolates were incubated for 90 min with each antibiotic alone and in combination with Tris-EDTA at concentrations of 0.075 μg/mL to 5 mg/mL for marbofloxacin, 0.001 μg/mL to 10 mg/mL for gentamicin and 17.8:4.7 to 0.14:0.04 mg/mL for Tris-EDTA. Positive and negative controls were included. Aliquots of each antibiotic and/or Tris-EDTA concentration were subsequently transferred to sheep blood agar to determine the MBCs, and tryptone soy broth was added to the remaining suspensions to determine the MICs. Tris-EDTA alone was bacteriostatic but not bactericidal at any concentration. The addition of Tris-EDTA significantly reduced the median MBC (from 625 to 468.8 μg/mL; P < 0.001) and MIC (from 29.3 to 2.4 μg/mL; P = 0.008) of marbofloxacin, and the median MBC (from 625 to 39.1 μg/mL) and MIC (from 19.5 to 1.2 μg/mL) of gentamicin (both P < 0.001). Tris-EDTA significantly reduced the MBCs and MICs of marbofloxacin and gentamicin for multidrug-resistant P. aeruginosa in vitro. This may be of use to clinicians managing these infections in dogs. © 2013 ESVD and ACVD.

Molecularly imprinted sensor based on Russian Matryoshka structured molecules for enhanced specific identification and double amplification in ultra-trace Tb3+ determination.

PubMed

Li, Jianping; Yang, Bin; Pan, Hongcheng; Xu, Guobao

2018-06-30

The selective and sensitive detection of rare earth elements is thought to be difficult because the concentration of those elements in the sample is commonly at a low level and they normally have severe mutual interference which is caused by homologous chemical properties. In this study, a novel molecularly imprinted polymer (MIP) sensor was fabricated for highly sensitive and selective determination of ultra-trace Tb 3+ . The Tb 3+ -ethylenediaminetetraacetic acid complex (Tb-EDTA) as the template molecule was incorporated into mono-6-mercapto-β-cyclodextrin (mono-6-SH-β-CD) to form a Russian Matryoshka (RM)-structured molecule (CD/Tb-EDTA). Titanium isopropoxide was utilized in vapor sol-gel polymerization to construct MIP membrane. Moreover, the selectivity of the RM MIP sensor was remarkably enhanced by the "triple-selectivity" recognition of EDTA-to-Tb 3+ , β-CD-to-(Tb-EDTA), and 3D cavity-to-(CD/Tb-EDTA), while the sensitivity of the MIP sensor was significantly improved by ECL signal enhancement based on double amplification, in other words, the electrochemiluminescence resonance energy transfer (ECL-RET) between the ECL donor of CD/Tb-EDTA and the ECL acceptor of Ru(bpy) 3 2+ , and the ECL enhancement by the co-reactant of CD/Tb-EDTA on Ru(bpy) 3 Cl 2 . When the imprinted cavities were occupied by Tb-EDTA during rebinding, the host-guest inclusion structured complex was formed and the ECL intensities produced by the Ru(bpy) 3 Cl 2 ECL system increased with increasing concentration of Tb-EDTA. The proposed sensor was used for quantitative analysis of Tb 3+ with concentrations ranging from 8.00 × 10 -13 mol/L to 4.00 × 10 -9 mol/L and successfully applied to detect Tb 3+ in seawater samples. The detection limit of the sensor was found to be 3.90 × 10 -13 mol/L (DL = 3δ b /K), which is lower than previously reported values. Thus, the fabricated sensor is feasible for practical applications. Copyright © 2018 Elsevier B.V. All rights reserved.

Quantitative studies of bone using (18)F-fluoride and (99m)Tc-methylene diphosphonate: evaluation of renal and whole-blood kinetics.

PubMed

Park-Holohan, S J; Blake, G M; Fogelman, I

2001-09-01

We report a study of the renal and whole-blood kinetics of (18)F-fluoride and (99m)Tc-methylene diphosphonate ((99m)Tc-MDP) and their effect on the evaluation of the skeletal kinetics of the two bone tracers. Data were obtained during an investigation of postmenopausal women taking hormone replacement therapy who were compared with untreated, age-matched controls. After intravenous injection of 18F-fluoride (1 MBq), (99m)Tc-MDP (1 MBq), (51)Cr-ethylenediaminetetraacetic acid (51Cr-EDTA) (3 MBq) and (125)I-human serum albumin ((125)I-HSA) (0.25 MBq), multiple blood samples and urine collections were taken between 0 and 4 h after injection. (51)Cr-EDTA data were used to evaluate the glomerular filtration rate (GFR) and the completeness of each timed urine collection. (125)I-HSA data were used to evaluate the plasma volume and the red cell uptake of the other three tracers. At 4 h, the cumulative urine excretions (and standard deviations, SDs) were: (99m)Tc-MDP, 58.2% (4.8%); (18)F-fluoride, 36.1% (5.7%); (51)Cr-EDTA, 81.5% (4.5%). Plots of the renal clearance of (18)F-fluoride against urine volume showed that urine flow rates greater than 5 ml.min-1 were necessary to ensure a constant renal clearance of (18)F and hence stable conditions for the evaluation of bone tracer kinetics. In contrast, a low urine flow rate had no effect on the renal kinetics of (99m)Tc-MDP. For MDP, the evaluation of skeletal kinetics requires data on protein binding so that calculations can be performed for free MDP. In the present study, protein binding of MDP was evaluated from the ratio of total (99m)Tc-MDP renal clearance to GFR based on the principle that free (99m)Tc-MDP is a GFR tracer. Between 0 and 4 h after injection, the fractional protein binding of MDP increased linearly with time, changing from 21+/-5% immediately after injection to 58+/-5% at 4 h. Although red cell uptake of (99m)Tc-MDP was negligible, for (18)F-fluoride around 30% of circulating tracer was transported in red cells. In view of the data showing the rapid transport of (18)F-fluoride across the red cell membrane, bone kinetic data for (18)F are more accurately reported as whole-blood clearance rather than plasma clearance.

Cryopreservation for delayed circulating tumor cell isolation is a valid strategy for prognostic association of circulating tumor cells in gastroesophageal cancer.

PubMed

Brungs, Daniel; Lynch, David; Luk, Alison Ws; Minaei, Elahe; Ranson, Marie; Aghmesheh, Morteza; Vine, Kara L; Carolan, Martin; Jaber, Mouhannad; de Souza, Paul; Becker, Therese M

2018-02-21

To demonstrate the feasibility of cryopreservation of peripheral blood mononuclear cells (PBMCs) for prognostic circulating tumor cell (CTC) detection in gastroesophageal cancer. Using 7.5 mL blood samples collected in EDTA tubes from patients with gastroesopheagal adenocarcinoma, CTCs were isolated by epithelial cell adhesion molecule based immunomagnetic capture using the IsoFlux platform. Paired specimens taken during the same blood draw ( n = 15) were used to compare number of CTCs isolated from fresh and cryopreserved PBMCs. Blood samples were processed within 24 h to recover the PBMC fraction, with PBMCs used for fresh analysis immediately processed for CTC isolation. Cryopreservation of PBMCs lasted from 2 wk to 25.2 mo (median 14.6 mo). CTCs isolated from pre-treatment cryopreserved PBMCs ( n = 43) were examined for associations with clinicopathological variables and survival outcomes. While there was a significant trend to a decrease in CTC numbers associated with cryopreserved specimens (mean number of CTCs 34.4 vs 51.5, P = 0.04), this was predominately in samples with a total CTC count of > 50, with low CTC count samples less affected ( P = 0.06). There was no significant association between the duration of cryopreservation and number of CTCs. In cryopreserved PBMCs from patient samples prior to treatment, a high CTC count (> 17) was associated with poorer overall survival (OS) ( n = 43, HR = 4.4, 95%CI: 1.7-11.7, P = 0.0013). In multivariate analysis, after controlling for sex, age, stage, ECOG performance status, and primary tumor location, a high CTC count remained significantly associated with a poorer OS (HR = 3.7, 95%CI: 1.2-12.4, P = 0.03). PBMC cryopreservation for delayed CTC isolation is a valid strategy to assist with sample collection, transporting and processing.

Responses of artificially reared cat fleas Ctenocephalides felis felis (Bouché, 1835) to different mammalian bloods.

PubMed

Kernif, T; Stafford, K; Coles, G C; Bitam, I; Papa, K; Chiaroni, J; Raoult, D; Parola, P

2015-06-01

The cat flea, Ctenocephalides felis felis (Bouche, 1835) (Siphonaptera: Pulicidae), which is found worldwide and which parasitizes many species of wild and domestic animal, is a vector and/or reservoir of bacteria, protozoa and helminths. To aid in the study of the physiology and behaviour of fleas and of their transmission of pathogens, it would be of value to improve the laboratory rearing of pathogen-free fleas. The conditions under which artificially reared fleas at the University of Bristol (U.K.) and the Rickettsial Diseases Institute (France) are maintained were studied, with different ratios of male to female fleas per chamber (25 : 50, 50 : 100, 100 : 100, 200 : 200). The fleas were fed with bovine, ovine, caprine, porcine or human blood containing the anticoagulants sodium citrate or EDTA. Egg production was highest when fleas were kept in chambers with a ratio of 25 males to 100 females. In addition, the use of EDTA as an anticoagulant rather than sodium citrate resulted in a large increase in the number of eggs produced per female; however, the low percentage of eggs developing through to adult fleas was lower with EDTA. The modifications described in our rearing methods will improve the rearing of cat fleas for research. © 2015 The Royal Entomological Society.

Enhancement of Phloem Exudation from Fraxinus uhdei Wenz. (Evergreen Ash) using Ethylenediaminetetraacetic Acid 1

PubMed Central

Costello, L. R.; Bassham, James A.; Calvin, Melvin

1982-01-01

Ethylenediaminetetraacetic acid (EDTA) enhanced the exudation of 14C-labeled assimilates from excised leaflets and whole plant specimens of Fraxinus uhdei Wenz. A 2 millimolar EDTA concentration was found to be most effective in promoting exudation from excised leaflets, while 10 millimolar EDTA was most effective in whole plants experiments. Exudation rate reached a maximum after 24 hours in both experiments. The continuous presence of EDTA throughout the treatment period was required for maximum exudation from excised leaflets. Stachyose, raffinose, verbascose, and sucrose were the principal compounds found to occur in exudate samples. These compounds are typically transported in sieve elements of various Fraxinus species suggesting the exudate was of phloem origin. Electron microscope studies of petiolule sieve plate pores from excised leaflets showed substantially less callose appearing after treatment with EDTA than after H2O treatment. It is suggested that EDTA enhances phloem exudation by inhibiting or reducing callose formation in sieve plate pores. The exudation enhancement technique described for whole plant specimens is suggested as a useful means of collecting phloem sap and studying translocation in woody plants. Images PMID:16662189

Leukodepletion as a Point-of-Care Method for Monitoring HIV-1 Viral Load in Whole Blood

PubMed Central

Titchmarsh, Logan; Zeh, Clement; Verpoort, Thierry; Allain, Jean-Pierre

2014-01-01

In order to limit the interference of HIV-1 cellular nucleic acids in estimating viral load (VL), the feasibility of leukodepletion of a small whole-blood (WB) volume to eliminate only leukocyte cell content was investigated, using a selection of filters. The efficacy of leukocyte filtration was evaluated by counting, CD45 quantitative PCR, and HIV-1 DNA quantification. Plasma HIV-1 was tested by real-time reverse transcription (RT)-PCR. A specific, miniaturized filter was developed and tested for leukocyte and plasma virus retention, WB sample dilution, and filtration parameters in HIV-1-spiked WB samples. This device proved effective to retain >99.9% of white blood cells in 100 μl of WB without affecting plasma VL. The Samba sample preparation chemistry was adapted to use a leukodepleted WB sample for VL monitoring using the point-of-care Samba-1 semiautomated system. The clinical performance of the assay was evaluated by testing 207 consecutive venous EDTA WB samples from HIV-1-infected patients attending a CD4 testing clinic. Most patients were on antiretroviral treatment (ART), but their VL status was unknown. Compared to the Roche Cobas AmpliPrep/Cobas TaqMan HIV-1 test, the new Samba assay had a concordance of 96.5%. The use of the Samba system with a VL test for WB might contribute to HIV-1 ART management and reduce loss-to-follow-up rates in resource-limited settings. PMID:25428162

Assessment of the body burden of chelatable lead: a model and its application to lead workers.

PubMed Central

Araki, S; Ushio, K

1982-01-01

A hypothetical model was introduced to estimate the body burden of chelatable lead from the mobilisation yield of lead by calcium disodium ethylenediamine tetra-acetate (CaEDTA). It was estimated that, on average, 14 and 19% of the body burden was mobilised into the urine during the 24 hours after an injection of 53.4 mumol (20 mg) and 107 mumol (40 mg) CaEDTA per kg bodyweight, respectively. The body burden of chelatable lead ranged from 4 mumol (0.8 mg) to 120 mumol (24.9 mg) (mean 37 mumol (7.7 mg) in lead workers with blood lead concentrations of 0.3-2.9 mumol/kg (6-60 microgram/100 g) (mean 1.4 mumol/kg (29 microgram/100 g)). There were linear relationships between blood lead concentrations and body burden of chelatable lead on a log scale. PMID:6802167

Inhibition on Candida albicans biofilm formation using divalent cation chelators (EDTA).

PubMed

Ramage, Gordon; Wickes, Brian L; López-Ribot, José L

2007-12-01

Candida albicans can readily form biofilms on both inanimate and biological surfaces. In this study we investigated a means of inhibiting biofilm formation using EDTA (Ethylenediaminetetra-acetic acid), a divalent cation chelating agent, which has been shown to affect C. albicans filamentation. Candida albicans biofilms were formed in 96-well microtitre plates. Cells were allowed to adhere for 1, 2, and 4 h at 37 degrees C, washed in PBS, and then treated with different concentrations of EDTA (0, 2.5, 25, and 250 mM). EDTA was also added to the standardized suspension prior to adding to the microtiter plate and to a preformed 24 h biofilm. All plates were then incubated at 37 degrees C for an additional 24 h to allow for biofilm formation. The extent and characteristics of biofilm formation were then microscopically assessed and with a semi-quantitative colorimetric technique based on the use of an XTT-reduction assay. Northern blot analysis of the hyphal wall protein (HWP1) expression was also monitored in planktonic and biofilm cells treated with EDTA. Microscopic analysis and colorimetric readings revealed that filamentation and biofilm formation were inhibited by EDTA in a concentration dependent manner. However, preformed biofilms were minimally affected by EDTA (maximum of 31% reduction at 250 mM). The HWP1 gene expression was reduced in EDTA-treated planktonic and biofilm samples. These results indicate that EDTA inhibits C. albicans biofilm formation are most likely through its inhibitory effect on filamentation and indicates the potential therapeutic effects of EDTA. This compound may serve a non-toxic means of preventing biofilm formation on infections with a C. albicans biofilm etiology.

Prevalence and effect of Theileria orientalis infection in homebred calves in the Gloucester region of New South Wales, Australia.

PubMed

Swilks, E; Jenkins, C; Poynting, A; Collins, D; Krebs, G L

2017-06-01

To examine the prevalence of Theileria orientalis in the Gloucester area of New South Wales and its effect on individual animals. Blood samples (EDTA and clotted blood) were collected from a total of 55 calves and their dams from 6 properties over a 16-week period. A total of 202 and 190 blood samples were collected from the calves and dams, respectively, and were examined via blood film for the presence of intraerythrocytic T. orientalis piroplasms. Packed cell volume (PCV) was measured to determine infection resulting in anaemia. The presence of antibodies against the T. orientalis major piroplasm surface protein (MPSP) was tested using ELISA. The overall prevalence of T. orientalis infection in both dams and calves from all herds examined was 95%. Mean peak parasitaemia was observed in calves between 6 and 9 weeks of age (P = 0.051), coinciding with a decline in mean PCV. Only 3 (6%) of the blood samples collected from the dams were positive for Theileria-associated antibodies and no significant relationship (P > 0.05) was found between the presence of antibodies in the dams and PCV levels in the calves. There was no evidence that passive transfer of antibodies from dams to calves protected the calves against a decline in PCV. This study confirmed a high prevalence of low-level Theileria infection, but low MPSP seroconversion rates, in dams and calves in an area where the disease has been endemic for a number of years. © 2017 Australian Veterinary Association.

Qualitative Time-of-Flight Secondary Ion Mass Spectrometry Analysis of Root Dentin Irrigated with Sodium Hypochlorite, EDTA, or Chlorhexidine.

PubMed

Kolosowski, Kamil P; Sodhi, Rana N S; Kishen, Anil; Basrani, Bettina R

2015-10-01

Sodium hypochlorite (NaOCl), chelating agents, and chlorhexidine (CHX), which are commonly used irrigants during endodontic treatment, have the potential to alter the physical and chemical properties of the dentin structure. The aim of this study was to use time-of-flight secondary ion mass spectrometry to qualitatively evaluate the chemical characteristics of dentin surface and compare it with dentin exposed to NaOCl, EDTA, or CHX. Four blocks of dentin from a root of a human maxillary molar were embedded in resin and trimmed with a microtome to expose the dentin. Samples were randomly assigned to 4 treatment groups: (1) no irrigation treatment (sample A), (2) 2.5% NaOCl (sample B), (3) 17% EDTA (sample C), and (4) 2% CHX (sample D). Dentin surfaces were analyzed by time-of-flight secondary ion mass spectrometry, which allowed characterization of dentin surface chemistry by both imaging and mass spectroscopic analysis obtained in high mass and spatial resolution modes. Sample A revealed intense peaks characteristics of hydroxyapatite in addition to Na(+), K(+), CH4N(+), CN(-), CNO(-), Mg(+), F(-), and HCO2(-) peaks. Sample B showed severely decreased CH4N(+) and increased intensity of Cl(-). Sample C lacked Ca(+) and Mg(+) and showed decreased PO2(-) and PO3(-). Sample D exhibited a distinct presence of CHX. The spectral image of sample A displayed even distribution of Na(+) and Ca(+) on a smeared surface. The surfaces of samples B and D had patent dentinal tubules, whereas sample D showed an intense CHX signal. Sample C had some patent dentinal tubules and lacked Ca(+). NaOCl removed protein components from the dentin matrix, EDTA removed calcium and magnesium ions from the dentin, and CHX formed an adsorbed layer on the dentin surface. Copyright © 2015 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

The effect of root surface conditioning on smear layer removal in periodontal regeneration (a scanning electron microscopic study)

NASA Astrophysics Data System (ADS)

Fidyawati, D.; Soeroso, Y.; Masulili, S. L. C.

2017-08-01

The role of root surface conditioning treatment on smear layer removal of human teeth is affected by periodontitis in periodontal regeneration. The objective of this study is to analyze the smear layer on root surface conditioned with 2.1% minocycline HCl ointment (Periocline), and 24% EDTA gel (Prefgel). A total of 10 human teeth indicated for extraction due to chronic periodontitis were collected and root planed. The teeth were sectioned in thirds of the cervical area, providing 30 samples that were divided into three groups - minocycline ointment treatment, 24% EDTA gel treatment, and saline as a control. The samples were examined by scanning electron microscope. No significant differences in levels of smear layer were observed between the minocycline group and the EDTA group (p=0.759). However, there were significant differences in the level of smear layer after root surface treatment in the minocycline and EDTA groups, compared with the control group (p=0.00). There was a relationship between root surface conditioning treatment and smear layer levels following root planing.

Stability of BDNF in Human Samples Stored Up to 6 Months and Correlations of Serum and EDTA-Plasma Concentrations.

PubMed

Polyakova, Maryna; Schlögl, Haiko; Sacher, Julia; Schmidt-Kassow, Maren; Kaiser, Jochen; Stumvoll, Michael; Kratzsch, Jürgen; Schroeter, Matthias L

2017-06-03

Brain-derived neurotrophic factor (BDNF), an important neural growth factor, has gained growing interest in neuroscience, but many influencing physiological and analytical aspects still remain unclear. In this study we assessed the impact of storage time at room temperature, repeated freeze/thaw cycles, and storage at -80 °C up to 6 months on serum and ethylenediaminetetraacetic acid (EDTA)-plasma BDNF. Furthermore, we assessed correlations of serum and plasma BDNF concentrations in two independent sets of samples. Coefficients of variations (CVs) for serum BDNF concentrations were significantly lower than CVs of plasma concentrations ( n = 245, p = 0.006). Mean serum and plasma concentrations at all analyzed time points remained within the acceptable change limit of the inter-assay precision as declared by the manufacturer. Serum and plasma BDNF concentrations correlated positively in both sets of samples and at all analyzed time points of the stability assessment ( r = 0.455 to r s = 0.596; p < 0.004). In summary, when considering the acceptable change limit, BDNF was stable in serum and in EDTA-plasma up to 6 months. Due to a higher reliability, we suggest favoring serum over EDTA-plasma for future experiments assessing peripheral BDNF concentrations.

Rapid Bedside Inactivation of Ebola Virus for Safe Nucleic Acid Tests.

PubMed

Rosenstierne, Maiken Worsøe; Karlberg, Helen; Bragstad, Karoline; Lindegren, Gunnel; Stoltz, Malin Lundahl; Salata, Cristiano; Kran, Anne-Marte Bakken; Dudman, Susanne Gjeruldsen; Mirazimi, Ali; Fomsgaard, Anders

2016-10-01

Rapid bedside inactivation of Ebola virus would be a solution for the safety of medical and technical staff, risk containment, sample transport, and high-throughput or rapid diagnostic testing during an outbreak. We show that the commercially available Magna Pure lysis/binding buffer used for nucleic acid extraction inactivates Ebola virus. A rapid bedside inactivation method for nucleic acid tests is obtained by simply adding Magna Pure lysis/binding buffer directly into vacuum blood collection EDTA tubes using a thin needle and syringe prior to sampling. The ready-to-use inactivation vacuum tubes are stable for more than 4 months, and Ebola virus RNA is preserved in the Magna Pure lysis/binding buffer for at least 5 weeks independent of the storage temperature. We also show that Ebola virus RNA can be manually extracted from Magna Pure lysis/binding buffer-inactivated samples using the QIAamp viral RNA minikit. We present an easy and convenient method for bedside inactivation using available blood collection vacuum tubes and reagents. We propose to use this simple method for fast, safe, and easy bedside inactivation of Ebola virus for safe transport and routine nucleic acid detection. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

All-integrated and highly sensitive paper based device with sample treatment platform for Cd2+ immunodetection in drinking/tap waters.

PubMed

López Marzo, Adaris M; Pons, Josefina; Blake, Diane A; Merkoçi, Arben

2013-04-02

Nowadays, the development of systems, devices, or methods that integrate several process steps into one multifunctional step for clinical, environmental, or industrial purposes constitutes a challenge for many ongoing research projects. Here, we present a new integrated paper based cadmium (Cd(2+)) immunosensing system in lateral flow format, which integrates the sample treatment process with the analyte detection process. The principle of Cd(2+) detection is based on competitive reaction between the cadmium-ethylenediaminetetraacetic acid-bovine serum albumin-gold nanoparticles (Cd-EDTA-BSA-AuNP) conjugate deposited on the conjugation pad strip and the Cd-EDTA complex formed in the analysis sample for the same binding sites of the 2A81G5 monoclonal antibody (mAb), specific to Cd-EDTA but not Cd(2+) free, which is immobilized onto the test line. This platform operates without any sample pretreatment step for Cd(2+) detection thanks to an extra conjugation pad that ensures Cd(2+) complexation with EDTA and interference masking through ovalbumin (OVA). The detection and quantification limits found for the device were 0.1 and 0.4 ppb, respectively, these being the lowest limits reported up to now for metal sensors based on paper. The accuracy of the device was evaluated by addition of known quantities of Cd(2+) to different drinking water samples and subsequent Cd(2+) content analysis. Sample recoveries ranged from 95 to 105% and the coefficient of variation for the intermediate precision assay was less than 10%. In addition, the results obtained here were compared with those obtained with the well-established inductively coupled plasma emission spectroscopy (ICPES) and the analysis of certificate standard samples.

Assessing matrix, interferences and comparability between the Abbott Diagnostics and the Beckman Coulter high-sensitivity cardiac troponin I assays.

PubMed

Kavsak, Peter A; Malinowski, Paul; Roy, Chantele; Clark, Lorna; Lamers, Shana

2018-03-13

Analytical evaluation of high-sensitivity cardiac troponin (hs-cTn) assays, with particular attention to imprecision, interferences and matrix effects, at normal cTn concentrations, is of utmost importance as many different clinical algorithms use concentration cutoffs <10 ng/L for decision-making. The objective for the present analytical study was to compare the new Beckman Coulter hs-cTnI assay (Access hsTnI) to Abbott's hs-cTnI assay in different matrices and for different interferences, with a focus on concentrations <10 ng/L. The limit of blank (LoB) and the limit of detection (LoD) were determined in different matrices for the Beckman hs-cTnI assay. Passing-Bablok regression and difference plots were determined for 200 matched lithium heparin and EDTA plasma samples for the Beckman assay and 200 lithium heparin samples for the Abbott assay. Both EDTA and heparin plasma samples were also evaluated for stability under refrigerated conditions, for endogenous alkaline phosphatase interference and for hemolysis and icterus. The Beckman hs-cTnI assay LoB was 0.5 ng/L with the following range of LoDs=0.8-1.2 ng/L, with EDTA plasma yielding lower concentrations as compared to lithium heparin plasma (mean difference=-14.9%; 95% CI=-16.9 to 12.9). Below 10 ng/L, lithium heparin cTnI results from the Beckman assay were on average 1.1 ng/L (95% CI=0.7 to 1.5) higher than the Abbott results, with no difference between the methods when using EDTA plasma (mean difference =-0.1 ng/L; 95% CI=-0.3 to 0.2). Low cTnI concentrations were less effected by interferences in EDTA plasma. The Access hsTnI method can reliably detect normal cTnI concentrations with both lithium heparin and EDTA plasma being suitable matrices.

[Selenium deficiency in an organic extensive water buffalo farm].

PubMed

Große, Reinhard; Binici, Cagri; Pieper, Robert; Müller, Kerstin E

2018-06-01

This case report presents investigations of muscle problems in three male water buffaloes (1-2 years) kept extensively (loose housing, pasture). The bulls were presented because of listlessness and increased lying periods. They displayed difficulties to stand up, a stilted gait, and tremor in the legs. The determination of the selenium concentration by the measurement of glutathione peroxidase activity in whole blood samples (EDTA) demonstrated selenium deficiency in all three buffaloes. This confirmed the tentative diagnosis of nutritive myodystrophy due to selenium deficiency. Following a single injection of 1500 mg all-rac-alpha-tocopherol acetate and 11 mg sodium selenite, the bulls recovered clinically. The whole blood samples taken subsequently from seven adult water buffaloes on the farm showed selenium deficiency in all animals. Consequently, slow-release multi-trace element boluses were administered once orally - as far as possible - to all adult animals of the herd. After 1 year, a good to very good selenium supply was observed in all these buffaloes, except for one cow, in which bolus application had failed. Schattauer GmbH.

Chemical and thermal studies on esterification of EDTA with raw cellulose and mercerized cellulose EFB

NASA Astrophysics Data System (ADS)

Azamkamal, Fatihah; Zakaria, Sarani; Gan, Sinyee; Kaco, Hatika

2018-04-01

Oil palm empty fruit bunch fibre (EFB) was bleached using four stages bleaching sequences (DEED) where D was a bleaching process composed of 1.7 wt% NaClO2 and buffer solution while E was composed of NaOH solution. Raw cellulose and mercerized cellulose which treated with 3.5 N sodium hydroxide were used as a raw material for esterification with ethylenediaminetetraacetic acid (EDTA) and enhancement with acetic acid. The samples of raw cellulose and mercerized cellulose were observed using optical microscope. The thermal properties of raw cellulose and mercerized cellulose esterified with EDTA were studied. The effect of mercerized cellulose on esterification process of EDTA was investigated. The studies suggested that the mercerization process affect the thermal stability of the cellulose. The transmittance of FTIR band showed that raw cellulose gave better esterification product compared to mercerized cellulose. Hence, the mercerization process of cellulose does not improve the esterification of cellulose with EDTA.

Effect of NaFeEDTA-fortified soy sauce on zinc absorption in children.

PubMed

Li, Min; Wu, Jinghuan; Ren, Tongxiang; Wang, Rui; Li, Weidong; Piao, Jianhua; Wang, Jun; Yang, Xiaoguang

2015-03-01

NaFeEDTA has been applied in many foods as an iron fortificant and is used to prevent iron deficiency in Fe-depleted populations. In China, soy sauce is fortified with NaFeEDTA to control iron deficiency. However, it is unclear whether Fe-fortified soy sauce affects zinc absorption. To investigate whether NaFeEDTA-fortified soy sauce affects zinc absorption in children, sixty children were enrolled in this study and randomly assigned to three groups (10 male children and 10 female children in each group). All children received daily 3 mg of (67)Zn and 1.2 mg of dysprosium orally, while the children in the three groups were supplemented with NaFeEDTA-fortified soy sauce (6 mg Fe, NaFeEDTA group), FeSO₄-fortified soy sauce (6 mg Fe, FeSO₄ group), and no iron-fortified soy sauce (control group), respectively. Fecal samples were collected during the experimental period and analyzed for the Zn content, (67)Zn isotope ratio and dysprosium content. The Fe intake from NaFeEDTA-fortified and FeSO₄-fortified groups was significantly higher than that in the control group (P < 0.0001). The daily total Zn intake was not significantly different among the three groups. There were no significant differences in fractional Zn absorption (FZA) (P = 0.3895), dysprosium recovery (P = 0.7498) and Zn absorption (P = 0.5940) among the three groups. Therefore, NaFeEDTA-fortified soy sauce does not affect Zn bioavailability in children.

Simplifying sampling for African swine fever surveillance: Assessment of antibody and pathogen detection from blood swabs.

PubMed

Carlson, J; Zani, L; Schwaiger, T; Nurmoja, I; Viltrop, A; Vilem, A; Beer, M; Blome, S

2018-02-01

African swine fever (ASF) is a notifiable disease with serious socio-economic consequences that has been present in wild boar in the Baltic States and Poland since 2014. An introduction of ASF is usually accompanied by increased mortality, making fallen wild boar and hunted animals with signs of disease the main target for early warning and passive surveillance. It is difficult, however, to encourage hunters and foresters to report and take samples from these cases. A pragmatic and easy sampling approach with quick-drying swabs could facilitate this. In this study, we further evaluated the use of dry blood swabs for the detection of ASFV antibody and genome with samples from animal trials and diagnostic submissions (blood, bone and organs) from Estonia. Compared to serum samples, dried blood swabs yielded 93.1% (95% confidence interval: [83.3, 98.1]) sensitivity and 100% [95.9, 100.0] specificity in a commercial ASFV antibody ELISA. Similarly, the swabs gave a sensitivity of 98.9% [93.4, 100.0] and a specificity of 98.1% [90.1, 100.0] for genome detection by a standard ASFV p72 qPCR when compared to EDTA blood. The same swabs were tested in a VP72-antibody lateral flow device, with a sensitivity of 94.7% [85.4, 98.9] and specificity of 96.1% [89.0, 99.2] compared to the serum ELISA. When GenoTube samples tested in ELISA and LFD were compared, the sensitivity was 96.3% [87.3, 99.5] and the specificity was 93.8% [86.0, 97.9]. This study demonstrates reliable detection of ASFV antibody and genome from swabs. A field test of the swabs with decomposed wild boar carcasses in an endemic area in Estonia also gave promising results. Thus, this technique is a practical approach for surveillance of ASF in both free and endemic areas. © 2017 Blackwell Verlag GmbH.

Bluetongue virus surveillance in the Islamic Republic of Mauritania: Is serotype 26 circulating among cattle and dromedaries?

PubMed

Lorusso, Alessio; Baba, Doumbia; Spedicato, Massimo; Teodori, Liana; Bonfini, Barbara; Marcacci, Maurilia; Di Provvido, Andrea; Isselmou, Katia; Marini, Valeria; Carmine, Irene; Scacchia, Massimo; Di Sabatino, Daria; Petrini, Antonio; Bezeid, Beyatt Ahmed; Savini, Giovanni

2016-06-01

In March 2013, EDTA-blood and serum samples were collected from 119 cattle and 159 dromedaries at the slaughterhouse of Nouakchott, the capital city of the Islamic Republic of Mauritania. Serum samples were screened for the presence of Bluetongue (BT) antibodies by competitive ELISA (cELISA). Positive samples were then tested by serum-neutralization (SN) to determine BTV serotype. RNA from blood samples was first tested by a genus-specific quantitative RT-PCR assay which is able to detect all 27 existing BTV serotypes (RT-qPCR1-27). Positive samples were further screened by a RT-qPCR assay which, instead, is able to detect the classical 24 BTV serotypes only (RT-qPCR1-24). Of the 278 serum samples tested, 177 (mean=63.7%; 95% CI: 57.9%-69.1%) resulted positive by cELISA. Of these, 69 were from cattle (mean=58.0%; 95% CI: 49.0%-66.5%) and 108 from dromedaries (mean=67.9%; 95% CI: 60.3%-74.7%). BTV-26 neutralizing antibodies were by far the most frequently found as they were detected in 146 animals with titres ranging from 1:10 to 1:80. Out of 278 blood samples, 25 (mean=9.0%; 95% CI: 6.2%-12.9%) were found positive for BTV by RT-qPCR1-27, 20 (mean=16.8%; 95% CI: 11.2%-24.6%) were from cattle and 5 (mean=3.1%; 95% CI: 1.4%-7.1%) from dromedaries. When tested by RT-qPCR1-24 the 25 BTV positive samples were negative. Unfortunately, no genetic information by molecular typing or by next generation sequencing has been obtained as for the very low levels of RNA in the blood samples. Copyright © 2016 Elsevier B.V. All rights reserved.

DETECTION OF HEAVY METALS BY IMMUNOASSAY: OPTIMIZATION AND VALIDATION OF A RAPID, PORTABLE ASSAY FOR IONIC CADMIUM (R824029)

EPA Science Inventory

An immunoassay is described that measured Cd(II) in aqueous samples at
concentrations from approximately 7 to 500 ppb. The assay utilized a monoclonal
antibody that bound tightly to a cadmium-ethylenediaminetetraacetic acid (EDTA)
complex but not to metal-free EDTA...

Nisin, rosemary, and ethylenediaminetetraacetic acid affect the growth of Listeria monocytogenes on ready-to-eat turkey ham stored at four degrees Celsius for sixty-three days.

PubMed

Ruiz, A; Williams, S K; Djeri, N; Hinton, A; Rodrick, G E

2009-08-01

The objectives of this study were to determine the anti-Listeria and general antimicrobial properties of nisin, rosemary, and EDTA alone and in combination on Listeria monocytogenes inoculated on ready-to-eat vacuum-packaged diced turkey ham and to ascertain the effects of the treatments on pH and objective color. The turkey hams were cut into 0.5-cm pieces, inoculated with a L. monocytogenes cocktail containing 5 strains of the bacterium, and treated with either no treatment and no inoculum (negative control), inoculum only (positive control), 0.5% nisin, 20 mM EDTA, 1% rosemary, 0.5% nisin + 20 mM EDTA, 0.5% nisin + 1% rosemary, 0.5% nisin + 20 mM EDTA + 1% rosemary, or 20 mM EDTA + 1% rosemary. All samples were vacuum-packaged, stored for 63 d at 4 degrees C +/- 1 degrees C, and analyzed at 1-wk intervals for total aerobes, L. monocytogenes, lactic acid organisms, pH, and objective color. Nisin, nisin with rosemary, nisin with EDTA, and nisin with rosemary and EDTA treatments reduced (P < 0.05) L. monocytogenes counts by 4.42, 4.20, 3.73, and 4.11 log cfu/g when compared with the positive control, respectively, on d 0. Listeria monocytogenes counts remained less than 2.75 log cfu/g for all hams treated with nisin. The EDTA and rosemary treatments alone and in combination were ineffective in inhibiting growth of L. monocytogenes. Although none of the treatments completely eliminated L. monocytogenes, the results indicated that ready-to-eat turkey ham can have significantly decreased L. monocytogenes when treated with nisin alone or in combination with rosemary or EDTA, or both.

A comparative evaluation of smear layer removal by using edta, etidronic acid, and maleic acid as root canal irrigants: An in vitro scanning electron microscopic study

PubMed Central

Kuruvilla, Aby; Jaganath, Bharath Makonahalli; Krishnegowda, Sahadev Chickmagaravalli; Ramachandra, Praveen Kumar Makonahalli; Johns, Dexton Antony; Abraham, Aby

2015-01-01

Aim: The purpose of this study is to evaluate and compare the efficacy of 17% EDTA, 18% etidronic acid, and 7% maleic acid in smear layer removal using scanning electron microscopic image analysis. Materials and Methods: Thirty, freshly extracted mandibular premolars were used. The teeth were decoronated to obtain working length of 17mm and instrumentation up to 40 size (K file) with 2.5% NaOCl irrigation between each file. The samples were divided into Groups I (17% ethylenediaminetetraacetic acid (EDTA)), II (18% etidronic acid), and III (7% maleic acid) containing 10 samples each. Longitudinal sectioning of the samples was done. Then the samples were observed under scanning electron microscope (SEM) at apical, middle, and coronal levels. The images were scored according to the criteria: 1. No smear layer, 2. moderate smear layer, and 3 heavy smear layer. Statistical Analysis: Data was analyzed statistically using Kruskal–Wallis analysis of variance (ANOVA) followed by Mann-Whitney U test for individual comparisons. The level for significance was set at 0.05. Results: The present study showed that all the three experimental irrigants removed the smear layer from different tooth levels (coronal, middle, and apical). Final irrigation with 7% maleic acid is more efficient than 17% EDTA and 18% etidronic acid in the removal of smear layer from the apical third of root canal. PMID:26069414

Evaluation of on-line chelant addition to PWR steam generators. Steam generator cleaning project

DOE Office of Scientific and Technical Information (OSTI.GOV)

Tvedt, T.J.; Wallace, S.L.; Griffin, F. Jr.

1983-09-01

The investigation of chelating agents for continuous water treatment of secondary loops of PWR steam generators were conducted in two general areas: the study of the chemistry of chelating agents and the study of materials compatability with chelating agents. The thermostability of both EDTA and HEDTA metal chelates in All Volatile Treatment (AVT) water chemistry were shown to be greater than or equal to the thermostability of EDTA metal chelates in phosphate-sulfite water chemistry. HEDTA metal chelates were shown to have a much greater stability than EDTA metal chelates. Using samples taken from the EDTA metal chelate thermostability study andmore » from the Commonwealth Research Corporation (CRC) model steam generators (MSG), EDTA decomposition products were determined. Active metal surfaces were shown to become passivated when exposed to EDTA and HEDTA concentrations as high as 0.1% w/w in AVT. Trace amounts of iron in the water were found to increase the rate of passivation. Material balance and visual inspection data from CRC model steam generators showed that metal was transported through and cleaned from the MSG's. The Inconel 600 tubes of the salt water fouled model steam generators experienced pitting corrosion. Results of this study demonstrates the feasibility of EDTA as an on-line water treatment additive to maintain nuclear steam generators in a clean condition.« less

Automation of a flocculation test for syphilis on Groupamatic equipment.

PubMed Central

Garretta, M; Paris-Hamelin, A; Gener, J; Muller, A; Matte, C; Vaisman, A

1975-01-01

A flocculation reaction employing a cardiolipid antigen was used for syphilis screening on Groupamatic equipment in parallel with conventional screening reactions: Kolmer CF, RPCF, Kahn, Kline, and RPR. The positive samples were confirmed by FTA-200, FTA-ABS, TPI, and in some cases by TPHA. There were 5,212 known samples which had already been tested by all methods and of which 1,648 were positive, and 58,636 screened samples including 65 positives. Half of the samples in the first series were taken without anticoagulant; the remainder were collected in potassium EDTA. The percentage of false positives with the Groupamatic was about 1-4 per cent. The percentage of false negatives among positve (greater than or equal+) samples varied from 0-18 to 1-3 per cent.; on the other hand the sensitivity was less good for samples giving doubtful and/or dissociated reactions in conventional screening reactions. The specificity and sensitivity of this technique are acceptable for a blood transfusion centre. The reproducibility is excellent and the automatic reading of results accurate. Additional advantages are rapidity (340 samples processed per hour); simultaneous performance of eleven other immunohaematological reactions; no contamination between samples; automatic reading, interpretation, and print-out of results; and saving of time because samples are not filed sequentially and are automatically identified when the results are obtained. Although the importance of syphilis in blood transfusion seems small, estimates of the risk are difficult and further investigations are planned. Images PMID:1098731

Diagnosis of bacteremia in pediatric oncologic patients by in-house real-time PCR.

PubMed

Quiles, Milene Gonçalves; Menezes, Liana Carballo; Bauab, Karen de Castro; Gumpl, Elke Kreuscher; Rocchetti, Talita Trevizani; Palomo, Flavia Silva; Carlesse, Fabianne; Pignatari, Antonio Carlos Campos

2015-07-23

Infections are the major cause of morbidity and mortality in children with cancer. Gaining a favorable prognosis for these patients depends on selecting the appropriate therapy, which in turn depends on rapid and accurate microbiological diagnosis. This study employed real-time PCR (qPCR) to identify the main pathogens causing bloodstream infection (BSI) in patients treated at the Pediatric Oncology Institute IOP-GRAACC-UNIFESP-Brazil. Antimicrobial resistance genes were also investigated using this methodology. A total of 248 samples from BACTEC® blood culture bottles and 99 whole-blood samples collected in tubes containing EDTA K2 Gel were isolated from 137 patients. All samples were screened by specific Gram probes for multiplex qPCR. Seventeen sequences were evaluated using gender-specific TaqMan probes and the resistance genes bla SHV, bla TEM, bla CTX, bla KPC, bla IMP, bla SPM, bla VIM, vanA, vanB and mecA were detected using the SYBR Green method. Positive qPCR results were obtained in 112 of the blood culture bottles (112/124), and 90 % agreement was observed between phenotypic and molecular microbial detection methods. For bacterial and fungal identification, the performance test showed: sensitivity 87 %; specificity 91 %; NPV 90 %; PPV 89 % and accuracy of 89 % when compared with the phenotypic method. The mecA gene was detected in 37 samples, extended-spectrum β-lactamases were detected in six samples and metallo-β-lactamase coding genes in four samples, with 60 % concordance between the two methods. The qPCR on whole blood detected eight samples possessing the mecA gene and one sample harboring the vanB gene. The bla KPC, bla VIM, bla IMP and bla SHV genes were not detected in this study. Real-time PCR is a useful tool in the early identification of pathogens and antimicrobial resistance genes from bloodstream infections of pediatric oncologic patients.

Small and cheap: accurate differential blood count with minimal sample volume by laser scanning cytometry (LSC)

NASA Astrophysics Data System (ADS)

Mittag, Anja; Lenz, Dominik; Smith, Paul J.; Pach, Susanne; Tarnok, Attila

2005-04-01

Aim: In patients, e.g. with congenital heart diseases, a differential blood count is needed for diagnosis. To this end by standard automatic analyzers 500 μl of blood is required from the patients. In case of newborns and infants this is a substantial volume, especially after operations associated with blood loss. Therefore, aim of this study was to develop a method to determine a differential blood picture with a substantially reduced specimen volume. Methods: To generate a differential blood picture 10 μl EDTA blood were mixed with 10 μl of a DRAQ5 solution (500μM, Biostatus) and 10 μl of an antibody mixture (CD45-FITC, CD14-PE, diluted with PBS). 20 μl of this cell suspension was filled into a Neubauer counting chamber. Due to the defined volume of the chamber it is possible to determine the cell count per volume. The trigger for leukocyte counting was set on DRAQ5 signal in order to be able to distinguish nucleated white blood cells from erythrocytes. Different leukocyte subsets could be distinguished due to the used fluorescence labeled antibodies. For erythrocyte counting cell suspension was diluted another 150 times. 20 μl of this dilution was analyzed in a microchamber by LSC with trigger set on forward scatter signal. Results: This method allows a substantial decrease of blood sample volume for generation of a differential blood picture (10 μl instead of 500μl). There was a high correlation between our method and the results of routine laboratory (r2=0.96, p<0.0001 n=40). For all parameters intra-assay variance was less than 7 %. Conclusions: In patients with low blood volume such as neonates and in critically ill infants every effort has to be taken to reduce the blood volume needed for diagnostics. With this method only 2% of standard sample volume is needed to generate a differential blood picture. Costs are below that of routine laboratory. We suggest this method to be established in paediatric cardiology for routine diagnostics and for resource poor settings.

Redistribution of fractions of zinc, cadmium, nickel, copper, and lead in contaminated calcareous soils treated with EDTA.

PubMed

Jalali, Mohsen; Khanlari, Zahra V

2007-11-01

Effect of ethylene diamine tetraacetic acid (EDTA) on the fractionation of zinc (Zn), cadmium (Cd), nickel (Ni), copper (Cu), and lead (Pb) in contaminated calcareous soils was investigated. Soil samples containing variable levels of contamination, from 105.9 to 5803 mg/kg Zn, from 2.2 to 1361 mg/kg Cd, from 31 to 64.0 mg/kg Ni, from 24 to 84 mg/kg Cu, and from 109 to 24,850 mg/kg Pb, were subjected to EDTA treatment at different dosages of 0, 1.0, and 2.0 g/kg. Metals in the incubated soils were fractionated after 5 months by a sequential extraction procedure, in which the metal fractions were experimentally defined as exchangeable (EXCH), carbonate (CARB), Mn oxide (MNO), Fe oxide (FEO), organic matter (OM), and residual (RES) fractions. In contaminated soils without EDTA addition, Zn, Ni, Cu, and Pb were predominately present in the RES fraction, up to 60.0%, 32.3%, 41.1%, and 36.8%, respectively. In general, with the EDTA addition, the EXCH and CARB fractions of these metals increased dramatically while the OM fraction decreased. The Zn, Ni, Cu, and Pb were distributed mostly in RES, OM, FEO, and CARB fractions in contaminated soils, but Cd was found predominately in the CARB, MNO, and RES fractions. The OM fraction decreased with increasing amounts of EDTA. In the contaminated soils, EDTA removed some Pb, Zn, Cu, and Ni from MNO, FEO, and OM fractions and redistributed them into CARB and EXCH fractions. Based on the relative percent in the EXCH and CARB fractions, the order of solubility was Cd > Pb > Ni > Cu > Zn for contaminated soils, before adding of EDTA, and after adding of EDTA, the order of solubility was Pb > Cd > Zn > Ni > Cu. The risk of groundwater contamination will increase after applying EDTA and it needed to be used very carefully.

How the reference values for serum parathyroid hormone concentration are (or should be) established?

PubMed

Souberbielle, J-C; Brazier, F; Piketty, M-L; Cormier, C; Minisola, S; Cavalier, E

2017-03-01

Well-validated reference values are necessary for a correct interpretation of a serum PTH concentration. Establishing PTH reference values needs recruiting a large reference population. Exclusion criteria for this population can be defined as any situation possibly inducing an increase or a decrease in PTH concentration. As recommended in the recent guidelines on the diagnosis and management of asymptomatic primary hyperparathyroidism, PTH reference values should be established in vitamin D-replete subjects with a normal renal function with possible stratification according to various factors such as age, gender, menopausal status, body mass index, and race. A consensus about analytical/pre-analytical aspects of PTH measurement is also needed with special emphasis on the nature of the sample (plasma or serum), the time and the fasting/non-fasting status of the blood sample. Our opinion is that blood sample for PTH measurement should be obtained in the morning after an overnight fast. Furthermore, despite longer stability of the PTH molecule in EDTA plasma, we prefer serum as it allows to measure calcium, a prerequisite for a correct interpretation of a PTH concentration, on the same sample. Once a consensus is reached, we believe an important international multicentre work should be performed to recruit a very extensive reference population of apparently healthy vitamin D-replete subjects with a normal renal function in order to establish the PTH normative data. Due to the huge inter-method variability in PTH measurement, a sufficient quantity of blood sample should be obtained to allow measurement with as many PTH kits as possible.

Detection of thalassemia genes using smeared blood film or leukocytes adhering to polysthylene fibers.

PubMed

Tatsumi, N; Yokota, M; Shindoh, K; Funahara, Y; Nathalang, O; Sukpanichnant, S; Bunyaratvej, A; Fucharoen, S

1997-01-01

Presently genetic analyses for thalassemia types require relatively large amounts of heparinized blood (5 to 10 ml), and transport as well as degeneration of these sample is a problem in the developing world. We have developed a new method to simplify this procedure and obtain DNAs from small specimens. As experimental materials, thinly smeared blood on a glass slide or blood filtered with and adhered on polysthylene telephtalate (PST) fibers were used. These materials could be safely stored without interfering with DNA extraction for up to 3 months. The slide materials were digested with proteinase K, and DNA was extracted with Tris-EDTA-phenol:chloroform and precipitated with absolute ethanol. The PST specimens were washed with physiologic saline and treated in the same manner as described above. Products were easily amplified by PCR and digested with restriction endonucleases for beta thalassemia typing as well as for HLA-DQA1 gene typing. Results obtained by this method correlated well with previously reported incidences for thalassemia and HLA-DQA1 types in Thailand. This method can be used in the routine laboratory because it allows for stable and biosafe genetic analyses.

Preserving the distribution of inorganic arsenic species in groundwater and acid mine drainage samples

USGS Publications Warehouse

Bednar, A.J.; Garbarino, J.R.; Ranville, J.F.; Wildeman, T.R.

2002-01-01

The distribution of inorganic arsenic species must be preserved in the field to eliminate changes caused by metal oxyhydroxide precipitation, photochemical oxidation, and redox reactions. Arsenic species sorb to iron and manganese oxyhydroxide precipitates, and arsenite can be oxidized to arsenate by photolytically produced free radicals in many sample matrices. Several preservatives were evaluated to minimize metal oxyhydroxide precipitation, such as inorganic acids and ethylenediaminetetraacetic acid (EDTA). EDTA was found to work best for all sample matrices tested. Storing samples in opaque polyethylene bottles eliminated the effects of photochemical reactions. The preservation technique was tested on 71 groundwater and six acid mine drainage samples. Concentrations in groundwater samples reached 720 ??g-As/L for arsenite and 1080 ??g-As/L for arsenate, and acid mine drainage samples reached 13 000 ??g-As/L for arsenite and 3700 ??g-As/L for arsenate. The arsenic species distribution in the samples ranged from 0 to 90% arsenite. The stability of the preservation technique was established by comparing laboratory arsenic speciation results for samples preserved in the field to results for subsamples speciated onsite. Statistical analyses indicated that the difference between arsenite and arsenate concentrations for samples preserved with EDTA in opaque bottles and field speciation results were analytically insignificant. The percentage change in arsenite:arsenate ratios for a preserved acid mine drainage sample and groundwater sample during a 3-month period was -5 and +3%, respectively.

Evaluation of Blood Cell Attachment on Er:Yag Laser Applied Root Surface Using Scanning Electron Microscopy

PubMed Central

CEKICI, Ali; MADEN, Ilay; YILDIZ, Sercan; SAN, Tangul; ISIK, Gulden

2013-01-01

Background: Periodontal regeneration is dependent on the uninterrupted adhesion, maturation and absorption of fibrin clots to a periodontally compromised root surface. The modification of the root surface with different agents has been used for better fibrin clot formation and blood cell attachment. It is known that Er:YAG laser application on dentin removes the smear layer succesfully. Aim: The aim of this study is to observe blood cell attachment and fibrin network formation following ER:YAG laser irradiation on periodontally compromised root surfaces in comparison to chemical root conditioning techniques in vitro. Materials and methods: 40 dentin blocks prepared from freshly extracted periodontally compromised hopeless teeth. Specimens were divided in 5 groups; those applied with PBS, EDTA, Citric acid and Er:YAG. They were further divided into two groups: those which had received these applications, and the control group. The specimens were evaluated with scanning electron microscope and micrographs were taken. Smear layer and blood cell attachment scoring was performed. Results: In the Er:YAG laser applied group, smear layer were totally removed. In the blood applied specimens, better fibrin clot formation and blood cell attachment were observed in the Er:YAG group. In the group that had been applied with citric acid, the smear layer was also removed. The smear layer could not be fully removed in the EDTA group. Conclusion: Er:YAG laser application on the root dentin seems to form a suitable surface for fibrin clot formation and blood cell attachment. Further clinical studies to support these results are necessitated. PMID:23533017

Impact of iron particles in groundwater on the UV inactivation of bacteriophages MS2 and T4.

PubMed

Templeton, M R; Andrews, R C; Hofmann, R

2006-09-01

To investigate the impact of iron particles in groundwater on the inactivation of two model viruses, bacteriophages MS2 and T4, by 254-nm ultraviolet (UV) light. One-litre samples of groundwater with high iron content (from the Indianapolis Water Company, mean dissolved iron concentration 1.3 mg l(-1)) were stirred vigorously while exposed to air, which oxidized and precipitated the dissolved iron. In parallel samples, ethylenediaminetetra-acetic acid (EDTA) was added to chelate the iron and prevent formation of iron precipitate. The average turbidity in the samples without EDTA (called the 'raw' samples) after 210 min of stirring was 2.7 +/- 0.1 NTU while the average turbidity of the samples containing EDTA (called the 'preserved' samples) was 1.0 +/- 0.1 NTU. 'Raw' and 'preserved' samples containing bacteriophage MS2 were exposed to 254-nm UV light at doses of 20, 40, or 60 mJ (cm(2))(-1), while samples containing bacteriophage T4 were exposed to 2 or 5 mJ (cm(2))(-1), using a low pressure UV collimated beam. The UV inactivation of both phages in the 'raw' groundwater was lower than in the EDTA-'preserved' groundwater to a statistically significant degree (alpha = 0.05), due to the association of phage with the UV-absorbing iron precipitate particles. A phage elution technique confirmed that a large fraction of the phage that survived the UV exposures were particle-associated. Phages that are associated with iron oxide particles in groundwater are shielded from UV light to a measurable and statistically significant degree at a turbidity level of 2.7 NTU when the phage particle association is induced under experimental conditions. While the particle association of the phage in this study was induced experimentally, the findings provide further evidence that certain particles in natural waters and wastewaters (e.g. iron oxide particles) may have the potential to shield viruses from UV light.

Evaluation of a simple method for storage of blood samples that enables isolation of circulating tumor cells 96 h after sample collection.

PubMed

Apostolou, Panagiotis; Ntanovasilis, Dimitrios-Athanasios; Papasotiriou, Ioannis

2017-12-01

Minimizing the effects of transportation on the properties of biological material is a major challenge for the scientific community. The viability of cells is important in cases where their study is urgent for evaluation of treatment response or for the study of cancer progression. Circulating tumor cells (CTCs) constitute a cell subpopulation with great importance for oncologists, because of their prognostic value. Detection and isolation of CTCs from blood samples is a routine activity in many laboratories, but concerns exist with regard to the maintenance of the cells during transportation. In this study, experiments were conducted to determine the stability of gene and protein expression in CTCs over a period of 96 h. Blood samples collected from healthy individuals and patients with cancer were each divided into five aliquots, which were stored at 2-8 °C and analyzed after 0, 24, 48, 72 and 96 h of storage. CTCs from patients and CD45-negative cells from healthy individuals were isolated each day using enrichment protocols, and qPCR was performed to determine expression levels of genes encoding specific biological markers. In addition, cells from breast and colon cancer cell lines were spiked into blood samples from healthy individuals, and these samples were stored and analyzed over a period of 96 h by PCR and by flow cytometry. The markers that were studied included housekeeping genes and genes associated with the response to chemotherapy, as well as genes encoding transcription factors. The results demonstrated that the expression profiles of specific genes and proteins in CTCs were not significantly affected by 72 h of storage. After 96 h of storage, expression of some genes was altered. The transportation of blood at low temperature (2-8 °C) in the presence of the anticoagulant EDTA can protect CTCs from alteration of gene and protein expression for at least 72 h. Furthermore, under these conditions, CTCs can be detected and isolated 96 h after blood collection.

EDTA functionalized magnetic nanoparticle as a multifunctional adsorbent for Congo red dye from contaminated water

NASA Astrophysics Data System (ADS)

Sahoo, Jitendra Kumar; Rath, Juhi; Dash, Priyabrat; Sahoo, Harekrushna

2017-05-01

The present work reports the applicability of magnetite iron nanoparticles (Fe3O4) functionalized with ethylenediaminetetraacetic acid (EDTA) as an efficient adsorbent for the removal of Congo red (CR) dye from contaminated water. Magnetic nanoparticles (Fe3O4) are prepared by chemical precipitation method in which Fe2+ and Fe3+ salt from aqueous solution were reacted in presence of ammonia solution. The surface of Fe3O4 nanoparticle was first coated with (3-aminopropyl) triethoxy silane (APTES) by a salinization reaction and then linked with EDTA via reaction between -NH2 and -COOH to form well dispersed surface functionalised biocompatible magnetic nanoparticles. The obtained EDTA functionalized magnetic nanoparticles are characterized in terms of their morphological, XRD, BET surface area analysis, Fourier transform infrared spectroscopy (FT-IR) and Vibrating sample magnetometer (VSM). The adsorption of CR on Fe3O4-APTES-EDTA nanocomposite corresponds well to the Langmuir model and the Freundlich model respectively. The adsorption processes for CR followed the pseudo-second-order model.

Highly sensitive measurement of whole blood chromium by inductively coupled plasma mass spectrometry.

PubMed

Cieslak, Wendy; Pap, Kathleen; Bunch, Dustin R; Reineks, Edmunds; Jackson, Raymond; Steinle, Roxanne; Wang, Sihe

2013-02-01

Chromium (Cr), a trace metal element, is implicated in diabetes and cardiovascular disease. A hypochromic state has been associated with poor blood glucose control and unfavorable lipid metabolism. Sensitive and accurate measurement of blood chromium is very important to assess the chromium nutritional status. However, interferents in biological matrices and contamination make the sensitive analysis challenging. The primary goal of this study was to develop a highly sensitive method for quantification of total Cr in whole blood by inductively coupled plasma mass spectrometry (ICP-MS) and to validate the reference interval in a local healthy population. This method was developed on an ICP-MS with a collision/reaction cell. Interference was minimized using both kinetic energy discrimination between the quadrupole and hexapole and a selective collision gas (helium). Reference interval was validated in whole blood samples (n=51) collected in trace element free EDTA tubes from healthy adults (12 males, 39 females), aged 19-64 years (38.8±12.6), after a minimum of 8 h fasting. Blood samples were aliquoted into cryogenic vials and stored at -70 °C until analysis. The assay linearity was 3.42 to 1446.59 nmol/L with an accuracy of 87.7 to 99.8%. The high sensitivity was achieved by minimization of interference through selective kinetic energy discrimination and selective collision using helium. The reference interval for total Cr using a non-parametric method was verified to be 3.92 to 7.48 nmol/L. This validated ICP-MS methodology is highly sensitive and selective for measuring total Cr in whole blood. Copyright © 2012 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved. Published by Elsevier Inc. All rights reserved.

An evaluation of different soil washing solutions for remediating arsenic-contaminated soils.

PubMed

Wang, Yiwen; Ma, Fujun; Zhang, Qian; Peng, Changsheng; Wu, Bin; Li, Fasheng; Gu, Qingbao

2017-04-01

Soil washing is a promising way to remediate arsenic-contaminated soils. Most research has mostly focused on seeking efficient extractants for removing arsenic, but not concerned with any changes in soil properties when using this technique. In this study, the removal of arsenic from a heavily contaminated soil employing different washing solutions including H 3 PO 4 , NaOH and dithionite in EDTA was conducted. Subsequently, the changes in soil physicochemical properties and phytotoxicity of each washing technique were evaluated. After washing with 2 M H 3 PO 4 , 2 M NaOH or 0.1 M dithionite in 0.1 M EDTA, the soil samples' arsenic content met the clean-up levels stipulated in China's environmental regulations. H 3 PO 4 washing decreased soil pH, Ca, Mg, Al, Fe, and Mn concentrations but increased TN and TP contents. NaOH washing increased soil pH but decreased soil TOC, TN and TP contents. Dithionite in EDTA washing reduced soil TOC, Ca, Mg, Al, Fe, Mn and TP contents. A drastic color change was observed when the soil sample was washed with H 3 PO 4 or 0.1 M dithionite in 0.1 M EDTA. After adjusting the soil pH to neutral, wheat planted in the soil sample washed by NaOH evidenced the best growth of all three treated soil samples. These results will help with selecting the best washing solution when remediating arsenic-contaminated soils in future engineering applications. Copyright © 2017 Elsevier Ltd. All rights reserved.

Comparison of phosphorus forms in three extracts of dairy feces by solution 31P NMR analysis

USDA-ARS?s Scientific Manuscript database

Using solution 31P NMR spectroscopy, we compared three extractants, deionized water, sodium acetate buffer (pH 5.0) with fresh sodium dithionite (NaAc-SD), and 0.25 M NaOH-0.05 M EDTA (NaOH-EDTA), for the profile of P compounds in two dairy fecal samples. Phosphorus extracted was 35% for water, and...

Dissolution of bovine pulp tissue in solutions consisting of varying NaOCl concentrations and combined with EDTA.

PubMed

Irala, Luis Eduardo Duarte; Grazziotin-Soares, Renata; Salles, Alexandre Azevedo; Munari, Aline Zen; Pereira, Joseani Santos

2010-01-01

This in vitro study evaluated (1) the dissolution of bovine pulp tissue in solutions consisting of varying NaOCl concentrations and combined with EDTA; and (2) the pH of these solutions before and after the experiment. The independent variables were the concentration and the volume of the solution. Thirty bovine pulps were divided in equal fragments, resulting in 90 fragments of pulp tissue. Each fragment was immersed in one of the following solutions: 1% NaOCl (4 ml), 2.5% NaOCl (4 ml), 1% NaOCl + 17% EDTA (2 ml : 2 ml), 1% NaOCl + 17% EDTA (1 ml : 3 ml), 2.5% NaOCl + 17% EDTA (2 ml : 2 ml), and 2.5% NaOCl + 17% EDTA (1 ml : 3 ml). The test solutions were dichotomized as either able or not able to dissolve the tissue, the latter being attributed when the dissolution of the pulp tissue was not complete within 48 hours. When the samples were able to dissolve the tissue, the time required for complete tissue dissolution was submitted to statistical analysis. The pH of the solutions was measured before and after the experiment. The pH variable was dichotomized as either changed or unchanged. The results demonstrated that the NaOCl solutions combined with 17% EDTA were not able to dissolve the tissue. The t-test revealed that the 2.5% NaOCl solution presented a lower mean dissolution time than the 1% NaOCl solution (p < 0.001). The pH of the solutions with equal volumes of NaOCl and EDTA decreased in 48 hours.

Severe anaemia associated with Plasmodium falciparum infection in children: consequences for additional blood sampling for research.

PubMed

Kuijpers, Laura Maria Francisca; Maltha, Jessica; Guiraud, Issa; Kaboré, Bérenger; Lompo, Palpouguini; Devlieger, Hugo; Van Geet, Chris; Tinto, Halidou; Jacobs, Jan

2016-06-02

Plasmodium falciparum infection may cause severe anaemia, particularly in children. When planning a diagnostic study on children suspected of severe malaria in sub-Saharan Africa, it was questioned how much blood could be safely sampled; intended blood volumes (blood cultures and EDTA blood) were 6 mL (children aged <6 years) and 10 mL (6-12 years). A previous review [Bull World Health Organ. 89: 46-53. 2011] recommended not to exceed 3.8 % of total blood volume (TBV). In a simulation exercise using data of children previously enrolled in a study about severe malaria and bacteraemia in Burkina Faso, the impact of this 3.8 % safety guideline was evaluated. For a total of 666 children aged >2 months to <12 years, data of age, weight and haemoglobin value (Hb) were available. For each child, the estimated TBV (TBVe) (mL) was calculated by multiplying the body weight (kg) by the factor 80 (ml/kg). Next, TBVe was corrected for the degree of anaemia to obtain the functional TBV (TBVf). The correction factor consisted of the rate 'Hb of the child divided by the reference Hb'; both the lowest ('best case') and highest ('worst case') reference Hb values were used. Next, the exact volume that a 3.8 % proportion of this TBVf would present was calculated and this volume was compared to the blood volumes that were intended to be sampled. When applied to the Burkina Faso cohort, the simulation exercise pointed out that in 5.3 % (best case) and 11.4 % (worst case) of children the blood volume intended to be sampled would exceed the volume as defined by the 3.8 % safety guideline. Highest proportions would be in the age groups 2-6 months (19.0 %; worst scenario) and 6 months-2 years (15.7 %; worst case scenario). A positive rapid diagnostic test for P. falciparum was associated with an increased risk of violating the safety guideline in the worst case scenario (p = 0.016). Blood sampling in children for research in P. falciparum endemic settings may easily violate the proposed safety guideline when applied to TBVf. Ethical committees and researchers should be wary of this and take appropriate precautions.

Detection of Total Phenol in Green and Black Teas by Flow Injection System and Unmodified Screen Printed Electrode

PubMed Central

de Mattos, Ivanildo Luiz; Zagal, José Heraclito

2010-01-01

A flow injection system using an unmodified gold screen-printed electrode was employed for total phenol determination in black and green teas. In order to avoid passivation of the electrode surface due to the redox reaction, preoxidation of the sample was realized by hexacyanoferrate(III) followed by addition of an EDTA solution. The complex formed in the presence of EDTA minimizes or avoids polymerization of the oxidized phenols. The previously filtered tea sample and hexacyanoferrate(III) reagent were introduced simultaneously into two-carrier streams producing two reproducible zones. At confluence point, the pre-oxidation of the phenolic compounds occurs while this zone flows through the coiled reactor and receives the EDTA solution before phenol detection. The consumption of ferricyanide was monitorized at 360 mV versus Ag/AgCl and reflected the total amount of phenolic compounds present in the sample. Results were reported as gallic acid equivalents (GAEs). The proposed system is robust, versatile, environmentally-friendly (since the reactive is used only in the presence of the sample), and allows the analysis of about 35–40 samples per hour with detection limit = 1 mg/L without the necessity for surface cleaning after each measurement. Precise results are in agreement with those obtained by the Folin-Ciocalteu method. PMID:21461407

The effects of CaEDTA injection on lead, zinc, copper and ALAD in erythrocyte, plasma and urine in lead-exposed workers: a 24-h observation.

PubMed

Aono, H; Araki, S

1984-01-01

To evaluate the effects of calcium disodium ethylenediamine tetraacetate (CaEDTA) on the concentrations of lead, zinc and copper in plasma, erythrocyte and urine, and the delta-aminolevulinic acid dehydratase (ALAD) activity in erythrocyte, we administered CaEDTA in 1-h intravenous infusion to ten male gun metal founders with blood-lead concentration of 39 to 64 micrograms/dl (mean 49 micrograms/dl). We found that the plasma concentration of lead, following a rapid rise within the first 3 h, fell temporarily to the level significantly lower than the initial level 19 h after start of the infusion. The plasma concentration of zinc fell to the minimal level 5 h after the infusion; and the erythrocyte concentration of zinc and the ALAD activity concurrently rose to the maximal level 5 h after the infusion. By contrast, no significant alteration was observed in the concentrations of copper in plasma and erythrocyte. The maximal level of urinary metal excretion was attained during the period between 1 and 2 h after start of CaEDTA infusion for lead; within 2 h for zinc; and between 2 and 4 h for copper. The urinary metal excretion returned to the initial level 14 to 24 h after infusion for zinc and copper; but lead excretion was still higher than the initial level during this period. The difference in the kinetics of the three metals following CaEDTA injection is discussed in the light of these findings.

Cryopreservation for delayed circulating tumor cell isolation is a valid strategy for prognostic association of circulating tumor cells in gastroesophageal cancer

PubMed Central

Brungs, Daniel; Lynch, David; Luk, Alison WS; Minaei, Elahe; Ranson, Marie; Aghmesheh, Morteza; Vine, Kara L; Carolan, Martin; Jaber, Mouhannad; de Souza, Paul; Becker, Therese M

2018-01-01

AIM To demonstrate the feasibility of cryopreservation of peripheral blood mononuclear cells (PBMCs) for prognostic circulating tumor cell (CTC) detection in gastroesophageal cancer. METHODS Using 7.5 mL blood samples collected in EDTA tubes from patients with gastroesopheagal adenocarcinoma, CTCs were isolated by epithelial cell adhesion molecule based immunomagnetic capture using the IsoFlux platform. Paired specimens taken during the same blood draw (n = 15) were used to compare number of CTCs isolated from fresh and cryopreserved PBMCs. Blood samples were processed within 24 h to recover the PBMC fraction, with PBMCs used for fresh analysis immediately processed for CTC isolation. Cryopreservation of PBMCs lasted from 2 wk to 25.2 mo (median 14.6 mo). CTCs isolated from pre-treatment cryopreserved PBMCs (n = 43) were examined for associations with clinicopathological variables and survival outcomes. RESULTS While there was a significant trend to a decrease in CTC numbers associated with cryopreserved specimens (mean number of CTCs 34.4 vs 51.5, P = 0.04), this was predominately in samples with a total CTC count of > 50, with low CTC count samples less affected (P = 0.06). There was no significant association between the duration of cryopreservation and number of CTCs. In cryopreserved PBMCs from patient samples prior to treatment, a high CTC count (> 17) was associated with poorer overall survival (OS) (n = 43, HR = 4.4, 95%CI: 1.7-11.7, P = 0.0013). In multivariate analysis, after controlling for sex, age, stage, ECOG performance status, and primary tumor location, a high CTC count remained significantly associated with a poorer OS (HR = 3.7, 95%CI: 1.2-12.4, P = 0.03). CONCLUSION PBMC cryopreservation for delayed CTC isolation is a valid strategy to assist with sample collection, transporting and processing. PMID:29467551

Evaluation of red blood cell and platelet antigen genotyping platforms (ID CORE XT/ID HPA XT) in routine clinical practice.

PubMed

Finning, Kirstin; Bhandari, Radhika; Sellers, Fiona; Revelli, Nicoletta; Villa, Maria Antonietta; Muñiz-Díaz, Eduardo; Nogués, Núria

2016-03-01

High-throughput genotyping platforms enable simultaneous analysis of multiple polymorphisms for blood group typing. BLOODchip® ID is a genotyping platform based on Luminex® xMAP technology for simultaneous determination of 37 red blood cell (RBC) antigens (ID CORE XT) and 18 human platelet antigens (HPA) (ID HPA XT) using the BIDS XT software. In this international multicentre study, the performance of ID CORE XT and ID HPA XT, using the centres' current genotyping methods as the reference for comparison, and the usability and practicality of these systems, were evaluated under working laboratory conditions. DNA was extracted from whole blood in EDTA with Qiagen methodologies. Ninety-six previously phenotyped/genotyped samples were processed per assay: 87 testing samples plus five positive controls and four negative controls. Results were available for 519 samples: 258 with ID CORE XT and 261 with ID HPA XT. There were three "no calls" that were either caused by human error or resolved after repeating the test. Agreement between the tests and reference methods was 99.94% for ID CORE XT (9,540/9,546 antigens determined) and 100% for ID HPA XT (all 4,698 alleles determined). There were six discrepancies in antigen results in five RBC samples, four of which (in VS, N, S and Do(a)) could not be investigated due to lack of sufficient sample to perform additional tests and two of which (in S and C) were resolved in favour of ID CORE XT (100% accuracy). The total hands-on time was 28-41 minutes for a batch of 16 samples. Compared with the reference platforms, ID CORE XT and ID HPA XT were considered simpler to use and had shorter processing times. ID CORE XT and ID HPA XT genotyping platforms for RBC and platelet systems were accurate and user-friendly in working laboratory settings.

Occurrence of selected pharmaceutical and non-pharmaceutical compounds, and stable hydrogen and oxygen isotope ratios in a riverbank filtration study, Platte River, Nebraska, 2002 to 2005, Volume 2

USGS Publications Warehouse

Vogel, J.R.; Barber, L.B.; Furlong, E.T.; Coplen, T.B.; Verstraeten, Ingrid M.; Meyer, M.T.

2005-01-01

This document is the second volume of a data series report that describes the data collected during a study conducted during 2001 through 2005 by the U.S. Geological Survey, in cooperation with the U.S. Environmental Protection Agency and the City of Lincoln, at an established riverbank-filtration well field with horizontal collector wells and vertical wells. The data were collected as part of a study designed to help researchers better understand the efficiency of riverbank filtration with respect to endocrine disrupting compounds and to evaluate the use of riverbank filtration as an effective means of drinking-water treatment. This study provides information that will be useful for (1) increased understanding of the processes and factors controlling the transport of endocrine disrupters, such as pesticides and pharmaceuticals during riverbank filtration, (2) better understanding of the physical and chemical processes that affect riverbank-filtration efficiency, and (3) managing the water resources of the eastern Platte River Basin. This report presents analytical methods and additional data for pharmaceuticals, dissolved organic carbon (DOC), ultraviolet absorbance at 254 nanometer (nm) wavelength (UV254), specific ultraviolet absorbance (SUVA), nitrilotriacetic acid (NTA), ethylenediaminetetraacetic acid (EDTA), nonylphenol ethoxycarboxylates (NPECs), and stable hydrogen and oxygen isotope ratios that were not available at the time of publication of Volume 1 in the data series. Data are presented as generalized statistics and in figures showing temporal variations. Sites from which water-quality samples were collected for this study included wastewater sites (a cattle feedlot lagoon, a hog confinement lagoon, and wastewater-treatment plant effluent), surface-water sites (Platte River, Salt Creek, and Loup Power Canal), ground-water sites (one collector well and five vertical wells), and drinking-water sites (raw and finished). Field water-quality properties also were measured in samples from these sites. Pharmaceuticals detected at least once in samples collected from the Platte River included 1,7-dimethylxanthine, acetaminophen, caffeine, carbamazapine, and cotinine. Among the ground-water samples, pharmaceutical compounds detected at low concentrations in at least one sample included 1,7-dimethylxanthine, acetaminophen, carbamazapine, and trimethoprim. When analyzing for non-pharmaceutical compounds in samples from the wastewater sites, the wastewater-treatment plant effluent samples had the highest concentrations of each of NTA, EDTA, and NPECs compounds. Surface-water samples from Salt Creek had higher concentrations of EDTA and NPECs than samples from the Platte River. NTA was not detected in any samples from the ground-water sites. EDTA was detected in all samples from all wells. Detectable concentrations of EDTA were also observed in all samples from the raw water and finished water.

Extracellular distribution volumes of hydrophilic solutes used to measure the glomerular filtration rate: comparison between chromium-51-EDTA and iohexol.

PubMed

Bird, Nicholas J; Peters, Christina; Michell, A Robert; Peters, A Michael

2007-02-01

Extracellular fluid volume (ECV) is larger when measured with Tc-99m-DTPA ( approximately 500 Da) than inulin (6 kDa). As part of an assessment of the suitability of the non-radioactive marker, iohexol, against the gold standard tracer, Cr-51-EDTA, for measurement of the glomerular filtration rate (GFR) based on a postal service, we took the opportunity to determine if this volume dependence is present for diffusible markers less disparate in size than inulin and Tc-99m-DTPA. Cr-51-EDTA ( approximately 400 Da) and iohexol ( approximately 900 Da) were administered into the opposite arms of 20 normal volunteers (fasting and non-fasting) and 60 patients (non-fasting), including 36 diabetics, 10 cancer patients and 13 dermatology patients. Blood was obtained from both arms 20, 40, 60, 120, 180 and 240 min after injection and assayed for a marker injected contra-laterally. The glomerular filtration rate (GFR) and mean indicator transit time, T, were measured from the bi-exponential clearance curves. ECV, the product of GFR and T, was subdivided into V(1) (administered indicator divided by the sum of zero-time intercepts of the two exponentials) and V(2) (the difference between V(1) and ECV). Variables were scaled to 1.73 m(2). For all 100 studies, the mean GFR from Cr-51-EDTA was 3 ml min(-1) higher than iohexol (p < 0.01). ECV was 0.41 L higher (p < 0.02) and V(1) 0.65 L higher (p < 0.001) from Cr-51-EDTA but V(2) was 0.33 L lower (p < 0.02). V(1)/ECV was 0.031 higher from Cr-51-EDTA (p < 0.01). ECV and V(2) from Cr-51-EDTA were both higher in diabetics (15.1 [1.7] and 5.0 [0.095] L, respectively) compared with normal non-fasting subjects (13.7 [1.5] and 4.3 [1.0]; p < 0.01). ECV and the volumes of its sub-compartments are different between markers that are less than an order of magnitude different in size.

Multispecies reactive tracer test in an aquifer with spatially variable chemical conditions

USGS Publications Warehouse

Davis, J.A.; Kent, D.B.; Coston, J.A.; Hess, K.M.; Joye, J.L.

2000-01-01

A field investigation of multispecies reactive transport was conducted in a well‐characterized, sand and gravel aquifer on Cape Cod, Massachusetts. The aquifer is characterized by regions of differing chemical conditions caused by the disposal of secondary sewage effluent. Ten thousand liters of groundwater with added tracers (Br, Cr(VI), and EDTA complexed with Pb, Zn, Cu, and Ni) were injected into the aquifer and distributions of the tracers were monitored for 15 months. Most of the tracers were transported more than 200 m; transport was quantified using spatial moments computed from the results of a series of synoptic samplings. Cr(VI) transport was retarded relative to Br; the retardation factor varied from 1.1 to 2.4 and was dependent on chemical conditions. At 314 days after the injection, dissolved Cr(VI) mass in the tracer cloud had decreased 85%, with the likely cause being reduction to Cr(III) in a suboxic region of the aquifer. Transport of the metal‐EDTA complexes was affected by aqueous complexation, adsorption, and dissolution‐precipitation reactions of Fe oxyhydroxide minerals in the aquifer sediments. Dissolved Pb‐EDTA complexes disappeared from the tracer cloud within 85 days, probably due to metal exchange reactions with Fe and adsorbed Zn (present prior to the injection from contamination by the sewage effluent). About 30% of the Cu‐EDTA complexes remained within the tracer cloud 314 days after injection, even though the thermodynamic stability of the Pb‐EDTA complex is greater than Cu‐EDTA. It is hypothesized that stronger adsorption of Pb2+ to the aquifer sediments causes the Pb‐EDTA complex to disassociate to a greater degree than the Cu‐EDTA complex. The mass of dissolved Zn‐EDTA increased during the first 175 days of the tracer test to 140% of the mass injected, with the increase due to desorption of sewage‐derived Zn. Dissolved Ni‐EDTA mass remained nearly constant throughout the tracer test, apparently only participating in reversible adsorption reactions. The results of the field experiment provide a chemically complex data set that can be used in the testing of reactive transport models of flow coupled with chemical reactions.

A case report describing detection of Rhodoturola minuta fungemia in an ewe lamb

USDA-ARS?s Scientific Manuscript database

An eight-month-old crossbred ewe that was normal upon physical examination was humanely euthanized for tissue collection. Prior to euthanasia, whole blood was collected via jugular venipuncture into 60-ml syringes containing EDTA anticoagulant. After sacrifice, the brain was removed and the choroi...

Determination of branched chain amino acids, methionine, phenylalanine, tyrosine and alpha-keto acids in plasma and dried blood samples using HPLC with fluorescence detection.

PubMed

Kand'ár, Roman; Záková, Pavla; Jirosová, Jana; Sladká, Michaela

2009-01-01

The determination of branched chain amino acids [BCAA; valine (Val), leucine (Leu), isoleucine (Ile)], alpha-keto acids derived from BCAA [BCKA; alpha-ketoisovaleric acid (KIV), alpha-ketoisocaproic acid (KIC), alpha-ketomethylvaleric acid (KMV)], methionine (Met), phenylalanine (Phe) and tyrosine (Tyr) is currently the most reliable approach for the diagnosis of maple syrup urine disease (MSUD), hypermethioninemia, phenylketonuria (PKU) and tyrosinemia. The aim of this study was to develop rapid and simple HPLC methods for measurement of BCAA, Met, Phe, Tyr and BCKA in plasma and dried blood samples. Samples of peripheral venous blood with EDTA as anticoagulant were obtained from a group of healthy blood donors (n=70, 35 females, 27-41 years of age and 35 males, 28-43 years of age). Blood-spot samples from a group of newborns (n=80, 40 girls and 40 boys 3-5 days of age) were collected onto #903 Specimen Collection Paper and allowed to dry for at least 24 h before analysis. Prior to separation, the amino acids (AA) were derivatized with o-phthaldialdehyde (OPA) and BCKA with o-phenylenediamine (OPD). Reverse phase column chromatography (LiChroCart 125-4 Purospher RP-18e, 5 microm) was used for separation and fluorescence detection used to monitoring of effluent. For AA analysis, 25 mmol/L sodium hydrogenphosphate-methanol (90:10, v/v), pH 6.5+/-0.1 was used as mobile phase A and 100% methanol was used as mobile phase B. Measurement of BCKA used a mixture of methanol and deionized water (55:45, v/v) as mobile phase A and mobile phase B consisted of 100% methanol. Analytical performance of these methods was satisfactory for the determination of all AA and BCKA. The intra-assay and inter-assay coefficients of variation were below 10% and recovery ranged from 90%-110%. We have developed simple, rapid and selective HPLC methods with fluorescence detection for the determination of BCAA, Met, Phe, Tyr and BCKA in plasma and dried blood samples.

Digital-Direct-RT-PCR: a sensitive and specific method for quantification of CTC in patients with cervical carcinoma.

PubMed

Pfitzner, Claudia; Schröder, Isabel; Scheungraber, Cornelia; Dogan, Askin; Runnebaum, Ingo Bernhard; Dürst, Matthias; Häfner, Norman

2014-02-05

The detection of circulating tumour cells (CTC) in cancer patients may be useful for therapy monitoring and prediction of relapse. A sensitive assay based on HPV-oncogene transcripts which are highly specific for cervical cancer cells was established. The Digital-Direct-RT-PCR (DD-RT-PCR) combines Ficoll-separation, ThinPrep-fixation and one-step RT-PCR in a low-throughput digital-PCR format enabling the direct analysis and detection of individual CTC without RNA isolation. Experimental samples demonstrated a sensitivity of one HPV-positive cell in 500,000 HPV-negative cells. Spike-in experiments with down to 5 HPV-positive cells per millilitre EDTA-blood resulted in concordant positive results by PCR and immunocytochemistry. Blood samples from 3 of 10 CxCa patients each contained a single HPV-oncogene transcript expressing CTC among 5 to 15*10(5) MNBC. Only 1 of 7 patients with local but 2 of 3 women with systemic disease had CTC. This highly sensitive DD-RT-PCR for the detection of CTC may also be applied to other tumour entities which express tumour-specific transcripts.

Prevalence of Theileria annulata in dairy cattle in Nyala, South Darfur State, Sudan.

PubMed

Abaker, Ismail A; Salih, Diaeldin A; Haj, Lima M El; Ahmed, Rawia E; Osman, Manal M; Ali, Awadia M

2017-12-01

This study was conducted in dairy cattle in Nyala, South Darfur State, during the period from June to September 2015, to study the prevalence of bovine tropical theileriosis. Apparently, healthy cattle of different age groups, different breeds, and from both sexes were randomly selected from seven locations. Three age groups of cattle were selected, group one <1 year old, group two 1-3 years old, and group three older than 3 years. These cattle were indigenous and cross (Friesian X zebu). A total of 150 blood samples were collected for blood smears, blood in EDTA tubes, and serum samples as well as ticks infesting cattle. Three diagnostic techniques were used such as blood smear, indirect fluorescent antibody test (IFAT), and polymerase chain reaction (PCR). Of 150 samples, 11 (7.3%, 95% confidence interval [CI]: 9.1-5.5) were positive for Theileria spp. piroplasms in the blood smears, 70 (46.7%, 95% CI: 35.7-57.7) were positive for Theileria annulata antibodies in the IFAT, and of 100 samples, 39 (39%, 95% CI: 46.6-31.4) were positive for T. annulata using PCR. The prevalence of T. annulata was higher in indigenous breed than cross cattle by the three diagnostic techniques. The highest prevalence of T. annulata was recorded among cattle older than 3 years old. There were three genera and ten species of ticks found feeding on cattle. These were Rhipicephalus evertsi evertsi, Rhipicephalus decoloratus, Rhipicephalus annulatus, Hyalomma dromedrii, Hyalomma impeltatum, Hyalomma rufipes, Hyalomma anatolicum, Hyalomma truncatum, Amblyomma variegatum , and Amblyomma lepidum . The study concluded that tropical theileriosis is prevalent among dairy cattle in Nyala. H. anatolicum was found in very low numbers, suggesting other ticks may play a role in the transmission of the disease. Molecular characterization of T. annulata is recommended for accurate mapping of the disease and evaluates the magnitude problem of tropical theileriosis in South Darfur region.

Prevalence of Theileria annulata in dairy cattle in Nyala, South Darfur State, Sudan

PubMed Central

Abaker, Ismail A.; Salih, Diaeldin A.; Haj, Lima M. El; Ahmed, Rawia E.; Osman, Manal M.; Ali, Awadia M.

2017-01-01

Aim: This study was conducted in dairy cattle in Nyala, South Darfur State, during the period from June to September 2015, to study the prevalence of bovine tropical theileriosis. Materials and Methods: Apparently, healthy cattle of different age groups, different breeds, and from both sexes were randomly selected from seven locations. Three age groups of cattle were selected, group one <1 year old, group two 1-3 years old, and group three older than 3 years. These cattle were indigenous and cross (Friesian X zebu). A total of 150 blood samples were collected for blood smears, blood in EDTA tubes, and serum samples as well as ticks infesting cattle. Three diagnostic techniques were used such as blood smear, indirect fluorescent antibody test (IFAT), and polymerase chain reaction (PCR). Results: Of 150 samples, 11 (7.3%, 95% confidence interval [CI]: 9.1-5.5) were positive for Theileria spp. piroplasms in the blood smears, 70 (46.7%, 95% CI: 35.7-57.7) were positive for Theileria annulata antibodies in the IFAT, and of 100 samples, 39 (39%, 95% CI: 46.6-31.4) were positive for T. annulata using PCR. The prevalence of T. annulata was higher in indigenous breed than cross cattle by the three diagnostic techniques. The highest prevalence of T. annulata was recorded among cattle older than 3 years old. There were three genera and ten species of ticks found feeding on cattle. These were Rhipicephalus evertsi evertsi, Rhipicephalus decoloratus, Rhipicephalus annulatus, Hyalomma dromedrii, Hyalomma impeltatum, Hyalomma rufipes, Hyalomma anatolicum, Hyalomma truncatum, Amblyomma variegatum, and Amblyomma lepidum. Conclusion: The study concluded that tropical theileriosis is prevalent among dairy cattle in Nyala. H. anatolicum was found in very low numbers, suggesting other ticks may play a role in the transmission of the disease. Molecular characterization of T. annulata is recommended for accurate mapping of the disease and evaluates the magnitude problem of tropical theileriosis in South Darfur region. PMID:29391689

Measurement by ICP-MS of lead in plasma and whole blood of lead workers and controls.

PubMed Central

Schütz, A; Bergdahl, I A; Ekholm, A; Skerfving, S

1996-01-01

OBJECTIVES: To test a simple procedure for preparing samples for measurement of lead in blood plasma (P-Pb) and whole blood (B-Pb) by inductively coupled plasma mass spectrometry (ICP-MS), to measure P-Pb and B-Pb in lead workers and controls, and to evaluate any differences in the relation between B-Pb and P-Pb between people. METHODS: P-Pb and B-Pb were measured by ICP-MS in 43 male lead smelter workers and seven controls without occupational exposure to lead. For analysis, plasma and whole blood were diluted 1 in 4 and 1 in 9, respectively, with a diluted ammonia solution containing Triton-X 100 and EDTA. The samples were handled under routine laboratory conditions, without clean room facilities. RESULTS: P-Pb was measured with good precision (CV = 5%) even at concentrations present in the controls. Freeze storage of the samples had no effect on the results. The detection limit was 0.015 microgram/l. The P-Pb was 0.15 (range 0.1-0.3) microgram/l in controls and 1.2 (0.3-3.6) micrograms/l in lead workers, although the corresponding B-Pbs were 40 (24-59) micrograms/l and 281 (60-530) micrograms/l (1 microgram Pb/I = 4.8 nmol/l). B-Pb was closely associated with P-Pb (r = 0.90). The association was evidently non-linear; the ratio B-Pb/P-Pb decreased with increasing P-Pb. CONCLUSIONS: By means of ICP-MS and a simple dilution procedure, P-Pb may be measured accurately and with good precision down to concentrations present in controls. Contamination of blood at sampling and analysis is no major problem. With increasing P-Pb, the percentage of lead in plasma increases. In studies of lead toxicity, P-Pb should be considered as a complement to current indicators of lead exposure and risk. PMID:9038796

Impact of Ficoll density gradient centrifugation on major and trace element concentrations in erythrocytes and blood plasma.

PubMed

Lu, Ying; Ahmed, Sultan; Harari, Florencia; Vahter, Marie

2015-01-01

Ficoll density gradient centrifugation is widely used to separate cellular components of human blood. We evaluated the suitability to use erythrocytes and blood plasma obtained from Ficoll centrifugation for assessment of elemental concentrations. We determined 22 elements (from Li to U) in erythrocytes and blood plasma separated by direct or Ficoll density gradient centrifugation, using inductively coupled plasma mass spectrometry. Compared with erythrocytes and blood plasma separated by direct centrifugation, those separated by Ficoll had highly elevated iodine and Ba concentration, due to the contamination from the Ficoll-Paque medium, and about twice as high concentrations of Sr and Mo in erythrocytes. On the other hand, the concentrations of Ca in erythrocytes and plasma were markedly reduced by the Ficoll separation, to some extent also Li, Co, Cu, and U. The reduced concentrations were probably due to EDTA, a chelator present in the Ficoll medium. Arsenic concentrations seemed to be lowered by Ficoll, probably in a species-specific manner. The concentrations of Mg, P, S, K, Fe, Zn, Se, Rb, and Cs were not affected in the erythrocytes, but decreased in plasma. Concentrations of Mn, Cd, and Pb were not affected in erythrocytes, but in plasma affected by EDTA and/or pre-analytical contamination. Ficoll separation changed the concentrations of Li, Ca, Co, Cu, As, Mo, I, Ba, and U in erythrocytes and blood plasma, Sr in erythrocytes, and Mg, P, S, K, Fe, Zn, Se, Rb and Cs in blood plasma, to an extent that will invalidate evaluation of deficiencies or excess intakes. Copyright © 2014 Elsevier GmbH. All rights reserved.

Advanced zinc-doped adhesives for high performance at the resin-carious dentin interface.

PubMed

Toledano, Manuel; Osorio, Raquel; Osorio, Estrella; García-Godoy, Franklin; Toledano-Osorio, Manuel; Aguilera, Fátima S

2016-09-01

The purpose of this study was to evaluate the remineralization ability of an etch-and-rinse Zn-doped resin applied on caries-affected dentin (CAD). CAD surfaces were subjected to: (i) 37% phosphoric acid (PA) or (ii) 0.5M ethylenediaminetetraacetic acid (EDTA). 10wt% ZnO nanoparticles or 2wt% ZnCl2 were added into the adhesive Single Bond (SB), to create the following groups: PA+SB, PA+SB-ZnO, PA+SB-ZnCl2, EDTA+SB, EDTA+SB-ZnO, EDTA+SB-ZnCl2. Bonded interfaces were submitted to mechanical loading or stored during 24h. Remineralization of the bonded interfaces was studied by AFM nano-indentation (hardness and Young׳s modulus), Raman spectroscopy [mapping with principal component analysis (PCA), and hierarchical cluster analysis (HCA)] and Masson׳s trichrome staining technique. Dentin samples treated with PA+SB-ZnO attained the highest values of nano-mechanical properties. Load cycling increased both mineralization and crystallographic maturity at the interface; this effect was specially noticed when using ZnCl2-doped resin in EDTA-treated carious dentin. Crosslinking attained higher frequencies indicating better conformation and organization of collagen in specimens treated with PA+SB-ZnO, after load cycling. Trichrome staining technique depicted a deeper demineralized dentin fringe that became reduced after loading, and it was not observable in EDTA+SB groups. Multivariate analysis confirmed de homogenizing effect of load cycling in the percentage of variances, traces of centroids and distribution of clusters, especially in specimens treated with EDTA+SB-ZnCl2. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effects of chelating agent and acids on Biodentine.

PubMed

Ballal, V; Marques, J N; Campos, C N; Lima, C O; Simão, R A; Prado, M

2018-06-01

To evaluate the effect of distilled water, ethylenediaminetetraacetic acid (EDTA), phosphoric acid and maleic acid on Biodentine regarding surface topography, microhardness and push-out bond strength (POBS). Fifty-two cylindrical shaped Biodentine specimens were divided into groups: control (distilled water); EDTA (17% EDTA); PA (37% phosphoric acid); and MA (7% maleic acid). Surfaces were evaluated by topographic analysis and Vickers microhardness test. Topographic changes were evaluated qualitatively and microhardness was statistically analyzed by Kruskal-Wallis test. Forty mandibular molars were used to simulate clinical conditions. The crowns were removed and a perforation was created at the furcal floor. The Biodentine was packed into the root perforations and the roots were divided into four groups (DW, EDTA, PA, MA). Samples were stored and subjected to interfacial analysis. POBS data were analyzed by Kruskal-Wallis and Dunn tests. Ethylenediaminetetraacetic acid, MA and PA changed the morphology of the Biodentine surface. PA showed microhardness similar to distilled water (P > 0.05), while MA and EDTA demonstrated reduced values when compared with PA (P < 0.05). PA improved the POBS of Biodentine in comparison with the control. Changes in the topography, microhardness and POBS of Biodentine are associated with irrigant agent used. © 2018 Australian Dental Association.

Decalcification using ethylenediaminetetraacetic acid for clear microstructure imaging of cochlea through optical coherence tomography

NASA Astrophysics Data System (ADS)

Lee, Jaeyul; Kim, Kanghae; Wijesinghe, Ruchire Eranga; Jeon, Doekmin; Lee, Sang Heun; Jeon, Mansik; Jang, Jeong Hun

2016-08-01

The aim of this study was to analyze the effectiveness of decalcification using ethylenediaminetetraacetic acid (EDTA) as an optical clearing method to enhance the depth visibility of internal soft tissues of cochlea. Ex vivo mouse and guinea pig cochlea samples were soaked in EDTA solutions for decalcification, and swept source optical coherence tomography (OCT) was used as imaging modality to monitor the decalcified samples consecutively. The monitored noninvasive cross-sectional images showed that the mouse and guinea pig cochlea samples had to be decalcified for subsequent 7 and 14 days, respectively, to obtain the optimal optical clearing results. Using this method, difficulties in imaging of internal cochlea microstructures of mice could be evaded. The obtained results verified that the depth visibility of the decalcified ex vivo samples was enhanced.

A method for reducing the sloughing of thick blood films for malaria diagnosis.

PubMed

Norgan, Andrew P; Arguello, Heather E; Sloan, Lynne M; Fernholz, Emily C; Pritt, Bobbi S

2013-07-08

The gold standard for malaria diagnosis is the examination of thick and thin blood films. Thick films contain 10 to 20 times more blood than thin films, correspondingly providing increased sensitivity for malaria screening. A potential complication of thick film preparations is sloughing of the blood droplet from the slide during staining or rinsing, resulting in the loss of sample. In this work, two methods for improving thick film slide adherence ('scratch' (SCM) and 'acetone dip' (ADM) methods) were compared to the 'standard method' (SM) of thick film preparation. Standardized blood droplets from 26 previously examined EDTA whole blood specimens (22 positive and four negative) were concurrently spread on glass slides using the SM, ADM, and SCM. For the SM and ADM prepared slides, the droplet was gently spread to an approximate 22 millimeters in diameter spot on the slide using the edge of a second glass slide. For the SCM, the droplet was spread by carefully grinding (or scratching) it into the slide with the point of a second glass slide. Slides were dried for one hour in a laminar flow hood. For the ADM, slides were dipped once in an acetone filled Coplin jar and allowed to air dry. All slides were then Giemsa-stained and examined in a blinded manner. Adherence was assessed by blinded reviewers. No significant or severe defects were observed for slides prepared with the SCM. In contrast, 8 slides prepared by the ADM and 3 prepared using the SM displayed significant or severe defects. Thick films prepared by the three methods were microscopically indistinguishable and concordant results (positive or negative) were obtained for the three methods. Estimated parasitaemia of the blood samples ranged from 25 to 429,169 parasites/μL of blood. The SCM is an inexpensive, rapid, and simple method that improves the adherence of thick blood films to standard glass slides without altering general slide preparation, microscopic appearance or interpretability. Using the SCM, thick films can be reliably examined less than two hours after sample receipt. This represents a significant diagnostic improvement over protocols requiring extended drying periods.

Microwave assisted extraction for trace element analysis of plant materials by ICP-AES.

PubMed

Borkowska-Burnecka, J

2000-11-01

Application of microwave assisted extraction for the decomposition and dissolution of plant samples for trace metal determination by ICP-AES was examined. Dried onion, leaves of spinach beet and three reference materials CTA-OTL-1, CTA-VTL-2 and CL-1 were analyzed. Water, EDTA and hydrochloric acid (0.01, 0.10 and 1.0 M, respectively) were used as leaching solutions. The extraction efficiency was investigated by comparison of the results with those obtained after microwave wet digestion. HCl was found to be very suitable for quantitative extraction of B, Ba, Cd, Cu, Mn, Ni, Pb, Sr and Zn from the samples. For reference materials, the measured concentrations are well consistent with the certified values. The use of EDTA led to a complete extraction of B, Cd, Ni, Pb, Sr and Zn. Water was found to be a good leaching solution for boron. For extraction with HCl and EDTA, the RSD values for the concentrations measured were below 8% for most of the elements.

Digital-Direct-RT-PCR: a sensitive and specific method for quantification of CTC in patients with cervical carcinoma

PubMed Central

Pfitzner, Claudia; Schröder, Isabel; Scheungraber, Cornelia; Dogan, Askin; Runnebaum, Ingo Bernhard; Dürst, Matthias; Häfner, Norman

2014-01-01

The detection of circulating tumour cells (CTC) in cancer patients may be useful for therapy monitoring and prediction of relapse. A sensitive assay based on HPV-oncogene transcripts which are highly specific for cervical cancer cells was established. The Digital-Direct-RT-PCR (DD-RT-PCR) combines Ficoll-separation, ThinPrep-fixation and one-step RT-PCR in a low-throughput digital-PCR format enabling the direct analysis and detection of individual CTC without RNA isolation. Experimental samples demonstrated a sensitivity of one HPV-positive cell in 500,000 HPV-negative cells. Spike-in experiments with down to 5 HPV-positive cells per millilitre EDTA-blood resulted in concordant positive results by PCR and immunocytochemistry. Blood samples from 3 of 10 CxCa patients each contained a single HPV-oncogene transcript expressing CTC among 5 to 15*105 MNBC. Only 1 of 7 patients with local but 2 of 3 women with systemic disease had CTC. This highly sensitive DD-RT-PCR for the detection of CTC may also be applied to other tumour entities which express tumour-specific transcripts. Abbreviations: CTC – circulating tumour cells, CxCa – cervical cancer, DD-RT-PCR – Digital-Direct Reverse Transcriptase PCR, HPV – Human Papilloma Virus, MNBC – mononuclear blood cells, ICC – immunocytochemistry. PMID:24496006

Accuracy of the Precision® point-of-care ketone test examined by liquid chromatography tandem-mass spectrometry (LC-MS/MS) in the same fingerstick sample.

PubMed

Janssen, Marcel J W; Hendrickx, Ben H E; Habets-van der Poel, Carin D; van den Bergh, Joop P W; Haagen, Anton A M; Bakker, Jaap A

2010-12-01

The Precision(®) (Abbott Diabetes Care) point-of-care biosensor test strips are widely used by patients with diabetes and clinical laboratories for measurement of plasma β-hydroxybutyrate (β-HB) concentrations in capillary blood samples obtained by fingerstick. In the literature, this procedure has been validated only against the enzymatic determination of β-HB in venous plasma, i.e., the method to which the Precision(®) has been calibrated. In this study, the Precision(®) Xceed was compared to a methodologically different and superior procedure: determination of β-HB by liquid chromatography tandem-mass spectrometry (LC-MS/MS) in capillary blood spots. Blood spots were obtained from the same fingerstick sample from out of which Precision(®) measurements were performed. Linearity was tested by adding varying amounts of standard to an EDTA venous whole blood matrix. The Precision(®) was in good agreement with LC-MS/MS within the measuring range of 0.0-6.0 mmol/L (Passing and Bablok regression: slope=1.20 and no significant intercept, R=0.97, n=59). Surprisingly, the Precision(®) showed non-linearity and full saturation at concentrations above 6.0 mmol/L, which were confirmed by a standard addition experiment. Results obtained at the saturation level varied between 3.0 and 6.5 mmol/L. The Precision(®) β-HB test strips demonstrate good comparison with LC-MS/MS. Inter-individual variation around the saturation level, however, is large. Therefore, we advise reporting readings above 3.0 as >3.0 mmol/L. The test is valid for use in the clinically relevant range of 0.0-3.0 mmol/L.

Adhesive blood microsampling systems for steroid measurement via LC-MS/MS in the rat.

PubMed

Heussner, Kirsten; Rauh, Manfred; Cordasic, Nada; Menendez-Castro, Carlos; Huebner, Hanna; Ruebner, Matthias; Schmidt, Marius; Hartner, Andrea; Rascher, Wolfgang; Fahlbusch, Fabian B

2017-04-01

Liquid Chromatography Tandem Mass Spectrometry (LC-MS/MS) allows for the direct analysis of multiple hormones in a single probe with minimal sample volume. Rodent-based animal studies strongly rely on microsampling, such as the dry blood spot (DBS) method. However, DBS suffers the drawback of hematocrit-dependence (non-volumetric). Hence, novel volumetric microsampling techniques were introduced recently, allowing sampling of fixed accurate volumes. We compared these methods for steroid analysis in the rat to improve inter-system comparability. We analyzed steroid levels in blood using the absorptive microsampling devices Whatman® 903 Protein Saver Cards, Noviplex™ Plasma Prep Cards and the Mitra™ Microsampling device and compared the obtained results to the respective EDTA plasma levels. Quantitative steroid analysis was performed via LC-MS/MS. For the determination of the plasma volume factor for each steroid, their levels in pooled blood samples from each human adults and rats (18weeks) were compared and the transferability of these factors was evaluated in a new set of juvenile (21days) and adult (18weeks) rats. Hematocrit was determined concomitantly. Using these approaches, we were unable to apply one single volume factor for each steroid. Instead, plasma volume factors had to be adjusted for the recovery rate of each steroid and device individually. The tested microsampling systems did not allow the use of one single volume factor for adult and juvenile rats based on an unexpectedly strong hematocrit-dependency and other steroid specific (pre-analytic) factors. Our study provides correction factors for LC-MS/MS steroid analysis of volumetric and non-volumetric microsampling systems in comparison to plasma. It argues for thorough analysis of chromatographic effects before the use of novel volumetric systems for steroid analysis. Copyright © 2017 Elsevier Inc. All rights reserved.

Usefulness of real-time PCR for lytA, ply, and Spn9802 on plasma samples for the diagnosis of pneumococcal pneumonia.

PubMed

Abdeldaim, G; Herrmann, B; Mölling, P; Holmberg, H; Blomberg, J; Olcén, P; Strålin, K

2010-08-01

In the present study, we evaluated rapid real-time PCR assays for ply, Spn9802, and lytA applied to plasma samples for the detection of Streptococcus pneumoniae in patients with community-acquired pneumonia (CAP). In a prospective study of CAP aetiology, an EDTA plasma sample was collected together with blood culture in 92 adult CAP patients and 91 adult controls. Among the 92 CAP patients, lytA PCR was positive in eight (9%), Spn9802 PCR was positive in 11 (12%) and ply PCR was positive in 19 (21%) cases. Of 91 controls, the ply PCR was positive in eight cases (9%), but no positive cases were noted by Spn9802 or lytA PCRs. Ten CAP patients had pneumococcal bacteraemia. Compared to blood culture, PCR for lytA, Spn9802 and ply had sensitivities of 70% (7/10), 60% (6/10) and 70% (7/10), and specificities of 96% (79/82), 94% (77/82) and 85% (70/82) respectively. With blood culture and/or culture of representative sputum, and/or urinary antigen detection, S. pneumoniae was identified in 31 CAP patients. Compared to these tests in combination, PCR for lytA, Spn9802 and ply showed sensitivities of 26% (8/31), 32% (10/31) and 42% (13/31), and specificities of 100% (61/61), 98% (60/61) and 90% (55/61) respectively. We conclude that Spn9802 and lytA PCRs may be useful for the rapid detection of bacteraemic pneumococcal pneumonia, whereas ply PCR is not specific enough for routine use and blood PCR with small plasma volumes is not useful for the detection of nonbacteraemic pneumococcal pneumonia.

[Clinical research of EDTA pretreatment on the bonding strength of resin].

PubMed

Li, Ting-ting; Sun, Min-min; Kang, Jun-tang; Sun, Zhe

2015-10-01

This study was to evaluate clinical results of adhesives to sclerotic dentin in non-carious cervical lesions pretreated with ethylene diamine tetraacetic acid (EDTA). Twelve patients who had at least one pair of teeth with typical cervical wedge-shaped defects and class III sclerotic dentin were selected. The teeth were randomly divided into experimental and control groups. In the experimental group, EDTA was used to pretreat dentin surface first and then self-etching resin adhesive; In the control group, only self-etching resin adhesive was used. Modified USPHS rating system was applied in baseline, 6 months and 12 months after treatment. SAS 8.02 software package was used for two-sample rank sum test. At the time point of 6-month and 12-month, the scores in the experimental group were significantly better than the control group (P<0.05). EDTA pretreatment of the sclerotic dentin of cervical wedge-shaped defects could improve the resin bonding strength, marginal discoloration, marginal form and surface quality.

Lead poisoning in whooper and tundra swans.

PubMed

Nakade, Tetsuya; Tomura, Yoshihiro; Jin, Kazuo; Taniyama, Hiroyuki; Yamamoto, Mutsuki; Kikkawa, Aya; Miyagi, Kunitaro; Uchida, Eiji; Asakawa, Mitsuhiko; Mukai, Takeshi; Shirasawa, Masahiko; Yamaguchi, Mamoru

2005-01-01

Six weak whooper swans (Cygnus cygnus) and two weak tundra swans (Cygnus columbianus) were found at Swamp Miyajima (Hokkaido, Japan) in May 1998. Anorexia, depression, green watery feces, pale conjunctiva, and anemia were observed. Radiographs showed from six to 38 suspected lead pellets in the gizzard. Blood lead concentrations were 2.5-6.7 microg/g (mean+/-SD=4.6+/-1.14 microg/g) on day 1. After blood collection, the birds were treated with calcium disodium ethylenediaminetetraacetate (CaEDTA) given intravenously and force fed. Despite treatment, seven birds died the next day. Green, bile-stained livers and pale or green kidneys were observed on necropsy. Microscopically, bile pigment was widespread in the liver and acid-fast intranuclear inclusion bodies were observed in renal tubular epithelium. Lead concentrations in livers and kidneys were 14.0-30.4 microg/g and 30.2-122 microg/g wet weight, respectively. Only one bird survived and this whooper swan continued to be treated with CaEDTA and activated charcoal. No lead shot was observed in the proventriculus and gizzard by radiography on day 64 and the blood lead concentration decreased from 2.9 microg/g to 0.09 microg/g during that same period. After 4 mo of rehabilitation, the whooper swan was returned to the wild. Lead intoxication continues to be a problem at Swamp Miyajima.

Mössbauer studies on some Argentinian soil: Mollisols from Bahia Blanca

NASA Astrophysics Data System (ADS)

Saragovi, C.; Labenski, F.; Duhalde, S. M.; Acebal, S.; Venegas, R.

1994-12-01

Clay fractions of a Mollisol sample as is, treated with ammonium oxalate (AO), with dithionite-citrate-bicarbonate (DCB) and with dithionite-ethilene-diamine-tetraacetic acid (D-EDTA) methods, were studied. Illite-montmorillonites together with hematites, goethites and maghemites, all of the AI-substituted and with a wide range of sizes, were identified. It is found that the AO attack extracts little iron, whereas the other two attacks extract the magnetic signal. Furthermore, the DCB attack facilitates the reduction of the Fe3+ ions, while the D-EDTA method does not. Instead, this attack extracts more clay mineral Fe ions. A comparison with large grain soil samples is made.

Silver Nanoparticle-Enhanced Resonance Raman Sensor of Chromium(III) in Seawater Samples.

PubMed

Ly, Nguyễn Hoàng; Joo, Sang-Woo

2015-04-29

Tris(hydroxymethyl)aminomethane ethylenediaminetetraacetic acid (Tris-EDTA), upon binding Cr(III) in aqueous solutions at pH 8.0 on silver nanoparticles (AgNPs), was found to provide a sensitive and selective Raman marker band at ~563 cm-1, which can be ascribed to the metal-N band. UV-Vis absorption spectra also supported the aggregation and structural change of EDTA upon binding Cr(III). Only for Cr(III) concentrations above 500 nM, the band at ~563 cm-1 become strongly intensified in the surface-enhanced Raman scattering spectra. This band, due to the metal-EDTA complex, was not observed in the case of 50 mM of K+, Cd2+, Mg2+, Ca2+, Mn2+, Co2+, Na+, Cu2+, NH4+, Hg2+, Ni2+, Fe3+, Pb2+, Fe2+, and Zn2+ ions. Seawater samples containing K, Mg, Ca, and Na ion concentrations higher than 8 mM also showed the characteristic Raman band at ~563 cm-1 above 500 nM, validating our method. Our approach may be useful in detecting real water samples by means of AgNPs and Raman spectroscopy.

Is automated kinetic measurement superior to end-point for advanced oxidation protein product?

PubMed

Oguz, Osman; Inal, Berrin Bercik; Emre, Turker; Ozcan, Oguzhan; Altunoglu, Esma; Oguz, Gokce; Topkaya, Cigdem; Guvenen, Guvenc

2014-01-01

Advanced oxidation protein product (AOPP) was first described as an oxidative protein marker in chronic uremic patients and measured with a semi-automatic end-point method. Subsequently, the kinetic method was introduced for AOPP assay. We aimed to compare these two methods by adapting them to a chemistry analyzer and to investigate the correlation between AOPP and fibrinogen, the key molecule responsible for human plasma AOPP reactivity, microalbumin, and HbA1c in patients with type II diabetes mellitus (DM II). The effects of EDTA and citrate-anticogulated tubes on these two methods were incorporated into the study. This study included 93 DM II patients (36 women, 57 men) with HbA1c levels > or = 7%, who were admitted to the diabetes and nephrology clinics. The samples were collected in EDTA and in citrate-anticoagulated tubes. Both methods were adapted to a chemistry analyzer and the samples were studied in parallel. In both types of samples, we found a moderate correlation between the kinetic and the endpoint methods (r = 0.611 for citrate-anticoagulated, r = 0.636 for EDTA-anticoagulated, p = 0.0001 for both). We found a moderate correlation between fibrinogen-AOPP and microalbumin-AOPP levels only in the kinetic method (r = 0.644 and 0.520 for citrate-anticoagulated; r = 0.581 and 0.490 for EDTA-anticoagulated, p = 0.0001). We conclude that adaptation of the end-point method to automation is more difficult and it has higher between-run CV% while application of the kinetic method is easier and it may be used in oxidative stress studies.

A combinatory approach for analysis of protein sets in barley sieve-tube samples using EDTA-facilitated exudation and aphid stylectomy.

PubMed

Gaupels, Frank; Knauer, Torsten; van Bel, Aart J E

2008-01-01

This study investigated advantages and drawbacks of two sieve-tube sap sampling methods for comparison of phloem proteins in powdery mildew-infested vs. non-infested Hordeum vulgare plants. In one approach, sieve tube sap was collected by stylectomy. Aphid stylets were cut and immediately covered with silicon oil to prevent any contamination or modification of exudates. In this way, a maximum of 1muL pure phloem sap could be obtained per hour. Interestingly, after pathogen infection exudation from microcauterized stylets was reduced to less than 40% of control plants, suggesting that powdery mildew induced sieve tube-occlusion mechanisms. In contrast to the laborious stylectomy, facilitated exudation using EDTA to prevent calcium-mediated callose formation is quick and easy with a large volume yield. After two-dimensional (2D) electrophoresis, a digital overlay of the protein sets extracted from EDTA solutions and stylet exudates showed that some major spots were the same with both sampling techniques. However, EDTA exudates also contained large amounts of contaminative proteins of unknown origin. A combinatory approach may be most favourable for studies in which the protein composition of phloem sap is compared between control and pathogen-infected plants. Facilitated exudation may be applied for subtractive identification of differentially expressed proteins by 2D/mass spectrometry, which requires large amounts of protein. A reference gel loaded with pure phloem sap from stylectomy may be useful for confirmation of phloem origin of candidate spots by digital overlay. The method provides a novel opportunity to study differential expression of phloem proteins in monocotyledonous plant species.

Platelets in blood stored in untreated and siliconed glass bottles and plastic bags

PubMed Central

Kissmeyer-Nielsen, F.; Madsen, C. B.; Nedergaard, Jytte

1961-01-01

Platelet survival was determined using untreated and siliconed glass bottles and plastic bags (Fenwal) for collecting and storing blood. The platelets were tagged in vivo with P32 in six polycythaemic patients undergoing treatment with P32. The results showed that fresh ACD blood collected in untreated glass, siliconed glass, and plastic gave the same recovery of platelets in the recipients. The use of EDTA (Fenwal formula) as anticoagulant gave results inferior to those obtained with blood using ACD as anticoagulant. Even after storage up to 24 hours in untreated glass bottles (ordinary bank blood) a satisfactory recovery of platelets was observed. After storage for 72 hours the recovery was less but not negligible. PMID:14456481

Collection and analysis of peritoneal fluid from healthy llamas and alpacas.

PubMed

Cebra, Christopher K; Tornquist, Susan J; Reed, Shannon K

2008-05-01

To describe a technique for abdominocentesis in camelids and report peritoneal fluid biochemical and cytologic findings from healthy llamas and alpacas. Prospective study. Animals-17 adult llamas and 5 adult alpacas. Right paracostal abdominocentesis was performed. Peritoneal fluid was collected by gravity flow into tubes containing potassium-EDTA for cell count and cytologic evaluation and lithium heparin for biochemical analysis. Blood samples were collected via jugular venipuncture into heparinized tubes at the same time. Cytologic components were quantified. Fluid pH and concentrations of total carbon dioxide, sodium, potassium, chloride, lactate, and glucose were compared between peritoneal fluid and venous blood. All but 3 camelids had peritoneal fluid cell counts of < 3,000 nucleated cells/microL, with < 2,000 neutrophils/microL and < 1,040 large mononuclear cells/microL. All but 1 had peritoneal fluid protein concentrations of > or = 2.5 g/dL. Peritoneal fluid of camelids generally contained slightly less glucose, lactate, and sodium and roughly equal concentrations of potassium and chloride as venous blood. Peritoneal fluid was collected safely from healthy camelids. Compared with blood, peritoneal fluid usually had a low cell count and protein concentration, but some individuals had higher values. Electrolyte concentrations resembled those found in blood. High cell counts and protein concentrations found in peritoneal fluid of some healthy camelids may overlap with values found in diseased camelids, complicating interpretation of peritoneal fluid values.

Clinical efficacy of EDTA ultrasonic activation in the reduction of endotoxins and cultivable bacteria.

PubMed

Herrera, D R; Martinho, F C; de-Jesus-Soares, A; Zaia, A A; Ferraz, C C R; Almeida, J F A; Gomes, B P F A

2017-10-01

This clinical study was conducted to investigate the influence of 17% ethylenediaminetetraacetic acid (EDTA) ultrasonic activation after chemomechanical preparation (CMP) on eliminating/reducing oral bacterial lipopolysaccharides (known as endotoxins) and cultivable bacteria in teeth with pulp necrosis and apical periodontitis. Samples were taken from 24 root canals at several clinical periods: S1 - before CMP; S2 - after CMP; S3 - after EDTA: G1 - with ultrasonic activation (n = 12) and G2 - without ultrasonic activation (n = 12). Root canals were instrumented using Mtwo rotary files. Culture techniques were used to determine the number of colony-forming units (CFU). Limulus amebocyte lysate (LAL) was used to measure endotoxin levels. Friedman's and Wilcoxon signed-rank tests were used to compare the amount of bacteria and endotoxin levels in each period (P < 0.05). Endotoxins and cultivable bacteria were recovered in 100% of the initial samples (S1). CMP was effective in reducing endotoxins and bacterial load (all with P < 0.05). Higher values of endotoxin reduction were achieved with EDTA ultrasonic activation [G1, 0.02 EU mL -1 (range 0.01-0.75)] compared with the no activation group [G2, 1.13 EU mL -1 (range 0.01-8.34)] (P < 0.05). Regarding bacterial reduction, no statistically significant difference was found in S3, regardless of the group (G1, G2, P > 0.05). Chemomechanical preparation was effective in reducing bacteria and endotoxins, but could not completely eliminate them. The ultrasonic activation of EDTA was effective in further reducing endotoxin levels in the root canals of teeth with pulp necrosis and apical periodontitis. © 2016 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Biomarkers and Molecular Analysis to Improve Bloodstream Infection Diagnostics in an Emergency Care Unit

PubMed Central

Loonen, Anne J. M.; de Jager, Cornelis P. C.; Tosserams, Janna; Kusters, Ron; Hilbink, Mirrian; Wever, Peter C.; van den Brule, Adriaan J. C.

2014-01-01

Molecular pathogen detection from blood is still expensive and the exact clinical value remains to be determined. The use of biomarkers may assist in preselecting patients for immediate molecular testing besides blood culture. In this study, 140 patients with ≥ 2 SIRS criteria and clinical signs of infection presenting at the emergency department of our hospital were included. C-reactive protein (CRP), neutrophil-lymphocyte count ratio (NLCR), procalcitonin (PCT) and soluble urokinase plasminogen activator receptor (suPAR) levels were determined. One ml EDTA blood was obtained and selective pathogen DNA isolation was performed with MolYsis (Molzym). DNA samples were analysed for the presence of pathogens, using both the MagicPlex Sepsis Test (Seegene) and SepsiTest (Molzym), and results were compared to blood cultures. Fifteen patients had to be excluded from the study, leaving 125 patients for further analysis. Of the 125 patient samples analysed, 27 presented with positive blood cultures of which 7 were considered to be contaminants. suPAR, PCT, and NLCR values were significantly higher in patients with positive blood cultures compared to patients without (p < 0.001). Receiver operating characteristic curves of the 4 biomarkers for differentiating bacteremia from non-bacteremia showed the highest area under the curve (AUC) for PCT (0.806 (95% confidence interval 0.699–0.913)). NLCR, suPAR and CRP resulted in an AUC of 0.770, 0.793, and 0.485, respectively. When compared to blood cultures, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for SepsiTest and MagicPlex Sepsis Test were 11%, 96%, 43%, 80%, and 37%, 77%, 30%, 82%, respectively. In conclusion, both molecular assays perform poorly when one ml whole blood is used from emergency care unit patients. NLCR is a cheap, fast, easy to determine, and rapidly available biomarker, and therefore seems most promising in differentiating BSI from non-BSI patients for subsequent pathogen identification using molecular diagnostics. PMID:24475269

Biomarkers and molecular analysis to improve bloodstream infection diagnostics in an emergency care unit.

PubMed

Loonen, Anne J M; de Jager, Cornelis P C; Tosserams, Janna; Kusters, Ron; Hilbink, Mirrian; Wever, Peter C; van den Brule, Adriaan J C

2014-01-01

Molecular pathogen detection from blood is still expensive and the exact clinical value remains to be determined. The use of biomarkers may assist in preselecting patients for immediate molecular testing besides blood culture. In this study, 140 patients with ≥ 2 SIRS criteria and clinical signs of infection presenting at the emergency department of our hospital were included. C-reactive protein (CRP), neutrophil-lymphocyte count ratio (NLCR), procalcitonin (PCT) and soluble urokinase plasminogen activator receptor (suPAR) levels were determined. One ml EDTA blood was obtained and selective pathogen DNA isolation was performed with MolYsis (Molzym). DNA samples were analysed for the presence of pathogens, using both the MagicPlex Sepsis Test (Seegene) and SepsiTest (Molzym), and results were compared to blood cultures. Fifteen patients had to be excluded from the study, leaving 125 patients for further analysis. Of the 125 patient samples analysed, 27 presented with positive blood cultures of which 7 were considered to be contaminants. suPAR, PCT, and NLCR values were significantly higher in patients with positive blood cultures compared to patients without (p < 0.001). Receiver operating characteristic curves of the 4 biomarkers for differentiating bacteremia from non-bacteremia showed the highest area under the curve (AUC) for PCT (0.806 (95% confidence interval 0.699-0.913)). NLCR, suPAR and CRP resulted in an AUC of 0.770, 0.793, and 0.485, respectively. When compared to blood cultures, the sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) for SepsiTest and MagicPlex Sepsis Test were 11%, 96%, 43%, 80%, and 37%, 77%, 30%, 82%, respectively. In conclusion, both molecular assays perform poorly when one ml whole blood is used from emergency care unit patients. NLCR is a cheap, fast, easy to determine, and rapidly available biomarker, and therefore seems most promising in differentiating BSI from non-BSI patients for subsequent pathogen identification using molecular diagnostics.

Extracting natural dyes from wool--an evaluation of extraction methods.

PubMed

Manhita, Ana; Ferreira, Teresa; Candeias, António; Dias, Cristina Barrocas

2011-05-01

The efficiency of eight different procedures used for the extraction of natural dyes was evaluated using contemporary wool samples dyed with cochineal, madder, woad, weld, brazilwood and logwood. Comparison was made based on the LC-DAD peak areas of the natural dye's main components which had been extracted from the wool samples. Among the tested methods, an extraction procedure with Na(2)EDTA in water/DMF (1:1, v/v) proved to be the most suitable for the extraction of the studied dyes, which presented a wide range of chemical structures. The identification of the natural dyes used in the making of an eighteenth century Arraiolos carpet was possible using the Na(2)EDTA/DMF extraction of the wool embroidery samples and an LC-DAD-MS methodology. The effectiveness of the Na(2)EDTA/DMF extraction method was particularly observed in the extraction of weld dye components. Nine flavone derivatives previously identified in weld extracts could be identified in a single historical sample, confirming the use of this natural dye in the making of Arraiolos carpets. Indigo and brazilwood were also identified in the samples, and despite the fact that these natural dyes were referred in the historical recipes of Arraiolos dyeing, it is the first time that the use of brazilwood is confirmed. Mordant analysis by ICP-MS identified the widespread use of alum in the dyeing process, but in some samples with darker hues, high amounts of iron were found instead.

A nationwide multicentre study in Turkey for establishing reference intervals of haematological parameters with novel use of a panel of whole blood

PubMed Central

Ozarda, Yesim; Ichihara, Kiyoshi; Bakan, Ebubekir; Polat, Harun; Ozturk, Nurinnisa; Baygutalp, Nurcan K.; Taneli, Fatma; Guvenc, Yesim; Ormen, Murat; Erbayraktar, Zubeyde; Aksoy, Nurten; Sezen, Hatice; Demir, Meltem; Eskandari, Gulcin; Polat, Gurbuz; Mete, Nuriye; Yuksel, Hatice; Vatansev, Husamettin; Gun, Fatma; Akin, Okhan; Ceylan, Ozlem; Noyan, Tevfik; Gozlukaya, Ozgul; Aliyazicioglu, Yuksel; Kahraman, Sevim; Dirican, Melahat; Tuncer, Gul Ozlem; Kimura, Shogo; Eker, Pinar

2017-01-01

Introduction A nationwide multicentre study was conducted to establish well-defined reference intervals (RIs) of haematological parameters for the Turkish population in consideration of sources of variation in reference values (RVs). Materials and methods K2-EDTA whole blood samples (total of 3363) were collected from 12 laboratories. Sera were also collected for measurements of iron, UIBC, TIBC, and ferritin for use in the latent abnormal values exclusion (LAVE) method. The blood samples were analysed within 2 hours in each laboratory using Cell Dyn and Ruby (Abbott), LH780 (Beckman Coulter), or XT-2000i (Sysmex). A panel of freshly prepared blood from 40 healthy volunteers was measured in common to assess any analyser-dependent bias in the measurements. The SD ratio (SDR) based on ANOVA was used to judge the need for partitioning RVs. RIs were computed by the parametric method with/without applying the LAVE method. Results Analyser-dependent bias was found for basophils (Bas), MCHC, RDW and MPV from the panel test results and thus those RIs were derived for each manufacturer. RIs were determined from all volunteers’ results for WBC, neutrophils, lymphocytes, monocytes, eosinophils, MCV, MCH and platelets. Gender-specific RIs were required for RBC, haemoglobin, haematocrit, iron, UIBC and ferritin. Region-specific RIs were required for RBC, haemoglobin, haematocrit, UIBC, and TIBC. Conclusions With the novel use of a freshly prepared blood panel, manufacturer-specific RIs’ were derived for Bas, Bas%, MCHC, RDW and MPV. Regional differences in RIs were observed among the 7 regions of Turkey, which may be attributed to nutritional or environmental factors, including altitude. PMID:28694726

Evaluation of two portable meters for determination of blood triglyceride concentration in dogs.

PubMed

Kluger, Elissa K; Dhand, Navneet K; Malik, Richard; Ilkin, William J; Snow, David H; Govendir, Merran

2010-02-01

To evaluate agreement between 2 portable triglyceride meters and a veterinary laboratory for measurement of blood triglyceride concentrations in dogs and evaluate effects of Hct and blood volume analyzed. 97 blood samples collected from 60 dogs. Triglyceride concentrations were measured in blood by use of 2 meters and compared with serum triglyceride concentrations determined by a veterinary laboratory. Within- and between-day precision, accuracy, and effects of blood volume and Hct were analyzed. Accuracy of both meters varied with triglyceride concentration, although both accurately delineated dogs with triglyceride concentrations < 180 mg/dL versus > or = 180 mg/dL. One meter had results with excellent overall correlation with results of the standard laboratory method, with a concordance correlation coefficient of 0.94 and mean difference of 20.3 mg/dL. The other meter had a good overall concordance correlation coefficient of 0.86 with a higher absolute mean difference of -27.7 mg/dL. Results were only affected by blood volume; triglyceride concentrations determined via both meters were significantly lower when 7 microL of EDTA-anticoagulated blood was used, compared with larger volumes. 1 meter had greater accuracy in the range of 140 to 400 mg/dL and was therefore well suited to detect hypertriglyceridemia. The other meter was accurate with triglyceride values < 140 mg/dL and yielded results similar to those of the veterinary laboratory in the range of 140 to 400 mg/dL, therefore being suitable for determination of triglyceride concentrations in nonfed dogs and dogs with mildly high concentrations.

First external quality assurance program for bloodstream Real-Time PCR monitoring of treatment response in clinical trials of Chagas disease.

PubMed

Ramírez, Juan C; Parrado, Rudy; Sulleiro, Elena; de la Barra, Anabelle; Rodríguez, Marcelo; Villarroel, Sandro; Irazu, Lucía; Alonso-Vega, Cristina; Alves, Fabiana; Curto, María A; García, Lineth; Ortiz, Lourdes; Torrico, Faustino; Gascón, Joaquim; Flevaud, Laurence; Molina, Israel; Ribeiro, Isabela; Schijman, Alejandro G

2017-01-01

Real-Time PCR (qPCR) testing is recommended as both a diagnostic and outcome measurement of etiological treatment in clinical practice and clinical trials of Chagas disease (CD), but no external quality assurance (EQA) program provides performance assessment of the assays in use. We implemented an EQA system to evaluate the performance of molecular biology laboratories involved in qPCR based follow-up in clinical trials of CD. An EQA program was devised for three clinical trials of CD: the E1224 (NCT01489228), a pro-drug of ravuconazole; the Sampling Study (NCT01678599), that used benznidazole, both conducted in Bolivia; and the CHAGASAZOL (NCT01162967), that tested posaconazole, conducted in Spain. Four proficiency testing panels containing negative controls and seronegative blood samples spiked with 1, 10 and 100 parasite equivalents (par. eq.)/mL of four Trypanosoma cruzi stocks, were sent from the Core Lab in Argentina to the participating laboratories located in Bolivia and Spain. Panels were analyzed simultaneously, blinded to sample allocation, at 4-month intervals. In addition, 302 random blood samples from both trials carried out in Bolivia were sent to Core Lab for retesting analysis. The analysis of proficiency testing panels gave 100% of accordance (within laboratory agreement) and concordance (between laboratory agreement) for all T. cruzi stocks at 100 par. eq./mL; whereas their values ranged from 71 to 100% and from 62 to 100% at 1 and 10 par. eq./mL, respectively, depending on the T. cruzi stock. The results obtained after twelve months of preparation confirmed the stability of blood samples in guanidine-EDTA buffer. No significant differences were found between qPCR results from Bolivian laboratory and Core Lab for retested clinical samples. This EQA program for qPCR analysis of CD patient samples may significantly contribute to ensuring the quality of laboratory data generated in clinical trials and molecular diagnostics laboratories of CD.

Mobilized plasma lead as an index of lead body burden and its relation to the heme-related indices.

PubMed

Sakai, T; Ushio, K; Ikeya, Y

1998-07-01

Plasma lead (Pb-P) from workers were distributed in two main fractions: a protein bound fraction and low molecular weight fractions. Lead mobilized into plasma by CaEDTA was mainly observed in the low molecular weight fraction corresponding to lead disodium ethylenediamine tetraacetic acid (PbEDTA). The peak levels of Pb-P was attained around 1.5 and 2.5 hours after the start of CaEDTA injection. Pb-P and blood lead levels (Pb-B) at 2 h after the injection were 4.26 (+/- 2.84) and 0.96 (+/- 0.27) fold of the initial levels just before the injection. Pb-P concentrations at 2 hours after the start of CaEDTA injection (MPb-P) were well correlated (r = 0.740) with amounts of lead excreted in urine for 24 h thereafter (MPb-U). log MPb-P as well as log MPb-U were correlated with Pb-B (r = 0.765 and 0.817, respectively). Correlation coefficients of lead body burden (MPb-P or MPb-U) vs the logarithms of the effect indices (delta-aminolevulinic acid (ALA) dehydratase, ALA in urine, coproporphyrin in urine, and erythrocyte zinc protoporphyrin) were higher than the correlation coefficients of exposure indices (Pb-B or Pb-U) vs the logarithms of the effect indices. Thus the biological effect monitoring is significant and reliable for evaluating the functional components of lead body burden (MPb-P or MPb-U).

Reexamination of the ORAC assay: effect of metal ions.

PubMed

Nkhili, E; Brat, P

2011-05-01

The oxygen radical absorbance capacity (ORAC) assay method has been employed extensively in the field of antioxidant and oxidative stress. It uses fluorescein as probe for oxidation by peroxyl radical. Hundreds of reports have been published on the use of this method to determine antioxidant capacity in food and biological samples. The question is whether the results of all these reports are influenced by antioxidant autoxidation, which occurs during the ORAC test. Indeed, the presence of metal ions in the studied matrix will influence antioxidant stability, thereby leading to the underestimation of their antioxidant properties. Ethylenediaminetetraacetic acid hydrate (EDTA) can be used as a metal complexation agent. This paper examines the effect of the addition of EDTA on the ORAC values of pure compounds (quercetin, ascorbic, and dehydroascorbic acid) and five food juices (kiwi, orange, tomato, red grape, and apple). Metal complexation by EDTA (80 μM) clearly increased the ORAC values, given that the antioxidant was protected against rapid autoxidation incited by trace metal ions within samples and then by free radicals. Our finding also undoubtedly demonstrated that the number of literature values is potentially underestimated.

Fully automated, internally controlled quantification of hepatitis B Virus DNA by real-time PCR by use of the MagNA Pure LC and LightCycler instruments.

PubMed

Leb, Victoria; Stöcher, Markus; Valentine-Thon, Elizabeth; Hölzl, Gabriele; Kessler, Harald; Stekel, Herbert; Berg, Jörg

2004-02-01

We report on the development of a fully automated real-time PCR assay for the quantitative detection of hepatitis B virus (HBV) DNA in plasma with EDTA (EDTA plasma). The MagNA Pure LC instrument was used for automated DNA purification and automated preparation of PCR mixtures. Real-time PCR was performed on the LightCycler instrument. An internal amplification control was devised as a PCR competitor and was introduced into the assay at the stage of DNA purification to permit monitoring for sample adequacy. The detection limit of the assay was found to be 200 HBV DNA copies/ml, with a linear dynamic range of 8 orders of magnitude. When samples from the European Union Quality Control Concerted Action HBV Proficiency Panel 1999 were examined, the results were found to be in acceptable agreement with the HBV DNA concentrations of the panel members. In a clinical laboratory evaluation of 123 EDTA plasma samples, a significant correlation was found with the results obtained by the Roche HBV Monitor test on the Cobas Amplicor analyzer within the dynamic range of that system. In conclusion, the newly developed assay has a markedly reduced hands-on time, permits monitoring for sample adequacy, and is suitable for the quantitative detection of HBV DNA in plasma in a routine clinical laboratory.

The morphological changes of optically cleared cochlea using optical coherence tomography (Conference Presentation)

NASA Astrophysics Data System (ADS)

Lee, Jaeyul; Song, Jaewon; Jeon, Mansik; Kim, Jeehyun

2017-02-01

In this study, we monitored the optical clearing effects by immersing ex vivo guinea pig cochlea samples in ethylenediaminetetraacetic acid (EDTA) to study the internal microstructures in the morphology of guinea pig cochlea. The imaging limitations due to the guinea pig cochlea structures were overcome by optical clearing technique. Subsequently, the study was carried out to confirm the required approximate immersing duration of cochlea in EDTA-based optical clearing to obtain the best optimal depth visibility for guinea pig cochlea samples. Thus, we implemented a decalcification-based optical clearing effect to guinea pig cochlea samples to enhance the depth visualization of internal microstructures using swept source optical coherence tomography (OCT). The obtained nondestructive two-dimensional OCT images successfully illustrated the feasibility of the proposed method by providing clearly visible microstructures in the depth direction as a result of decalcification. The most optimal clearing outcomes for the guinea pig cochlea were obtained after 14 consecutive days. The quantitative assessment results verified the increase of the intensity as well as the thickness measurements of the internal microstructures. Following this method, difficulties in imaging of internal cochlea microstructures of guinea pigs could be avoided. The obtained results verified that the depth visibility of the decalcified ex vivo guinea pig cochlea samples was enhanced. Therefore, the proposed EDTA-based optical clearing method for guinea pig can be considered as a potential application for depth-enhanced OCT visualization.

Dissolution of Fe(III) (hydr) oxides by metal-EDTA complexes

NASA Astrophysics Data System (ADS)

Ngwack, Bernd; Sigg, Laura

1997-03-01

The dissolution of Fe(III)(hydr)oxides (goethite and hydrous ferric oxide) by metal-EDTA complexes occurs by ligand-promoted dissolution. The process is initiated by the adsorption of metal-EDTA complexes to the surface and is followed by the dissociation of the complex at the surface and the release of Fe(III)EDTA into solution. The dissolution rate is decreased to a great extent if EDTA is complexed by metals in comparison to the uncomplexed EDTA. The rate decreases in the order EDTA CaEDTA ≫ PbEDTA > ZnEDTA > CuEDTA > Co(II)EDTA > NiEDTA. Two different rate-limiting steps determine the dissolution process: (1) detachment of Fe(III) from the oxide-structure and (2) dissociation of the metal-EDTA complexes. In the case of goethite, step 1 is slower than step 2 and the dissolution rates by various metals are similar. In the case of hydrous ferric oxide, step 2 is rate-limiting and the effect of the complexed metal is very pronounced.

[Study of methods of decalcification for making united slices of tooth and affiliated periodontic tissues].

PubMed

Wang, Yu; Mu, Ya-bing; Miao, Lei-ying; Sun, Hong-chen; Li, Cheng-ku

2007-03-01

To study the methods of decalcification for making united slices of tooth and affiliated periodontic tissues. Twenty-one samples containing dog molars and affiliated periodontic tissues were divided into seven mean groups. The pH value of solution, time of decalcification, weight and volume of samples, and content of decalcified calcium were detected. The slices were observed by HE, specific, and immunohistochemical stain. The velocity of decalcification increased with decrease of solution pH. The weight of samples lightened by 37.61%, the volume reduced by 25.97% on average, and calcium decalcified was 174.49 mg per gram humid samples. The EDTA decalcification was slowest, but it was best. Decalcification was fast in Plank-Rycho solution while the section was worst, and faster in the formyl solution containing aluminium chloride than in EDTA, and the section was better. The 50% formyl solution containing aluminium chloride is an ideal decalcifying solution.

Interaction between lidocaine hydrochloride (with and without adrenaline) and various irrigants: A nuclear magnetic resonance analysis

PubMed Central

Vidhya, Nirmal; Karthikeyan, Balasubramanian Saravana; Velmurugan, Natanasabapathy; Abarajithan, Mohan; Nithyanandan, Sivasankaran

2014-01-01

Background: Interaction between local anesthetic solution, lidocaine hydrochloride (with and without adrenaline), and root canal irrigants such as sodium hypochlorite (NaOCl), ethylene diamine tetra-acetic acid (EDTA), and chlorhexidine (CHX) has not been studied earlier. Hence, the purpose of this in vitro study was to evaluate the chemical interaction between 2% lidocaine hydrochloride (with and without adrenaline) and commonly used root canal irrigants, NaOCl, EDTA, and CHX. Materials and Methods: Samples were divided into eight experimental groups: Group I-Lidocaine hydrochloride (with adrenaline)/3% NaOCl, Group II-Lidocaine hydrochloride (with adrenaline)/17% EDTA, Group III- Lidocaine hydrochloride (with adrenaline)/2% CHX, Group IV-Lidocaine hydrochloride (without adrenaline)/3% NaOCl, Group V-Lidocaine hydrochloride (without adrenaline)/17% EDTA, Group VI-Lidocaine hydrochloride (without adrenaline)/2% CHX, and two control groups: Group VII-Lidocaine hydrochloride (with adrenaline)/deionized water and Group VIII-Lidocaine hydrochloride (without adrenaline)/deionized water. The respective solutions of various groups were mixed in equal proportions (1 ml each) and observed for precipitate formation. Chemical composition of the formed precipitate was then analysed by nuclear magnetic resonance spectroscopy (NMR) and confirmed with diazotation test. Results: In groups I and IV, a white precipitate was observed in all the samples on mixing the respective solutions, which showed a color change to reddish brown after 15 minutes. This precipitate was then analysed by NMR spectroscopy and was observed to be 2,6-xylidine, a reported toxic compound. The experimental groups II, III, V, and VI and control groups VII and VIII showed no precipitate formation in any of the respective samples, until 2 hours. Conclusion: Interaction between lidocaine hydrochloride (with and without adrenaline) and NaOCl showed precipitate formation containing 2,6-xylidine, a toxic compound. PMID:25097652

Identification of phlebotomine sand fly blood meals by real-time PCR.

PubMed

Sales, Kamila Gaudêncio da Silva; Costa, Pietra Lemos; de Morais, Rayana Carla Silva; Otranto, Domenico; Brandão-Filho, Sinval Pinto; Cavalcanti, Milena de Paiva; Dantas-Torres, Filipe

2015-04-16

Phlebotomine sand flies are blood-feeding insects of great medical and veterinary significance acting as vectors of Leishmania parasites. Studying the blood-feeding pattern of these insects may help in the understanding of their interactions with potential reservoir hosts of Leishmania parasites. In this study, we developed real time PCR assays for the identification of sand fly blood meal. Six pairs of primers were designed based on cytochrome b gene sequences available in GenBank of the following potential hosts: dog, cat, horse, chicken, black rat, and human. Firstly, SYBR Green-based real time PCR assays were conducted using a standard curve with eight different concentrations (i.e., 10 ng, 1 ng, 100 pg, 10 pg, 1 pg, 100 fg, 10 fg and 1 fg per 2 μl) of DNA samples extracted from EDTA blood samples from each target animal. Then, DNA samples extracted from field-collected engorged female sand flies belonging to three species (i.e., Lutzomyia longipalpis, L. migonei and L. lenti) were tested by the protocols standardized herein. Additionally, female sand flies were experimentally fed on a black rat (Rattus rattus) and used for evaluating the time course of the detection of the protocol targeting this species. The protocols performed well with detection limits of 10 pg to 100 fg. Field-collected female sand flies were fed on blood from humans (73%), chickens (23%), dogs (22%), horses (15%), black rats (11%) and cats (2%). Interestingly, 76.1% of the L. longipalpis females were positive for human blood. In total, 48% of the tested females were fed on single sources, 31% on two and 12% on three. The analysis of the time course showed that the real time PCR protocol targeting the black rat DNA was able to detect small amounts of the host DNA up to 5 days after the blood meal. The real time PCR assays standardized herein successfully detected small amounts of host DNA in female sand flies fed on different vertebrate species and, specifically for the black rats, up to 5 days after the blood meal. These assays represent promising tools for the identification of blood meal in field-collected female sand flies.

Analysis and stability of retinol in plasma.

PubMed

Peng, Y M; Xu, M J; Alberts, D S

1987-01-01

A simple, precise, and specific high-performance liquid chromatography (HPLC) method was developed for the simultaneous measurement of retinol (ROH), 13-cis-retinoic acid (13-cRA), and 4-oxo-13-cRA. The average recovery of ROH from serum or plasma was 95%, and the precision of the assay was less than 5%. With this HPLC method, a series of studies was carried out to evaluate the stability of ROH in various matrices. ROH was stable under our HPLC assay conditions as well as in plasma- and in serum-enriched culture media; however, ROH was not stable in aqueous matrices. Serum or heparinized plasma may be routinely used for measurement of ROH concentrations, providing EDTA, oxalate, and citrate are not used as anticoagulants. Because of ROH stability, blood samples can be kept on ice in the dark for at least 24 hours prior to separation of plasma. In addition, plasma samples containing ROH can be stored for up to 1 year at -20 degrees C without loss of stability.

The effect of anticoagulants on the distribution of chromium VI in blood fractions.

PubMed

Afolaranmi, Grace A; Tettey, Justice N A; Murray, Helen M; Meek, R M Dominic; Grant, M Helen

2010-01-01

Metal-on-metal resurfacing arthroplasty is associated with elevated circulating levels of cobalt and chromium ions. To establish the long-term safety of metal-on-metal resurfacing arthroplasty, it has been recommended that during clinical follow-up of these patients, the levels of these metal ions in blood be monitored. In this article, we provide information on the distribution of chromium VI ions (the predominant form of chromium released by cobalt-chrome alloys in vivo and in vitro) in blood fractions. Chromium VI is predominantly partitioned into red blood cells compared with plasma (analysis of variance, P < .05). The extent of accumulation in red blood cells is influenced by the anticoagulant used to collect the blood, with EDTA giving a lower partitioning into red cells compared with sodium citrate and sodium heparin. 2010 Elsevier Inc. All rights reserved.

PCR detection of Anaplasma phagocytophilum in goat flocks in an area endemic for tick-borne fever in Switzerland.

PubMed

Silaghi, C; Scheuerle, M C; Friche Passos, L M; Thiel, C; Pfister, K

2011-02-01

Central Switzerland is a highly endemic region for tick-borne fever (TBF) in cattle, however, little is known about A. phagocytophilum in goats. In the present study, 72 animals from six goat flocks (373 EDTA blood-samples) in Central Switzerland were analysed for A. phagocytophilum DNA. A real-time PCR targeting the msp2 gene of A. phagocytophilum was performed and in positive samples the partial 165 rRNA, groEL and msp4 gene were amplified for sequence analysis. Four DNA extracts were positive. Different sequence types on basis of the amplified genes were found. For comparison, sequences of A. phagocytophilum from 12 cattle (originating from Switzerland and Southern Germany) were analysed. The 165 rRNA gene sequences from cattle were all identical amongst each other, but the groEL and msp4 gene differed depending on the origin of the cattle samples and differed from the variants from goats. This study clearly provides molecular evidence for the presence of different types of A. phagocytophilum in goat flocks in Switzerland, a fact which deserves more thorough attention in clinical studies.

Detection of tetracosactide in plasma by enzyme-linked immunosorbent assay (ELISA).

PubMed

Martin, Laurent; Chaabo, Ayman; Lasne, Françoise

2015-06-01

As a synthetic analogue of adrenocorticotropic hormone (ACTH), tetracosactide is prohibited in sport by the World Anti-Doping Agency (WADA). An enzyme-linked immunosorbent assay (ELISA) method is proposed for detection of this drug in plasma. Since its structure corresponds to the 24 N-terminal of the 39 amino acids of the natural endogenous peptide ACTH, tetracosactide can be detected with a commercial ELISA kit for ACTH that uses antibodies, the epitopes of which are located in the 1-24 part of ACTH. However, an essential condition for detection specificity is the preliminary total clearance of endogenous ACTH in the plasma samples. This is achieved by a preparative step based on cation-exchange chromatography before ELISA. The method is specific and sensitive (LOD: 30 pg/mL) and may be used as a screening analysis in anti-doping control. The pre-analytical conditions are shown to be of the upmost importance and recommendations for blood collection (EDTA tubes), sample transport (4 °C) and plasma sample storage (-20 °C) are presented. Copyright © 2014 John Wiley & Sons, Ltd.

Dissolution of root canal sealers in EDTA and NaOCl solutions.

PubMed

Keleş, Ali; Köseoğlu, Mustafa

2009-01-01

Solutions of ethylenediaminetetraacetic acid (EDTA) and sodium hypochlorite (NaOCl) have been used as canal irrigants in endodontic treatment. The authors conducted a study to compare the ability of these solutions to dissolve sealers. The authors assessed the solubility of six sealers-calcium hydroxide, polyketone, zinc oxide-eugenol, silicone and two epoxy resins-in EDTA and two concentrations of NaOCl (2.5 percent and 5.0 percent). They immersed standardized samples (n = 5) of each sealer for two minutes and 10 minutes. They obtained the mean values of sealer dissolution in solutions by calculating the difference between the original preimmersion and postimmersion weights to determine the amount of sealer removed. They compared the values via factorial analysis of variance. They analyzed differences between the six sealers with respect to their solubility in EDTA or NaOCl solutions at two minutes and 10 minutes by using a one-way analysis of variance (P < .05). In comparison with NaOCl solutions, EDTA was markedly superior in dissolving root canal sealers (P < .05). There were no significant differences between the two concentrations of NaOCl. The two epoxy resins and the silicone-based sealer were of low solubility. The zinc oxide-eugenol-based sealer was significantly more soluble than were the epoxy resins and the silicone-based sealers. Polyketone and calcium hydroxide-based sealers were the most soluble sealers (P < .05). The results of this study indicate that during nonsurgical endodontic re-treatment, EDTA and NaOCl solutions used for removing smear layer aided in the retreatment by dissolving some root canal sealers.

Comparison of the Antimicrobial Efficacy of the EndoVac System and Conventional Needle Irrigation in Primary Molar Root Canals.

PubMed

Buldur, Burak; Kapdan, Arife

The purpose of this study was to compare the antimicrobial efficacy of the EndoVac system and conventional needle irrigation to eliminate E faecalis in primary molar root canals. 60 extracted human primary second molar roots were instrumented up to an apical size .04/35 and randomly divided into two groups; Group 1: conventional needle (n=30) and Group 2: EndoVac (n=30), and four subgroups (two experimental subgroups; (a) 2.5% sodium hypochlorite (NaOCl) + ethylenediaminetetraacetic acid (EDTA) (n=20), (b) ozonated water (OW) + EDTA (n=20), and control groups (c) 5.25% NaOCl (n=10) and (d) saline (n=10). All roots were sterilized and then inoculated with E.faecalis. Before and after final irrigation procedures, root canals were sampled and the grown colony forming units (CFUs) were counted. Data were analyzed by Kruskall-Wallis and Mann-Whitney U tests using a 0.05 significance level. The EndoVac reduced more bacteria than the conventional needle did but it was not statistically significant (p>0.05). NaOCl alone or followed by EDTA totally eliminated bacteria. OW + EDTA showed higher reduction of bacteria but could not totally eliminate bacterias. In the context of bacterial elimination, the EndoVac was not significantly better than the conventional needle. Although, there were fewer CFU/mg when using EndoVac, there was not any statistically significant superiority to conventional needle irrigation. An OW+EDTA regimen showed antibacterial effect in the primary molar root canals but it was significantly less effective than NaOCl+EDTA.

Simultaneous speciation and preservation of aqueous As, Sb and Se redox couples

NASA Astrophysics Data System (ADS)

Wu, D.; Pichler, T.

2014-12-01

We developed a new method for the simultaneous speciation analysis of inorganic arsenic (III, V), antimony (III, V) and selenium (IV, VI) in water samples via double-focusing sector field-inductively coupled plasma-mass spectrometry (SF-ICP-MS) coupled to high performance liquid chromatography (HPLC). A Hamilton PRX-X100 anion exchange column with EDTA (pH of 4.7) and 3% methanol as mobile phase was used for species separation. The flow rate was set to 1.5 mL min-1 and a solvent gradient (linear ramp from 5 mM to 30 mM) was applied. The overall analysis time for all six desired species was 11 minutes. The detection limits for As(III), As(V), Sb(III), Sb(V), Se(VI) and Se(IV) were 0.02 μg L-1, 0.06 μg L-1, 0.2 μg L-1, 0.02 μg L-1, 0.2 μg L-1 and 0.4 μg L-1 respectively. The retention times for As(III), As(V), Sb(III), Sb(V), Se(IV) and Se(VI) were 1.70, 2.94, 7.14, 2.28, 3.38 and 9.36 min, respectively. Subsequently, the stability of inorganic As(III, V), Sb(III, V) and Se(IV, VI) species in different water samples (groundwater, lake water and river water) was studied over a time scale of 11 weeks. High concentrations of Fe (25.0 mg/L) and Mn (25.0 mg/L) were added to different matrices to simulate Fe and Mn rich environments. All samples were spiked with 5.0 μg/L As(III, V) and Sb(III, V) and 15.0 μg/L Se(IV, VI).. We investigated several strategies for species preservation, i.e., EDTA only, EDTA combined with acidification (HCl, HNO3, formic acid and acetic acid). The preserved samples were stored at 4 °C in the dark. For comparison, another subsample without any preservation was stored at room temperature in the presence of light. The results showed that a combination EDTA acidified to pH of 3 can be used to preserve all species for at least 11 weeks. While EDTA only (pH = 6) failed to preserve As and Sb species, although Se species were preserved.

Transport of EDTA into cells of the EDTA-degrading bacterial strain DSM 9103.

PubMed

Witschel, M; Egli, T; Zehnder, A J; Wehrli, E; Spycher, M

1999-04-01

In the bacterial strain DSM 9103, which is able to grow with the complexing agent EDTA as the sole source of carbon, nitrogen and energy, the transport of EDTA into whole cells was investigated. EDTA uptake was found to be dependent on speciation: free EDTA and metal-EDTA complexes with low stability constants were readily taken up, whereas those with stability constants higher than 1016 were not transported. In EDTA-grown cells, initial transport rates of CaEDTA showed substrate-saturation kinetics with a high apparent affinity for CaEDTA (affinity constant Kt= 0.39 microM). Several uncouplers had an inhibitory effect on CaEDTA transport. CaEDTA uptake was also significantly reduced in the presence of an inhibitor of ATPase and the ionophore nigericin, which dissipates the proton gradient. Valinomycin, however, which affects the electrical potential, had little effect on uptake, indicating that EDTA transport is probably driven by the proton gradient. Of various structurally related compounds tested only Ca2+-complexed diethylenetriaminepentaacetate (CaDTPA) competitively inhibited CaEDTA transport. Uptake in fumarate-grown cells was low compared to that measured in EDTA-grown bacteria. These results strongly suggest that the first step in EDTA degradation by strain DSM 9103 consists of transport by an inducible energy-dependent carrier. Uptake experiments with 45Ca2+ in the presence and absence of EDTA indicated that Ca2+ is transported together with EDTA into the cells. In addition, these transport studies and electron-dispersive X-ray analysis of electron-dense intracellular bodies present in EDTA-grown cells suggest that two mechanisms acting simultaneously allow the cells to cope with the large amounts of metal ions taken up together with EDTA. In one mechanism the metal ions are excreted, in the other they are inactivated intracellularly in polyphosphate granules.

Purification and Characterization of EDTA Monooxygenase from the EDTA-Degrading Bacterium BNC1

PubMed Central

Payne, Jason W.; Bolton, Harvey; Campbell, James A.; Xun, Luying

1998-01-01

The synthetic chelating agent EDTA can mobilize radionuclides and heavy metals in the environment. Biodegradation of EDTA should reduce this mobilization. Although several bacteria have been reported to mineralize EDTA, little is known about the biochemistry of EDTA degradation. Understanding the biochemistry will facilitate the removal of EDTA from the environment. EDTA-degrading activities were detected in cell extracts of bacterium BNC1 when flavin mononucleotide (FMN), NADH, and O2 were present. The degradative enzyme system was separated into two different enzymes, EDTA monooxygenase and an FMN reductase. EDTA monooxygenase oxidized EDTA to glyoxylate and ethylenediaminetriacetate (ED3A), with the coconsumption of FMNH2 and O2. The FMN reductase provided EDTA monooxygenase with FMNH2 by reducing FMN with NADH. The FMN reductase was successfully substituted in the assay mixture by other FMN reductases. EDTA monooxygenase was purified to greater than 95% homogeneity and had a single polypeptide with a molecular weight of 45,000. The enzyme oxidized both EDTA complexed with various metal ions and uncomplexed EDTA. The optimal conditions for activity were pH 7.8 and 35°C. Kms were 34.1 μM for uncomplexed EDTA and 8.5 μM for MgEDTA2−; this difference in Km indicates that the enzyme has greater affinity for MgEDTA2−. The enzyme also catalyzed the release of glyoxylate from nitrilotriacetate and diethylenetriaminepentaacetate. EDTA monooxygenase belongs to a small group of FMNH2-utilizing monooxygenases that attack carbon-nitrogen, carbon-sulfur, and carbon-carbon double bonds. PMID:9683478

Immunoassay and antibody microarray analysis of the HUPO Plasma Proteome Project reference specimens: Systematic variation between sample types and calibration of mass spectrometry data

DOE Office of Scientific and Technical Information (OSTI.GOV)

Haab, Brian B.; Geierstanger, Bernhard H.; Michailidis, George

2005-08-01

Four different immunoassay and antibody microarray methods performed at four different sites were used to measure the levels of a broad range of proteins (N = 323 assays; 39, 88, 168, and 28 assays at the respective sites; 237 unique analytes) in the human serum and plasma reference specimens distributed by the Plasma Proteome Project (PPP) of the HUPO. The methods provided a means to (1) assess the level of systematic variation in protein abundances associated with blood preparation methods (serum, citrate-anticoagulated-plasma, EDTA-anticoagulated-plasma, or heparin-anticoagulated-plasma) and (2) evaluate the dependence on concentration of MS-based protein identifications from data sets usingmore » the HUPO specimens. Some proteins, particularly cytokines, had highly variable concentrations between the different sample preparations, suggesting specific effects of certain anticoagulants on the stability or availability of these proteins. The linkage of antibody-based measurements from 66 different analytes with the combined MS/MS data from 18 different laboratories showed that protein detection and the quality of MS data increased with analyte concentration. The conclusions from these initial analyses are that the optimal blood preparation method is variable between analytes and that the discovery of blood proteins by MS can be extended to concentrations below the ng/mL range under certain circumstances. Continued developments in antibody-based methods will further advance the scientific goals of the PPP.« less

Optimal cut points of plasma and urine neutrophil gelatinase-associated lipocalin for the prediction of acute kidney injury among critically ill adults: retrospective determination and clinical validation of a prospective multicentre study.

PubMed

Tecson, Kristen M; Erhardtsen, Elisabeth; Eriksen, Peter M; Gaber, A Osama; Germain, Michael; Golestaneh, Ladan; Lavoria, Maria de Los Angeles; Moore, Linda W; McCullough, Peter A

2017-07-10

To determine the optimal threshold of blood and urine neutrophil gelatinase-associated lipocalin (NGAL) to predict moderate to severe acute kidney injury (AKI) and persistent moderate to severe AKI lasting at least 48 consecutive hours, as defined by an adjudication panel. A multicentre prospective observational study enrolled intensive care unit (ICU) patients and recorded daily ethylenediaminetetraacetic acid (EDTA) plasma, heparin plasma and urine NGAL. We used natural log-transformed NGAL in a logistic regression model to predict stage 2/3 AKI (defined by Kidney Disease International Global Organization). We performed the same analysis using the NGAL value at the start of persistent stage 2/3 AKI. Of 245 subjects, 33 (13.5%) developed stage 2/3 AKI and 25 (10.2%) developed persistent stage 2/3 AKI. Predicting stage 2/3 AKI revealed the optimal NGAL cutoffs in EDTA plasma (142.0 ng/mL), heparin plasma (148.3 ng/mL) and urine (78.0 ng/mL) and yielded the following decision statistics: sensitivity (SN)=78.8%, specificity (SP)=73.0%, positive predictive value (PPV)=31.3%, negative predictive value (NPV)=95.7%, diagnostic accuracy (DA)=73.8% (EDTA plasma); SN=72.7%, SP=73.8%, PPV=30.4%, NPV=94.5%, DA=73.7% (heparin plasma); SN=69.7%, SP=76.8%, PPV=32.9%, NPV=94%, DA=75.8% (urine). The optimal NGAL cutoffs to predict persistent stage 2/3 AKI were similar: 148.3 ng/mL (EDTA plasma), 169.6 ng/mL (heparin plasma) and 79.0 ng/mL (urine) yielding: SN=84.0%, SP=73.5%, PPV=26.6%, NPV=97.6, DA=74.6% (EDTA plasma), SN=84%, SP=76.1%, PPV=26.8%, NPV=96.5%, DA=76.1% (heparin plasma) and SN=75%, SP=75.8%, PPV=26.1, NPV=96.4%, DA=75.7% (urine). Blood and urine NGAL predicted stage 2/3 AKI, as well as persistent 2/3 AKI in the ICU with acceptable decision statistics using a single cut point in each type of specimen. © Article author(s) (or their employer(s) unless otherwise stated in the text of the article) 2017. All rights reserved. No commercial use is permitted unless otherwise expressly granted.

The effect of vacuum packaging, EDTA, oregano and thyme oils on the microbiological quality of chicken's breast.

PubMed

Pavelková, Adriana; Kačániová, Miroslava; Horská, Elena; Rovná, Katarína; Hleba, Lukáš; Petrová, Jana

2014-10-01

The effect of ethylenediaminetetraacetate (EDTA), oregano (Origanum vulgare) and thyme (Thymus vulgaris) oils, on the chicken breast fillets was examined in this study. The chicken breast fillets were stored under vacuum packaging (VP), at 4 ± 0.5 °C for a period of 18 days. There were used the following treatments of chicken breast fillets: Air-packaged (AC, control samples), vacuum-packaged (VPC, control samples), VP with EDTA solution 1.50% w/w (VPEC, control samples), VP with oregano oil 0.20% v/w (VP + O) and VP with thyme oil 0.20% v/w, (VP + T). The quality assessment for vacuum packaging of the product in accordance with the terms above and EDTA treatment, oregano and thyme oil was established by microbiological analyzes. The microbiological properties as the total viable counts on Plate Count Agar, after incubation for 2 days at 37 °C and coliform bacteria on Violet Red Bile Glucose agar incubated at 37 °C for 24 h, lactobacilli on Rogosa and Sharpe agar after incubation 48-78 h at 37 °C in an aerobic atmosphere supplemented with carbon dioxide (5% CO2) and Pseudomonas aeruginosa on Pseudomonas Isolation agar (PIA, Oxoid, UK) after incubation at 48 h at 35 °C were monitored. The using of oregano, thyme oil and EDTA with combination of vacuum packaging has significant effects to reduction of all followed groups of microorganisms compared with control group without vacuum packaging and untreated control group. The natural preservatives can be used as alternatives to chemical additives which could extend the meat and meat products shelf life. The knowledge about them can have an important economic feedback by reducing losses attributed to spoilage and by allowing the products to reach distant and new markets. This study shows how using of natural antimicrobials can extend the shelf-life of the meat product. Copyright © 2013 Elsevier Ltd. All rights reserved.

Effect of EDTA and Fe-EDTA complex concentration on TCF Kraft mill effluent degradability. Batch and continuous treatments.

PubMed

Diez, M C; Pouleurs, D; Navia, R; Vidal, G

2005-09-01

The effect of ethylenediaminetetracetic acid (EDTA) and Fe-EDTA complex on synthetic totally chlorine-free (TCF) effluent degradability in batch and continuously operating reactors was evaluated. Under batch treatment, the addition of EDTA and Fe-EDTA complex was studied in the range of 80 to 320 mg l(-1). Under continuously operated reactors, the Fe-EDTA complex concentration varied from 20 to 80 mg l(-1), and the hydraulic retention time (HRT) varied from 48 to 24 h. Sludge oxygen uptake rate (OUR) and chemical oxygen demand (COD) removal decreased when EDTA concentration increased in the influent under batch treatment; however, this inhibitory effect was reduced by the addition of Fe-EDTA complex. Without the addition of EDTA, COD removal decreased from 71% to 8%. The most efficient EDTA removal treatment (almost 10%) was the treatment of 80 mg l(-1) Fe-EDTA. Under continuously operated reactors, COD removal was greater than 57% in the synthetic TCF effluent with a Fe-EDTA concentration that varied from 20 to 80 mg l(-1); however, EDTA removal was lower than 25% in all cases. Synthetic TCF effluent with a Fe -EDTA concentration higher than 80 mg l(-1) could not be treated by the activated sludge treatment due to EDTA's inhibitory effect on the sludge.

Biodegradation of PuEDTA and Impacts on Pu Mobility

DOE Office of Scientific and Technical Information (OSTI.GOV)

Bolton, H., Jr.; Rai, D.; Xun, L.

The contamination of many DOE sites by Pu presents a long-term problem because of its long half-life (240,000 yrs) and the low drinking water standard (<10{sup -12} M). EDTA was co-disposed with radionuclides (e.g., Pu, {sup 60}Co), formed strong complexes, and enhanced radionuclide transport at several DOE sites. Biodegradation of EDTA should decrease Pu mobility. One objective of this project was to determine the biodegradation of EDTA in the presence of PuEDTA complexes. The aqueous system investigated at pH 7 (10{sup -4} M EDTA and 10{sup -6} M Pu) contained predominantly Pu(OH){sub 2}EDTA{sup 2-}. The EDTA was degraded at amore » faster rate in the presence of Pu. As the total concentration of both EDTA and PuEDTA decreased (i.e., 10{sup -5} M EDTA and 10{sup -7} M PuEDTA), the presence of Pu decreased the biodegradation rate of the EDTA. It is currently unclear why the concentration of Pu directly affects the increase/decrease in rate of EDTA biodegradation. The soluble Pu concentration decreased, in agreement with thermodynamic predictions, as the EDTA was biodegraded, indicating that biodegradation of EDTA will decrease Pu mobility when the Pu is initially present as Pu(IV)EDTA. A second objective was to investigate how the presence of competing metals, commonly encountered in geologic media, will influence the speciation and biodegradation of Pu(IV)-EDTA. Studies on the solubilities of Fe(OH){sub 3}(s) and of Fe(OH){sub 3}(s) plus PuO{sub 2}(am) in the presence of EDTA and as a function of pH showed that Fe(III) out competes the Pu(IV) for the EDTA complex, thereby showing that Pu(IV) will not form stable complexes with EDTA for enhanced transport of Pu in Fe(III) dominated subsurface systems. A third objective is to investigate the genes and enzymes involved in EDTA biodegradation. BNC1 can use EDTA and another synthetic chelating agent nitrilotriacetate (NTA) as sole carbon and nitrogen sources. The same catabolic enzymes are responsible for both EDTA and NTA degradation except that additional enzymes are required for EDTA degradation. When the catabolic genes were cloned and sequenced, the gene cluster also contained genes encoding a hypothetical ABC-type transporter. RT-PCR analysis showed that the transporter genes and EDTA monooxygenase gene (emoA) are co-transcribed. EppA is one of the transporter genes, and it codes for a periplasmic binding protein responsible for binding to the substrate before transport across the membrane can occur. EppA was cloned, expressed, and purified in Escherichia coli and found to bind, MgEDTA, CaEDTA, Fe(III)EDTA, MgNTA, CaNTA, and Fe(III)NTA. Our data also suggest that BNC1 uses the same ABC-type transporter for both EDTA and NTA uptake. Results from these studies can provide mechanistic understanding and approaches to assist in the bioremediate PuEDTA and other radionuclide-EDTA complexes at DOE sites.« less

Alternative sampling strategies for passive classical and African swine fever surveillance in wild boar.

PubMed

Petrov, Anja; Schotte, Ulrich; Pietschmann, Jana; Dräger, Carolin; Beer, Martin; Anheyer-Behmenburg, Helena; Goller, Katja V; Blome, Sandra

2014-10-10

In view of the fact that African swine fever (ASF) was recently introduced into the wild boar population of the European Union and that classical swine fever (CSF) keeps reoccurring, targeted surveillance is of utmost importance for early detection. Introduction of both diseases is usually accompanied by an increased occurrence of animals found dead. Thus, fallen wild boar are the main target for passive surveillance. However, encouraging reporting by hunters and sampling of these animals is difficult. Partly, these problems could be solved by providing a pragmatic sampling approach. For this reason, we assessed the applicability of three different dry/semi-dry blood swabs, namely a cotton swab, a flocked swab, and a forensic livestock swab, for molecular swine fever diagnosis. After nucleic acid extraction using manual and automated systems, routine quantitative real-time polymerase chain reactions (qPCR) were carried out. Results obtained from swabs or their fragments were compared to results generated from EDTA blood. It was shown that reliable detection of both pathogens was possible by qPCR. Shifts in genome copy numbers were observed, but they did not change the qualitative results. In general, all swabs were suitable, but the forensic swab showed slight advantages, especially in terms of cutting and further storage. Robustness of the method was confirmed by the fact that different extraction methods and protocols as well as storage at room temperature did not have an influence on the final outcome. Taken together, swab samples could be recommended as a pragmatic approach to sample fallen wild boar. Copyright © 2014 Elsevier B.V. All rights reserved.

Influence of the collection tube on metabolomic changes in serum and plasma.

PubMed

López-Bascón, M A; Priego-Capote, F; Peralbo-Molina, A; Calderón-Santiago, M; Luque de Castro, M D

2016-04-01

Major threats in metabolomics clinical research are biases in sampling and preparation of biological samples. Bias in sample collection is a frequently forgotten aspect responsible for uncontrolled errors in metabolomics analysis. There is a great diversity of blood collection tubes for sampling serum or plasma, which are widely used in metabolomics analysis. Most of the existing studies dealing with the influence of blood collection on metabolomics analysis have been restricted to comparison between plasma and serum. However, polymeric gel tubes, which are frequently proposed to accelerate the separation of serum and plasma, have not been studied. In the present research, samples of serum or plasma collected in polymeric gel tubes were compared with those taken in conventional tubes from a metabolomics perspective using an untargeted GC-TOF/MS approach. The main differences between serum and plasma collected in conventional tubes affected to critical pathways such as the citric acid cycle, metabolism of amino acids, fructose and mannose metabolism and that of glycerolipids, and pentose and glucuronate interconversion. On the other hand, the polymeric gel only promoted differences at the metabolite level in serum since no critical differences were observed between plasma collected with EDTA tubes and polymeric gel tubes. Thus, the main changes were attributable to serum collected in gel and affected to the metabolism of amino acids such as alanine, proline and threonine, the glycerolipids metabolism, and two primary metabolites such as aconitic acid and lactic acid. Therefore, these metabolite changes should be taken into account in planning an experimental protocol for metabolomics analysis. Copyright © 2016 Elsevier B.V. All rights reserved.

Erythrocyte Alloimmunization and Autoimmunization among Blood Donors and Recipients visiting a Tertiary Care Hospital.

PubMed

Kaur, Daljit; Bains, Lovenish; Kandwal, Manoj; Parmar, Indu

2017-03-01

The ultimate aim of pretransfusion testing is the acceptable survival of donor red cells in recipient's body and antibody detection plays a critical role in achieving the same. The cornerstone of antibody detection method is detecting an unexpected antibody as against the expected antibodies of ABO blood group system. Autoantibodies can also interfere with the detection of clinically significant alloantibodies. To study the frequency of alloantibodies and autoantibodies in the healthy blood donors and patient population visiting our hospital. The Column Agglutination Technology (CAT) was used for ABO RhD blood grouping, Direct Antiglobulin Test (DAT), Autocontrol (AC), Indirect Antiglobulin Test (IAT) and red cell antibody screening and the unexpected reactions in any of these tests were recorded for further evaluation. Ethylene Diamine Tetra Acetic Acid (EDTA) blood samples were used for all these tests for both blood donors and admitted patients. The CAT was exercised for the blood grouping (using ABD-Reverse Diluent cassettes) and antibody screening (using 0.8% Surgiscreen, Ortho Clinical Diagnostics Limited, USA and Low Ionic Strength Saline Ortho BLISS with AHG cassettes) on the automated immunohaematology platform ORTHO AutoVue ® Innova system (Ortho Clinical Diagnostics Limited, USA). Among all blood donors (n=6350), seven (0.11%) donors had showed unexpected reaction. Of these, four had positive antibody screen (three having naturally occuring antibodies 2=anti-M, 1=anti-Le a and 1=inconclusive) and the other three had positive DAT. Of all the patient samples (n=6136) screened for irregular red cell antibodies, four (0.06%) patients were found to have unexpected reaction revealing one (0.02%) with anti-M antibody and the other three (0.05%) had autoantibodies in their serum. The combined prevalence for both blood donor and recipient population (n=12,486) was found to be 0.11% at our center. The alloimmunisation among patient population was found to be lower than many other studies worldwide as our hospital does not cater to multitransfused or transfusion dependant patients with haematological disorders and majorly elective surgery patients with no history of previous blood transfusions visit our hospital.

Erythrocyte Alloimmunization and Autoimmunization among Blood Donors and Recipients visiting a Tertiary Care Hospital

PubMed Central

Bains, Lovenish; Kandwal, Manoj; Parmar, Indu

2017-01-01

Introduction The ultimate aim of pretransfusion testing is the acceptable survival of donor red cells in recipient’s body and antibody detection plays a critical role in achieving the same. The cornerstone of antibody detection method is detecting an unexpected antibody as against the expected antibodies of ABO blood group system. Autoantibodies can also interfere with the detection of clinically significant alloantibodies. Aim To study the frequency of alloantibodies and autoantibodies in the healthy blood donors and patient population visiting our hospital. Materials and Methods The Column Agglutination Technology (CAT) was used for ABO RhD blood grouping, Direct Antiglobulin Test (DAT), Autocontrol (AC), Indirect Antiglobulin Test (IAT) and red cell antibody screening and the unexpected reactions in any of these tests were recorded for further evaluation. Ethylene Diamine Tetra Acetic Acid (EDTA) blood samples were used for all these tests for both blood donors and admitted patients. The CAT was exercised for the blood grouping (using ABD-Reverse Diluent cassettes) and antibody screening (using 0.8% Surgiscreen, Ortho Clinical Diagnostics Limited, USA and Low Ionic Strength Saline Ortho BLISS with AHG cassettes) on the automated immunohaematology platform ORTHO AutoVue® Innova system (Ortho Clinical Diagnostics Limited, USA). Results Among all blood donors (n=6350), seven (0.11%) donors had showed unexpected reaction. Of these, four had positive antibody screen (three having naturally occuring antibodies 2=anti-M, 1=anti-Lea and 1=inconclusive) and the other three had positive DAT. Of all the patient samples (n=6136) screened for irregular red cell antibodies, four (0.06%) patients were found to have unexpected reaction revealing one (0.02%) with anti-M antibody and the other three (0.05%) had autoantibodies in their serum. Conclusion The combined prevalence for both blood donor and recipient population (n=12,486) was found to be 0.11% at our center. The alloimmunisation among patient population was found to be lower than many other studies worldwide as our hospital does not cater to multitransfused or transfusion dependant patients with haematological disorders and majorly elective surgery patients with no history of previous blood transfusions visit our hospital. PMID:28511387

Mixed nano/micro-sized calcium phosphate composite and EDTA root surface etching improve availability of graft material in intrabony defects: an in vivo scanning electron microscopy evaluation.

PubMed

Gamal, Ahmed Y; Iacono, Vincent J

2013-12-01

The use of nanoparticles of graft materials may lead to breakthrough applications for periodontal regeneration. However, due to their small particle size, nanoparticles may be eliminated from periodontal defects by phagocytosis. In an attempt to improve nanoparticle retention in periodontal defects, the present in vivo study uses scanning electron microscopy (SEM) to evaluate the potential of micrograft particles of β-tricalcium phosphate (β-TCP) to enhance the binding and retention of nanoparticles of hydroxyapatite (nHA) on EDTA-treated and non-treated root surfaces in periodontal defects after 14 days of healing. Sixty patients having at least two hopeless periodontally affected teeth designated for extraction were randomly divided into four treatment groups (15 patients per group). Patients in group 1 had selected periodontal intrabony defects grafted with nHA of particle size 10 to 100 nm. Patients in group 2 were treated in a similar manner but had the affected roots etched for 2 minutes with a neutral 24% EDTA gel before grafting of the associated vertical defects with nHA. Patients in group 3 had the selected intrabony defects grafted with a composite graft consisting of equal volumes of nHA and β-TCP (particle size 63 to 150 nm). Patients in group 4 were treated as in group 3 but the affected roots were etched with neutral 24% EDTA as in group 2. For each of the four groups, one tooth was extracted immediately, and the second tooth was extracted after 14 days of healing for SEM evaluation. Fourteen days after surgery, all group 1 samples were devoid of any nanoparticles adherent to the root surfaces. Group 2 showed root surface areas 44.7% covered by a single layer of clot-blended grafted particles 14 days following graft application. After 14 days, group 3 samples appeared to retain fibrin strands devoid of grafted particles. Immediately extracted root samples of group 4 had adherent graft particles that covered a considerable area of the root surfaces (88.6%). Grafted particles appeared to cover all samples in a multilayered pattern. After 14 days, the group 4 extracted samples showed multilayered fibrin-covered nano/micro-sized graft particles adherent to the root surfaces (78.5%). The use of a composite graft consisting of nHA and microsized β-TCP after root surface treatment with 24% EDTA may be a suitable method to improve nHA retention in periodontal defects with subsequent graft bioreactivity.

Comparison effects and electron spin resonance studies of α-Fe2O4 spinel type ferrite nanoparticles.

PubMed

Bayrakdar, H; Yalçın, O; Cengiz, U; Özüm, S; Anigi, E; Topel, O

2014-11-11

α-Fe2O4 spinel type ferrite nanoparticles have been synthesized by cetyltrimethylammonium bromide (CTAB) and ethylenediaminetetraacetic acid (EDTA) assisted hydrothermal route by using NaOH solution. Electron spin resonance (ESR/EPR) measurements of α-Fe2O4 nanoparticles have been performed by a conventional x-band spectrometer at room temperature. The comparison effect of nanoparticles prepared by using CTAB and EDTA in different α-doping on the structural and morphological properties have been investigated in detail. The effect of EDTA-assisted synthesis for α-Fe2O4 nanoparticles are refined, and thus the spectroscopic g-factor are detected by using ESR signals. These samples can be considered as great benefits for magnetic recording media, electromagnetic and drug delivery applications. Copyright © 2014 Elsevier B.V. All rights reserved.

Anaerobic Biotransformation and Mobility of Pu and of Pu-EDTA

DOE Office of Scientific and Technical Information (OSTI.GOV)

Xun, Luying

2009-11-20

The enhanced mobility of radionuclides by co-disposed chelating agent, ethylenediaminetetraacetate (EDTA), is likely to occur only under anaerobic conditions. Our extensive effort to enrich and isolate anaerobic EDTA-degrading bacteria has failed. Others has tried and also failed. To explain the lack of anaerobic biodegradation of EDTA, we proposed that EDTA has to be transported into the cells for metabolism. A failure of uptake may contribute to the lack of EDTA degradation under anaerobic conditions. We demonstrated that an aerobic EDTA-degrading bacterium strain BNC1 uses an ABC-type transporter system to uptake EDTA. The system has a periplasmic binding protein that bindmore » EDTA and then interacts with membrane proteins to transport EDTA into the cell at the expense of ATP. The bind protein EppA binds only free EDTA with a Kd of 25 nM. The low Kd value indicates high affinity. However, the Kd value of Ni-EDTA is 2.4 x 10^(-10) nM, indicating much stronger stability. Since Ni and other trace metals are essential for anaerobic respiration, we conclude that the added EDTA sequestrates all trace metals and making anaerobic respiration impossible. Thus, the data explain the lack of anaerobic enrichment cultures for EDTA degradation. Although we did not obtain an EDTA degrading culture under anaerobic conditions, our finding may promote the use of certain metals that forms more stable metal-EDTA complexes than Pu(III)-EDTA to prevent the enhanced mobility. Further, our data explain why EDTA is the most dominant organic pollutant in surface waters, due to the lack of degradation of certain metal-EDTA complexes.« less

A comparison of serum and plasma cytokine values using a multiplexed assay in cats.

PubMed

Gruen, Margaret E; Messenger, Kristen M; Thomson, Andrea E; Griffith, Emily H; Paradise, Hayley; Vaden, Shelly; Lascelles, B D X

2016-12-01

Degenerative joint disease (DJD) is highly prevalent in cats, and pain contributes to morbidity. In humans, alterations of cytokine concentrations have been associated with joint deterioration and pain. Similar changes have not been investigated in cats. Cytokine concentrations can be measured using multiplex technology with small samples of serum or plasma, however, serum and plasma are not interchangeable for most bioassays. Correlations for cytokine concentrations between serum and plasma have not been evaluated in cats. To evaluate the levels of detection and agreement between serum and plasma samples in cats. Paired serum and plasma samples obtained from 38 cats. Blood was collected into anti-coagulant free and EDTA Vacutainer ® tubes, serum or plasma extracted, and samples frozen at -80°C until testing. Duplicate samples were tested using a 19-plex feline cytokine/chemokine magnetic bead panel. Agreement between serum and plasma for many analytes was high, however correlation coefficients ranged from -0.01 to 0.97. Results from >50% of samples were below the lower limit of quantification for both serum and plasma for nine analytes, and for an additional three analytes for plasma only. While serum and plasma agreement was generally good, detection was improved using serum samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Assessing bioavailability levels of metals in effluent-affected rivers: effect of Fe(III) and chelating agents on the distribution of metal speciation.

PubMed

Han, Shuping; Naito, Wataru; Masunaga, Shigeki

To assess the effects of Fe(III) and anthropogenic ligands on the bioavailability of Ni, Cu, Zn, and Pb, concentrations of bioavailable metals were measured by the DGT (diffusive gradients in thin films) method in some urban rivers, and were compared with concentrations calculated by a chemical equilibrium model (WHAM 7.0). Assuming that dissolved Fe(III) (<0.45 μm membrane filtered) was in equilibrium with colloidal iron oxide, the WHAM 7.0 model estimated that bioavailable concentrations of Ni, Cu, and Zn were slightly higher than the corresponding values estimated assuming that dissolved Fe(III) was absent. In contrast, lower levels of free Pb were predicted by the WHAM 7.0 model when dissolved Fe(III) was included. Estimates showed that most of the dissolved Pb was present as colloidal iron-Pb complex. Ethylene-diamine-tetra-acetic acid (EDTA) concentrations at sampling sites were predicted from the relationship between EDTA and the calculated bioavailable concentration of Zn. When both colloidal iron and predicted EDTA concentrations were included in the WHAM 7.0 calculations, dissolved metals showed a strong tendency to form EDTA complexes, in the order Ni > Cu > Zn > Pb. With the inclusion of EDTA, bioavailable concentrations of Ni, Cu, and Zn predicted by WHAM 7.0 were different from those predicted considering only humic substances and colloidal iron.

Simulation of the mobility of metal - EDTA complexes in groundwater: The influence of contaminant metals

USGS Publications Warehouse

Friedly, J.C.; Kent, D.B.; Davis, J.A.

2002-01-01

Reactive transport simulations were conducted to model chemical reactions between metal - EDTA (ethylenediaminetetraacetic acid) complexes during transport in a mildly acidic quartz - sand aquifer. Simulations were compared with the results of small-scale tracer tests wherein nickel-, zinc-, and calcium - EDTA complexes and free EDTA were injected into three distinct chemical zones of a plume of sewage-contaminated groundwater. One zone had a large mass of adsorbed, sewage-derived zinc; one zone had a large mass of adsorbed manganese resulting from mildly reducing conditions created bythe sewage plume; and one zone had significantly less adsorbed manganese and negligible zinc background. The chemical model assumed that the dissolution of iron(III) from metal - hydroxypolymer coatings on the aquifer sediments by the metal - EDTA complexes was kinetically restricted. All other reactions, including metal - EDTA complexation, zinc and manganese adsorption, and aluminum hydroxide dissolution were assumed to reach equilibrium on the time scale of transport; equilibrium constants were either taken from the literature or determined independently in the laboratory. A single iron(III) dissolution rate constant was used to fit the breakthrough curves observed in the zone with negligible zinc background. Simulation results agreed well with the experimental data in all three zones, which included temporal moments derived from breakthrough curves at different distances downgradient from the injections and spatial moments calculated from synoptic samplings conducted at different times. Results show that the tracer cloud was near equilibrium with respect to Fe in the sediment after 11 m of transport in the Zn-contaminated region but remained far from equilibrium in the other two zones. Sensitivity studies showed that the relative rate of iron(III) dissolution by the different metal - EDTA complexes was less important than the fact that these reactions are rate controlled. Results suggest that the published solubility for ferrihydrite reasonably approximates the Fe solubility of the hydroxypolymer coatings on the sediments. Aluminum may be somewhat more soluble than represented by the equilibrium constant for gibbsite, and its dissolution may be rate controlled when reacting with Ca - EDTA complexes.

Phytoextraction of Pb and Cu contaminated soil with maize and microencapsulated EDTA.

PubMed

Xie, Zhiyi; Wu, Longhua; Chen, Nengchang; Liu, Chengshuai; Zheng, Yuji; Xu, Shengguang; Li, Fangbai; Xu, Yanling

2012-09-01

Chelate-assisted phytoextraction using agricultural crops has been widely investigated as a remediation technique for soils contaminated with low mobility potentially toxic elements. Here, we report the use of a controlled-release microencapsulated EDTA (Cap-EDTA) by emulsion solvent evaporation to phytoremediate soil contaminated with Pb and Cu. Incubation experiments were carried out to assess the effect of Cap- and non-microencapsulated EDTA (Ncap-EDTA) on the mobility of soil metals. Results showed EDTA effectively increased the mobility of Pb and Cu in the soil solution and Cap-EDTA application provided lower and more constant water-soluble concentrations of Pb and Cu in comparison with. Phytotoxicity may be alleviated and plant uptake of Pb and Cu may be increased after the incorporation of Cap-EDTA. In addition phytoextraction efficiencies of maize after Cap- and Ncap-EDTA application were tested in a pot experiment. Maize shoot concentrations of Pb and Cu were lower with Cap-EDTA application than with Ncap-EDTA. However, shoot dry weight was significantly higher with Cap-EDTA application. Consequently, the Pb and Cu phytoextraction potential of maize significantly increased with Cap-EDTA application compared with the control and Ncap-EDTA application.

Frequency of DEA 1 antigen in 1037 mongrel and PUREBREED dogs in ITALY.

PubMed

Carli, E; Carminato, A; Ravagnan, S; Capello, K; Antognoni, M T; Miglio, A; Furlanello, T; Proverbio, D; Spada, E; Stefani, A; Mutinelli, F; Vascellari, M

2017-11-29

The prevalence of dog erythrocyte antigen (DEA 1) in canine population is approximately 40-60%. Often data are limited to a small number of breeds and/or dogs. The aims of this study were to evaluate frequency of DEA 1 in a large population of purebred and mongrel dogs including Italian native breeds and to recognize a possible association between DEA 1 and breed, sex, and genetic and phenotypical/functional classifications of breeds. Frequencies of DEA 1 blood group collected from screened/enrolled blood donors and from healthy and sick dogs were retrospectively evaluated. The breed and the sex were recorded when available. DEA 1 blood typing was assessed by immunocromatographic test on K3EDTA blood samples. The prevalence of DEA 1 antigen was statistically related to breed, gender, Fédération Cynologique Internationale (FCI) and genotypic grouping. Sixty-two per cent dogs resulted DEA 1+ and 38% DEA 1-. DEA 1- was statistically associated with Dogo Argentino, Dobermann, German Shepherd, Boxer, Corso dogs, the molossian dogs, the FCI group 1, 2 and 3 and the genetic groups "working dogs" and "mastiff". DEA 1+ was statistically associated with Rottweiler, Briquet Griffon Vendéen, Bernese mountain dog, Golden Retriever, the hunting breeds, the FCI group 4, 6, 7 and 8 and the genetic groups "scent hounds" and "retrievers". No gender association was observed. Data obtained by this work may be clinically useful to drive blood donor enrollment and selection among different breeds.

Iron bioavailability in corn-masa tortillas is improved by the addition of disodium EDTA.

PubMed

Walter, Tomás; Pizarro, Fernando; Olivares, Manuel

2003-10-01

Corn-masa flour flat bread tortillas are the main staple of Mexican and Central American populations. Due to high concentrations of inhibitors of iron absorption, the bioavailability from this matrix is unknown. We wanted to determine the most suitable fortificant that would efficaciously improve iron bioavailability. In tortillas prepared with commercial precooked, lime-treated, corn-masa flour, we examined the in vitro solubility of the following forms of iron: native iron with and without Na2EDTA, elemental reduced iron plus Na2EDTA, ferrous fumarate with and without Na2EDTA, bisglycine iron, ferrous sulfate and NaFeEDTA. We also examined the in vivo bioavailability in humans with double radioiron erythrocyte incorporation of ferrous fumarate with and without Na2EDTA, bisglycine iron, NaFeEDTA and native iron plus Na2EDTA, beans and rice. In vitro, solubility ranged from 1% in iron forms without Na2EDTA to 19.4% for NaFeEDTA. Forms of iron with Na2EDTA had intermediate values. In vivo radioiron studies showed that iron forms without Na2EDTA also had low bioavailability (< or =1%). NaFeEDTA had the highest bioavailability (5.3%). The bioavailability of all iron forms improved significantly when tested with Na2EDTA (<0.05). Adding Na2EDTA to ferrous fumarate increased bioavailability from 0.87% to 2.9% (P < 0.001). We conclude that NaFeEDTA is the form of iron best absorbed, but alternatively, ferrous fumarate plus Na2EDTA comprises a feasible option as a fortificant.

Verification of an immunoturbidimetric assay for heart-type fatty acid-binding protein (H-FABP) on a clinical chemistry platform and establishment of the upper reference limit.

PubMed

Da Molin, Simona; Cappellini, Fabrizio; Falbo, Rosanna; Signorini, Stefano; Brambilla, Paolo

2014-11-01

Heart-type fatty acid-binding protein (H-FABP) is an early biomarker of cardiac injury. Randox Laboratories developed an immunoturbidimetric H-FABP assay for non-proprietary automated clinical chemistry analysers that could be useful in the emergency department. We verified the analytical performances claimed by Randox Laboratories on Roche Cobas 6000 clinical chemistry platform in use in our laboratory, and we defined our own 99th percentile upper reference limit for H-FABP. For the verification of method performances, we used pools of spared patient samples from routine and two levels of quality control material, while samples for the reference value study were collected from 545 blood donors. Following CLSI guidelines we verified limit of blank (LOB), limit of detection (LOD), limit of quantitation (LOQ), repeatability and within-laboratory precision, trueness, linearity, and the stability of H-FABP in EDTA over 24h. The LOQ (3.19 μg/L) was verified with a CV% of 10.4. The precision was verified for the low (mean 5.88 μg/L, CV=6.7%), the medium (mean 45.28 μg/L, CV=3.0%), and the high concentration (mean 88.81 μg/L, CV=4.0%). The trueness was verified as well as the linearity over the indicated measurement interval of 0.747-120 μg/L. The H-FABP in EDTA samples is stable throughout 24h both at room temperature and at 4 °C. The H-FABP 99th percentile upper reference limit for all subjects (3.60 μg/L, 95% CI 3.51-3.77) is more appropriate than gender-specific ones that are not statistically different. Copyright © 2014 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Clinical Relevance of Pathogens Detected by Multiplex PCR in Blood of Very-Low-Birth Weight Infants with Suspected Sepsis - Multicentre Study of the German Neonatal Network.

PubMed

Tröger, Birte; Härtel, Christoph; Buer, Jan; Dördelmann, Michael; Felderhoff-Müser, Ursula; Höhn, Thomas; Hepping, Nico; Hillebrand, Georg; Kribs, Angela; Marissen, Janina; Olbertz, Dirk; Rath, Peter-Michael; Schmidtke, Susanne; Siegel, Jens; Herting, Egbert; Göpel, Wolfgang; Steinmann, Joerg; Stein, Anja

2016-01-01

In the German Neonatal Network (GNN) 10% of very-low-birth weight infants (VLBWI) suffer from blood-culture confirmed sepsis, while 30% of VLBWI develop clinical sepsis. Diagnosis of sepsis is a difficult task leading to potential over-treatment with antibiotics. This study aims to investigate whether the results of blood multiplex-PCR (SeptiFast®) for common sepsis pathogens are relevant for clinical decision making when sepsis is suspected in VLBWI. We performed a prospective, multi-centre study within the GNN including 133 VLBWI with 214 episodes of suspected late onset sepsis (LOS). In patients with suspected sepsis a multiplex-PCR (LightCycler SeptiFast MGRADE-test®) was performed from 100 μl EDTA blood in addition to center-specific laboratory biomarkers. The attending neonatologist documented whether the PCR-result, which was available after 24 to 48 hrs, had an impact on the choice of antibiotic drugs and duration of therapy. PCR was positive in 110/214 episodes (51%) and blood culture (BC) was positive in 55 episodes (26%). Both methods yielded predominantly coagulase-negative staphylococci (CoNS) followed by Escherichia coli and Staphylococcus aureus. In 214 BC-PCR paired samples concordant results were documented in 126 episodes (59%; n = 32 were concordant pathogen positive results, n = 94 were negative in both methods). In 65 episodes (30%) we found positive PCR results but negative BCs, with CoNS being identified in 43 (66%) of these samples. Multiplex-PCR results influenced clinical decision making in 30% of episodes, specifically in 18% for the choice of antimicrobial therapy and in 22% for the duration of antimicrobial therapy. Multiplex-PCR results had a moderate impact on clinical management in about one third of LOS-episodes. The main advantage of multiplex-PCR was the rapid detection of pathogens from micro-volume blood samples. In VLBWI limitations include risk of contamination, lack of resistance testing and high costs. The high rate of positive PCR results in episodes of negative BC might lead to overtreatment of infants which is associated with risk of mortality, antibiotic resistance, fungal sepsis and NEC.

Circulating C5L2 gene polymorphism is associated with type 2 diabetes mellitus in Saudi population.

PubMed

Alharbi, Khalid K; Khan, Imran Ali; Syed, Rabbani

2013-11-01

The aim of the present study was to examine the relationship between the novel single nucleotide polymorphism, 698C>T that causes an amino acid change from proline to leucine at codon 233 and type 2 diabetes mellitus (T2DM) in the Saudi population. From the general population in the Saudi Arabia a total of 551 samples were collected and categorized them as T2DM (n = 376) and healthy controls (n = 175). Five ml of the blood sample was collected and used for the Biochemical and Molecular analysis. With the help of serum sample lipid profile: Fasting blood sugar (FBS), Total Cholesterol (TC), Triglycerides (TG), High Density Lipoprotein Cholesterol (HDL-C), Low Density Lipoprotein Cholesterol (LDL-C) and VLDL were performed. PCR–RFLP was performed after separating the genomic DNA from the EDTA blood. The genotype distribution of C698T polymorphism was performed by the Chi square test with SPSS version 16.0 software for comparing T2DM subjects and healthy controls. In our study, genotypic distributions of C5L2 C698T polymorphism and allele frequency of patients and controls were found to be significant difference in the allele and the genotypic distribution. [For T Vs C; p = 0.01; Odds ratio = 3.594 (95 % CI; 1.256–10.28); and CT+TT Vs CC; p = 0.009; Odds ratio = 3.707 (95 % CI; 1.285–10.69)]. TT genotype was completely absent in both the cases and the controls. In conclusion, our study indicates that 698C>T polymorphism of C5L2 gene is associated with the T2DM in individuals of Saudi population which was found to be similar with other studies.

Analytical Performance of a Multiplex Real-Time PCR Assay Using TaqMan Probes for Quantification of Trypanosoma cruzi Satellite DNA in Blood Samples

PubMed Central

Abate, Teresa; Cayo, Nelly M.; Parrado, Rudy; Bello, Zoraida Diaz; Velazquez, Elsa; Muñoz-Calderon, Arturo; Juiz, Natalia A.; Basile, Joaquín; Garcia, Lineth; Riarte, Adelina; Nasser, Julio R.; Ocampo, Susana B.; Yadon, Zaida E.; Torrico, Faustino; de Noya, Belkisyole Alarcón; Ribeiro, Isabela; Schijman, Alejandro G.

2013-01-01

Background The analytical validation of sensitive, accurate and standardized Real-Time PCR methods for Trypanosoma cruzi quantification is crucial to provide a reliable laboratory tool for diagnosis of recent infections as well as for monitoring treatment efficacy. Methods/Principal Findings We have standardized and validated a multiplex Real-Time quantitative PCR assay (qPCR) based on TaqMan technology, aiming to quantify T. cruzi satellite DNA as well as an internal amplification control (IAC) in a single-tube reaction. IAC amplification allows rule out false negative PCR results due to inhibitory substances or loss of DNA during sample processing. The assay has a limit of detection (LOD) of 0.70 parasite equivalents/mL and a limit of quantification (LOQ) of 1.53 parasite equivalents/mL starting from non-boiled Guanidine EDTA blood spiked with T. cruzi CL-Brener stock. The method was evaluated with blood samples collected from Chagas disease patients experiencing different clinical stages and epidemiological scenarios: 1- Sixteen Venezuelan patients from an outbreak of oral transmission, 2- Sixty three Bolivian patients suffering chronic Chagas disease, 3- Thirty four Argentinean cases with chronic Chagas disease, 4- Twenty seven newborns to seropositive mothers, 5- A seronegative receptor who got infected after transplantation with a cadaveric kidney explanted from an infected subject. Conclusions/Significance The performing parameters of this assay encourage its application to early assessment of T. cruzi infection in cases in which serological methods are not informative, such as recent infections by oral contamination or congenital transmission or after transplantation with organs from seropositive donors, as well as for monitoring Chagas disease patients under etiological treatment. PMID:23350002

The nanoparticle protein corona formed in human blood or human blood fractions.

PubMed

Lundqvist, Martin; Augustsson, Cecilia; Lilja, Malin; Lundkvist, Kristoffer; Dahlbäck, Björn; Linse, Sara; Cedervall, Tommy

2017-01-01

The protein corona formed around nanoparticles in protein-rich fluids plays an important role for nanoparticle biocompatibility, as found in several studies during the last decade. Biological fluids have complex compositions and the molecular components interact and function together in intricate networks. Therefore, the process to isolate blood or the preparation of blood derivatives may lead to differences in the composition of the identified protein corona around nanoparticles. Here, we show distinct differences in the protein corona formed in whole blood, whole blood with EDTA, plasma, or serum. Furthermore, the ratio between particle surface area to protein concentration influences the detected corona. We also show that the nanoparticle size per se influences the formed protein corona due to curvature effects. These results emphasize the need of investigating the formation and biological importance of the protein corona in the same environment as the nanoparticles are intended for or released into.

Effect of soil organic matter on antimony bioavailability after the remediation process.

PubMed

Nakamaru, Yasuo Mitsui; Martín Peinado, Francisco José

2017-09-01

We evaluated the long-term (18 year) and short-term (4 weeks) changes of Sb in contaminated soil with SOM increase under remediation process. In the Aznalcóllar mine accident (1998) contaminated area, the remediation measurement implemented the Guadiamar Green Corridor, where residual pollution is still detected. Soils of the re-vegetated area (O2) with high pH and high SOM content, moderately re-vegetated area (O1) and unvegetated area (C) were sampled. Soil pH, CEC, SOM amount and soil Sb forms were evaluated. Soil Sb was measured as total, soluble, exchangeable, EDTA extractable, acid oxalate extractable, and pyro-phosphate extractable fractions. Further, the short-term effect of artificial organic matter addition was also evaluated with incubation study by adding compost to the sampled soil from C, O1 and O2 areas. After 4 weeks of incubation, soil chemical properties and Sb forms were evaluated. In re-vegetated area (O2), soil total Sb was two times lower than in unvegetated area (C); however, soluble, exchangeable, and EDTA extractable Sb were 2-8 times higher. The mobile/bioavailable Sb increase was also observed after 4 weeks of incubation with the addition of compost. Soluble, exchangeable, and EDTA extractable Sb was increased 2-4 times by compost addition. By the linear regression analysis, the significantly related factors for soluble, exchangeable, and EDTA extractable Sb values were pH, CEC, and SOM, respectively. Soluble Sb increase was mainly related to pH rise. Exchangeable Sb should be bound by SOM-metal complex and increased with CEC. EDTA extractable fraction should be increased with increase of SOM as SOM-Fe associated Sb complex. From these results, it was shown that increase of SOM under natural conditions or application of organic amendment under remediation process should increase availability of Sb to plants. Copyright © 2017 Elsevier Ltd. All rights reserved.

Unraveling the Fate and Transport of SrEDTA-2 and Sr+2 in Hanford Sediments

NASA Astrophysics Data System (ADS)

Pace, M. N.; Mayes, M. A.; Jardine, P. M.; Mehlhorn, T. L.; Liu, Q. G.; Yin, X. L.

2004-12-01

Accelerated migration of strontium-90 has been observed in the vadose zone beneath the Hanford tank farm. The goal of this paper is to provide an improved understanding of the hydrogeochemical processes that contribute to strontium transport in the far-field Hanford vadose zone. Laboratory scale batch, saturated packed column experiments, and an unsaturated transport experiment in an undisturbed core were conducted to quantify geochemical and hydrological processes controlling Sr+2 and SrEDTA-2 sorption to Hanford flood deposits. After experimentation, the undisturbed core was disassembled and samples were collected from different bedding units as a function of depth. Sequential extractions were then performed on the samples. It has been suggested that organic chelates such as EDTA may be responsible for the accelerated transport of strontium due to the formation of stable anionic complexes. Duplicate batch and column experiments performed with Sr+2 and SrEDTA-2 suggested that the SrEDTA-2 complex was not stable in the presence of soil and rapid dissociation allowed strontium to be transported as a divalent cation. Batch experiments indicated a decrease in sorption with increasing rock:water ratios, whereas saturated packed column experiments indicated equal retardation in columns of different lengths. This difference between the batch and column experiments is primarily due to the difference between equilibrium conditions where dissolution of cations may compete for sorption sites versus flowing conditions where any dissolved cations are flushed through the system minimizing competition for sorption sites. Unsaturated transport in the undisturbed core resulted in significant Sr+2 retardation despite the presence of physical nonequilibrium. Core disassembly and sequential extractions revealed the mass wetness distribution and reactive mineral phases associated with strontium in the core. Overall, results indicated that strontium will most likely be transported through the Hanford far-field vadose zone as a divalent cation.

Effects of different lasers on organic/inorganic ratio of radicular dentin.

PubMed

Lopes, F C; Roperto, R; Akkus, A; Akkus, O; Souza-Gabriel, A E; Sousa-Neto, M D

2016-04-01

The aim of this study was to evaluate the chemical stability of endodontic-treated root dentin after different laser irradiations through Raman spectroscopy. Fifty maxillary canines were selected and prepared with K3 system. Roots were randomly distributed into five groups (n = 10) according to the surface treatment: GI (water), GII (NaOCl + EDTA), GIII (NaOCl + EDTA + 980 nm Diode laser), GIV (NaOCl + EDTA+ 1064 nm Nd: YAG laser) and GV (NaOCl + EDTA+ 2780 nm Er,Cr: YSGG laser). Lasers were applied for 20 s. Samples were bisected, and the organic and inorganic content of dentin was analyzed by Raman spectroscopy. Data were submitted to ANOVA and Tukey tests (p < 0.05). None of the surface treatments alter the inorganic content (cts) (p = 0.183). Roots irradiated with Er,Cr: YSGG laser had a reduced collagen content (GV-290.7 ± 41.7) compared with the water-treated roots (GI-328.3 ± 63.5) and those treated with NaOCl + EDTA (GII-333.9 ± 55.8). Roots irradiated with Er,Cr: YSGG laser also showed a higher inorganic/organic ratio (GV-9.5 ± 1.1) than roots treated with water (GI-7.7 ± 1.5), NaOCl + EDTA (GII-8.0 ± 1.4) and diode laser (GIII-8.2 ± 1.6). Both organic and inorganic contents increased from cervical to apical thirds in all groups. None of the surface treatments were able to promote changes in the inorganic content of the root dentin; treatment with NaOCl + EDTA combined with Er,Cr: YSGG altered collagen.

High sensitive gold-nanoparticle based lateral flow Immunodevice for Cd2+ detection in drinking waters.

PubMed

López Marzo, Adaris M; Pons, Josefina; Blake, Diane A; Merkoçi, Arben

2013-09-15

In this work for first time a lateral flow immunosensor device (LFID) for Cd(2+) determination in drinking and tap waters using the Cd-EDTA-BSA-AuNP conjugate as signal producer tool is introduced. The principle of working is based on competitive reaction between the Cd-EDTA-BSA-AuNP conjugate deposited on the conjugation pad strip and the Cd-EDTA complex formed in the analysis sample for the same binding sites of the 2A81G5 monoclonal antibody, specific to Cd-EDTA but not Cd(2+) free, which is immobilized onto the test line. The device has a large response range within 0.4-2000ppb, being the linear response between 0.4 and 10ppb. The quantification and detection limits of 0.4 and 0.1ppb, respectively, represent the lowest ones reported so far for paper based metal sensors. The obtained detection limit is 50 times lower than the maximum contamination level required for drinking water. Here we also show a new option for increasing the sensibility in the LFDs with competitive format, through the decreasing in concentrations of the Cd-EDTA-BSA-AuNP conjugate deposited in the conjugation strip and the mAbs deposited in the test and control zones until to reach optimized concentrations. It is an important result take into account that the increase in sensibility is one of the challenges in the field of LFD sensors, where are focused many of the ongoing researches. In addition, a specificity study of the device for several metal interferences, where potential metal interferences are masked with the use of the EDTA and OVA optimized concentrations, is presented too. Copyright © 2013 Elsevier B.V. All rights reserved.

The investigation of interspecies diversity of erythrocyte aggregation properties by two different photometric methods in four animal species.

PubMed

Kiss, F; Toth, E; Peto, K; Miko, I; Nemeth, N

2015-12-01

Among the haemorheological parameters, red blood cell (RBC) aggregation shows the largest interspecies diversity, and often controversial data can be found in the literature, besides the methodology-dependent issues. In this present investigation, we compared four experimental/laboratory animal species' RBC aggregation by two different photometric methods for better revealing the differences. Blood samples (K3-EDTA, 1.5 mg/ml) were taken from female animals: 16 inbred mice (Mus musculus, cardiac puncture), 15 outbred rats (Rattus norvegicus, caudal caval vein puncture), 15 beagle dogs (Canis canis, cephalic vein) and 23 juvenile pigs (Sus scrofa domesticus, medial saphenous vein). Haematological parameters (microcell counter) and RBC aggregation (light transmission and syllectometry-laser backscatter methods) were determined within 2 h after sampling. Describing the first 5-10 s of the aggregation process, additional parameters were calculated out of the syllectometric raw data. Standardized difference was calculated to determine the sensitivity of the two devices. Parameters describing the extent and magnitude of red blood cell aggregation showed the lowest values in the rat and the highest in the pig and canine blood. In turn, parameters describing the kinetics of aggregation showed the lowest values in the mouse and the highest in the rat. The standardized difference values for the laser backscattering method were 2-4 times larger vs. the light transmission one. The magnitude of the differences was not consequent in the aggregation parameters. These comparative results show that the laser backscattering method can detect the RBC aggregation differences between the investigated species more sensitively than the light transmission method. Journal of Animal Physiology and Animal Nutrition © 2015 Blackwell Verlag GmbH.

Reliable noninvasive prenatal testing by massively parallel sequencing of circulating cell-free DNA from maternal plasma processed up to 24h after venipuncture.

PubMed

Buysse, Karen; Beulen, Lean; Gomes, Ingrid; Gilissen, Christian; Keesmaat, Chantal; Janssen, Irene M; Derks-Willemen, Judith J H T; de Ligt, Joep; Feenstra, Ilse; Bekker, Mireille N; van Vugt, John M G; Geurts van Kessel, Ad; Vissers, Lisenka E L M; Faas, Brigitte H W

2013-12-01

Circulating cell-free fetal DNA (ccffDNA) in maternal plasma is an attractive source for noninvasive prenatal testing (NIPT). The amount of total cell-free DNA significantly increases 24h after venipuncture, leading to a relative decrease of the ccffDNA fraction in the blood sample. In this study, we evaluated the downstream effects of extended processing times on the reliability of aneuploidy detection by massively parallel sequencing (MPS). Whole blood from pregnant women carrying normal and trisomy 21 (T21) fetuses was collected in regular EDTA anti-coagulated tubes and processed within 6h, 24 and 48h after venipuncture. Samples of all three different time points were further analyzed by MPS using Z-score calculation and the percentage of ccffDNA based on X-chromosome reads. Both T21 samples were correctly identified as such at all time-points. However, after 48h, a higher deviation in Z-scores was noticed. Even though the percentage of ccffDNA in a plasma sample has been shown previously to significantly decrease 24h after venipuncture, the percentages based on MPS results did not show a significant decrease after 6, 24 or 48h. The quality and quantity of ccffDNA extracted from plasma samples processed up to 24h after venipuncture are sufficiently high for reliable downstream NIPT analysis by MPS. Furthermore, we show that it is important to determine the percentage of ccffDNA in the fraction of the sample that is actually used for NIPT, as downstream procedures might influence the fetal or maternal fraction. © 2013.

Pathway of FeEDTA transformation and its impact on performance of NOx removal in a chemical absorption-biological reduction integrated process

PubMed Central

Li, Wei; Zhao, Jingkai; Zhang, Lei; Xia, Yinfeng; Liu, Nan; Li, Sujing; Zhang, Shihan

2016-01-01

A novel chemical absorption-biological reduction (CABR) integrated process, employing ferrous ethylenediaminetetraacetate (Fe(II)EDTA) as a solvent, is deemed as a potential option for NOx removal from the flue gas. Previous work showed that the Fe(II)EDTA concentration was critical for the NOx removal in the CABR process. In this work, the pathway of FeEDTA (Fe(III)/Fe(II)-EDTA) transformation was investigated to assess its impact on the NOx removal in a biofilter. Experimental results revealed that the FeEDTA transformation involved iron precipitation and EDTA degradation. X-ray photoelectron spectroscopy analysis confirmed the iron was precipitated in the form of Fe(OH)3. The iron mass balance analysis showed 44.2% of the added iron was precipitated. The EDTA degradation facilitated the iron precipitation. Besides chemical oxidation, EDTA biodegradation occurred in the biofilter. The addition of extra EDTA helped recover the iron from the precipitation. The transformation of FeEDTA did not retard the NO removal. In addition, EDTA rather than the iron concentration determined the NO removal efficiency. PMID:26743930

[Effectiveness of social mobilization and social marketing in promoting NaFeEDTA-fortified soya sauce in adult women].

PubMed

Wang, Bo; Chen, Junshi; Zhan, Siyan; Sun, Jing; Li, Liming

2011-05-01

To assess the effectiveness of social mobilization and social marketing in promoting NaFeEDTA-fortified soy sauce in an iron deficient population. This study was an uncontrolled, community-based, before-after study, which was implemented in three counties of Shijiazhuang Municipality. The intervention was a social mobilization and social marketing strategy. Adult women older than 20 years of age participated in the evaluation protocol. The main outcomes included KAP relevant to IDA. Cross-sectional samples were used to assess the outcomes at baseline and 1 year later. Knowledge and attitudes of adult women had changed positively, and the percentage of women who had adopted NaFeEDTA-fortified soy sauce increased from 8.9% to 36.6% (P < 0.001). Social mobilization and social marketing had a positive impact on the KAP of adult women in the iron deficient population.

Laser Monitoring Of Phytoextraction Enhancement Of Lead Contaminated Soil Adopting EDTA And EDDS

NASA Astrophysics Data System (ADS)

Hassan, M.; Abdelhamied, M.; Hanafy, A. H.; Fantoni, R.; Harith, M. A.

2011-09-01

Removal of heavy metals (HMs) such as Pb from soil, wastewater, and air is essential for environment and human health. Phytoremediation is a well established technology based on the use of certain green plants for contaminants removal from soil, wastewater as well as air. Scented geranium, Pelargonium zonal, is a flowering plant recently used in HMs removal from contaminated soil. In the present work, EDTA (ethylenediaminetetraacetic acid) and EDDS (S, S-ethylenediaminedisuccinic acid) were used as chemical assistants providing higher Pb availability for extraction by plant roots. Lead was artificially added to the planting media, peatmoss, at different concentrations. Laser induced breakdown spectroscopy (LIBS) was used to follow up Pb relative concentrations in peatmoss as well as plant shoots, at different sampling times during the experiment period. Laser induced chlorophyll fluorescence (LICF), has been also used to evaluate chlorophyll formation and photosynthetic apparatus status in geranium plants. Such measurements were performed on geranium plants grown under various Pb levels, as well as EDTA and EDDS combinations. The combined effect of EDTA and EDDS was found to enhance Pb extraction with time. Good correlation was found between LICF results and chlorophyll (a) (Chl.a) concentrations in plant tissues extracted by chemical analysis.

A new spectrophotometric method for determination of EDTA in water using its complex with Mn(III)

NASA Astrophysics Data System (ADS)

Andrade, Carlos Eduardo O.; Oliveira, André F.; Neves, Antônio A.; Queiroz, Maria Eliana L. R.

2016-11-01

EDTA is an important ligand used in many industrial products as well as in agriculture, where it is employed to assist in phytoextraction procedures and the absorption of nutrients by plants. Due to its intensive use and recalcitrance, it is now considered an emerging pollutant in water, so there is great interest in techniques suitable for its monitoring. This work proposes a method based on formation of the Mn(III)-EDTA complex after oxidation of the Mn(II)-EDTA complex by PbO2 immobilized on cyanoacrylate spheres. A design of experiments (DOE) based on the Doehlert matrix was used to determine the optimum conditions of the method, and the influence of the variables was evaluated using a multiple linear regression (MLR) model. The optimized method presented a linear response in the range from 0.77 to 100.0 μmol L- 1, with analytical sensitivity of 7.7 × 103 L mol- 1, a coefficient of determination of 0.999, and a limit of detection of 0.23 μmol L- 1. The method was applied using samples fortified at different concentration levels, and the recoveries achieved were between 97.0 and 104.9%.

Multifunctional adhesive polymers: Preactivated thiolated chitosan-EDTA conjugates.

PubMed

Netsomboon, Kesinee; Suchaoin, Wongsakorn; Laffleur, Flavia; Prüfert, Felix; Bernkop-Schnürch, Andreas

2017-02-01

The aim of this study was to synthesis preactivated thiolated chitosan-EDTA (Ch-EDTA-cys-2MNA) conjugates exhibiting in particular high mucoadhesive, cohesive and chelating properties. Thiol groups were coupled with chitosan by carbodiimide reaction and further preactivated by attachment with 2-mercaptonicotinic acid (2MNA) via disulfide bond formation. Determinations of primary amino and sulfhydryl groups were performed by TNBS and Ellman's tests, respectively. Cytotoxicity was screened by resazurin assay in Caco-2 cells. Mucoadhesive properties and bivalent cation binding capacity with Mg 2+ and Ca 2+ in comparison to chitosan-EDTA (Ch-EDTA) and thiolated Ch-EDTA (Ch-EDTA-cys) were evaluated. Determination of 2MNA and total sulfhydryl groups indicated that 80% of thiol groups were preactivated. The results from cytotoxicity studies demonstrated that Ch-EDTA-cys and Ch-EDTA-cys-2MNA were not toxic to the cells at the polymer test concentration of 0.25% (w/v) while cell viability decreased by increasing the concentration of Ch-EDTA. Although EDTA molecule was modified by thiolation and preactivation, approximately 50% of chelating properties of the conjugates were maintained compared to Ch-EDTA. Ch-EDTA-cys-2MNA adhered on freshly excised porcine intestinal mucosa up to 6h while Ch-EDTA adhered for just 1h. According to the combination of mucoadhesive and chelating properties of the conjugates synthesized in this study, Ch-EDTA-cys-2MNA might be useful for various mucosal drug delivery systems. Copyright © 2016 Elsevier B.V. All rights reserved.

Soft chelating irrigation protocol optimizes bonding quality of Resilon/Epiphany root fillings.

PubMed

De-Deus, Gustavo; Namen, Fátima; Galan, João; Zehnder, Matthias

2008-06-01

This study was designed to test the impact of either a strong (MTAD) or a soft (1-hydroxyethylidene-1, 1-bisphosphonate [HEPB]) chelating solution on the bond strength of Resilon/Epiphany root fillings. Both 17% EDTA and the omission of a chelator in the irrigation protocol were used as reference treatments. Forty extracted human upper lateral incisors were prepared using different irrigation protocols (n = 10): G1: NaOCl, G2: NaOCl + 17% EDTA, G3: NaOCl + BioPure MTAD (Dentsply/Tulsa, Tulsa, OK), and G4: NaOCl + 18% HEPB. The teeth were obturated and then prepared for micropush-out assessment using root slices of 1 mm thickness. Loading was performed on a universal testing machine at a speed of 0.5 mm/min. One-way analysis of variance and Tukey multiple comparisons were used to compare the results among the experimental groups. EDTA- and MTAD-treated samples revealed intermediate bond strength (0.3-3.6 MPa). The lowest bond strengths were achieved in NaOCl-treated samples (0.3-1.2 MPa, p < 0.05). The highest bond strength was reached in the HEBP-treated samples (3.1-6.1 MPa, p < 0.05). Under the present in vitro conditions, the soft chelating irrigation protocol (18% HEBP) optimized the bonding quality of Resilon/Epiphany (Resilon Research LLC, Madison, CT) root fillings.

Examining Radiation-Induced In Vivo and In Vitro Gene Expression Changes of the Peripheral Blood in Different Laboratories for Biodosimetry Purposes: First RENEB Gene Expression Study.

PubMed

Abend, M; Badie, C; Quintens, R; Kriehuber, R; Manning, G; Macaeva, E; Njima, M; Oskamp, D; Strunz, S; Moertl, S; Doucha-Senf, S; Dahlke, S; Menzel, J; Port, M

2016-02-01

The risk of a large-scale event leading to acute radiation exposure necessitates the development of high-throughput methods for providing rapid individual dose estimates. Our work addresses three goals, which align with the directive of the European Union's Realizing the European Network of Biodosimetry project (EU-RENB): 1. To examine the suitability of different gene expression platforms for biodosimetry purposes; 2. To perform this examination using blood samples collected from prostate cancer patients (in vivo) and from healthy donors (in vitro); and 3. To compare radiation-induced gene expression changes of the in vivo with in vitro blood samples. For the in vitro part of this study, EDTA-treated whole blood was irradiated immediately after venipuncture using single X-ray doses (1 Gy/min(-1) dose rate, 100 keV). Blood samples used to generate calibration curves as well as 10 coded (blinded) samples (0-4 Gy dose range) were incubated for 24 h in vitro, lysed and shipped on wet ice. For the in vivo part of the study PAXgene tubes were used and peripheral blood (2.5 ml) was collected from prostate cancer patients before and 24 h after the first fractionated 2 Gy dose of localized radiotherapy to the pelvis [linear accelerator (LINAC), 580 MU/min, exposure 1-1.5 min]. Assays were run in each laboratory according to locally established protocols using either microarray platforms (2 laboratories) or qRT-PCR (2 laboratories). Report times on dose estimates were documented. The mean absolute difference of estimated doses relative to the true doses (Gy) were calculated. Doses were also merged into binary categories reflecting aspects of clinical/diagnostic relevance. For the in vitro part of the study, the earliest report time on dose estimates was 7 h for qRT-PCR and 35 h for microarrays. Methodological variance of gene expression measurements (CV ≤10% for technical replicates) and interindividual variance (≤twofold for all genes) were low. Dose estimates based on one gene, ferredoxin reductase (FDXR), using qRT-PCR were as precise as dose estimates based on multiple genes using microarrays, but the precision decreased at doses ≥2 Gy. Binary dose categories comprising, for example, unexposed compared with exposed samples, could be completely discriminated with most of our methods. Exposed prostate cancer blood samples (n = 4) could be completely discriminated from unexposed blood samples (n = 4, P < 0.03, two-sided Fisher's exact test) without individual controls. This could be performed by introducing an in vitro-to-in vivo correction factor of FDXR, which varied among the laboratories. After that the in vitro-constructed calibration curves could be used for dose estimation of the in vivo exposed prostate cancer blood samples within an accuracy window of ±0.5 Gy in both contributing qRT-PCR laboratories. In conclusion, early and precise dose estimates can be performed, in particular at doses ≤2 Gy in vitro. Blood samples of prostate cancer patients exposed to 0.09-0.017 Gy could be completely discriminated from pre-exposure blood samples with the doses successfully estimated using adjusted in vitro-constructed calibration curves.

The Effects of EDTA (Ethylenediaminetetraacetic Acid) and Sonication on the Disaggregation of Oral Bacteria,

DTIC Science & Technology

1984-01-01

Virginia) and 0.5 g/ml menadione . The plates were c,,ltured for 72 hours anaerobically (<-140 my) at 37C A . and the number of colony forming units (CFU) per...CONTROL Immol lommol lOOmmol L DISPERSED pVVORTEX ~Thuwuuuuiii~FOR 30 SECONDS4 1 4 4 * MENADIONE SUPPLEMENTED SHAEDLER BLOOD AGAR PLATES lw- 2.50 U2.45

The Effect of Heparin on Fibrinolytic Activity and Platelet Function

DTIC Science & Technology

1995-09-16

or e- aminocaproic acid (38). For example, aprotinin administration prior to bypass has been shown to prevent the formation of FDP and the...Stephenson, and K. Rooney. The role of e- aminocaproic acid in reducing bleeding after cardiac operation: a double-blind randomized study. ;. Thorac...Specimens: Acquisition and Assays Blood was collected into tripotassium ethylenediaminetetraacetic acid (EDTA) for the measurement of hematocrit

Impact of Storage Time on Hepatitis B Virus DNA Stability in Clinical Specimens Determined by Quantitative Real-time PCR.

PubMed

Zhang, Xiaolian; Yang, Dongmei; Lu, Yu; Lao, Xianjun; Qin, Xue; Li, Shan

2016-01-01

Detecting blood levels of hepatitis B virus (HBV) DNA must be accurate and credible. Shipment and storage conditions of clinical samples affect the quality of nucleic acids and can interfere with HBV DNA analysis. The aim of our study was to compare HBV DNA stability in plasma specimens at 4 degrees C for different storage periods. Blood samples from 30 hepatitis B surface antigen (HBsAg) positive patients were collected in tubes containing EDTA-K2. Each sample was divided into eight aliquots, one of which was measured immediately for the initial viral load. The remaining aliquots were then stored at 4 degrees C and assessed after 1, 2, 3, 7, 14, 21, and 30 days of storage. Quantification of HBV DNA was performed by real-time polymerase chain reaction (RT-PCR), and the difference in HBV DNA concentrations between two different time points was analysed with a paired-samples t-test. HBV DNA was measured in a range of 2.00 - 8.00 IU/mL, with low within-run and between-run coefficients of variation (< 10%). Storing plasma for one month at 4 degrees C revealed no significant decrease in HBV DNA level (p = 0.231), and no trend was evident to indicate continued reduction over a 3-week storage period. Based on the results of this study, storing plasma for up to one month at 4 degrees C does not affect the stability of HBV DNA, regardless of the initial viral load.

Comparative analysis of endodontic smear layer removal efficacy of 17% ethylenediaminetetraacetic acid, 7% maleic acid, and 2% chlorhexidine using scanning electron microscope: An in vitro study.

PubMed

Attur, Kailash; Joy, Mathew T; Karim, Riyas; Anil Kumar, V J; Deepika, C; Ahmed, Haseena

2016-08-01

The aim of the present study was to evaluate the efficiency of different endodontic irrigants in the removal of smear layer through scanning electron microscopic image analysis. The present in vitro study was carried out on 45 single-rooted extracted human mandibular premolar teeth with single canal and complete root formation. Teeth were randomly assigned to three groups with 15 teeth in each group. Group I samples were irrigated with 17% ethylenediaminetetraacetic (EDTA) irrigation, Group II with 7% maleic acid irrigation, and Group III with 2% chlorhexidine irrigation. Scanning electron microscope evaluation was done for the assessment of smear layer removal in the coronal, middle, and apical thirds. Comparison of the smear layer removal between the three different groups was done by Kruskal-Wallis test, followed by Mann-Whitney U test for comparing individual groups. A P value less than 0.05 was considered to be statistically significant. Statistically significant difference was seen between the two test groups (17% EDTA vs. 7% maleic acid and 17% EDTA vs. 2% chlorhexidine) in smear layer removal at coronal, middle, and apical thirds of the root canal. The most efficient smear layer removal was seen in Group I with 17% EDTA irrigation compared with other groups (P < 0.05) and the least by 2% chlorhexidine. The present study shows that 17% EDTA efficiently removes the smear layer from root canal walls.

Process dependent thermoelectric properties of EDTA assisted bismuth telluride

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kulsi, Chiranjit; Banerjee, Dipali, E-mail: dipalibanerjeebesu@gmail.com; Kargupta, Kajari

2016-04-13

Comparison between the structure and thermoelectric properties of EDTA (Ethylene-diamine-tetra-acetic acid) assisted bismuth telluride prepared by electrochemical deposition and hydrothermal route is reported in the present work. The prepared samples have been structurally characterized by high resolution X-ray diffraction spectra (HRXRD), field emission scanning electron microscopy (FESEM) and high resolution transmission electron microscopic images (HRTEM). Crystallite size and strain have been determined from Williamson-Hall plot of XRD which is in conformity with TEM images. Measurement of transport properties show sample in the pellet form (S{sub 1}) prepared via hydrothermal route has higher value of thermoelectric power (S) than the electrodepositedmore » film (S{sub 2}). But due to a substantial increase in the electrical conductivity (σ) of the film (S{sub 2}) over the pellet (S{sub 1}), the power factor and the figure of merit is higher for sample S{sub 2} than the sample S{sub 1} at room temperature.« less

EDTA aggregates induce SYPRO orange-based fluorescence in thermal shift assay

PubMed Central

Kroeger, Tobias; Frieg, Benedikt; Zhang, Tao; Hansen, Finn K.; Marmann, Andreas; Proksch, Peter; Nagel-Steger, Luitgard; Groth, Georg; Smits, Sander H. J.

2017-01-01

Ethylenediaminetetraacetic acid (EDTA) is widely used in the life sciences as chelating ligand of metal ions. However, formation of supramolecular EDTA aggregates at pH > 8 has been reported, which may lead to artifactual assay results. When applied as a buffer component at pH ≈ 10 in differential scanning fluorimetry (TSA) using SYPRO Orange as fluorescent dye, we observed a sharp change in fluorescence intensity about 20°C lower than expected for the investigated protein. We hypothesized that this change results from SYPRO Orange/EDTA interactions. TSA experiments in the presence of SYPRO Orange using solutions that contain EDTA-Na+ but no protein were performed. The TSA experiments provide evidence that suggests that at pH > 9, EDTA4- interacts with SYPRO Orange in a temperature-dependent manner, leading to a fluorescence signal yielding a “denaturation temperature” of ~68°C. Titrating Ca2+ to SYPRO Orange and EDTA solutions quenched fluorescence. Ethylene glycol tetraacetic acid (EGTA) behaved similarly to EDTA. Analytical ultracentrifugation corroborated the formation of EDTA aggregates. Molecular dynamics simulations of free diffusion of EDTA-Na+ and SYPRO Orange of in total 27 μs suggested the first structural model of EDTA aggregates in which U-shaped EDTA4- arrange in an inverse bilayer-like manner, exposing ethylene moieties to the solvent, with which SYPRO Orange interacts. We conclude that EDTA aggregates induce a SYPRO Orange-based fluorescence in TSA. These results make it relevant to ascertain that future TSA results are not influenced by interference between EDTA, or EDTA-related molecules, and the fluorescent dye. PMID:28472107

Deaths associated with hypocalcemia from chelation therapy--Texas, Pennsylvania, and Oregon, 2003-2005.

PubMed

2006-03-03

Chelating agents bind lead in soft tissues and are used in the treatment of lead poisoning to enhance urinary and biliary excretion of lead, thus decreasing total lead levels in the body. During the past 30 years, environmental and dietary exposures to lead have decreased substantially, resulting in a considerable decrease in population blood lead levels (BLLs) and a corresponding decrease in the number of patients requiring chelation therapy. Chelating agents also increase excretion of other heavy metals and minerals, such as zinc and, in certain cases, calcium. This report describes three deaths associated with chelation-therapy--related hypocalcemia that resulted in cardiac arrest. Several drugs are used in the treatment of lead poisoning, including edetate disodium calcium (CaEDTA), dimercaperol (British anti-Lewisite), D-penicillamine, and meso-2,3-dimercaptosuccinic acid (succimer). Health-care providers who are unfamiliar with chelating agents and are considering this treatment for lead poisoning should consult an expert in the chemotherapy of lead poisoning. Hospital pharmacies should evaluate whether continued stocking of Na2EDTA is necessary, given the established risk for hypocalcemia, the availability of less toxic alternatives, and an ongoing safety review by the Food and Drug Administration (FDA). Health-care providers and pharmacists should ensure that Na2EDTA is not administered to children during chelation therapy.

Circulating angiotensin-converting enzyme 2 activity in patients with chronic kidney disease without previous history of cardiovascular disease.

PubMed

Anguiano, Lidia; Riera, Marta; Pascual, Julio; Valdivielso, José Manuel; Barrios, Clara; Betriu, Angels; Mojal, Sergi; Fernández, Elvira; Soler, María José

2015-07-01

Patients with cardiovascular (CV) disease have an increased circulating angiotensin-converting enzyme 2 (ACE2) activity, but there is little information about changes in ACE2 in chronic kidney disease (CKD) patients without history of CV disease. We examined circulating ACE2 activity in CKD patients at stages 3-5 (CKD3-5) and in dialysis (CKD5D) without any history of CV disease. Circulating ACE2 activity was measured in human ethylenediamine-tetraacetic acid (EDTA)-plasma samples from the NEFRONA study (n = 2572): control group (CONT) (n = 568), CKD3-5 (n = 1458) and CKD5D (n = 546). Different clinical and analytical variables such as gender; age; history of diabetes mellitus (DM), dyslipidemia and hypertension; glycaemic, renal, lipid and anaemia profiles; vitamin D analogues treatment and antihypertensive treatments (angiotensin-converting enzyme inhibitor and angiotensin receptor blockade) were analysed. Circulating ACE2 and ACE activities were measured using modified fluorimetric assay for EDTA-plasma samples, where zinc chloride was added to recover enzymatic activity. In CKD3-5 and CKD5D, significant decrease in circulating ACE2 activity was observed when compared with CONT, but no differences were found between CKD3-5 and CKD5 when performing paired case-control studies. By multivariate linear regression analysis, male gender and advanced age were identified as independent predictors of ACE2 activity in all groups. Diabetes was identified as independent predictor of ACE2 activity in CKD3-5. Significant increase in the activity of circulating ACE was found in CKD3-5 and CKD5D when compared with CONT and in CKD5D when compared with CKD3-5. By multiple regression analysis, female gender and younger age were identified as independent predictors of ACE activity in CONT and CKD3-5. Diabetes was also identified as an independent predictor of ACE activity in CKD3-5 patients. Circulating ACE2 and ACE activities can be measured in human EDTA-plasma samples with zinc added to recover enzymatic activity. In a CKD population without previous history of CV disease, ACE2 activity from human EDTA-plasma samples directly correlated with the classical CV risk factors namely older age, diabetes and male gender. Our data suggest that circulating ACE2 is altered in CKD patients at risk for CV event. © The Author 2015. Published by Oxford University Press on behalf of ERA-EDTA. All rights reserved.

Stability and reproducibility of proteomic profiles measured with an aptamer-based platform.

PubMed

Kim, Claire H; Tworoger, Shelley S; Stampfer, Meir J; Dillon, Simon T; Gu, Xuesong; Sawyer, Sherilyn J; Chan, Andrew T; Libermann, Towia A; Eliassen, A Heather

2018-05-30

The feasibility of SOMAscan, a multiplex, high sensitivity proteomics platform, for use in studies using archived plasma samples has not yet been assessed. We quantified 1,305 proteins from plasma samples donated by 16 Nurses' Health Study (NHS) participants, 40 NHSII participants, and 12 local volunteers. We assessed assay reproducibility using coefficients of variation (CV) from duplicate samples and intra-class correlation coefficients (ICC) and Spearman correlation coefficients (r) of samples processed (i.e., centrifuged and aliquoted into separate components) immediately, 24, and 48 hours after collection, as well as those of samples collected from the same individuals 1 year apart. CVs were <20% for 99% of proteins overall and <10% for 92% of proteins in heparin samples compared to 66% for EDTA samples. We observed ICC or Spearman r (comparing immediate vs. 24-hour delayed processing) ≥0.75 for 61% of proteins, with some variation by anticoagulant (56% for heparin and 70% for EDTA) and protein class (ranging from 49% among kinases to 83% among hormones). Within-person stability over 1 year was good (ICC or Spearman r ≥ 0.4) for 91% of proteins. These results demonstrate the feasibility of SOMAscan for analyses of archived plasma samples.

Quality testing of an innovative cascade separation system for multiple cell separation

NASA Astrophysics Data System (ADS)

Pierzchalski, Arkadiusz; Moszczynska, Aleksandra; Albrecht, Bernd; Heinrich, Jan-Michael; Tarnok, Attila

2012-03-01

Isolation of different cell types from mixed samples in one separation step by FACS is feasible but expensive and slow. It is cheaper and faster but still challenging by magnetic separation. An innovative bead-based cascade-system (pluriSelect GmbH, Leipzig, Germany) relies on simultaneous physical separation of different cell types. It is based on antibody-mediated binding of cells to beads of different size and isolation with sieves of different mesh-size. We validated pluriSelect system for single parameter (CD3) and simultaneous separation of CD3 and CD15 cells from EDTA blood-samples. Results were compared with those obtained by MACS (Miltenyi-Biotech) magnetic separation (CD3 separation). pluriSelect separation was done in whole blood, MACS on Ficoll gradient isolated leukocytes, according to the manufacturer's protocols. Isolated and residual cells were immunophenotyped (7-color 8-antibody panel (CD3; CD16/56; CD4; CD8; CD14; CD19; CD45; HLADR) on a CyFlowML flow cytometer (Partec GmbH). Cell count (Coulter), purity, yield and viability (7-AAD exclusion) were determined. There were no significant differences between both systems regarding purity (92-98%), yield (50-60%) and viability (92-98%) of isolated cells. PluriSelect separation was slightly faster than MACS (1.15 h versus 1.5h). Moreover, no preenrichment steps were necessary. In conclusion, pluriSelect is a fast, simple and gentle system for efficient simultaneous separation of two cell subpopulation directly from whole blood and can provide a simple alternative to FACS. The isolated cells can be used for further research applications.

Chelant-enhanced heavy metals uptake by Eucalyptus trees under controlled deficit irrigation

NASA Astrophysics Data System (ADS)

Fine, Pinchas; Rathod, Paresh; Beriozkin, Anna; Ein-Gal, Oz; Hass, Amir

2014-05-01

Enhancement of phytoremediation of heavy metal polluted soils employs organic ligands, aimed to solubilize, phytoextract and translocate metals into the canopy. The use of more persistent chelants (e.g. EDTA) is phasing out due to concerns over their role in the environment. We tested the hypothesis that controlled deficit irrigation (CDI) of the fast growing, salinity resistant Eucalyptus camaldulensis coupled with timely EDTA application enhances sediment phytoremediation while minimizing leaching of metal complexes below the root-zone. This was tested in 220-L lysimeters packed with sand mixed with metals polluted biosolids. One year old trees were brought under CDI with tap or RO water for two growing seasons. EDTA, EDDS and citric acid fertigation at 2 mM started in each May for 2.5-3.5 months, and prescribed soil leaching and sampling of tree leaves started thereafter. While all 3 chelants solubilized biosolids metal in batch extraction (EDDS often being the more efficient), EDTA was the only to increased metal concentrations both in the soil solution and in the Eucalyptus leaves. The average concentrations in the soil solution and in the leaves, in the EDTA vs. control (chelant-free) treatments, all respectively, were: Cd - 200 mg L-1 vs. 1.0, and 67 vs. 21 mg kg-1; Cu: 90 vs. 1.5 mg L-1, and 17 vs. 3.0 mg kg-1; Cr: 4.0 vs. 1.4 mg L-1, and 3.0 vs. 1.0 mg kg-1; Ni: 60 mg L-1 vs. 14, and 20 vs. 6.0 mg kg-1; Pb: >44 vs. 0.1 mg L-1, and 9.0 vs. 1.0 mg kg-1; and Zn: 650 vs. 4.0 mg L-1 and 200 vs. 70 mg kg-1. While EDDS was undetectable in all the leachates, EDTA concentrated to up to 100 mM. At 10 mM soil solution concentration, EDDS half-life in acclimated lysimeter media was 5-11 days and that of EDTA was ≥27-d. The study suggests that sustainable phytostabilization and phytoextraction of heavy metals are achievable under CDI with EDTA augmentation at low dose. This was yet futile with the biodegradable EDDS and citric acid. CDI with RO water further widened the scope of this remediation technique

Biodegradation of PuEDTA and Impacts on Pu Mobility

DOE Office of Scientific and Technical Information (OSTI.GOV)

Xun, Luying; Bolton, Jr., Harvey

Ethylenediaminetetraacetate (EDTA) and nitrilotriacetate (NTA) are synthetic chelating agents, which can form strong water-soluble complexes with radionuclides and metals and has been used to decontaminate and process nuclear materials. Synthetic chelating agents were co-disposed with radionuclides (e.g., 60Co, Pu) and heavy metals enhancing their transport in the subsurface. An understanding of EDTA biodegradation is essential to help mitigate enhanced radionuclide transport by EDTA. The objective of this research is to develop fundamental data on factors that govern the biodegradation of radionuclide-EDTA. These factors include the dominant EDTA aqueous species, the biodegradation of various metal-EDTA complexes, the uptake of various metal-EDTAmore » complexes into the cell, the distribution and mobility of the radionuclide during and after EDTA biodegradation, and the enzymology and genetics of EDTA biodegradation.« less

Hemodynamic and permeability characteristics of acute experimental necrotizing enterocolitis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Miller, M.J.; Adams, J.; Gu, X.A.

1990-10-01

We examined the local hemodynamic response of intestinal loops during acute necrotizing enterocolitis (NEC) in anesthetized rabbits. NEC was induced in ileal loops by transmural injection of a solution containing casein (10 mg/ml) and calcium gluconate (50 mg/ml) acidified to pH 4.0 with propionic or acetic acid. Control loops received casein only (pH 5.0). Mucosal damage was quantified by the blood-to-lumen movement of (51Cr)EDTA, fluid shifts into the lumen, and histology. Mean arterial pressure and loop blood flow were steady over the 3-hr period, loop fluid volume decreased, and there was no evidence of necrosis or epithelial damage. In loopsmore » receiving acidified casein and calcium gluconate, there was an immediate dramatic increase in loop blood flow that returned to baseline by 50 min. In addition, loop fluid volume was dramatically increased, necrosis was noted in the form of blunting and loss of villi, and sevenfold increase in (51Cr)EDTA permeability was evident. Administration of CV 1808 (30 mg/kg/hr), a selective adenosine2 agonist, which maintained and elevated loop blood flow throughout the 3 hr protocol, failed to alter the changes in loop fluid volume or prevent necrosis. Histamine levels in loop fluid levels were significantly elevated 20-30 min after NEC induction when compared to saline controls, indicating an early activation of mucosal defenses with this luminal insult. Thus, this model of NEC is characterized by a transient, acute hyperemia, increased intestinal permeability, and histamine release. As mucosal damage was independent of ischemia and could not be prevented by vasodilatory therapy, this model supports the clinical findings that NEC is correlated with luminal factors related to feeding and independent of cardiovascular stress.« less

[Evaluation of initial results of treatment of lead poisoning with EDTA].

PubMed

Petkova, V; Adjarov, D; Pavlova, S; Naydenova, E; Kerimova, M; Kuneva, T

1994-01-01

The results of EDTA therapy were studied in 37 workers of a battery factory consisting of males with varying degrees of occupational lead poisoning (low exposure: 10 subjects, blood lead levels (PbB) lower than 400 micrograms/l with slight alterations in heme biosynthesis; beyond limit of effect: 5 subjects, PbB > 400 micrograms/l; slight intoxication: 19 subjects, with marked alterations in heme synthesis and preclinical signs of intoxication; average degree of intoxication: 3 subjects with clinical signs of intoxication. Clinical symptoms and the following parameters were investigated: blood lead (PbB), delta-aminolevulinic acid dehydratase in erythrocytes (ALA-D), zinc protoporphyrin (PP) in erythrocytes and delta-aminolevulinic acid (ALA) in 24-hour urine before and after EDTA chelating therapy. Simultaneous measurement of ALA-D and PP showed high diagnostic sensitivity in detecting lead poisoning in occupationally exposed subjects. In view of the high interindividual variability of the results, these indices did not, however, permit a useful differentiation to be made of the different degrees of intoxication at individual level, even though a good correlation was observed between PbB and porphyrin metabolism indices. From the alterations observed in ALA-D and PP values it was not possible to establish an association between degree of alteration and types of clinical symptoms in the different intoxication studies. At the end of EDTA treatment, a clinical improvement was observed in all cases studied but only in 5 cases was a reduction in PbB observed, to levels below 1.20 mol/l, which is accepted as a permissible limit for the general population; in 17 cases PbB remained at levels above the critical value for occupational lead poisoning (400 micrograms/l), although there was a decrease after treatment. The improvement observed in the indices of porphyrin metabolism at the end of treatment was only slight: significant variations were measured only for PbB. After treatment no association was observed between ALA-D and PP variations in erythrocytes and improvement in clinical symptoms; measurement of these indices therefore seems to be of little use in assessing the efficacy of the treatment. In spite of its limited diagnostic sensitivity during intoxication, measurement of ALA in urine could be useful to assess the efficacy of chelating therapy in subjects in whom the values are initially altered.

Zirconium(IV) functionalized magnetic nanocomposites for extraction of organophosphorus pesticides from environmental water samples.

PubMed

Jiang, Li; Huang, Tengjun; Feng, Shun; Wang, Jide

2016-07-22

The widespread use of organophosphate pesticides (OPPs) in agriculture leads to residue accumulation in the environment which is dangerous to human health and disrupts the ecological balance. In this work, one nanocomposite immobilized zirconium (Zr, IV) was prepared and used as the affinity probes to quickly and selectively extract organophosphorus pesticides (OPPs) from water samples. The Fe3O4-ethylenediamine tetraacetic acid (EDTA)@Zr(IV) nanocomposites (NPs) were prepared by simply mixing Zr(IV) ions with Fe3O4-EDTA NPs synthesized by one-pot chemical co-precipitation method. The immobilized Zr(IV) ions were further utilized to capture OPPs based on their high affinity for the phosphate moiety in OPPs. Coupled with GC-MS, four OPPs were used as models to demonstrate the feasibility of this approach. Under the optimum conditions, the limits of detection for target OPPs were in the range of 0.10-10.30ngmL(-1) with relative standard deviations (RSDs) of 0.61-4.40% (n=3), respectively. The linear ranges were over three orders of magnitudes (correlation coefficients, R(2)>0.9995). The Fe3O4-EDTA@Zr(IV) NPs were successfully applied to extract OPPs samples with recoveries of 86.95-112.60% and RSDs of 1.20-10.42% (n=3) from two spiked real water. By the proposed method, the matrix interference could be effectively eliminated. We hope our finding can provide a promising alternative for the fast extraction of OPPs from complex real samples. Copyright © 2016 Elsevier B.V. All rights reserved.

Unexpected effect of dry olive leaf extract on the level of DNA damage in lymphocytes of lead intoxicated workers, before and after CaNa2EDTA chelation therapy.

PubMed

Čabarkapa, Andrea; Dekanski, Dragana; Živković, Lada; Milanović-Čabarkapa, Mirjana; Bajić, Vladan; Topalović, Dijana; Giampieri, Francesca; Gasparrini, Massimiliano; Battino, Maurizio; Spremo-Potparević, Biljana

2017-08-01

The CaNa 2 EDTA chelation therapy is often practiced with antioxidant supplementation. Dry olive leaf extract (DOLE) is natural product with antioxidant and DNA protective properties. The effects of DOLE on the levels of DNA damage were investigated ex vivo in peripheral blood lymphocytes (PBLs) of 19 workers occupationally exposed to lead (Pb), before and after CaNa 2 EDTA chelation therapy. DOLE demonstrated pronounced radical scavenging activity in concentrations ≥1 mg/mL, and showed no genotoxicity per se, in concentrations 0.125-1 mg/mL. The level of DNA damage in PBLs of workers before chelation therapy was elevated (24.21 ± 14.26) compared to controls (6.0 ± 3.37). The incubation of PBLs before chelation therapy with selected concentration of DOLE lead to a severe increase of DNA damage (64.03 ± 20.96), exhibiting prooxidant rather than antioxidant effect. After the five-day CaNa 2 EDTA chelation regimen, DNA damage in PBLs of workers decreased (8.26 ± 4.62) significantly compared to baseline. Treatment of PBLs with DOLE after chelation, again produced high level of damage (41.82 ± 23.17) and the acute prooxidant effects of DOLE remained, but, DNA damage was less severe than before chelation. The DOLE exhibits prooxidant effect in presence of Pb in lymphocytes of exposed workers, and its effect is less pronounced following the removal of Pb after standard chelation therapy. Copyright © 2016 Elsevier Ltd. All rights reserved.

Flow Cytometry Method Analysis of Apoptosis: No Significant Difference Between EDTA and EDTA-free Trypsin Treatment Procedure.

PubMed

Xu, Xiao-yan; Nie, Xiao-cui; Ma, Hai-ying; Song, Guo-qing; Zhang, Xiao-tong; Jin, Yu-nan; Yu, Yan-qiu

2015-04-01

Flow cytometry method (FCM) is a generally accepted tool to analyze apoptosis. Although apoptosis assay kit was applied by many companies, the manufacturers were not consistent with whether using Trypsin with EDTA to collect the adherent cells. In another words, the influence of EDTA on apoptotic ratio is not clear. In this work, we compared the proportion of apoptotic cells with EDTA or EDTA-free Trypsin treatment by FCM. We concluded that Trypsin with or without EDTA has little influence on the proportion of apoptotic cells. In addition, we found that the ratio of necrosis and apoptosis was different in cells collected by scraping. WAVE2 protein was analyzed as a typical example for movement related protein. WAVE2 expression is elevated in the EDTA Trypsin treated group, compared with EDTA-free Trypsin treatment and scrapping group. © The Author(s) 2014.

Isolation and growth characteristics of an EDTA-degrading member of the alpha-subclass of Proteobacteria.

PubMed

Weilenmann, Hans-Ueli; Engeli, Barbara; Bucheli-Witschel, Margarete; Egli, Thomas

2004-10-01

A Gram-negative, ethylenediaminetetraacetic acid (EDTA)-degrading bacterium (deposited at the German Culture Collection as strain DSM 9103) utilising EDTA as the only source of carbon, energy and nitrogen was isolated from a mixed EDTA-degrading population that was originally enriched in a column system from a mixture of activated sludge and soil. Chemotaxonomic analysis of quinones, polar lipids and fatty acids allowed allocation of the isolate to the alpha-subclass of Proteobacteria. 16S rDNA sequencing and phylogenetic analysis revealed highest similarity to the Mesorhizobium genus followed by the Aminobacter genus. However, the EDTA-degrading strain apparently forms a new branch within the Phyllobacteriaceae/Mesorhizobia family. Growth of the strain was rather slow not only on EDTA (micro(max) = 0.05 h(-1)) but also on other substrates. Classical substrate utilisation testing in batch culture suggested a quite restricted carbon source spectrum with only lactate, glutamate, and complexing agents chemically related to EDTA (nitrilotriacetate, iminodiacetate and ethylenediaminedisuccinate) supporting growth. However, when EDTA-limited continuous cultures of strain DSM 9103 were pulsed with fumarate, succinate, glucose or acetate, these substrates were assimilated immediately. Apparently, the strain can use a broader spectrum than indicated by traditional substrate testing techniques. The EDTA species CaEDTA and MgEDTA served as growth substrates of the strain because in the mineral medium employed EDTA was predicted to be mainly present in the form of these two complexes. The bacterium was not able to degrade Fe3+-complexed EDTA.

Speciation and preservation of inorganic arsenic in drinking water sources using EDTA with IC separation and ICP-MS detection.

PubMed

Gallagher, P A; Schwegel, C A; Wei, X; Creed, J T

2001-08-01

The native distribution of As(III) and As(v) in drinking water supplies can influence the treatment removal strategy. The stability of As(III) and As(v) in iron-rich drinking waters can be affected by the formation of Fe precipitates (Fe oxides and/or hydroxides designated by "FeOOH"). These precipitates (ppts) can form during the transport of the sample to the laboratory for arsenic speciation analysis. The analysis of the ppt indicates considerable loss of the aqueous arsenic species (As(aq)) to the solid phase "FeOOH" ppt. Studies of laboratory reagent water containing both As(III) and Fe(III) indicate that the resulting "FeOOH" ppt contained a mixture of As(III) and As(v) with near quantitative removal of the As(aq) in 18 hr. The corresponding aqueous fraction after filtration through a 0.45 microm filter was composed primarily of As(v). The formation of "FeOOH" ppt and the loss of As(aq) to the ppt can be virtually eliminated by the use of EDTA, which sequesters the FeIII). Reagent water fortified with Fe(III), As(III) and EDTA produced less than a 1 ppb change in the As(III)aq concentration over 16 d. The EDTA treatment was also tested on three well waters with different native As(III )/As(v) ratios. The native distribution of As(III)/As(v) was stabilized over a period of 10 d with a worst case conversion of As(III) to As(v) of 2 ppb over a 30 d period. All well waters not treated with EDTA had dramatic losses (a factor of 2-5) of As(aq) in less than 1 d. These results indicated that EDTA preservation treatment can be used to preserve As(aq) in waters where the predominant species is the reduced form [As(III)] or in waters which the predominant species is the oxidized form [As(v)]. This preliminary investigation of EDTA to preserve As species in Fe-rich waters indicates stability can be achieved for greater than 14 d.

Complexometric determination of some toxic mixtures of ions using bromo-cresol orange with visual endpoint indication.

PubMed

Hafez, M A; Khalifa, M E

1997-05-01

A rapid and simple general complexometric method was presented for the determination of lead, cadmium and thallium or mercury or arsenic(V) in laboratory synthesized mixtures similar to those of some ores, minerals and alloys of such metals. The precision and accuracy attainable in successive titrations of Pb(2+), Cd(2+) and Tl(3+) or Hg(2+) or AsO(3-)(4) (As(5+)) with 0.05 and/or 0.01 mol 1(-1) solutions of disodium ethylenediaminetetraacetate (Na(2)EDTA) and standard Pb(NO(3))(2) of the same concentration using Bromo-Cresol Orange (BCO) as a new metallochromic indicator with visual endpoint indication were studied. For the analysis of a three component mixtures of the aforementioned ions, Tl(3+) was at first directly titrated with Na(2)EDTA at pH 0.5-1 (HNO(3)) using BCO as indicator. At the thallium endpoint an excess of Na(2)EDTA was added and the pH was adjusted at pH approximately 4.8 using hexamine-HNO(3) buffer (solution A). The excess EDTA was back-titrated with standard solution of Pb(NO(3))(2). 1,10-Phenanthroline (1,10-phen) was added to release the EDTA combined with Cd(2+), while thiosemicarbazide (TSC) was used to liberate the EDTA from the mercury-EDTA chelate. To determine AsO(3-)(4) ion in such type of mixtures the pH of (solution A) was raised to a value of 10 using ammonia buffer. Excess standard Mg(2+) solution was added and the formed precipitate of MgNH(4)AsO(4) was separated, dissolved and its magnesium content equivalent to AsO(3-)(4) was determined complexometrically using Eriochrome Black-T (EBT) indicator. The interference caused by different anions, cations and organic acids was investigated. A comparison of the indicators BCO and Xylenol Orange (XO) for successive titration of the studied metal ions was carried out. The proposed successive titration method was applied successfully to some real samples of ores, minerals and alloys of the studied metal ions and the results were satisfactory and agreed with those obtained by AAS.

Increased Proportion of Hematopoietic Stem and Progenitor Cell Population in Cord Blood of Neonates Born to Mothers with Gestational Diabetes Mellitus.

PubMed

Hadarits, Orsolya; Zóka, András; Barna, Gábor; Al-Aissa, Zahra; Rosta, Klára; Rigó, János; Kautzky-Willer, Alexandra; Somogyi, Anikó; Firneisz, Gábor

2016-01-01

We assessed the hematopoietic stem and progenitor cell (HSPC) population in the cord blood of neonates born to mothers with gestational diabetes mellitus (GDM) in a hypothesis generating pilot study, due to that, neonatal polycythemia may be the consequence of GDM pregnancy. Forty-five pregnant women with GDM (last trimester mean HbA1C = 33.9 mmol/mol) and 42 (nondiabetic) control pregnant women were enrolled after their routine 75 g oral glucose tolerance test (OGTT) between the 24th and 28th gestational week (with expected differences in their mean routine clinical characteristics: plasma glucose at OGTT: 0' = 5.07 vs. 4.62 mM, 120' = 8.9 vs. 5.76 mM, age = 35.07 vs. 31.66 years, prepregnancy body mass index = 27.9 vs. 23.9 kg/m(2), GDM vs. control, respectively) on a voluntary basis after signing the informed consent. EDTA-treated cord blood samples were analyzed by flow cytometry and the software Kaluza1.2 using CD45 and CD34-specific fluorescent antibodies to identify the HSPC population (CD34(+) cells within the CD45(dim) blast gate). The proportion of CD34(+)CD45(dim) HSPCs among the nucleated cells was significantly (P < 0.05, statistical power = 60.8%) higher in the cord blood samples of neonates born to mothers with GDM (median 0.38%) compared to neonates born to nondiabetic mothers (median 0.32%) and according to treatment types (P < 0.05) median: control 0.32%, GDM-diet only 0.37%, GDM-on insulin 0.45%; control versus GDM on insulin (P < 0.05). The increased proportion of circulating CD34(+)CD45(dim) cells in the cord blood may possibly be related to altered fetal stem cell mobilization in GDM pregnancy, yet these results should be interpreted only as preliminary due to the small sample sizes.

Preservation of As(III) and As(V) in drinking water supply samples from across the United States using EDTA and acetic acid as a means of minimizing iron-arsenic coprecipitation.

PubMed

Gallagher, Patricia A; Schwegel, Carol A; Parks, Amy; Gamble, Bryan M; Wymer, Larry; Creed, John T

2004-05-15

Seven different treatment/storage conditions were investigated for the preservation of the native As(III)/As(V) found in 10 drinking water supplies from across the United States. These 10 waters were chosen because they have different As(III)/As(V) distributions; six of these waters contained enough iron to produce an iron precipitate during shipment. The waters were treated and stored under specific conditions and analyzed periodically over a span of approximately 75 days. Linear least squares (LLS) was used to estimate the change in As(III) and As(V) over the study period. Point estimates for the first and last analyses days and 95% confidence bounds were calculated from the LLS. The difference in the point estimates for the first and last day were then evaluated with respect to drinking water treatment decision making. Three primary treatments were evaluated: EDTA/AcOH-treatment and AcOH treatment as well as no treatment. The effect of temperature was explored for all treatments, while the effect of aeration was evaluated for only the EDTA/AcOH treated samples. The nontreated samples experienced a 0-40% reduction in the native arsenic concentration due to the formation of Fe/As precipitates. The Fe/As precipitates were resolubilized and shown to contain elevated concentrations of As(V) relative to the native distribution. Once this Fe/As precipitate was removed from solution using a 0.45 and 0.2 microm filter, the resulting arsenic concentration (As(III) + As(V)) was relatively constant (the largest LLS slope was -1.4 x 10(-2) (ng As g water(-1)) day(-1)). The AcOH treatment eliminated the formation of the Fe/As precipitate observed in the nontreated samples. However, two of the AcOH water samples produced analytically significant changes in the As(III) concentration. The LLS slopes for these two waters were -5.7 x 10(-2) (ng As(III) g water(-1)) day(-1) and -1.0 x 10(-1) (ng As(III) g water(-1)) day(-1). This corresponds to a -4.3 ng/g and a -7.8 ng/g change in the As(III) concentration over the study period, which is a 10% shift in the native distribution. The third and final treatment was EDTA/AcOH. This treatment eliminated the Fe/As precipitate that formed in the nontreated sample. The LLS slopes were less than -7.5 x 10(-3) (ng As(III) g water(-1)) day(-1) for the above-mentioned waters, corresponding to a 0.6 ng/g change over the study period. One of the EDTA/AcOH treated waters did indicate that using the 5 degrees C storage temperature minimized the rate of conversion relative to 20 degrees C storage.

Renal blood flow using arterial spin labelling MRI and calculated filtration fraction in healthy adult kidney donors Pre-nephrectomy and post-nephrectomy.

PubMed

Cutajar, Marica; Hilton, Rachel; Olsburgh, Jonathon; Marks, Stephen D; Thomas, David L; Banks, Tina; Clark, Christopher A; Gordon, Isky

2015-08-01

Renal plasma flow (RPF) (derived from renal blood flow, RBF) and glomerular filtration rate (GFR) allow the determination of the filtration fraction (FF), which may have a role as a non-invasive renal biomarker. This is a hypothesis-generating pilot study assessing the effect of nephrectomy on renal function in healthy kidney donors. Eight living kidney donors underwent arterial spin labelling (ASL) magnetic resonance imaging (MRI) and GFR measurement prior to and 1 year after nephrectomy. Chromium-51 labelled ethylenediamine tetraacetic acid ((51)Cr-EDTA) with multi-blood sampling was undertaken and GFR calculated. The RBF and GFR obtained were used to calculate FF. All donors showed an increase in single kidney GFR of 24 - 75 %, and all but two showed an increase in FF (-7 to +52 %) after nephrectomy. The increase in RBF, and hence RPF, post-nephrectomy was not as great as the increase in GFR in seven out of eight donors. As with any pilot study, the small number of donors and their relatively narrow age range are potential limiting factors. The ability to measure RBF, and hence RPF, non-invasively, coupled with GFR measurement, allows calculation of FF, a biomarker that might provide a sensitive indicator of loss of renal reserve in potential donors. • Non-invasive MRI measured renal blood flow and calculated renal plasma flow. • Effect of nephrectomy on blood flow and filtration in donors is presented. • Calculated filtration fraction may be a useful new kidney biomarker.

Specificity of EIA immunoassay for complement factor Bb testing.

PubMed

Pavlov, Igor Y; De Forest, Nikol; Delgado, Julio C

2011-01-01

During the alternative complement pathway activation, factor B is cleaved in two fragments, Ba and Bb. Concentration of those fragments is about 2 logs lower than of factor B present in the blood, which makes fragment detection challenging because of potential cross-reactivity. Lack of information on Bb assay cross-reactivity stimulated the authors to investigate this issue. We ran 109 healthy donor EDTA plasmas and 80 sera samples with both factor B immunodiffusion (The Binding Site) and Quidel Bb EIA assays. During the study it was shown that physiological concentrations of gently purified factor B demonstrated approximately 0.15% cross-reactivity in the Quidel Bb EIA assay. We also observed that Bb concentration in serum is higher than in plasma due to complement activation during clot formation which let us use sera as samples representing complement activated state. Our study demonstrated that despite the potential 0.15% cross-reactivity between endogenous factor B and cleaved Bb molecule, measuring plasma concentrations of factor Bb is adequate to evaluate the activation of the alternative complement pathway.

Seasonal influence on the hematological parameters in cultured Nile tilapia from southern Brazil.

PubMed

Jerônimo, G T; Laffitte, L V; Speck, G M; Martins, M L

2011-08-01

This study evaluated seasonality in hematological parameters of Nile tilapia cultured in the state of Santa Catarina, southern Brazil. A total of 240 fish were examined during four seasons between April 2007 and March 2008 in three different fish farms. After being anesthetised in a benzocaine solution, blood samples were withdrawn into syringes containing a drop of 10% EDTA for hematological analysis. The results were compared between fish farms and seasons, which are well delimited in southern Brazil. In a traditional fish farm in Joinville in the summer, there was an increase in the percentage of hematocrit and in the red blood cell count. The highest values of total leukocytes were found in fish from fee-fishing in Blumenau in the autumn while the lowest values occurred in those from swine consorted system in Ituporanga in the summer. Thrombocytosis was observed in the autumn, and lymphocytosis was found in both the autumn and winter in tilapia from all fish farms investigated. Neutrophilia was only observed in winter and autumn in fish from Blumenau and Ituporanga. This work demonstrated the influence of seasonality and the handling characteristics of each fish farm on certain hematological parameters in Nile tilapia.

An association study between CHEK2 gene mutations and susceptibility to breast cancer.

PubMed

Jalilvand, Manizheh; Oloomi, Mana; Najafipour, Reza; Alizadeh, Safar Ali; Saki, Najmaldin; Rad, Fatemeh Samiee; Shekari, Mohammad

2017-01-01

CHEK2 gene is known as a tumor suppressor gene in breast cancer (BC), which plays a role in DNA repair. The germ line mutations in CEHK2 have been associated with different types of cancer. The present study was aimed at studying the association between CHEK2 mutations and BC. Peripheral blood was collected from patients into a test tube containing EDTA, and DNA was extracted from blood samples. Then, we analyzed mutations including 1100delc, IVS2+1>A, del5395bp, and I157T within CHEK2 gene in patients with BC and 100 normal healthy controls according to PCR-RFLP, allelic specific PCR, and multiplex-PCR. Although IVS2+1G>A mutation within CHEK2 gene was found in two BC patients, other defined mutants were not detected. For the first time, we identified CHEK2 IVS2+1G>A mutation, one out of four different CHEK2 alterations in two Iranian BC patients (2%). Also, our results showed that CHEK2 1100elC, del5395bp, and I157T mutations are not associated with genetic susceptibility for BC among Iranian population.

Capillary Electrophoresis Analysis of Organic Amines and Amino Acids in Saline and Acidic Samples Using the Mars Organic Analyzer

NASA Astrophysics Data System (ADS)

Stockton, Amanda M.; Chiesl, Thomas N.; Lowenstein, Tim K.; Amashukeli, Xenia; Grunthaner, Frank; Mathies, Richard A.

2009-11-01

The Mars Organic Analyzer (MOA) has enabled the sensitive detection of amino acid and amine biomarkers in laboratory standards and in a variety of field sample tests. However, the MOA is challenged when samples are extremely acidic and saline or contain polyvalent cations. Here, we have optimized the MOA analysis, sample labeling, and sample dilution buffers to handle such challenging samples more robustly. Higher ionic strength buffer systems with pKa values near pH 9 were developed to provide better buffering capacity and salt tolerance. The addition of ethylaminediaminetetraacetic acid (EDTA) ameliorates the negative effects of multivalent cations. The optimized protocol utilizes a 75 mM borate buffer (pH 9.5) for Pacific Blue labeling of amines and amino acids. After labeling, 50 mM (final concentration) EDTA is added to samples containing divalent cations to ameliorate their effects. This optimized protocol was used to successfully analyze amino acids in a saturated brine sample from Saline Valley, California, and a subcritical water extract of a highly acidic sample from the Río Tinto, Spain. This work expands the analytical capabilities of the MOA and increases its sensitivity and robustness for samples from extraterrestrial environments that may exhibit pH and salt extremes as well as metal ions.

Capillary electrophoresis analysis of organic amines and amino acids in saline and acidic samples using the Mars organic analyzer.

PubMed

Stockton, Amanda M; Chiesl, Thomas N; Lowenstein, Tim K; Amashukeli, Xenia; Grunthaner, Frank; Mathies, Richard A

2009-11-01

The Mars Organic Analyzer (MOA) has enabled the sensitive detection of amino acid and amine biomarkers in laboratory standards and in a variety of field sample tests. However, the MOA is challenged when samples are extremely acidic and saline or contain polyvalent cations. Here, we have optimized the MOA analysis, sample labeling, and sample dilution buffers to handle such challenging samples more robustly. Higher ionic strength buffer systems with pK(a) values near pH 9 were developed to provide better buffering capacity and salt tolerance. The addition of ethylaminediaminetetraacetic acid (EDTA) ameliorates the negative effects of multivalent cations. The optimized protocol utilizes a 75 mM borate buffer (pH 9.5) for Pacific Blue labeling of amines and amino acids. After labeling, 50 mM (final concentration) EDTA is added to samples containing divalent cations to ameliorate their effects. This optimized protocol was used to successfully analyze amino acids in a saturated brine sample from Saline Valley, California, and a subcritical water extract of a highly acidic sample from the Río Tinto, Spain. This work expands the analytical capabilities of the MOA and increases its sensitivity and robustness for samples from extraterrestrial environments that may exhibit pH and salt extremes as well as metal ions.

Electrospun polyacrylonitrile nanofibers functionalized with EDTA for adsorption of ionic dyes

NASA Astrophysics Data System (ADS)

Chaúque, Eutilério F. C.; Dlamini, Langelihle N.; Adelodun, Adedeji A.; Greyling, Corinne J.; Ngila, J. Catherine

2017-08-01

The manipulation of nanofibers' surface chemistry could enhance their potential application toward the removal of ionic dyes in wastewater. For this purpose, surface modification of electrospun polyacrylonitrile (PAN) nanofibers with ethylenediaminetetraacetic acid (EDTA) and ethylenediamine (EDA) crosslinker was experimented. The functionalized EDTA-EDA-PAN nanofibers were characterized using Fourier transform infrared (FT-IR) spectroscopy, X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM) and Brunauer-Emmett-Teller (BET) technique. The impregnation of EDA and EDTA chelating agents on the surface of PAN changed the distribution of nanofibers as proximity is increased (accompanied by reduced softness), but the nanofibrous structure of the pristine PAN nanofibers was not substantially altered. Adsorption equilibrium studies were performed with Freundlich, Langmuir and Temkin isotherm models with the former providing better correlation to the experimental data. The modified PAN nanofibers showed efficient sorption of methyl orange (MO) and reactive red (RR) from aqueous synthetic samples, evinced by the maximum adsorption capacities (at 25 °C) of 99.15 and 110.0 mg g-1, respectively. The fabricated nanofibers showed appreciable removal efficiency of the target dye sorptives from wastewater. However, the presence of high metal ions content affected the overall extraction of dyes from wastewater due to the depletion of the adsorbent's active adsorptive sites.

Fluorescence turn-on sensing of trace cadmium ions based on EDTA-etched CdTe@CdS quantum dot.

PubMed

Wang, Si-Nan; Zhu, Jian; Li, Xin; Li, Jian-Jun; Zhao, Jun-Wu

2018-05-01

Cadmium-caused environmental pollution and diseases have always been worldwide problems. Thus it is extremely urgent to establish a cheap, rapid, simple and selective detection method for trace cadmium in drinking water. In this study, a fluorescence "turn-on" method based on ethylene diamine tetraacetic acid (EDTA)-etched CdTe@CdS quantum dots (QDs) was designed to detect Cd 2+ . High resolution transmission electron microscopy (HRTEM) and X-ray photoelectron spectroscopy (XPS) were utilized for chemical and structural characterization of the as-prepared QDs. Based on chemical etching of EDTA on the surface of CdTe@CdS QDs, specific Cd 2+ recognition sites were produced, and then results in fluorescence quenching. The introduction of Cd 2+ could identify these sites and restore the fluorescence of the EDTA-QDs system. Under the optimum conditions, the nanoprobe shows a linear response range from 0.05 to 9 μM with a very low detection limit of 0.032 μM. In addition, the reported fluorescence probe in this work displays a good selectivity for trace Cd 2+ over other metal ions and an admirable practicability in real water samples. Copyright © 2018 Elsevier B.V. All rights reserved.

A new spectrophotometric method for determination of EDTA in water using its complex with Mn(III).

PubMed

Andrade, Carlos Eduardo O; Oliveira, André F; Neves, Antônio A; Queiroz, Maria Eliana L R

2016-11-05

EDTA is an important ligand used in many industrial products as well as in agriculture, where it is employed to assist in phytoextraction procedures and the absorption of nutrients by plants. Due to its intensive use and recalcitrance, it is now considered an emerging pollutant in water, so there is great interest in techniques suitable for its monitoring. This work proposes a method based on formation of the Mn(III)-EDTA complex after oxidation of the Mn(II)-EDTA complex by PbO2 immobilized on cyanoacrylate spheres. A design of experiments (DOE) based on the Doehlert matrix was used to determine the optimum conditions of the method, and the influence of the variables was evaluated using a multiple linear regression (MLR) model. The optimized method presented a linear response in the range from 0.77 to 100.0μmolL(-1), with analytical sensitivity of 7.7×10(3)Lmol(-1), a coefficient of determination of 0.999, and a limit of detection of 0.23μmolL(-1). The method was applied using samples fortified at different concentration levels, and the recoveries achieved were between 97.0 and 104.9%. Copyright © 2016 Elsevier B.V. All rights reserved.

In vitro activity of an ear rinse containing tromethamine, EDTA, benzyl alcohol and 0.1% ketoconazole on Malassezia organisms from dogs with otitis externa.

PubMed

Cole, Lynette K; Luu, Dao H; Rajala-Schultz, Paivi J; Meadows, Cheyney; Torres, Audrey H

2007-04-01

The purpose of this study was to evaluate the in vitro activity of an ear rinse containing tromethamine, EDTA, benzyl alcohol and 0.1% ketoconazole in purified water on Malassezia organisms from dogs with otitis externa. Malassezia organisms were collected from ear swab samples from the external ear canal of 19 dogs with otitis externa plus one control strain of Malassezia pachydermatis. Three test solutions were evaluated: ER (EDTA, tromethamine, benzyl alcohol), ER + keto (EDTA, tromethamine, benzyl alcohol, ketoconazole), and H2O (purified water). Ten-millilitre aliquots of each test solution was transferred into 20 tubes and inoculated with one of the isolates (1 tube per isolate: 19 clinical and 1 control strain). Samples were retrieved from each tube at five time points (0, 15, 30, 45 and 60 min), transferred to Petri dishes, mixed with Sabouraud dextrose agar supplemented with 0.5% Tween 80 and incubated. Following incubation, the plates were examined for growth and colonies counted as colony-forming units per millilitre. The data were analysed using a repeated measures analysis, with pair-wise comparisons of solution-time combinations. There was a significant reduction in Malassezia growth in ER + keto at all time points (P < 0.0001) compared to time zero. Neither ER nor H2O had any effect on the growth of Malassezia. ER + keto was significantly more effective in reducing Malassezia growth (P < 0.0001) at all time points compared to both ER and H2O. ER + keto may be useful in the treatment of Malassezia otitis externa. Future studies should be performed to evaluate the in vivo efficacy of ER + keto as treatment for otic infections caused by Malassezia.

Performance and Value of IFN-Lambda3 and IFN-Lambda4 Genotyping in Patients with Chronic Hepatitis C (CHC) Genotype 2/3 in a Real World Setting

PubMed Central

Susser, Simone; Rogalska-Taranta, Magdalena; Petersen, Jörg; Böker, Klaus H. W.; Grigorian, Natalia; Link, Ralph; Naumann, Uwe; John, Christine; Lueth, Stefan; Malfertheiner, Peter; Manns, Michael P.; Wedemeyer, Heiner; Sarrazin, Christoph; Cornberg, Markus

2015-01-01

Background SNPs near the interferon lambda (IFNL) 3 gene are predictors for sustained virological response (SVR) in patients with chronic hepatitis C genotype (GT) 1. In addition, a dinucleotide frame shift in ss469415590 was described, which generates IFNL4. In this study, we compared the role of IFNL4 variants with IFNL3-(rs12979860) and IFNL3-(rs8099917) on response to pegylated (PEG)-IFN and Ribavirin (RBV) in patients with chronic hepatitis C GT2/3. Methods We recruited 1006 patients with chronic hepatitis C and GT2/3 in a large German registry. A treatment with PEG-IFN and Ribavirin was started by 959 patients. We performed genotyping of IFNL3 (rs12979860, n = 726; rs8099917, n = 687) and of IFNL4 (ss469415590; n = 631). Results Both preferable IFNL3 genotypes were associated with RVR (both p<0.0001) rather than with SVR (rs12979860: p = 0.251; rs8099917: p = 0.447). Only RVR was linked to SVR in univariate and multivariate analyzes (both p<0.001). Concordance of genotyping in patients with available serum samples and EDTA blood samples (n = 259) was more than 96% for both IFNL3 SNPs. IFNL3-(rs12979860) correlated with IFNL4: 99.2% of patients with IFNL3-(rs12979860)-CC were IFNL4-(ss469415590)-TT/TT. IFNL3-(rs12979860)-CT was linked with IFNL4-(ss469415590)-TT/ΔG (98.0%) and IFNL3-(rs12979860)-TT was associated with IFNL4-(ss469415590)-ΔG/ΔG (97.6%). Conclusion IFNL3 genotyping from serum was highly efficient and can be used as an alternative if EDTA whole blood is not available. In Caucasian GT2/3 patients genotyping for INFL4-(ss469415590) does not lead to additional information for the decision-making process. Importantly, IFNL3 SNPs were not associated with SVR but with RVR. Even in the era of new direct acting antiviral (DAA) therapies, IFNL3 testing may therefore still be considered for naïve GT2/3 patients to decide if dual Peg-IFN/RBV therapy is an option in resource limited regions. PMID:26699619

Performance and Value of IFN-Lambda3 and IFN-Lambda4 Genotyping in Patients with Chronic Hepatitis C (CHC) Genotype 2/3 in a Real World Setting.

PubMed

Wiegand, Steffen B; Heidrich, Benjamin; Susser, Simone; Rogalska-Taranta, Magdalena; Petersen, Jörg; Böker, Klaus H W; Grigorian, Natalia; Link, Ralph; Naumann, Uwe; John, Christine; Lueth, Stefan; Malfertheiner, Peter; Manns, Michael P; Wedemeyer, Heiner; Sarrazin, Christoph; Cornberg, Markus

2015-01-01

SNPs near the interferon lambda (IFNL) 3 gene are predictors for sustained virological response (SVR) in patients with chronic hepatitis C genotype (GT) 1. In addition, a dinucleotide frame shift in ss469415590 was described, which generates IFNL4. In this study, we compared the role of IFNL4 variants with IFNL3-(rs12979860) and IFNL3-(rs8099917) on response to pegylated (PEG)-IFN and Ribavirin (RBV) in patients with chronic hepatitis C GT2/3. We recruited 1006 patients with chronic hepatitis C and GT2/3 in a large German registry. A treatment with PEG-IFN and Ribavirin was started by 959 patients. We performed genotyping of IFNL3 (rs12979860, n = 726; rs8099917, n = 687) and of IFNL4 (ss469415590; n = 631). Both preferable IFNL3 genotypes were associated with RVR (both p<0.0001) rather than with SVR (rs12979860: p = 0.251; rs8099917: p = 0.447). Only RVR was linked to SVR in univariate and multivariate analyzes (both p<0.001). Concordance of genotyping in patients with available serum samples and EDTA blood samples (n = 259) was more than 96% for both IFNL3 SNPs. IFNL3-(rs12979860) correlated with IFNL4: 99.2% of patients with IFNL3-(rs12979860)-CC were IFNL4-(ss469415590)-TT/TT. IFNL3-(rs12979860)-CT was linked with IFNL4-(ss469415590)-TT/ΔG (98.0%) and IFNL3-(rs12979860)-TT was associated with IFNL4-(ss469415590)-ΔG/ΔG (97.6%). IFNL3 genotyping from serum was highly efficient and can be used as an alternative if EDTA whole blood is not available. In Caucasian GT2/3 patients genotyping for INFL4-(ss469415590) does not lead to additional information for the decision-making process. Importantly, IFNL3 SNPs were not associated with SVR but with RVR. Even in the era of new direct acting antiviral (DAA) therapies, IFNL3 testing may therefore still be considered for naïve GT2/3 patients to decide if dual Peg-IFN/RBV therapy is an option in resource limited regions.

Effective analysis of the use of peracetic acid after instrumentation of root canals contaminated with Enterococcus faecalis.

PubMed

Cord, Caroline Berwanger; Velasco, Rafael Vidal Cortez; Ribeiro Melo Lima, Laíla Fernanda; Rocha, Daniel Guimarães Pedro; da Silveira Bueno, Carlos Eduardo; Pinheiro, Sérgio Luiz

2014-08-01

The aim of this study was to evaluate the effectiveness of peracetic acid (PAA) in cleaning root canals contaminated with Enterococcus faecalis. Sixty first and second mandibular molars were used. Their mesiobuccal canals were prepared with the Reciproc System (VDW, Munich, Germany). The canals were irrigated with 10 mL saline during instrumentation. The teeth were randomly divided into 3 groups (n = 20), according to the irrigation solution to be used after instrumentation: group PAA (5 mL 1% PAA), group EDTA/sodium hypochlorite (NaOCl) (5 mL 17% EDTA followed by 5 mL 2.5% sodium hypochlorite), and group S (5 mL saline). Microbiological samples were collected before instrumentation and after final irrigation. Bacterial quantification was performed by counting the number of colony-forming units (CFUs/mL). The results were analyzed by the nonparametric Wilcoxon and Kruskal-Wallis tests. The 3 groups showed a significant reduction (P < .05) in CFUs/mL after final irrigation. PAA and NaOCl associated with EDTA produced a significantly higher reduction in CFUs/mL (P < .05) compared with saline. There was no statistically significant difference between PAA and EDTA + 2.5% NaOCl (P > .05). According to the results of the present study, the effectiveness of 1% PAA was similar to that of 17% EDTA + 2.5% NaOCl in cleaning curved root canals contaminated with E. faecalis. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Enzymes and chelating agent in cotton pretreatment.

PubMed

Csiszár, E; Losonczi, A; Szakács, G; Rusznák, I; Bezúr, L; Reicher, J

2001-08-23

Desized cotton fabric and cotton seed-coat fragments (impurities) have been treated with commercial cellulase (Celluclast 1.5 L), hemicellulase-pectinase (Viscozyme 120 L) and xylanase (Pulpzyme HC) enzymes. Seed-coat fragments hydrolyzed much faster than the cotton fabric itself. This relative difference in hydrolysis rates makes possible a direct enzymatic removal of seed-coat fragments from desized cotton fabric. Addition of chelating agents such as ethylenediamine-tetra-acetic acid (EDTA) markedly enhanced the directed enzyme action. Pretreatments carried out in acidic solution at pH 5 increased the lightness of seed-coat fragments, contrary to the samples treated in neutral medium at pH 7. Alkaline scouring resulted in darker seed-coat fragments except for the samples pretreated with Pulpzyme HC plus EDTA. This effect is similar to that observed in the biobleaching process in pulp and paper industry.

The Impact of Three Different Canal Lubricants on Fracture, Deformity and Metal Slivering of ProTaper Rotary Instruments

PubMed Central

Shantiaee, Yazdan; Dianat, Omid; Sharifi, Farnoud; Nahvi, Golnaz; Kolahi Ahari, Golbarg

2014-01-01

Introduction: The aim of this in vitro study was to evaluate the effect(s) of three canal lubricants i.e. sodium hypochlorite, RC-Prep as the paste form of ethylenediaminetetraacetic acid (EDTA) and aqueous EDTA on the occurrence/incidence of fracture, deformity and metal slivering of ProTaper rotary instruments. Methods: A total of 120 mesial canals (i.e. mesiobuccal and mesiolingual) of first mandibular molars or buccal canals (i.e. mesiobuccal and distobuccal) of first maxillary molars, with curvatures of 10-20 degrees were selected and randomly divided into three groups of forty samples each. These selected canals all had approximate 19-21 mm working length and apical diameter equal to a #15 K-file. In each group, the root canals were prepared using ProTaper rotary instruments with an electric motor using one of the three aforementioned irrigants. Subsequently, samples were compared to each other at different magnifications (16×, 20×, 40× and 57×) for any fracture, deformity or metal slivering, by the Cox regression analysis. Results: The fractures rate of samples in RC-Prep group was significantly higher compared to other groups (P=0.01). No evidence of instrument deformity was detected in any groups. A statistically significant reverse relation between metal slivering and instrument fracture was observed. Conclusions: Application of aqueous EDTA and/or sodium hypochlorite as intracanal lubricants caused less fracture of ProTaper instruments compared to canal lubrication with RC-Prep. PMID:24688582

Transfer of copper, lead and zinc in soil-grass ecosystem in aspect of soils properties, in Poland.

PubMed

Niesiobędzka, Krystyna

2012-04-01

The total metal concentrations in soil samples from polluted area (roadside soils) ranged from 13.87 to 195.76 mg/kg for Cu; 13.56-310.17 mg/kg for Pb and 18.43-894.11 mg/kg for Zn and they were, respectively about 5, 2 and 13 times above the corresponding values in soil samples from country area. The mean values of EDTA-extractable concentrations in soil samples at unpolluted sites were: 2.47 mg/kg for Cu, 6.33 mg/kg for Pb and 4.94 mg/kg for Zn. The highest concentrations of Cu, Pb and Zn in grass were measured in soils from polluted area. Higher values of proportions of EDTA-extractable metals (24% for Cu, 40% for Pb and 38% for Zn) indicate that anthropogenic metals were more mobile and bioavailable than the same metals in soils from unpolluted area (20, 16 and 20% for Cu, Pb and Zn, respectively). The availability of Cu, Pb and Zn are affected by soil properties such as pH, organic matter content and cation exchange capacity. Correlation between the EDTA-extractable forms concentrations of metals and the total concentration in the various soils was observed. The coefficients of determination (R(2)) varied between 0.809 for Cu; 0,709 for Pb and 0.930 for Zn in polluted soils and they are higher than corresponding values in unpolluted soils.

In situ observation of fluoride-ion-induced hydroxyapatite collagen detachment on bone fracture surfaces by atomic force microscopy

NASA Astrophysics Data System (ADS)

Kindt, J. H.; Thurner, P. J.; Lauer, M. E.; Bosma, B. L.; Schitter, G.; Fantner, G. E.; Izumi, M.; Weaver, J. C.; Morse, D. E.; Hansma, P. K.

2007-04-01

The topography of freshly fractured bovine and human bone surfaces was determined by the use of atomic force microscopy (AFM). Fracture surfaces from both kinds of samples exhibited complex landscapes formed by hydroxyapatite mineral platelets with lateral dimensions ranging from ~90 nm × 60 nm to ~20 nm × 20 nm. Novel AFM techniques were used to study these fracture surfaces during various chemical treatments. Significant topographical changes were observed following exposure to aqueous solutions of ethylenediaminetetraacetic acid (EDTA) or highly concentrated sodium fluoride (NaF). Both treatments resulted in the apparent loss of the hydroxyapatite mineral platelets on a timescale of a few seconds. Collagen fibrils situated beneath the overlying mineral platelets were clearly exposed and could be resolved with high spatial resolution in the acquired AFM images. Time-dependent mass loss experiments revealed that the applied agents (NaF or EDTA) had very different resulting effects. Despite the fact that the two treatments exhibited nearly identical results following examination by AFM, bulk bone samples treated with EDTA exhibited a ~70% mass loss after 72 h, whereas for the NaF-treated samples, the mass loss was only of the order of ~10%. These results support those obtained from previous mechanical testing experiments, suggesting that enhanced formation of superficial fluoroapatite dramatically weakens the protein-hydroxyapatite interfaces. Additionally, we discovered that treatment with aqueous solutions of NaF resulted in the effective extraction of noncollagenous proteins from bone powder.

Differential stability of therapeutic peptides with different proteolytic cleavage sites in blood, plasma and serum.

PubMed

Böttger, Roland; Hoffmann, Ralf; Knappe, Daniel

2017-01-01

Proteolytic degradation of peptide-based drugs is often considered as major weakness limiting systemic therapeutic applications. Therefore, huge efforts are typically devoted to stabilize sequences against proteases present in serum or plasma, obtained as supernatants after complete blood coagulation or centrifugation of blood supplemented with anticoagulants, respectively. Plasma and serum are reproducibly obtained from animals and humans allowing consistent for clinical analyses and research applications. However, the spectrum of active or activated proteases appears to vary depending on the activation of proteases and cofactors during coagulation (serum) or inhibition of such enzymes by anticoagulants (plasma), such as EDTA (metallo- and Ca2+-dependent proteases) and heparin (e.g. thrombin, factor Xa). Here, we studied the presumed effects on peptide degradation by taking blood via cardiac puncture of CD-1 mice using a syringe containing a peptide solution. Due to absence of coagulation activators (e.g. glass surfaces and damaged cells), visible blood clotting was prevented allowing to study peptide degradation for one hour. The remaining peptide was quantified and the degradation products were identified using mass spectrometry. When the degradation rates (half-life times) were compared to serum derived freshly from the same animal and commercial serum and plasma samples, peptides of three different families showed indeed considerably different stabilities. Generally, peptides were faster degraded in serum than in plasma, but surprisingly all peptides were more stable in fresh blood and the order of degradation rates among the peptides varied among the six different incubation experiments. This indicates, that proteolytic degradation of peptide-based therapeutics may often be misleading stimulating efforts to stabilize peptides at degradation sites relevant only in vitro, i.e., for serum or plasma stability assays, but of lower importance in vivo.

Detection of hemoplasma infection of goats by use of a quantitative polymerase chain reaction assay and risk factor analysis for infection.

PubMed

Johnson, Kathy A; do Nascimento, Naíla C; Bauer, Amy E; Weng, Hsin-Yi; Hammac, G Kenitra; Messick, Joanne B

2016-08-01

OBJECTIVE To develop and validate a real-time quantitative PCR (qPCR) assay for the detection and quantification of Mycoplasma ovis in goats and investigate the prevalence and risk factors for hemoplasma infection of goats located in Indiana. ANIMALS 362 adult female goats on 61 farms. PROCEDURES Primers were designed for amplification of a fragment of the dnaK gene of M ovis by use of a qPCR assay. Blood samples were collected into EDTA-containing tubes for use in total DNA extraction, blood film evaluation, and determination of PCV. Limit of detection, intra-assay variability, interassay variability, and specificity of the assay were determined. RESULTS Reaction efficiency of the qPCR assay was 94.45% (R(2), 0.99; slope, -3.4623), and the assay consistently detected as few as 10 copies of plasmid/reaction. Prevalence of infection in goats on the basis of results for the qPCR assay was 18.0% (95% confidence interval, 14% to 22%), with infected goats ranging from 1 to 14 years old, whereby 61% (95% confidence interval, 47% to 73%) of the farms had at least 1 infected goat. Bacterial load in goats infected with M ovis ranged from 1.05 × 10(3) target copies/mL of blood to 1.85 × 10(5) target copies/mL of blood; however, no bacteria were observed on blood films. Production use of a goat was the only risk factor significantly associated with hemoplasma infection. CONCLUSIONS AND CLINICAL RELEVANCE The qPCR assay was more sensitive for detecting hemoplasma infection than was evaluation of a blood film, and production use of a goat was a risk factor for infection.

Synergistic effect of EDTA and boric acid on corneal penetration of CS-088.

PubMed

Kikuchi, Takayuki; Suzuki, Masahiko; Kusai, Akira; Iseki, Ken; Sasaki, Hitoshi

2005-02-16

In order to investigate the effects of EDTA and boric acid (EDTA/boric acid) on the corneal penetration of CS-088, an ophthalmic agent, the apparent permeability coefficient of CS-088 in the presence of EDTA/boric acid across the isolated corneal membranes of rabbits was measured using an in vitro penetration chamber system. FITC-dextran (M.W. 4400) and an electrical method based on membrane resistance were used to provide a quantitative assessment of the enhancing effect of EDTA/boric acid. The corneal penetration of CS-088 was significantly enhanced in the presence of EDTA/boric acid by approximately 1.6-fold. The permeability-enhancing effect of EDTA/boric acid was apparently synergistic and concentration-dependent on both EDTA and boric acid. The penetration of FITC-dextran, a paracellular marker, and electrical resistance of corneal membranes were not affected in the presence of EDTA/boric acid. Furthermore, no enhancing effect of EDTA/boric acid was observed in de-epithelialized corneas, although de-epithelialized corneas exhibited a markedly higher permeability of CS-088 that was 24-fold greater than that for intact corneas. In conclusion, EDTA/boric acid synergistically enhances the transcellular permeability of CS-088 in the outer layer but not in the inner layers of the corneal membrane.

Effects of calcium binding and of EDTA and CaEDTA on the clotting of bovine fibrinogen by thrombin.

PubMed

Perizzolo, K E; Sullivan, S; Waugh, D F

1985-03-01

Studies were carried out at pH 7.0 and gamma/2 0.15 before addition of CaCl2 or EDTA. Clotting time, tau, at 3.03 microM fibrinogen and 0.91 u/ml thrombin was determined for equilibrium systems. With added Ca2+, tau decreases, from tau 0 at 0 added Ca2+ (mean, 29.7 +/- 3 s), by approximately 3 s at 5 mM added Ca2+. With added EDTA, tau increases sigmoidally from tau 0 at 0 EDTA to a maximum (mean tau m = 142 +/- 23 s) at approximately 200 microM EDTA. tau then decreases slightly to a minimum at approximately 1.3 mM and finally increases to infinity at approximately 10 mM EDTA. Between 0 and 1.3 mM EDTA, effects on clotting time are completely reversed by adding Ca2+ and, after equilibration at 400 microM EDTA, tau is independent of EDTA concentration. Thus, up to 400 microM EDTA, effects on clotting time are attributed to decreasing fibrinogen bound Ca2+. Between 5 mM Ca2+ and 200 microM EDTA it is assumed that an equilibrium distribution of fibrinogen species having 3, 2, 1, or 0 bound calcium ions is established and that a clotting time is determined by the sum of products of species fractional abundance and pure species clotting time. Analysis indicates that pure species clotting times increase proportionately with decreasing Ca2+ binding, binding sites are nearly independent, and the microscopic association constant for the first bound Ca2+ is approximately 4.9 X 10(6) M-1. Effects of adding Ca2+ at times t1 after thrombin addition to systems initially equilibrated at 200 microM EDTA were determined. Analysis of the relation between tau and t1 indicates that as Ca2+ binding decreases, rate constants for release of B peptides decrease less than those for release of A peptides. As EDTA concentration is increased above 1.3 mM, inhibitory effects of EDTA and CaEDTA progressively increase.

Effect of adsorbed metals ions on the transport of Zn- and Ni-EDTA complexes in a sand and gravel aquifer

USGS Publications Warehouse

Kent, D.B.; Davis, J.A.; Anderson, L.C.D.; Rea, B.A.; Coston, J.A.

2002-01-01

Adsorption, complexation, and dissolution reactions strongly influenced the transport of metal ions complexed with ethylenediaminetetraacetic acid (EDTA) in a predominantly quartz-sand aquifer during two tracer tests conducted under mildly reducing conditions at pH 5.8 to 6.1. In tracer test M89, EDTA complexes of zinc (Zn) and nickel (Ni), along with excess free EDTA, were injected such that the lower portion of the tracer cloud traveled through a region with adsorbed manganese (Mn) and the upper portion of the tracer cloud traveled through a region with adsorbed Zn. In tracer test S89, Ni- and Zn-EDTA complexes, along with excess EDTA complexed with calcium (Ca), were injected into a region with adsorbed Mn. The only discernable chemical reaction between Ni-EDTA and the sediments was a small degree of reversible adsorption leading to minor retardation. In the absence of adsorbed Zn, the injected Zn was displaced from EDTA complexes by iron(III) [Fe(III)] dissolved from the sediments. Displacement of Zn by Fe(III) on EDTA became increasingly thermodynamically favorable with decreasing total EDTA concentration. The reaction was slow compared to the time-scale of transport. Free EDTA rapidly dissolved aluminum (Al) from the sediments, which was subsequently displaced slowly by Fe. In the portion of tracer cloud M89 that traveled through the region contaminated with adsorbed Zn, little displacement of Zn complexed with EDTA was observed, and Al was rapidly displaced from EDTA by Zn desorbed from the sediments, in agreement with equilibrium calculations. In tracer test S89, desorption of Mn dominated over the more thermodynamically favorable dissolution of Al oxyhydroxides. Comparison with results from M89 suggests that dissolution of Al oxyhydroxides in coatings on these sediment grains by Ca-EDTA was rate-limited whereas that by free EDTA reached equilibrium on the time-scale of transport. Rates of desorption are much faster than rates of dissolution of Fe oxyhydroxides from sediment-grain surfaces and, therefore, adsorbed metal ions can strongly influence the speciation of ligands like EDTA in soils and sediments, especially over small temporal and spatial scales. Copyright ?? 2002 Elsevier Science Ltd.

A Scanning Electron Microscope Evaluation of Smear Layer Removal and Antimicrobial Action of Mixture of Tetracycline, Acid and Detergent, Sodium Hypochlorite, Ethylenediaminetetraacetic Acid, and Chlorhexidine Gluconate: An In vitro Study.

PubMed

Charlie, K M; Kuttappa, M A; George, Liza; Manoj, K V; Joseph, Bobby; John, Nishin K

2018-01-01

The main objective is to evaluate the efficiency in removal of smear layer of mixture of tetracycline, acid and detergent (MTAD), sodium hypochlorite (NaOCl), ethylenediaminetetraacetic acid (EDTA) and chlorhexidine gluconate by scanning electron microscope (SEM) evaluation and also to evaluate the antimicrobial action of the same irrigants against standard culture strains of Enterococcus faecalis . This study included 60 extracted permanent teeth with single root canal. The sample was categorized into five groups with 12 teeth in each group. Root canals were enlarged till size 40 with K-files. One group was kept as control and irrigated only with saline. Other four groups used 5% NaOCl as irrigant during instrumentation and MTAD, 5% NaOCl, 17% EDTA, and 2% chlorhexidine gluconate as final rinse. Teeth were split and examined under SEM. To test the antibacterial action, the zone of inhibition method using agar plates was used. Obtained data were statistically analyzed by SPSS version 17 (SPSS Inc., Chicago, IL, USA). MTAD and 17% EDTA removed smear layer from all regions of the root canals. About 5% NaOCl and 2% chlorhexidine gluconate were ineffective in removing the smear layer. The mean zone of inhibition formed by the irrigants was in the following order; MTAD (40.5 mm), 2% chlorhexidine gluconate (29.375 mm), 17% EDTA (24.125 mm), 5% NaOCl (22.125 mm), and saline (zero). MTAD showed high smear layer removal efficacy, but no significant difference was found to that of 17% EDTA. As the dimensions of the zones of inhibition showed, MTAD has got highest antibacterial action against E. faecalis , followed by 2% chlorhexidine gluconate, 17% EDTA, and 5% NaOCl. However, the exact correlation of in vitro study results to clinical conditions is impossible due to the variables involved.

Influence of variable chemical conditions on EDTA-enhanced transport of metal ions in mildly acidic groundwater

USGS Publications Warehouse

Kent, D.B.; Davis, J.A.; Joye, J.L.; Curtis, G.P.

2008-01-01

Adsorption of Ni and Pb on aquifer sediments from Cape Cod, Massachusetts, USA increased with increasing pH and metal-ion concentration. Adsorption could be described quantitatively using a semi-mechanistic surface complexation model (SCM), in which adsorption is described using chemical reactions between metal ions and adsorption sites. Equilibrium reactive transport simulations incorporating the SCMs, formation of metal-ion-EDTA complexes, and either Fe(III)-oxyhydroxide solubility or Zn desorption from sediments identified important factors responsible for trends observed during transport experiments conducted with EDTA complexes of Ni, Zn, and Pb in the Cape Cod aquifer. Dissociation of Pb-EDTA by Fe(III) is more favorable than Ni-EDTA because of differences in Ni- and Pb-adsorption to the sediments. Dissociation of Ni-EDTA becomes more favorable with decreasing Ni-EDTA concentration and decreasing pH. In contrast to Ni, Pb-EDTA can be dissociated by Zn desorbed from the aquifer sediments. Variability in adsorbed Zn concentrations has a large impact on Pb-EDTA dissociation.

Evaluation of the colour change in enamel and dentine promoted by the interaction between 2% chlorhexidine and auxiliary chemical solutions.

PubMed

Souza, Matheus; Cecchin, Doglas; Barbizam, Joao V B; Almeida, José F A; Zaia, Alexandre Augusto; Gomes, Brenda P F A; Ferraz, Caio C R

2013-12-01

To evaluate the colour change in enamel and dentine, promoted by interaction of 2% chlorhexidine gluconate (CHX) with 5.25% sodium hypochlorite (NaOCl) and 17% ethylenediaminetetraacetic acid (EDTA). Fragments containing enamel and dentine were obtained from the crowns of extracted bovine incisors. Before and after immersion of the samples in the substances, they were evaluated with reference to the colour of the enamel and dentine. The values obtained in numerical scores were subjected to statistical analysis using Wilcoxon test. A colour change in the enamel and dentine in groups treated with CHX gel + NaOCl and CHX gel + NaOCl + EDTA, and a change in colour only in the dentine in groups treated with CHX solution + NaOCl and CHX solution + NaOCl + EDTA. When used prior to NaOCl, CHX has the ability to induce a colour change in dental structures. © 2011 The Authors. Australian Endodontic Journal © 2011 Australian Society of Endodontology.

In vitro degradation of neurotensin in human plasma.

PubMed

Lee, Y C; Uttenthal, L O; Smith, H A; Bloom, S R

1986-01-01

To study the degradation of neurotensin in plasma in vitro, fresh human plasma was incubated with neurotensin in the presence and absence of the peptidase inhibitors pepstatin A, EDTA, PMSF and aprotinin. The half-time of disappearance of neurotensin at 37 degrees C was calculated to be 226 min in vitro as opposed to 1.4 min in vivo when measured by radioimmunoassay with a C-terminally directed neurotensin antiserum. Both gel filtration and reversed phase high-pressure liquid chromatography (HPLC) showed that the main degradation product of neurotensin in human plasma in vitro was chromatographically and immunologically identical to neurotensin 1-8 and HPLC also demonstrated the formation of neurotensin 1-11. The loss of neurotensin incubated in human plasma in vitro was greatly reduced by EDTA but not by the other peptidase inhibitors tested. In this respect peptidase(s) responsible for the degradation of neurotensin in plasma differ from those present in brain homogenates. EDTA may be of importance in the preservation of neurotensin in plasma samples.

Evaluation of the Microsemi CRP, an automated hematology analyzer for rapid 3-part WBC differential and CRP using whole blood.

PubMed

Nomura, N; Saito, K; Ikeda, M; Yuasa, S; Pastore, M; Chabert, C; Kono, E; Sakai, A; Tanaka, H; Ikemoto, T; Takubo, T

2015-08-01

We evaluated the basic performance of Microsemi CRP, an unique automated hematology analyzer which can simultaneously measure CBC including 3-part WBC differential (3-Diff) and CRP using whole blood treated with EDTA-2K anticoagulant. We found that it produced generally the acceptable results for all parameters performed (repeatability, reproducibility, linearity, interference effect, carry over, and correlation) using control materials, fresh human whole bloods, and serum samples. CBC data examined using Microsemi CRP showed the good correlation with the previous model, Micros CRP200 (r ≧ 0.9), and also those obtained using the routine analyzer, ADVIA 2120i (r ≧ 0.989). Concerning the 3-Diff, both GRA (%) and LYM (%) showed the excellent correlation coefficient between Microsemi CRP and Micros CRP200 (r ≧ 0.992) as well as ADVIA 2120i (r ≧ 0.957). MON (%) showed good correlation between Microsemi CRP and Micros CRP200 (r = 0.959), but lower correlation between Microsemi CRP and ADVIA 2120 i (r = 0.471). CRP data showed the good correlation with HITACHI7600 (r ≧ 0.997) and Micros CRP200 (r ≧ 0.997). From these findings, we concluded that Microsemi CRP seemed the convenient laboratory analyzer in the setting of point of care testing (POCT) especially at NICU or primary care unit. © 2014 John Wiley & Sons Ltd.

Gene expression profiling of immunomagnetically separated cells directly from stabilized whole blood for multicenter clinical trials

PubMed Central

2014-01-01

Background Clinically useful biomarkers for patient stratification and monitoring of disease progression and drug response are in big demand in drug development and for addressing potential safety concerns. Many diseases influence the frequency and phenotype of cells found in the peripheral blood and the transcriptome of blood cells. Changes in cell type composition influence whole blood gene expression analysis results and thus the discovery of true transcript level changes remains a challenge. We propose a robust and reproducible procedure, which includes whole transcriptome gene expression profiling of major subsets of immune cell cells directly sorted from whole blood. Methods Target cells were enriched using magnetic microbeads and an autoMACS® Pro Separator (Miltenyi Biotec). Flow cytometric analysis for purity was performed before and after magnetic cell sorting. Total RNA was hybridized on HGU133 Plus 2.0 expression microarrays (Affymetrix, USA). CEL files signal intensity values were condensed using RMA and a custom CDF file (EntrezGene-based). Results Positive selection by use of MACS® Technology coupled to transcriptomics was assessed for eight different peripheral blood cell types, CD14+ monocytes, CD3+, CD4+, or CD8+ T cells, CD15+ granulocytes, CD19+ B cells, CD56+ NK cells, and CD45+ pan leukocytes. RNA quality from enriched cells was above a RIN of eight. GeneChip analysis confirmed cell type specific transcriptome profiles. Storing whole blood collected in an EDTA Vacutainer® tube at 4°C followed by MACS does not activate sorted cells. Gene expression analysis supports cell enrichment measurements by MACS. Conclusions The proposed workflow generates reproducible cell-type specific transcriptome data which can be translated to clinical settings and used to identify clinically relevant gene expression biomarkers from whole blood samples. This procedure enables the integration of transcriptomics of relevant immune cell subsets sorted directly from whole blood in clinical trial protocols. PMID:25984272

Effects of EDTA and low molecular weight organic acids on soil solution properties of a heavy metal polluted soil.

PubMed

Wu, L H; Luo, Y M; Christie, P; Wong, M H

2003-02-01

A pot experiment was conducted to study the effects of EDTA and low molecular weight organic acids (LMWOA) on the pH, total organic carbon (TOC) and heavy metals in the soil solution in the rhizosphere of Brassica juncea grown in a paddy soil contaminated with Cu, Zn, Pb and Cd. The results show that EDTA and LMWOA have no effect on the soil solution pH. EDTA addition significantly increased the TOC concentrations in the soil solution. The TOC concentrations in treatments with EDTA were significantly higher than those in treatments with LMWOA. Adding 3 mmol kg(-1) EDTA to the soil markedly increased the total concentrations of Cu, Zn, Pb and Cd in the soil solution. Compared to EDTA, LMWOA had a very small effect on the metal concentrations. Total concentrations in the soil solution followed the sequence: EDTA > citric acid (CA) approximately oxalic acid (OA) approximately malic acid (MA) for Cu and Pb; EDTA > MA > CA approximately OA for Zn; and EDTA > MA > CA > OA for Cd. The labile concentrations of Cu, Zn, Pb and Cd showed similar trends to the total concentrations.

Teratogenic effect of calcium edetate (CaEDTA) in rats and the protective effect of zinc.

PubMed

Brownie, C F; Brownie, C; Noden, D; Krook, L; Haluska, M; Aronson, A L

1986-03-15

The calcium chelate of EDTA (CaEDTA) currently is the drug of choice in the treatment of lead intoxication. This study investigated the teratogenic potential of CaEDTA, administered parenterally during periods of organogenesis and determined if incorporating zinc into EDTA would protect against teratogenic effects. Four doses (2, 4, 6, and 8 mmol/m2/day) of CaEDTA, two concentrations (8 and 20 mmol/m2/day) of ZnEDTA and ZnCaEDTA (molar ratio 0.5:0.5:1) were used, and a saline control (0.9% NaCl). Timed-pregnant Long-Evans rats were assigned at random to the treatment groups, 20 per dose for each chelate and 30 to the saline control. Rats were injected with the chelate or saline solution sc, twice daily during the 11th through 15th days of gestation. Pups removed by cesarean section on the 21st day were processed for osseous and visceral examination. Additional animals per treatment group were used for maternal plasma and liver and fetal zinc determinations. Results showed increases in several abnormalities (submucous cleft, cleft palate, adactyly-syndactyly, curly tail, abnormal rib and vertebrae) with increasing amounts of CaEDTA. No malformations were seen with ZnEDTA at either dose or with ZnCaEDTA at 8 mmol/m2/day. However, submucous cleft was seen in 6 of 20 litters from the dams receiving the higher dose of ZnCaEDTA. It was concluded that CaEDTA is teratogenic in rats at concentrations which, except for decreased weight gain, produce no discernible toxicity to the dam, and which are comparable to the recommended therapeutic dosage in humans (1500 mg/m2/day corresponding to 4 mmol/m2/day). Protection is afforded by incorporating zinc in the chelate.

1064-nm Nd:YAG and 980-nm Diode Laser EDTA Agitation on the Retention of an Epoxy-Based Sealer to Root Dentin.

PubMed

Macedo, Helena Suleiman de; Messias, Danielle Cristine Furtado; Rached-Júnior, Fuad Jacob; Oliveira, Ligia Teixeira de; Silva-Sousa, Yara Teresinha Correa; Raucci-Neto, Walter

2016-01-01

Root canal irrigants are used to minimize the negative effects of smear layer on endodontic sealer retention. The aim of this study was to evaluate the efficacy of agitation of 17% ethylenediaminetetraacetic acid (EDTA) with ultrasonic, 1064-nm Nd:YAG and 980-nm diode laser on the retention of an epoxy-based sealer to the root canal walls. Forty single-rooted bovine teeth were instrumented with ProTaper rotary system and divided into four groups according to the final irrigation protocol (n = 10): (1) 17% EDTA (control); (2) 17% EDTA with 50-s ultrasonic agitation; (3) 17% EDTA with 50-s diode laser (2-W) agitation; and (4) 17% EDTA with 50-s Nd:YAG (1.5-W) laser agitation. After endodontic filling with gutta-percha F5 master cone and Sealer 26, the roots were sectioned at the cervical, middle, and apical root thirds to obtain 1.5-mm slices. Push-out tests were performed using a universal testing machine at a 1 mm/min crosshead speed. Data were analyzed using two-way ANOVA and Tukey's tests (α=0.05). Apical root thirds had significant higher retention values than cervical and middle thirds (p < 0.05). EDTA with 1064-nm Nd:YAG or 980-nm diode laser presented the highest retention values and was significantly different from EDTA with ultrasonic agitation and EDTA only (p < 0.05). Adhesive failures were predominant to EDTA only group. Mixed failures were predominant to all agitation groups. 1064-nm Nd:YAG and 980-nm diode laser EDTA agitation enhanced the retention of the epoxy-based sealer to the root canal walls compared with that due to EDTA only or EDTA with ultrasonic agitation.

Copper speciation and binding by organic matter in copper-contaminated streamwater

USGS Publications Warehouse

Breault, R.F.; Colman, J.A.; Aiken, G.R.; McKnight, D.

1996-01-01

Fulvic acid binding sites (1.3-70 ??M) and EDTA (0.0017-0.18 ??M) accounted for organically bound Cu in seven stream samples measured by potentiometric titration. Cu was 84-99% organically bound in filtrates with 200 nM total Cu. Binding of Cu by EDTA was limited by competition from other trace metals. Water hardness was inversely related to properties of dissolved organic carbon (DOC) that enhance fulvic acid binding: DOC concentration, percentage of DOC that is fulvic acid, and binding sites per fulvic acid carbon. Dissolved trace metals, stabilized by organic binding, occurred at increased concentration in soft water as compared to hard water.

Establishment of Protocols for Global Metabolomics by LC-MS for Biomarker Discovery.

PubMed

Saigusa, Daisuke; Okamura, Yasunobu; Motoike, Ikuko N; Katoh, Yasutake; Kurosawa, Yasuhiro; Saijyo, Reina; Koshiba, Seizo; Yasuda, Jun; Motohashi, Hozumi; Sugawara, Junichi; Tanabe, Osamu; Kinoshita, Kengo; Yamamoto, Masayuki

2016-01-01

Metabolomics is a promising avenue for biomarker discovery. Although the quality of metabolomic analyses, especially global metabolomics (G-Met) using mass spectrometry (MS), largely depends on the instrumentation, potential bottlenecks still exist at several basic levels in the metabolomics workflow. Therefore, we established a precise protocol initially for the G-Met analyses of human blood plasma to overcome some these difficulties. In our protocol, samples are deproteinized in a 96-well plate using an automated liquid-handling system, and conducted either using a UHPLC-QTOF/MS system equipped with a reverse phase column or a LC-FTMS system equipped with a normal phase column. A normalization protocol of G-Met data was also developed to compensate for intra- and inter-batch differences, and the variations were significantly reduced along with our normalization, especially for the UHPLC-QTOF/MS data with a C18 reverse-phase column for positive ions. Secondly, we examined the changes in metabolomic profiles caused by the storage of EDTA-blood specimens to identify quality markers for the evaluation of the specimens' pre-analytical conditions. Forty quality markers, including lysophospholipids, dipeptides, fatty acids, succinic acid, amino acids, glucose, and uric acid were identified by G-Met for the evaluation of plasma sample quality and established the equation of calculating the quality score. We applied our quality markers to a small-scale study to evaluate the quality of clinical samples. The G-Met protocols and quality markers established here should prove useful for the discovery and development of biomarkers for a wider range of diseases.

Prevalence and causes of abnormal PSA recovery.

PubMed

Lautenbach, Noémie; Müntener, Michael; Zanoni, Paolo; Saleh, Lanja; Saba, Karim; Umbehr, Martin; Velagapudi, Srividya; Hof, Danielle; Sulser, Tullio; Wild, Peter J; von Eckardstein, Arnold; Poyet, Cédric

2018-01-26

Prostate-specific antigen (PSA) test is of paramount importance as a diagnostic tool for the detection and monitoring of patients with prostate cancer. In the presence of interfering factors such as heterophilic antibodies or anti-PSA antibodies the PSA test can yield significantly falsified results. The prevalence of these factors is unknown. We determined the recovery of PSA concentrations diluting patient samples with a standard serum of known PSA concentration. Based on the frequency distribution of recoveries in a pre-study on 268 samples, samples with recoveries <80% or >120% were defined as suspect, re-tested and further characterized to identify the cause of interference. A total of 1158 consecutive serum samples were analyzed. Four samples (0.3%) showed reproducibly disturbed recoveries of 10%, 68%, 166% and 4441%. In three samples heterophilic antibodies were identified as the probable cause, in the fourth anti-PSA-autoantibodies. The very low recovery caused by the latter interference was confirmed in serum, as well as heparin- and EDTA plasma of blood samples obtained 6 months later. Analysis by eight different immunoassays showed recoveries ranging between <10% and 80%. In a follow-up study of 212 random plasma samples we found seven samples with autoantibodies against PSA which however did not show any disturbed PSA recovery. About 0.3% of PSA determinations by the electrochemiluminescence assay (ECLIA) of Roche diagnostics are disturbed by heterophilic or anti-PSA autoantibodies. Although they are rare, these interferences can cause relevant misinterpretations of a PSA test result.

Buffer substitution in malaria rapid diagnostic tests causes false-positive results

PubMed Central

2010-01-01

Background Malaria rapid diagnostic tests (RDTs) are kits that generally include 20 to 25 test strips or cassettes, but only a single buffer vial. In field settings, laboratory staff occasionally uses saline, distilled water (liquids for parenteral drugs dilution) or tap water as substitutes for the RDT kit's buffer to compensate for the loss of a diluent bottle. The present study assessed the effect of buffer substitution on the RDT results. Methods Twenty-seven RDT brands were run with EDTA-blood samples of five malaria-free subjects, who were negative for rheumatoid factor and antinuclear antibodies. Saline, distilled water and tap water were used as substitute liquids. RDTs were also run with distilled water, without adding blood. Results were compared to those obtained with the RDT kit's buffer and Plasmodium positive samples. Results Only eight cassettes (in four RDT brands) showed no control line and were considered invalid. Visible test lines occurred for at least one malaria-free sample and one of the substitutes in 20/27 (74%) RDT brands (saline: n = 16; distilled water: n = 17; and tap water: n = 20), and in 15 RDTs which were run with distilled water only. They occurred for all Plasmodium antigens and RDT formats (two-, three- and four-band RDTs). Clearance of the background of the strip was excellent except for saline. The aspects (colour, intensity and crispness) of the control and the false-positive test lines were similar to those obtained with the RDT kits' buffer and Plasmodium positive samples. Conclusion Replacement of the RDT kit's dedicated buffer by saline, distilled water and tap water can cause false-positive test results. PMID:20650003

Does long-term creatine supplementation impair kidney function in resistance-trained individuals consuming a high-protein diet?

PubMed Central

2013-01-01

Background The aim of this study was to determine the effects of creatine supplementation on kidney function in resistance-trained individuals ingesting a high-protein diet. Methods A randomized, double-blind, placebo-controlled trial was performed. The participants were randomly allocated to receive either creatine (20 g/d for 5 d followed by 5 g/d throughout the trial) or placebo for 12 weeks. All of the participants were engaged in resistance training and consumed a high-protein diet (i.e., ≥ 1.2 g/Kg/d). Subjects were assessed at baseline (Pre) and after 12 weeks (Post). Glomerular filtration rate was measured by 51Cr-EDTA clearance. Additionally, blood samples and a 24-h urine collection were obtained for other kidney function assessments. Results No significant differences were observed for 51Cr-EDTA clearance throughout the trial (Creatine: Pre 101.42 ± 13.11, Post 108.78 ± 14.41 mL/min/1.73m2; Placebo: Pre 103.29 ± 17.64, Post 106.68 ± 16.05 mL/min/1.73m2; group x time interaction: F = 0.21, p = 0.64). Creatinine clearance, serum and urinary urea, electrolytes, proteinuria, and albuminuria remained virtually unchanged. Conclusions A 12-week creatine supplementation protocol did not affect kidney function in resistance-trained healthy individuals consuming a high-protein diet; thus reinforcing the safety of this dietary supplement. Trial registration ClinicalTrials.gov NCT01817673 PMID:23680457

On the anomalous adsorption of [Pd(edta)]2- at the water/Goethite interface: spectroscopic evidence for two types of surface complexes.

PubMed

Kaplun, Marina; Nordin, Agneta; Persson, Per

2008-01-15

The structure of palladium(II) ethylenediaminetetraacetate (edta) in aqueous solutions and its adsorption on the surface of goethite (alpha-FeOOH) were studied using extended X-ray absorption fine structure spectroscopy and attenuated total reflection Fourier transform infrared spectroscopy. The obtained results show that in aqueous solutions, Pd-edta exists as a 1:1 complex, [Pd(edta)]2-, with edta acting as a quadridentate ligand. On the surface of goethite, [Pd(edta)]2- forms two different types of complexes over a pH range of 3.40-8.12. At pH < 5, [Pd(edta)]2- adsorbs as an outer-sphere species with possible hydrogen bonding. At higher pH values, the formation of inner-sphere complexes of the cation-type sets in after a cleavage of one glycinate ring and the formation of an (edta)Pd-O-Fe linkage.

Modulation of matrix metalloproteinase activity by EDTA prevents posterior capsular opacification

PubMed Central

Guha, Rajdeep; Jongkey, Geram; Palui, Himangshu; Mishra, Akhilesh; Vemuganti, Geeta K.; Basak, Samar K.; Mandal, Tapan Kumar; Konar, Aditya

2012-01-01

Purpose To evaluate the effect of ethylenediaminetetraacetic acid (EDTA) on posterior capsular opacification (PCO) of rabbits and to assess its effect on intraocular tissues. Methods Modulation of matrix metalloproteinase (MMP) activity in the aqueous following cataract surgery in rabbits and its prevention by different doses of EDTA was determined by zymography. For evaluation of PCO, lensectomized rabbits were intracamerally injected with single dose of either 5 mg EDTA or normal saline. After one month, the degree of PCO was determined by slitlamp biomicroscopy, Miyake-Apple view, and histology of the lens capsule. The effect of EDTA on intra ocular pressure (IOP), corneal endothelial cells, and the retina was evaluated by tonometry, specular microscopy and scanning electron microscopy, and electroretinography. The concentration of EDTA in the aqueous was determined by high performance liquid chromatography (HPLC) at different time points. Results The MMP activity was significantly increased in the aqueous of the operated eyes, and EDTA reduced the degree of increase in a dose-dependent manner. EDTA treatment significantly reduced the degree of PCO (p<0.05). Histopathology of the lens capsule showed a reduction in the number of proliferating and migrating cells as well as MMP2 expression in the EDTA-treated eyes. EDTA treatment did not change the IOP; density, morphology and ultrastructure of the corneal endothelial cells; and electroretinography (ERG). EDTA was detectable in the aqueous humor up to 72 h following a single intracameral injection. Conclusions EDTA reduces the degree of PCO by suppressing the MMP activity and it is not toxic to intra ocular structures at the concentration used. PMID:22815623

Biofilm Formation by Otopathogenic Strains of P. aeruginosa is not Consistently Inhibited by EDTA

PubMed Central

Zenga, Joseph; Gagnon, Patricia M.; Vogel, Joseph; Chole, Richard A.

2012-01-01

Hypothesis Biofilm formation in otopathogenic of P. aeruginosa (OPPA) strains is inhibited by ethylenediaminetetraacetic acid (EDTA). Background EDTA, a widely used chelating agent, has been shown to inhibit biofilm formation in a number of bacteria. Since EDTA may be a well-tolerated reagent to inhibit biofilm formation in cases of suppurative otitis media, we asked if it might be effective in all OPPA strains isolated from chronically infected cholesteatomas. Methods OPPA strains were isolated from patients with infected cholesteatomas. These strains were grown into log phase then were placed in minimal media with varying concentrations of EDTA, and incubated for varying periods. Biofilm production was measured colorimetrically by staining with crystal violet. Results Without added EDTA, most otopathogenic PA exhibited a distinct, but varying, time-course of biofilm formation and dissolution with peak production at 12–18 hours. Addition of 1 mM EDTA resulted in a delay in the time to peak biofilm formation for most strains, although the amount of biofilm was not decreased. In contrast, some strains showed greater biofilm production with 1 mM EDTA compared to the untreated bacteria. Addition of 10 mM EDTA resulted in a similar effect. Some strains increased biofilm production over controls. Moreover, EDTA inhibited planktonic growth of all OPPA strains at the concentrations studied. Conclusion Our hypothesis was disproven: EDTA tends to delay biofilm development while it consistently inhibits planktonic growth. Since EDTA does not cause suppression of biofilm production in all isolates of OPPA, usefulness as an antimicrobial is questioned. PMID:22772018

In vitro and in vivo activity of EDTA and antibacterial agents against the biofilm of mucoid Pseudomonas aeruginosa.

PubMed

Liu, Zhenqiu; Lin, Yaying; Lu, Qi; Li, Fang; Yu, Jialin; Wang, Zhengli; He, Yu; Song, Chao

2017-02-01

Refractory infection caused by bacterial biofilm is an important clinical problem. Pseudomonas aeruginosa is a common pathogen responsible for persistent and chronic biofilm infections. We aimed to explore the in vitro and in vivo activity of ethylenediamine tetraacetic acid (EDTA) in combination with antibacterial agents against mucoid P. aeruginosa biofilm. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration of ciprofloxacin, gentamicin, and ampicillin alone or with EDTA against P. aeruginosa were determined in vitro. Extracellular polysaccharides (EPS) and structural parameters of the biofilm were monitored. P. aeruginosa was aerosolized and delivered into the lungs of guinea pigs, which were treated with ciprofloxacin with or without EDTA. The colony-forming units (CFUs) of P. aeruginosa were determined from the lungs. EDTA reduced the MIC of ciprofloxacin and ampicillin by about 30-fold and that of gentamicin by twofold. EDTA reduced the biofilm EPS and the proportion of viable bacteria. The thickness, average diffusion distance, and textural entropy of EDTA-treated biofilm were significantly decreased. EDTA plus antibiotics reduced the colony counting from 10 7 to 10 3  CFU/mL. In vivo, EDTA plus ciprofloxacin had a significantly lower mean CFU/g of lung tissue (EDTA + ciprofloxacin 1.3 ± 0.19; EDTA 4.4 ± 0.57; ciprofloxacin 4.2 ± 0.47), and lung lesions were less severe compared with the single treatment groups. EDTA can destroy the biofilm structures of mucoid P. aeruginosa in vitro. Moreover, EDTA and ciprofloxacin had a significant bactericidal effect against biofilm in vivo.

The improved phytoextraction of lead (Pb) and the growth of maize (Zeamays L.): the role of plant growth regulators (GA3 and IAA) and EDTA alone and in combinations.

PubMed

Hadi, Fazal; Bano, Asghari; Fuller, Michael P

2010-06-01

This investigation was made to examine the role of gibberellic acid (GA(3)), indole-3-acetic acid (IAA) and EDTA in improving phytoextraction of the Pb and plant growth on Pb added soil. GA(3), IAA and EDTA were applied separately and in combinations. GA(3) and IAA were applied as foliar spray and seed soaking. EDTA was applied in single and split doses. Analysis of the Pb in different parts of plant was carried out using atomic absorption/flame spectrophotometer. EDTA significantly reduced the plant growth and dry biomass, whereas GA(3) and IAA foliar spray increased it significantly when compared with control (only Pb added soil). In combined treatments of EDTA+GA(3) and EDTA+IAA, the growth and biomass was restored, which shows that GA(3) and IAA did compensate the negative effect of EDTA on plant growth. The separate treatments of EDTA, GA(3) and IAA increased the Pb uptake and translocation significantly moreover in combine treatments, synergistic effect was found and remarkable increase in Pb uptake and translocation into shoot was observed. EDTA increased the Pb uptake but declined the biomass; subsequently the total Pb accumulation was decreased in plant. The maximum total Pb was found in combined treatment of EDTA+GA(3). These findings suggest more investigation to find a combination of GA(3) with a very low concentration of EDTA, as in high concentration it causes soil and ground water pollution.

Recovery of manganese from manganese oxide ores in the EDTA solution

NASA Astrophysics Data System (ADS)

Zhang, Chao; Wang, Shuai; Cao, Zhan-fang; Zhong, Hong

2018-04-01

A new process has been experimentally and theoretically established for the recovery of manganese from manganese oxide ores, mainly including the reductive leaching of manganese by ethylenediaminetetraacetic acid (EDTA), EDTA recovery, and manganese electrolysis. The experimental conditions for this process were investigated. Moderate leaching environment by EDTA with the pH in the range of 5-6 is of benefit to leach manganese from some manganese oxide ores with high-content impurities, such as iron and aluminum. Most of EDTA can be recovered by acidification. A small amount of the residual EDTA in the electrolyte can prevent the generation of anode mud. In addition, trimanganese tetroxide (Mn3O4) can be obtained by the roasting of the EDTA-Mn crystallized product.

Comparison between high concentration EDTA (24%) and low concentration EDTA (3%) with surfactant upon removal of smear layer after rotary instrumentation: a SEM study.

PubMed

Daghustani, Mohsen; Alhammadi, Ahmad; Merdad, Khalid; Ohlin, Johan; Erhardt, Fredrik; Ahlquist, Michael

2011-01-01

This in vitro study compare cleanliness of tooth canal walls regarding smear layer after final treatment with 24% ethylenediaminetetraaceticacid (EDTA) and 3% EDTA with or without surfactant. Sixty extracted teeth, randomly distributed into four groups, were prepared using ProFile instruments (DENTSPLY, Maillefer, Ballaigues, Switzerland), and subjected to different final irrigation solutions: group A, 24% EDTA; group B, 3% EDTA with surfactant; group C (positive control), 3% EDTA; and group D (negative control), 0.5% sodium hypochlorite. Roots were sectioned, examined and evaluated under scanning electron microscope; microphotographs were taken for the coronal, middle and apical third of each specimen. Statistical analysis showed no difference regarding presence of smear layer between test groups in the coronal and apical sections. They were cleaned in the coronal sections and uncleaned in the apical sections. In the middle section, group B was significantly cleaner (p < 0.05) than the other groups. In conclusion, surfactant in combination with EDTA did not improve root canal cleanliness and there is no difference between different EDTA concentrations in removing the smear layer.

The application of EDTA in drug delivery systems: doxorubicin liposomes loaded via NH4EDTA gradient

PubMed Central

Song, Yanzhi; Huang, Zhenjun; Song, Yang; Tian, Qingjing; Liu, Xinrong; She, Zhennan; Jiao, Jiao; Lu, Eliza; Deng, Yihui

2014-01-01

The applications of ethylenediaminetetraacetic acid (EDTA) have been expanded from the treatment of heavy metal poisoning to chelation therapies for atherosclerosis, heart disease, and cancers, in which EDTA reduces morbidity and mortality by chelating toxic metal ions. In this study, EDTA was used in a drug delivery system by adopting an NH4EDTA gradient method to load doxorubicin into liposomes with the goal of increasing therapeutic effects and decreasing drug-related cytotoxicity. The particle size of the optimum NH4EDTA gradient liposomes was 79.4±1.87 nm, and the entrapment efficiency was 95.54%±0.59%. In vitro studies revealed that liposomes prepared using an NH4EDTA gradient possessed long-term stability and delayed drug release. The in vivo studies also showed the superiority of the new doxorubicin formulation. Compared with an equivalent drug dose (5 mg/kg) prepared by (NH4)2SO4 gradient, NH4EDTA gradient liposomes showed no significant differences in tumor inhibition ratio, but cardiotoxicity and liposome-related immune organ damage were lower, and no drug-related deaths were observed. These results show that use of the NH4EDTA gradient method to load doxorubicin into liposomes could significantly reduce drug toxicity without influencing antitumor activity. PMID:25120359

Solution Preserves Nucleic Acids in Body-Fluid Specimens

NASA Technical Reports Server (NTRS)

Pierson, Duane L.; Stowe, Raymond P.

2004-01-01

A solution has been formulated to preserve deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) in specimens of blood, saliva, and other bodily fluids. Specimens of this type are collected for diagnostic molecular pathology, which is becoming the method of choice for diagnosis of many diseases. The solution makes it possible to store such specimens at room temperature, without risk of decomposition, for subsequent analysis in a laboratory that could be remote from the sampling location. Thus, the solution could be a means to bring the benefits of diagnostic molecular pathology to geographic regions where refrigeration equipment and diagnostic laboratories are not available. The table lists the ingredients of the solution. The functions of the ingredients are the following: EDTA chelates divalent cations that are necessary cofactors for nuclease activity. In so doing, it functionally removes these cations and thereby retards the action of nucleases. EDTA also stabilizes the DNA helix. Tris serves as a buffering agent, which is needed because minor contaminants in an unbuffered solution can exert pronounced effects on pH and thereby cause spontaneous degradation of DNA. SDS is an ionic detergent that inhibits ribonuclease activity. SDS has been used in some lysis buffers and as a storage buffer for RNA after purification. The use of the solution is straightforward. For example, a sample of saliva is collected by placing a cotton roll around in the subject's mouth until it becomes saturated, then the cotton is placed in a collection tube. Next, 1.5 mL of the solution are injected directly into the cotton and the tube is capped for storage at room temperature. The effectiveness of the solution has been demonstrated in tests on specimens of saliva containing herpes simplex virus. In the tests, the viral DNA, as amplified by polymerase chain reaction, was detected even after storage for 120 days.

In search of susceptibility genes for type 2 diabetes in West Africa: the design and results of the first phase of the AADM study.

PubMed

Rotimi, C N; Dunston, G M; Berg, K; Akinsete, O; Amoah, A; Owusu, S; Acheampong, J; Boateng, K; Oli, J; Okafor, G; Onyenekwe, B; Osotimehin, B; Abbiyesuku, F; Johnson, T; Fasanmade, O; Furbert-Harris, P; Kittles, R; Vekich, M; Adegoke, O; Bonney, G; Collins, F

2001-01-01

The purpose of this study is to map type 2 diabetes susceptibility genes in West African ancestral populations of African-Americans, through an international collaboration between West African and US investigators. Affected sib-pairs (ASP) along with unaffected spouse controls are being enrolled and examined in West Africa, with two sites established in Ghana (Accra and Kumasi) and three in Nigeria (Enugu, Ibadan, and Lagos). Eligible participants are invited to study clinics to obtain detailed epidemiologic, family, and medical history information. Blood samples are drawn from each participant to measure glucose, insulin, C-peptide, total cholesterol, LDL, HDL, triglycerides, albumin, creatinine, urea, uric acid, total calcium and to detect autoantibodies to glutamic acid decarboxylase (GAD). DNA is isolated from frozen white blood cells obtained from 20 ml of EDTA whole blood samples. With full informed consent, 162 individuals from 78 families have been enrolled and examined since the Africa America Diabetes Mellitus (AADM) study began in June of 1997. Logistics of field examinations and specimen shipping have been successfully established. At the end of the third year of field activity (September 2000) the AADM study will have enrolled and performed comprehensive examination on 400 ASP with type 2 diabetes, for a minimum of 800 cases and 200 controls from Ghana and Nigeria. At the current participation rate, the goal of 400 sib-pairs and 200 controls will be met before the scheduled closing date. The AADM study will create a comprehensive epidemiologic and genetic resource that will facilitate a powerful genome-wide search for West African susceptibility genes to type 2 diabetes.

Effect of PDGF-BB combined with EDTA gel on adhesion and proliferation to the root surface.

PubMed

Belal, Mahmoud Helmy; Watanabe, Hisashi; Ichinose, Shizuko; Ishikawa, Isao

2012-07-01

Periodontal regeneration using EDTA or PDGF showed promising results, but the effect of combined application was still unclear. This study aimed to verify the effect of EDTA and/or PDGF application on root adhesion and proliferation of PDL fibroblast cells. Eighty specimens were prepared from forty periodontitis teeth and made five groups: (1) diseased (untreated), (2) SRP (scaling root planing), (3) EDTA (24%), (4) PDGF (25 ng/ml) and (5) Combined application of EDTA and PDGF. Periodontal ligament cells were cultured on the above conditioned dentin plate, and SEM examination was preformed and cells were counted within a representative standard area for both cell morphology and density. All groups including untreated showed significantly increase of adhered cells from baseline to 7 days. Among them, rate of increase was much higher in EDTA, PDGF, and combined groups. ANOVA test indicated that the number of cells in PDGF and combined groups was significantly higher than diseased group at 1 day. On day 7, PDGF and combined groups showed significantly higher number of adhesion cells than that found in the diseased, SRP and EDTA groups. Thus, root conditioning with EDTA enhanced cell adhesion more than SRP alone. There was no significant difference of cell number between PDGF and combined group. Combined application of EDTA and PDGF increased significantly PDL cell adhesion than EDTA alone. PDGF alone, however, also showed comparable effect to combined application at all periods. Thus, synergistic effect between PDGF and EDTA was not observed.

Enhancing phosphorus release from waste activated sludge containing ferric or aluminum phosphates by EDTA addition during anaerobic fermentation process.

PubMed

Zou, Jinte; Zhang, Lili; Wang, Lin; Li, Yongmei

2017-03-01

The effect of ethylene diamine tetraacetic acid (EDTA) addition on phosphorus release from biosolids and phosphate precipitates during anaerobic fermentation was investigated. Meanwhile, the impact of EDTA addition on the anaerobic fermentation process was revealed. The results indicate that EDTA addition significantly enhanced the release of phosphorus from biosolids, ferric phosphate precipitate and aluminum phosphate precipitate during anaerobic fermentation, which is attributed to the complexation of metal ions and damage of cell membrane caused by EDTA. With the optimal EDTA addition of 19.5 mM (0.41 gEDTA/gSS), phosphorus release efficiency from biosolids was 82%, which was much higher than that (40%) without EDTA addition. Meanwhile, with 19.5 mM EDTA addition, almost all the phosphorus in ferric phosphate precipitate was released, while only 57% of phosphorus in aluminum phosphate precipitate was released. This indicates that phosphorus in ferric phosphate precipitate was much easier to be released than that in aluminum phosphate precipitate during anaerobic fermentation of sludge. In addition, proper EDTA addition facilitated the production of soluble total organic carbon and volatile fatty acids, as well as solid reduction during sludge fermentation, although methane production could be inhibited. Therefore, EDTA addition can be used as an alternative method for recovering phosphorus from waste activated sludge containing ferric or aluminum precipitates, as well as recovery of soluble carbon source. Copyright © 2016 Elsevier Ltd. All rights reserved.

Effect of super-oxidized water, sodium hypochlorite and EDTA on dentin microhardness.

PubMed

Ghisi, Alexandre Corrêa; Kopper, Patrícia Maria Poli; Baldasso, Flávia E R; Stürmer, Caroline P; Rossi-Fedele, Giampiero; Steier, Liviu; Figueiredo, José Antonio Poli de; Morgental, Renata Dornelles; Vier-Pelisser, Fabiana Vieira

2014-01-01

The present study aimed to evaluate the influence of the following irrigating solutions on the microhardness of root canal dentin: 2% sodium hypochlorite (2NaOCl), 5% sodium hypochlorite (5NaOCl), super-oxidized water (400 ppm Sterilox - Sx) and 17% EDTA (E). Eighty roots from bovine incisors were randomly divided into 8 groups (n=10): 2NaOCl, 5NaOCl, Sx, and 2NaOCl + E, 5NaOCl + E, Sx + E (associated with E as final irrigant for 5 min), E solely and distilled water (dH2O) as the negative control. Root canal preparation was performed by hand instruments, using one of the irrigation protocols for 30 min. Then, 5 mm of the cervical root third were cut out from each sample and subjected to the Vickers microhardness test, at two points, one at approximately 500-1000 µm from the root canal lumen (distance 1), and the other at approximately 500-1000 µm from the external root surface (distance 2). Data were analyzed by Wilcoxon and Kruskal-Wallis tests at 5% significance level. Microhardness values at distance 1 were significantly lower than those at distance 2 for all groups, except 5NaOCl and 5NaOCl + E groups (p>0.05). EDTA showed the lowest microhardness values. However, no statistically significant difference was detected among groups at distance 1 and EDTA was significantly different only from Sx at distance 2. In conclusion, all tested solutions showed lower microhardness at the most superficial root canal dentin layer compared to the one found near the external root surface, except 5NaOCl and 5NaOCl + E; EDTA promoted lower microhardness values in comparison to Sterilox at this site.

Potential endocrine disrupting organic chemicals in treated municipal wastewater and river water

USGS Publications Warehouse

Barber, L.B.; Brown, G.K.; Zaugg, S.D.

2000-01-01

Select endocrine disrupting organic chemicals were measured in treated wastewater from Chicago, IL, Minneapolis/St. Paul, MN, Detroit, MI, and Milwaukee, WI, and in the Des Plaines, Illinois, and Minnesota Rivers during the fall of 1997 and the spring of 1998. Emphasis was given to alkylphenolpolyethoxylate (APEO) derived compounds, although 17-??-estradiol, bisphenol A, caffeine, total organic carbon, ethylenediaminetetraacetic acid (EDTA), and other compounds also were measured. Contaminants were isolated by continuous liquid-liquid extraction (CLLE) with methylene chloride and analyzed by gas chromatography/mass spectrometry in full scan and selected ion monitoring modes. The extracts were derivatized to form the methyl esters of alkylphenolethoxycarboxylates (APEC), and EDTA was isolated by evaporation and derivatized to form the tetrapropyl ester. The mass spectra of nonylphenol (NP) and octylphenol (OP) compounds are complex and show variations among the different ethoxylate and carboxylate homologs, reflecting variations in the ethylene oxide chain length. Recoveries for target compounds and surrogate standards ranged from 20-130%, with relative standard deviations of 9.9-53%. Detection limits for the various compounds ranged from 0.06-0.35 ??g/L. Analysis of the wastewater effluents detected a number of compounds including NP, NPEO, OP, OPEO, NPEC, caffeine, and EDTA at concentrations ranging from <1-439 ??g/L, with EDTA and NPEC being most abundant. There was variability in compound distributions and concentrations between the various sewage treatment plants, indicating differences in treatment type and influent composition. Several wastewater-derived compounds were detected in the river samples, with EDTA and NPEC persisting for considerable distance downstream from wastewater discharges, and NP and NPEO being attenuated more rapidly.

Effects of Application of NTA and EDTA on Accumulation of Soil Heavy Metals in Chrysanthemum

NASA Astrophysics Data System (ADS)

Bai, Weiyang

2018-02-01

In order to find out the effect of non-bio chelating agent EDTA and bio-chelating agent NTA on soil heavy metal pollution, the effects of different ratio of chelating agent NTA and EDTA on soil heavy metals (Pb, Cu and Cd ), the effects of chelating on content of chlorophyll and vitamin C and the degree of soil nutrient loss were evaluated. The results showed: that the contents of Pb and Cd were the highest in the roots of Chrysanthemum in the treatment (EDTA / NTA = 2: 1). The treatment (EDTA / NTA = 1: 1) was the best chelating agent ratio for the synergistic effect, which can significantly promoted the Chrysanthemum on heavy metal Cu uptake and transport to aboveground. Chrysanthemum were inhibited by all chelating agents treatments, while the content of chlorophyll and vitamin C of the Chrysanthemum physiological indexes were reduced. In the treatment (EDTA = 1), chlorophyll SPAD, vitamin C content reached a minimum of 36 and 38mg · 100g-1, respectively. The nutrient element TN in the leachate were gradually decreased with the time, and the tenth day was significantly lower than the leaching rate of the first day (p <0.05) in the treatments (NTA = 1, EDTA / NTA = 1: 2). The nutrient element TN was decreased most, reaching 51.6%, and the activation effect was decreased significantly in the fifteenth day in treatment (NTA = 1), Treatment(EDTA / NTA = 1: 1)> treatment (EDTA = 1) >treatment (EDTA / NTA = 2: 1) >treatment (EDTA / NTA = 1: 2)> treatment (NTA = 1). Treatment (EDTA / NTA = 2: 1) was recommended for the chelating agent ratio with the better physiological parameters, the more heavy metal extraction and the less nitrogen and phosphorus loss

Effectiveness of Three Different Irrigants - 17% Ethylenediaminetetraacetic Acid, Q-MIX, and Phytic Acid in Smear Layer Removal: A Comparative Scanning Electron Microscope Study.

PubMed

Jagzap, Janhavi Balasaheb; Patil, Sanjay S; Gade, Vandana Jaykumar; Chandhok, Deepika J; Upagade, Madhura A; Thakur, Deepa A

2017-01-01

Removal of smear layer from the root canal walls is important for long-standing endodontic success. The aim of this study is to evaluate and compare smear layer removing ability among 17% ethylenediaminetetraacetic acid (EDTA), Q-MIX, and phytic acid by scanning electron microscopy (SEM). This in-vitro experimental study assessed smear layer removal using three different irrigants. Thirty single-rooted freshly extracted human permanent premolars were collected, disinfected, and decoronated to a standardized root length of 13 mm. Root canals were cleaned and shaped till F2 universal rotary protaper at working length 1 mm short of the apex. They were randomly divided into three groups, and final irrigation was done accordingly. Group 1 ( n = 10): with 1 ml of 17% EDTA, Group 2 ( n = 10): with 1 ml of Q-MIX, Group 3 ( n = 10): with 1 ml of phytic acid. Samples were then longitudinally sectioned and evaluated under SEM at coronal, middle, and apical levels. Two-way analysis of variance and Tukey's post hoc test were performed. The level of significance was set at 0.05. Smear layer removing ability among irrigants and sections in descending order: 17 EDTA > Q-MIX > phytic acid; coronal > middle > apical. 17% EDTA showed better and promising results followed by Q-MIX and then phytic acid.

Characterization of EDTA-soluble polysaccharides from the scape of Musa paradisiaca (banana).

PubMed

Raju, T S; Jagadish, R L; Anjaneyalu, Y V

2001-02-01

The polysaccharide components present in the scape of Musa paradisiaca (banana) were fractionated into water-soluble (WSP), EDTA-soluble (EDTA-SP), alkali-soluble (ASP) and alkali-insoluble (AISP) polysaccharide fractions [Anjaneyalu, Jagadish and Raju (1997) Glycoconj. J. 14, 507-512]. The EDTA-SP was further fractionated by iso-amyl alcohol into EDTA-SP-A and EDTA-SP-B. The homogeneity of these two polysaccharides was established by repeated precipitation with iso-amyl alcohol, gel-filtration chromatography and sedimentation analysis. The polysaccharides were characterized by monosaccharide composition analysis, methylation linkage analysis, iodine affinity, ferricyanide number, blue value, hydrolysis with alpha-amylase, gold-electron microscopy and X-ray diffraction spectroscopy. Data from all of these studies suggest that EDTA-SP-A is a branched amylose-type alpha-D-glucan and that EDTA-SP-B is a highly branched amylopectin-type polymer. The nature of the branching patterns of these polysaccharides suggests that they are unique to M. paradisiaca.

Smear layer removal evaluation of different protocol of Bio Race file and XP- endo Finisher file in corporation with EDTA 17% and NaOCl.

PubMed

Zand, Vahid; Mokhtari, Hadi; Reyhani, Mohammad-Frough; Nahavandizadeh, Neda; Azimi, Shahram

2017-11-01

The aim of the present study was to compare the amount of the smear layer remaining in prepared root canals with different protocols of Bio RaCe files and XP-endo Finisher file (XPF) in association with 17% EDTA and sodium hypochlorite solution. A total of 68 extracted single-rooted teeth were randomly divided into 4 experimental groups (n=14) and two control groups (n=6). The root canals were prepared with Bio RaCe files (FKG Dentaire, Switzerland) using the crown-down technique based on manufacturer's instructions and irrigated according to the following irrigation techniques: Group 1: XPF with 2 mL of 2.5% NaOCl for 1 minute. Group 2:, XPF with 1 mL of 17% EDTA for one minute. Group 3: XPF was used for 1 minute in association with normal saline solution. Group 4: XP-endo Finisher file for 30 seconds in association with 2.5% NaOCl and 17% EDTA for 30 seconds. The negative control group: NaOCl (2.5%) was used during root canal preparation, followed by irrigation with 17% EDTA at the end of root canal preparation. The positive control group: Normal saline solution was used for irrigation during root canal preparation. In all the groups, during preparation of the root canals with Bio RaCe file, 20 mL of 2.5% NaOCl was used for root canal irrigation and at the end of the procedural steps 20 mL of normal saline solution was used as a final irrigant. The samples were analyzed under SEM at ×1000‒2000 magnification and evaluated using Torabinejad scoring system. Data were analyzed with non-parametric Kruskal-Wallis test and post hoc Mann-Whitney U test, using SPSS. Statistical significant was defined at P <0.05. The results of the study showed the least amount of the smear layer at coronal, middle and apical thirds of the root canals in groups 2, which was not significantly different from the negative control group ( P <0.5). Under the limitations of the present study, use of a combination of NaOCl and EDTA in association with XPF exhibited the best efficacy for the removal of the smear layer. Key words: Smear layer, XP-endo Finisher file, EDTA, Sodium hypochlorite.

Effect of EDTA, sodium hypochlorite, and chlorhexidine gluconate with or without surface modifiers on dentin microhardness.

PubMed

Aslantas, Eda E; Buzoglu, Hatice Dogan; Altundasar, Emre; Serper, Ahmet

2014-06-01

This study aimed to evaluate the effects of root canal irrigants on the microhardness of root canal dentin in the presence and absence of surface-modifying agents. Forty-eight root halves were prepared by longitudinal splitting of the distal roots of 24 freshly extracted mandibular human third molars and embedded in autopolymerizing acrylic resin, leaving the dentin surface exposed. After polishing, the microhardness values of the untreated dentin surfaces were recorded by using Vickers tester at the mid-root level. The root halves were randomly assigned to 6 groups composed of 8 samples each and treated for 5 minutes with one of the following irrigants: 17% EDTA, REDTA, 2% chlorhexidine gluconate (CHX), 2% CHX with surface modifiers (CHX-Plus), 6% NaOCl, or 6% NaOCl with surface modifiers (Chlor-XTRA). After surface treatment, dentin microhardness values were recorded at close proximity to the initial indentation areas. Experimental data were statistically analyzed by using the t test and one-way analysis of variance, followed by Tukey honestly significant difference test at α = 0.05. EDTA, REDTA, NaOCl, and Chlor-XTRA significantly decreased the microhardness of root dentin compared with intact controls (P < .05). The addition of surface modifiers to the irrigants did not affect the microhardness of the samples. Copyright © 2014 American Association of Endodontists. Published by Elsevier Inc. All rights reserved.

Dual-wavelength excitation to reduce background fluorescence for fluorescence spectroscopic quantitation of erythrocyte zinc protoporphyrin-IX and protoporphyrin-IX from whole blood and oral mucosa

NASA Astrophysics Data System (ADS)

Hennig, Georg; Vogeser, Michael; Holdt, Lesca M.; Homann, Christian; Großmann, Michael; Stepp, Herbert; Gruber, Christian; Erdogan, Ilknur; Hasmüller, Stephan; Hasbargen, Uwe; Brittenham, Gary M.

2014-02-01

Erythrocyte zinc protoporphyrin-IX (ZnPP) and protoporphyrin-IX (PPIX) accumulate in a variety of disorders that restrict or disrupt the biosynthesis of heme, including iron deficiency and various porphyrias. We describe a reagent-free spectroscopic method based on dual-wavelength excitation that can measure simultaneously both ZnPP and PPIX fluorescence from unwashed whole blood while virtually eliminating background fluorescence. We further aim to quantify ZnPP and PPIX non-invasively from the intact oral mucosa using dual-wavelength excitation to reduce the strong tissue background fluorescence while retaining the faint porphyrin fluorescence signal originating from erythrocytes. Fluorescence spectroscopic measurements were made on 35 diluted EDTA blood samples using a custom front-face fluorometer. The difference spectrum between fluorescence at 425 nm and 407 nm excitation effectively eliminated background autofluorescence while retaining the characteristic porphyrin peaks. These peaks were evaluated quantitatively and the results compared to a reference HPLC-kit method. A modified instrument using a single 1000 μm fiber for light delivery and detection was used to record fluorescence spectra from oral mucosa. For blood measurements, the ZnPP and PPIX fluorescence intensities from the difference spectra correlated well with the reference method (ZnPP: Spearman's rho rs = 0.943, p < 0.0001; PPIX: rs = 0.959, p < 0.0001). In difference spectra from oral mucosa, background fluorescence was reduced significantly, while porphyrin signals remained observable. The dual-wavelength excitation method evaluates quantitatively the ZnPP/heme and PPIX/heme ratios from unwashed whole blood, simplifying clinical laboratory measurements. The difference technique reduces the background fluorescence from measurements on oral mucosa, allowing for future non-invasive quantitation of erythrocyte ZnPP and PPIX.

Induced Phytoextraction of Lead Through Chemical Manipulation of Switchgrass and Corn; Role of Iron Supplement.

PubMed

Johnson, Deayne M; Deocampo, Daniel M; El-Mayas, Hanan; Greipsson, Sigurdur

2015-01-01

The effects of combined chemical application of benomyl, ethylenedianinetetraacetate (EDTA), and iron (Fe) (foliar and root) on lead (Pb) phytoextraction by switchgrass (Panicum virgatum) and corn (Zea mays) was examined. Switchgrass was grown in Pb-contaminated urban topsoil with the following treatments: (C) Control, (B) benomyl, (E) EDTA, (F) foliar-Fe, (BE) benomyl + EDTA, (BF) benomyl + foliar-Fe, (FE) foliar-Fe + EDTA, (BFE) benomyl + foliar-Fe + EDTA. Corn was grown in sand-culture supplemented with Pb (500 mg kg(-1)) with the following treatments: (C) control, (B) benomyl, (E) EDTA, (F) root-Fe, (BE) benomyl + EDTA, (BF) benomyl + root-Fe, (FE) root-iron + EDTA, and, (BFE) benomyl + root-Fe + EDTA. All treatments were replicated three times and pots were arranged in a completely randomized design. Plants were analyzed for element concentration (Fe, Zn, P, and Pb) using either inductively coupled plasma (argon) atomic emission spectroscopy (ICP-AES) or graphite furnace atomic absorption spectrometer. Iron supplementation (foliar and root) affected Pb-translocation in plants. Foliar-Fe treatment increased translocation ratio of Pb (TF-Pb) significantly compared to other treatments with the exception of plants treated with benomyl and BF. Root-Fe treatment in combination with EDTA (FE) increased TF-Pb significantly compared to other treatments. Phytoextraction was improved by the combined chemical application; plants treated with BFE treatment increased Pb-total-phytoextraction by 424% compared to Control plants.

The influence of EDTA application on the interactions of cadmium, zinc, and lead and their uptake of rainbow pink (Dianthus chinensis).

PubMed

Lai, Hung-Yu; Chen, Zueng-Sang

2006-10-11

Soil used in this study was artificially contaminated with Cd, Zn, Pb, or applied in combinations (Cd-Zn, Cd-Pb, Zn-Pb, or Cd-Zn-Pb) to study the interactions of metals in soil contaminated with multiple metals. After planting rainbow pink (Dianthus chinensis) in these soils for 21 days, three different concentrations of ethylenedinitrilotetraacetic acid (EDTA) solutions were added to study the effect of applying EDTA on the interactions among these metals. The concentrations of Cd, Zn, and Pb in the soil solutions of different metals-treated soils increased significantly after applying 5 mmol EDTA kg(-1) soil (p<0.05). The potential of groundwater contamination will increase after applying EDTA and it is not recommended to be in situ used or have to use very carefully. The existence of Pb in the Cd-contaminated soil enhanced the uptake of Cd in rainbow pink in the treatments of control and 2 mmol EDTA kg(-1) soil. Cadmium inhibited the concentration of Zn without applying EDTA. However, whether the application of EDTA or not and the applied EDTA concentration had the greatest effect on the uptake of Pb when compared to Cd and Zn. After applying 5 mmol EDTA kg(-1) soil, Cd or Zn in the Pb-contaminated soil inhibited the uptake of Pb in rainbow pink, but there were no effect in other treatments.

Dual function of EDTA with silver nanoparticles for root canal treatment-A novel modification.

PubMed

Martinez-Andrade, Juan M; Avalos-Borja, Miguel; Vilchis-Nestor, Alfredo R; Sanchez-Vargas, Luis O; Castro-Longoria, Ernestina

2018-01-01

The chelating and antimicrobial capacity of a novel modification of 17% EDTA with silver nanoparticles (AgNPs) (EDTA-AgNPs) was evaluated in-vitro for root canal treatment (RCT). The EDTA-AgNPs solution was characterized by UV-Vis spectroscopy, ζ-potential and high-resolution transmission electron microscopy (HRTEM). Antimicrobial capacity was evaluated against Candida albicans and Staphylococcus aureus in planktonic and biofilm cells by broth macrodilution (24 h) and XTT assays, (1, 10 and 30 min) respectively. The chelating capacity of EDTA-AgNPs was assessed indirectly (smear layer removal) and directly (demineralizing effect) in bovine dentin at two silver concentrations, 16 and 512 μg/ml at 1 and 10 minutes of exposure time. Smear layer removal was evaluated by atomic force microscopy (AFM) and scanning electron microscopy (SEM). The demineralizing effect was determined by atomic absorption spectroscopy (AAS), microhardness test (MH) and X-ray diffractometer (XRD). Synthesized AgNPs were quasi-spherical in shape with an average size of 13.09 ± 8.05 nm. 17% EDTA-AgNPs was effective to inhibit C. albicans and S. aureus in planktonic and biofilm cultures. The smear layer removal and demineralizing effect were similar between 17% EDTA-AgNPs and 17% EDTA treatments. The 17% EDTA-AgNPs solution proved to be an effective antimicrobial agent, and has a similar chelating capacity to 17% EDTA alone. These in-vitro studies strongly suggest that EDTA-AgNPs could be used for effective smear layer removal, having an antimicrobial effect at the same time during RCT.

Dual function of EDTA with silver nanoparticles for root canal treatment–A novel modification

PubMed Central

Martinez-Andrade, Juan M.; Avalos-Borja, Miguel; Vilchis-Nestor, Alfredo R.; Sanchez-Vargas, Luis O.

2018-01-01

The chelating and antimicrobial capacity of a novel modification of 17% EDTA with silver nanoparticles (AgNPs) (EDTA-AgNPs) was evaluated in-vitro for root canal treatment (RCT). The EDTA-AgNPs solution was characterized by UV-Vis spectroscopy, ζ-potential and high-resolution transmission electron microscopy (HRTEM). Antimicrobial capacity was evaluated against Candida albicans and Staphylococcus aureus in planktonic and biofilm cells by broth macrodilution (24 h) and XTT assays, (1, 10 and 30 min) respectively. The chelating capacity of EDTA-AgNPs was assessed indirectly (smear layer removal) and directly (demineralizing effect) in bovine dentin at two silver concentrations, 16 and 512 μg/ml at 1 and 10 minutes of exposure time. Smear layer removal was evaluated by atomic force microscopy (AFM) and scanning electron microscopy (SEM). The demineralizing effect was determined by atomic absorption spectroscopy (AAS), microhardness test (MH) and X-ray diffractometer (XRD). Synthesized AgNPs were quasi-spherical in shape with an average size of 13.09 ± 8.05 nm. 17% EDTA-AgNPs was effective to inhibit C. albicans and S. aureus in planktonic and biofilm cultures. The smear layer removal and demineralizing effect were similar between 17% EDTA-AgNPs and 17% EDTA treatments. The 17% EDTA-AgNPs solution proved to be an effective antimicrobial agent, and has a similar chelating capacity to 17% EDTA alone. These in-vitro studies strongly suggest that EDTA-AgNPs could be used for effective smear layer removal, having an antimicrobial effect at the same time during RCT. PMID:29346398

Effects of calcium disodium EDTA and meso-2,3-dimercaptosuccinic acid on tissue concentrations of lead for use in treatment of calves with experimentally induced lead toxicosis.

PubMed

Meldrum, James B; Ko, Kam W

2003-06-01

To compare the efficacy of calcium disodium EDTA (CaNa2EDTA) and meso-2,3-dimercaptosuccinic acid (DMSA) in reducing concentrations of lead in selected tissues for use in treatment of calves with experimentally induced lead toxicosis. 19 sexually intact male Holstein calves that weighed 35 to 60 kg. Calves were randomly assigned to 1 of 5 treatment groups: group 1, control calves; group 2, lead only; group 3, lead and EDTA; group 4, lead and DMSA; and group 5, lead, EDTA, and DMSA. Calves in groups 2 to 5 were dosed daily with lead (5 mg/kg, PO) for 10 days. Doses of EDTA (100 mg/kg) and DMSA (25 mg/kg) were administered IV once daily for 4 consecutive days beginning on day 11. Effects of the chelators on lead concentrations in the liver, kidneys, testes, muscles, bones, and brain were compared among the various groups. Compared with the effects of EDTA, DMSA greatly reduced lead concentrations in renal and hepatic tissues. We did not detect significant differences for the effects of EDTA or DMSA on lead concentrations in the testes; there was an adverse interaction of EDTA with DMSA that caused an increase in lead concentrations in the testes. DMSA is much more effective than EDTA in removing lead from renal and hepatic tissues in calves. Use of DMSA in calves with lead intoxication appears to be a viable treatment option. Combining DMSA and EDTA as a treatment modality in calves did not offer any advantages.

Lead toxicity: consequences and interventions in an intensively managed (Gyps coprotheres) vulture colony.

PubMed

Naidoo, Vinny; Wolter, Kerri; Espie, Ian; Kotze, Antoinette

2012-09-01

The National Zoological Gardens of South Africa (NZG) is involved in the ex situ conservation of Gyps coprotheres, the Cape Griffon vulture (CGV) and houses 24 birds in a 100-yr-old aviary. Following the death of one vulture with high liver lead concentrations, an investigation was launched to ascertain the source(s) and consequences of lead toxicity in this breeding colony. Whole blood from 24 CGV, paint from the enclosure, water, and soil sampled at various locations within the enclosure were evaluated for their lead concentration, and data were gathered from NZG's medical records. The lead concentration in the paint, water, and enclosure soil was 5,100 microg/g, 0.5 microg/dl, and 72.48 +/- 21.83 microg/g, respectively. The whole-blood lead concentrations were 56.58 +/- 11 microg/dl. The breeding history of six pairs within the contaminated enclosure since 2002 showed 45 eggs laid, of which 44% were infertile and 24% successfully reared. The medical records revealed evidence of osteodystrophy despite adequate nutrition. As intervention measures, six birds were treated with Ca2+EDTA and the topsoil inside the enclosure was replaced. As a result, the lead concentration in the enclosure soil dropped to 14.74 +/- 14.39 microg/g, and the whole-blood lead concentrations declined to 42.75 +/- 11.64 microg/dl. It was concluded that lead concentrations in whole blood in excess of 100 microg/dl leads to clinical signs of lead toxicity in the CGV. Lower levels appear to interfere mainly with reproductive potential.

CD4 Lymphocyte Enumeration and Hemoglobin Assessment Aid for Priority Decisions: A Multisite Evaluation of the BD FACSPresto™ System

PubMed Central

Thakar, Madhuri; Angira, Francis; Pattanapanyasat, Kovit; Wu, Alan H.B.; O’Gorman, Maurice; Zeng, Hui; Qu, Chenxue; Mahajan, Bharati; Sukapirom, Kasama; Chen, Danying; Hao, Yu; Gong, Yan; Indig, Monika De Arruda; Graminske, Sharon; Orta, Diana; d’Empaire, Nicole; Lu, Beverly; Omana-Zapata, Imelda; Zeh, Clement

2017-01-01

Background: The BD FACSPresto™ system uses capillary and venous blood to measure CD4 absolute counts (CD4), %CD4 in lymphocytes, and hemoglobin (Hb) in approximately 25 minutes. CD4 cell count is used with portable CD4 counters in resource-limited settings to manage HIV/AIDS patients. A method comparison was performed using capillary and venous samples from seven clinical laboratories in five countries. The BD FACSPresto system was assessed for variability between laboratory, instrument/operators, cartridge lots and within-run at four sites. Methods: Samples were collected under approved voluntary consent. EDTA-anticoagulated venous samples were tested for CD4 and %CD4 T cells using the gold-standard BD FACSCalibur™ system, and for Hb, using the Sysmex® KX-21N™ analyzer. Venous and capillary samples were tested on the BD FACSPresto system. Matched data was analyzed for bias (Deming linear regression and Bland-Altman methods), and for concordance around the clinical decision point. The coefficient of variation was estimated per site, instrument/operator, cartridge-lot and between-runs. Results: For method comparison, 93% of the 720 samples were from HIV-positive and 7% from HIV-negative or normal subjects. CD4 and %CD4 T cells venous and capillary results gave slopes within 0.96–1.05 and R2 ≥0.96; Hb slopes were ≥1.00 and R2 ≥0.89. Variability across sites/operators gave %CV <5.8% for CD4 counts, <1.9% for %CD4 and <3.2% for Hb. The total %CV was <7.7% across instrument/cartridge lot. Conclusion: The BD FACSPresto system provides accurate, reliable, precise CD4/%CD4/Hb results compared to gold-standard methods, irrespective of venous or capillary blood sampling. The data showed good agreement between the BD FACSPresto, BD FACSCalibur and Sysmex systems. PMID:29290885

CD4 Lymphocyte Enumeration and Hemoglobin Assessment Aid for Priority Decisions: A Multisite Evaluation of the BD FACSPresto™ System.

PubMed

Thakar, Madhuri; Angira, Francis; Pattanapanyasat, Kovit; Wu, Alan H B; O'Gorman, Maurice; Zeng, Hui; Qu, Chenxue; Mahajan, Bharati; Sukapirom, Kasama; Chen, Danying; Hao, Yu; Gong, Yan; Indig, Monika De Arruda; Graminske, Sharon; Orta, Diana; d'Empaire, Nicole; Lu, Beverly; Omana-Zapata, Imelda; Zeh, Clement

2017-01-01

The BD FACSPresto ™ system uses capillary and venous blood to measure CD4 absolute counts (CD4), %CD4 in lymphocytes, and hemoglobin (Hb) in approximately 25 minutes. CD4 cell count is used with portable CD4 counters in resource-limited settings to manage HIV/AIDS patients. A method comparison was performed using capillary and venous samples from seven clinical laboratories in five countries. The BD FACSPresto system was assessed for variability between laboratory, instrument/operators, cartridge lots and within-run at four sites. Samples were collected under approved voluntary consent. EDTA-anticoagulated venous samples were tested for CD4 and %CD4 T cells using the gold-standard BD FACSCalibur ™ system, and for Hb, using the Sysmex ® KX-21N ™ analyzer. Venous and capillary samples were tested on the BD FACSPresto system. Matched data was analyzed for bias (Deming linear regression and Bland-Altman methods), and for concordance around the clinical decision point. The coefficient of variation was estimated per site, instrument/operator, cartridge-lot and between-runs. For method comparison, 93% of the 720 samples were from HIV-positive and 7% from HIV-negative or normal subjects. CD4 and %CD4 T cells venous and capillary results gave slopes within 0.96-1.05 and R 2 ≥0.96; Hb slopes were ≥1.00 and R 2 ≥0.89. Variability across sites/operators gave %CV <5.8% for CD4 counts, <1.9% for %CD4 and <3.2% for Hb. The total %CV was <7.7% across instrument/cartridge lot. The BD FACSPresto system provides accurate, reliable, precise CD4/%CD4/Hb results compared to gold-standard methods, irrespective of venous or capillary blood sampling. The data showed good agreement between the BD FACSPresto, BD FACSCalibur and Sysmex systems.

The importance of sample collection when using single cytokine levels and systemic cytokine profiles as biomarkers--a comparative study of serum versus plasma samples.

PubMed

Tvedt, Tor Henrik Anderson; Rye, Kristin Paulsen; Reikvam, Håkon; Brenner, Annette K; Bruserud, Øystein

2015-03-01

Cytokines, soluble adhesion molecules and metalloproteinases can be detected in human serum or plasma samples. Such systemic levels are widely used as biomarkers in epidemiological and clinical studies. We prepared serum samples and three types of plasma samples (EDTA, heparin, citric acid) from 20 healthy individuals. The levels of 31 cytokines, four soluble adhesion molecules and eight matrix metalloproteinases were analyzed by Luminex technology. Most mediators showed detectable levels in both plasma and serum. Several mediators that can be released by platelets showed increased serum levels, especially CCL5 and CD40L, but for the other mediators the serum levels did not correlate with peripheral blood platelet counts and for these last mediators serum and plasma levels often showed strong correlations. The use of bivalirudin for anticoagulation significantly increased and citric acid combined with platelet inhibitors (ticagrelor, acetylsalicylic acid plus prostaglandin E2) did not alter plasma levels of platelet-store mediators compared with citric acid alone. The impact of sample preparation differed between mediators; for many mediators strong correlations were seen between serum and plasma levels even when absolute levels differed. Soluble adhesion molecule levels showed only minor differences between samples. Unsupervised hierarchical clustering suggested that the effect of sampling/preparation was strongest for serum and heparin plasma samples. Careful standardization of sample preparation is usually necessary when analyzing systemic mediator levels, and differences caused by sample preparation should be considered as a possible explanation if studies show conflicting results. Copyright © 2015 Elsevier B.V. All rights reserved.

Manganese intoxication: the cause of an inexplicable epileptic syndrome in a 3 year old child.

PubMed

Herrero Hernandez, Elena; Discalzi, Gianluigi; Dassi, Patrizia; Jarre, Laura; Pira, Enrico

2003-08-01

Excess manganese (Mn) can cause several neurotoxic effects, however only a few studies have reported epileptic syndromes related to manganese intoxication. We describe an epileptic syndrome due to manganese intoxication in a 3 year old male child. His blood manganese was elevated, but no other abnormal values or toxic substances were found in blood or urine. The electroencephalogram (EEG) showed a picture of progressive encephalopathy, while brain magnetic resonance was normal. The patient's conditions rapidly worsened to epileptic status despite the use of antiepileptic drugs. Chelating treatment with CaNa(2)EDTA was initiated to remove excess manganese and promptly succeeded in reverting epileptic symptoms. Concurrently, manganese blood levels and electroencephalogram progressively normalized. Thereafter it has been possible to discontinue antiepileptic treatment, and the patient remains in excellent conditions without any treatment.

Sample storage conditions significantly influence faecal microbiome profiles

PubMed Central

Choo, Jocelyn M; Leong, Lex EX; Rogers, Geraint B

2015-01-01

Sequencing-based studies of the human faecal microbiota are increasingly common. Appropriate storage of sample material is essential to avoid the introduction of post-collection bias in microbial community composition. Rapid freezing to −80 °C is commonly considered to be best-practice. However, this is not feasible in many studies, particularly those involving sample collection in participants’ homes. We determined the extent to which a range of stabilisation and storage strategies maintained the composition of faecal microbial community structure relative to freezing to −80 °C. Refrigeration at 4 °C, storage at ambient temperature, and the use of several common preservative buffers (RNAlater, OMNIgene.GUT, Tris-EDTA) were assessed relative to freezing. Following 72 hours of storage, faecal microbial composition was assessed by 16 S rRNA amplicon sequencing. Refrigeration was associated with no significant alteration in faecal microbiota diversity or composition. However, samples stored using other conditions showed substantial divergence compared to −80 °C control samples. Aside from refrigeration, the use of OMNIgene.GUT resulted in the least alteration, while the greatest change was seen in samples stored in Tris-EDTA buffer. The commercially available OMNIgene.GUT kit may provide an important alternative where refrigeration and cold chain transportation is not available. PMID:26572876

Sample storage conditions significantly influence faecal microbiome profiles.

PubMed

Choo, Jocelyn M; Leong, Lex E X; Rogers, Geraint B

2015-11-17

Sequencing-based studies of the human faecal microbiota are increasingly common. Appropriate storage of sample material is essential to avoid the introduction of post-collection bias in microbial community composition. Rapid freezing to -80 °C is commonly considered to be best-practice. However, this is not feasible in many studies, particularly those involving sample collection in participants' homes. We determined the extent to which a range of stabilisation and storage strategies maintained the composition of faecal microbial community structure relative to freezing to -80 °C. Refrigeration at 4 °C, storage at ambient temperature, and the use of several common preservative buffers (RNAlater, OMNIgene.GUT, Tris-EDTA) were assessed relative to freezing. Following 72 hours of storage, faecal microbial composition was assessed by 16 S rRNA amplicon sequencing. Refrigeration was associated with no significant alteration in faecal microbiota diversity or composition. However, samples stored using other conditions showed substantial divergence compared to -80 °C control samples. Aside from refrigeration, the use of OMNIgene.GUT resulted in the least alteration, while the greatest change was seen in samples stored in Tris-EDTA buffer. The commercially available OMNIgene.GUT kit may provide an important alternative where refrigeration and cold chain transportation is not available.

Sample preservation for the analysis of antibiotics in water.

PubMed

Llorca, Marta; Gros, Meritxell; Rodríguez-Mozaz, Sara; Barceló, Damià

2014-11-21

This paper describes a stability study performed for 56 antibiotics belonging to 9 different groups--macrolides, tetracyclines, fluoroquinolones, quinolones, penicillins, cephalosporines, lincosamides, sulfonamides and nitroimidazole antibiotics--in purified water samples fortified with the selected compounds at 10 ng/ml. For this purpose, three different sample preservation modes were tested with the aim of avoiding biotic and abiotic degradation: (i) storage at -20°C, (ii) storage at -20°C with 0.1% of EDTA and (iii) pre-concentration in a solid phase extraction cartridge (SPE), which was afterwards stored at -20°C. Concentrations of antibiotics in the samples preserved using the different protocols were monitored after 0, 1, 2 and 12 weeks. The results showed that, for the accurate determination of all compounds they should be analyzed right after sampling. However, if this is not possible, most of the antibiotics can be analyzed within the 1st week after sampling and preservation at -20°C (with or without EDTA) or in a SPE cartridges at -20°C. Nonetheless, some antibiotics found extensively in the environment, such as sulfamethoxazole, ciprofloxacin, ofloxacin, erythromycin, azithromycin and clarithromycin exhibited low stability after 1 week preservation and, therefore, they should be analyzed within this time. Copyright © 2014 Elsevier B.V. All rights reserved.

Growth and characterization of EDTA assisted CBD-CdS

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kumarage, W. G. C.; Wijesundera, R. P.; Seneviratne, V. A.

2017-10-01

Chemical bath deposition of CdS (CBD-CdS) thin films with the assistance of a cationic surfactant, ethylenediamine tetraacetic acid (EDTA), is reported in this work. Also the EDTA treated CdS thin films are compared with that of conventional CBD-CdS. Fabricated thin, compact, uniform and adherent EDTA treated CdS films show enhanced effective surface area and roughness compared to conventional CBD-CdS. The grazing incidence x-ray diffraction analysis shows all the fabricated CdS films are hexagonally crystallized. EDTA-treated CdS films show excellent photo activity compared to conventional CBDCdS. The flat band potential (Vfb) value was found to be tunable with EDTA concentration.

Efficacy of sodium hypochlorite, ethylenediaminetetraacetic acid, citric acid and phosphoric acid in calcium hydroxide removal from the root canal: a microscopic cleanliness evaluation.

PubMed

da Silva, Juliana Melo; Silveira, Amanda; Santos, Elizandra; Prado, Laiìs; Pessoa, Oscar F

2011-12-01

Rooted molars were subjected to standardized canal instrumentation to a master apical file (MAF). The samples were dressed with Ca(OH)(2), and after 7 days, teeth were reopened and Ca(OH)(2) medication was removed by 1 of 4 different experimental procedures: 2.5% sodium hypochlorite (NaOCl) (n = 10); 17% EDTA-T (n = 10); 10% citric acid (n = 10); or 37% phosphoric acid (n = 10). This was followed by reinstrumentation with MAF plus 15 mL saline solution. The roots were prepared for scanning electron microscopic analysis of the cervical, middle, and apical thirds. Statistical analysis was performed with the Kruskal-Wallis test. EDTA-T and phosphoric acid gave the best results in the apical third, with significant statistical differences compared with other groups. NaOCl gave the worst results. Irrigation with 17% EDTA-T and 37% phosphoric acid is more effective than sodium hypochlorite and citric acid in the removal of calcium hydroxide from the apical third. Copyright © 2011 Mosby, Inc. All rights reserved.

Genetic diversity of piroplasmids species in equids from island of São Luís, northeastern Brazil.

PubMed

Braga, Maria do Socorro Costa de Oliveira; Costa, Francisca Neide; Gomes, Débora Regina Maia; Xavier, Daniele Rosa; André, Marcos Rogério; Gonçalves, Luiz Ricardo; Freschi, Carla Roberta; Machado, Rosangela Zacarias

2017-01-01

Equine piroplasmosisis, a tick-borne disease caused by the intra-erythrocytic protozoans Babesia caballi and Theileria equi, has economic importance due to the international trade and the increased movement of horses all over the world. The goal of this study was to evaluate the occurrence of phylogenetic diversity of T. equi and B. caballi genotypes among infected equids from São Luís Island, state of Maranhão, northeastern Brazil. Between December of 2011 and June of 2012, EDTA-blood and serum samples were collected from 139 equids (90 donkeys, 39 horses and 10 mules). From 139 serum samples submitted to ELISA assay, IgG antibodies to T. equi and B. caballi were detected in 19.4% (27/139) and 25.2% (35/139), respectively. Among sampled animals, 21.6% (30/139) and 55.4% (77/139) were positive for cPCR assays for T. equi and B. caballi, based on ema-1 and rap-1 genes, respectively. Overall, the T. equi sequences (n=7) submitted to Maximum Likelihood analysis (based on a 18S rRNA fragment of 1700 bp after alignment) grouped into three main groups, which were subdivided in eight clusters. The present work showed that different genotypes of T. equi and B. caballi circulate among equids in Brazil.

The assessment of the antibacterial and antifungal activities of aspirin, EDTA and aspirin-EDTA combination and their effectiveness as antibiofilm agents.

PubMed

Al-Bakri, A G; Othman, G; Bustanji, Y

2009-07-01

To evaluate the antimicrobial activities of aspirin, EDTA and an aspirin-EDTA (A-EDTA) combination against Pseudomonas aeruginosa, Escherichia coli and Candida albicans in planktonic and biofilm cultures. Minimal inhibitory concentrations (MIC) and minimal biocidal concentrations (MBC) were determined using twofold broth microdilution and viable counting methods, respectively. Aspirin's recorded MIC values ranged from 1.2 to 2.7 mg ml(-1). Checkerboard assay demonstrated a synergism in antimicrobial activity upon combination. Aspirin's minimal biofilm eradication concentration values (MBEC) against the established biofilms ranged between 1.35 and 3.83 mg ml(-1). A complete eradication of bacterial biofilms was achieved after a 4-h treatment with the A-EDTA combination. Both aspirin and EDTA possess broad-spectrum antimicrobial activity for both planktonic and biofilm cultures. Aspirin used at the MBEC for 24 h was successful in eradicating P. aeruginosa, E. coli and C. albicans biofilms established on abiotic surfaces. Moreover, the exposure to the A-EDTA combination (4 h) effected complete bacterial biofilm eradication. There is a continuous need for the discovery of new antimicrobial agents. Aspirin and EDTA are 'nonantibiotic drugs', the combination of which can be used successfully to treat and eradicate biofilms established on abiotic surfaces.

Targeted chelation therapy with EDTA-loaded albumin nanoparticles regresses arterial calcification without causing systemic side effects

PubMed Central

Lei, Yang; Nosoudi, Nasim; Vyavahare, Naren

2014-01-01

Background and aims Elastin-specific medial arterial calcification (MAC) is an arterial disease commonly referred as Monckeberg’s sclerosis. It causes significant arterial stiffness, and as yet, no clinical therapy exists to prevent or reverse it. We developed albumin nanoparticles (NPs) loaded with disodium ethylene diaminetetraacetic acid (EDTA) that were designed to target calcified elastic lamina when administrated by intravenous injection. Methods and Results We optimized NP size, charge, and EDTA-loading efficiency (150~200 nm, zeta potential of − 22.89 ~ − 31.72 mV, loading efficiency for EDTA ~20 %) for in vivo targeting in rats. These NPs released EDTA slowly for up to 5 days. In both ex-vivo study and in vivo study with injury-induced local abdominal aortic calcification, we showed that elastin antibody-coated and EDTA-loaded albumin NPs targeted the damaged elastic lamina while sparing healthy artery. Intravenous NP injections reversed elastin-specific MAC in rats after four injections over a 2-week period. EDTA-loaded albumin NPs did not cause the side effects observed in EDTA injection alone, such as decrease in serum calcium (Ca), increase in urine Ca, or toxicity to kidney. There was no bone loss in any treated groups. Conclusion We demonstrate that elastin antibody-coated and EDTA-loaded albumin NPs might be a promising nanoparticle therapy to reverse elastin-specific MAC and circumvent side effects associated with systemic EDTA chelation therapy. PMID:25285609

Iodide oxidation and iodine reduction mediated by horseradish peroxidase in the presence of ethylenediaminetetraacetic acid (EDTA): the superoxide effect.

PubMed

Chang, H C; Bumpus, J A

2001-04-01

Ethylenediaminetetraacetic acid (EDTA) is an inhibitor of iodide (I-) oxidation that is catalyzed by horseradish peroxidase (HRP). HRP-mediated iodine (I2) reduction and triiodide (I3+) disappearance occur in the presence of this inhibitor. It is interesting that in the presence of EDTA, HRP produces superoxide radical, a reactive oxygen species that is required for iodine reduction. Substitution of potassium superoxide (KO2) or a biochemical superoxide generating system (xanthine/xanthine oxidase) for HRP and H2O2 in the reaction mixture also can reduce iodine to iodide. Thus, iodine reduction mediated by HRP occurs because HRP is able to mediate the formation of superoxide in the presence of EDTA and H2O2. Although superoxide is able to mediate iodine reduction directly, other competing reactions appear to be more important. For example, high concentrations (mM range) of EDTA are required for efficient iodine reduction in this system. Under such conditions, the concentration (microM range) of contaminating EDTA-Fe(III) becomes catalytically important. In the presence of superoxide, EDTA-Fe(III) is reduced to EDTA-Fe(II), which is able to reduce iodine and form triiodide rapidly. Also of importance is the fact that EDTA-Fe(II) reacts with hydrogen peroxide to form hydroxyl radical. Hydroxyl radical involvement is supported by the fact that a wide variety of hydroxyl radical (OH) scavengers can inhibit HRP dependent iodine reduction in the presence of EDTA and hydrogen peroxide.

[Characterization and optimization of the NaOH-EDTA extracts for solution 31P-NMR analysis of organic phosphorus in river sediments].

PubMed

Zhang, Wen-Qiang; Shan, Bao-Qing; Zhang, Hong; Tang, Wen-Zhong

2014-01-01

Optimization and mechanism of NaOH-EDTA extraction solutions were studied in phosphorus (P) pollution river sediments, which were Fe, Al-rich sediment, by solution 31P nuclear magnetic resonance spectroscopy (31P-NMR). Different proportions of NaOH and EDTA showed different extraction efficiency on total P (TP) and organic P (Po) in the sediment. The concentration of Po in NaOH + EDTA extract was higher than that in NaOH extract. The mechanism was that the TP and Po were released under the conditions of EDTA chelating with Fe and Al. The concentration of TP and Po were the highest in 1.00 mol x L(-1) NaOH +75 mmol x L(-1) EDTA extract and 0.25 mol x L(-1) NaOH + 50 mmol x L(-1) EDTA extract, which were 3.88 mg x g(-1) and 0.24 mg x g(-1), respectively. The extractions of Fe, Mn, Ca, Mg, Al were increasing as the EDTA increased under the same NaOH concentration. Extraction efficiency of Fe, Mn, Ca showed negative correlation with the pH of the extracting solution (P < 0.01). Exponential relationship was found between the extraction of Al and the pH of the extraction solution (P < 0.01) because of the AlO2- and EDTA-Al complex. The quality of spectra of NaOH-EDTA extract was better than that of NaOH extract. Six P species were detected in different extractions, including phosphonates, orthophosphate, pyrophosphate, orthophosphate monoesters, phospholipids and deoxyribonucleic acids. Therefore, 0. 25 mol x L(-1) NaOH + 50 mmol x L(-1) EDTA was the optimization extraction solution for Po analysis in Fe and Al-rich river sediment by 31P-NMR.

High molecular weight hyaluronan decreases oxidative DNA damage induced by EDTA in human corneal epithelial cells

PubMed Central

Ye, J; Wu, H; Wu, Y; Wang, C; Zhang, H; Shi, X; Yang, J

2012-01-01

Purpose To investigate the toxic effects of ethylenediaminetetraacetic acid disodium salt (EDTA), a corneal penetration enhancer in topical ophthalmic formulations, on DNA in human corneal epithelial cells (HCEs), and to investigate whether the effect induced by EDTA can be inhibited by high molecular weight hyaluronan (HA). Methods Cells were exposed to EDTA in concentrations ranging from 0.00001 to 0.01% for 60 min, or 30 min high molecular weight HA pretreatment followed by EDTA treatment. The cell viability was measured by the MTT test. Cell apoptosis was determined with annexin V staining by flow cytometry. The DNA single- and double-strand breaks of HCEs were examined by alkaline comet assay and by immunofluorescence microscope detection of the phosphorylated form of histone variant H2AX (γH2AX) foci, respectively. Reactive oxygen species (ROS) production was assessed by the fluorescent probe, 2′, 7′-dichlorodihydrofluorescein diacetate. Results EDTA exhibited no adverse effect on cell viability and did not induce cell apoptosis in human corneal epithelial cells at concentrations lower than 0.01%. However, a significant increase of DNA single- and double-strand breaks was observed in a dose-dependent manner with all the concentrations of EDTA tested in HCEs. In addition, EDTA treatment led to elevated ROS generation. Moreover, 30 min preincubation with high molecular weight HA significantly decreased EDTA-induced ROS generation and DNA damage. Conclusions EDTA could induce DNA damage in HCEs, probably through oxidative stress. Furthermore, high molecular weight HA was an effective protective agent that had antioxidant properties and decreased DNA damage induced by EDTA. PMID:22595911

Comparison of EDTA- and citric acid-enhanced phytoextraction of heavy metals in artificially metal contaminated soil by Typha angustifolia.

PubMed

Muhammad, Dawood; Chen, Fei; Zhao, Jing; Zhang, Guoping; Wu, Feibo

2009-08-01

A pot experiment was conducted to study the performance of EDTA and citric acid (CA) addition in improving phytoextraction of Cd, Cu, Pb, and Cr from artificially contaminated soil by T. angustifolia. T. angustifolia showed the remarkable resistance to heavy metal toxicity with no visual toxic symptom including chlorosis and necrosis when exposed to metal stress. EDTA-addition significantly reduced plant height and biomass, compared with the control, and stunted plant growth, while 2.5 and 5 mM CA addition induced significant increases in root dry weight. EDTA, and 5 and 10 mM CA significantly increased shoot Cd, Pb, and Cr concentrations compared with the control, with EDTA being more effective. At final harvest, the highest shoot Cd, Cr, and Pb concentrations were recorded in the treatment of 5 mM EDTA addition, while maximal root Pb concentration was found at the 2.5 mM CA treatment. However, shoot Cd accumulation in the 10 mM CA treatment was 36.9% higher than that in 2.5 mM EDTA, and similar with that in 10 mM EDTA. Shoot Pb accumulation was lower in 10 mM CA than that in EDTA treatments. Further, root Cd, Cu, and Pb accumulation of CA treatments and shoot Cr accumulation in 5 or 10 mM CA treatments were markedly higher than that of control and EDTA treatments. The results also showed that EDTA dramatically increased the dissolution of Cu, Cr, Pb, and Cd in soil, while CA addition had less effect on water-soluble Cu, Cr, and Cd, and no effect on Pb levels. It is suggested that CA can be a good chelator candidate for T. angustifolia used for environmentally safe phytoextraction of Cd and Cr in soils.

Effects of iron electrovalence and species on growth and astaxanthin production of Haematococcus pluvialis

NASA Astrophysics Data System (ADS)

Cai, Minggang; Li, Zhe; Qi, Anxiang

2009-05-01

To increase the cell concentration and the accumulation of astaxanthin in the cultivation of Haematococcus pluvialis, effects of different iron electrovalencies (Fe2+-EDTA and Fe3+-EDTA) and species (Fe-EDTA, Fe(OH){x/32x} and FeC6H5O7) addition on cell growth and accumulation of astaxanthin were studied. Results show that different iron electrovalencies have various effects on cell growth and astaxanthin accumulation of H. pluvialis. Compared with Fe3+-EDTA, Fe2+-EDTA stimulate more effectively the formation of astaxanthin. The maximum astaxanthin content (30.70 mg/g biomass cell) was obtained under conditions of 18 μmol/L Fe2+-EDTA, despite the lower cell density (2.3×105 cell/ml) in such condition. Fe3+-EDTA is more effective than Fe2+-EDTA in improving the cell growth. Especially, the maximal steady-state cell density, 2.9×105 cell/ml was obtained at 18 μmol/L Fe3+-EDTA addition. On the other hand, all the various species of iron (EDTA-Fe, Fe(OH){x/32x}, FeC6H5O7) are capable to improve the growth of the algae and astaxanthin production. Among the three iron species, FeC6H5O7 performed the best. Under the condition of a higher concentration (36 μmol/L) of FeC6H5O7, the cell density and astaxanthin production is 2 and 7 times higher than those of iron-limited group, respectively. The present study demonstrates that the effects of the stimulation with different iron species increased in the order of FeC6H5O7, Fe(OH){x/32x} and EDTA-Fe.

Application of EDTA-functionalized bamboo activated carbon (BAC) for Pb(II) and Cu(II) removal from aqueous solutions

NASA Astrophysics Data System (ADS)

Lv, Dan; Liu, Yu; Zhou, Jiasheng; Yang, Kunlun; Lou, Zimo; Baig, Shams Ali; Xu, Xinhua

2018-01-01

In this study, a novel bamboo activated carbon (BAC) with ethylene diamine tetraacetic acid (EDTA) functionality was prepared by direct grafting in the presence of tetraethyl orthosilicate (TEOS) as a crosslinking agent. The BAC@SiO2-EDTA was characterized by SEM, TEM, TGA, FTIR, XPS and its adsorption property for removal of Pb(II) and Cu(II) under various experimental conditions was also investigated. The characterization results reflected that EDTA was successfully assembled on the surface of the BAC and average pore size increased from 4.10 to 4.83 nm as BAC grafted with EDTA. Adsorption data fitted very well in Langmuir isotherm model and pseudo-second-order kinetic model. As compared with the raw BAC, the maximum adsorption capacities of BAC@SiO2-EDTA for the Pb(II) and Cu(II) increased from 45.45 to 123.45 mg g-1 and from 6.85 to 42.19 mg g-1, since the existence of EDTA on modified BAC promoted the formation of chemical complex. The removal of heavy metal ions mainly depended on the complexation with EDTA and the electrostatic attractions with negatively charged surface of BAC@SiO2-EDTA. The adsorption of Pb(II)/Cu(II) on the BAC@SiO2-EDTA was pH dependent and pH 5-6 was considered an optimum. However, lower temperature favored the adsorption and the maximum adsorption was recorded at 20 °C. In addition, BAC@SiO2-EDTA had an excellent reusability with about 40% decline in the adsorption capacity for Pb(II) after fifth reuse. Insignificant influences of co-existing cations and natural organic matter (NOM) were found on the adsorption of Pb(II) and Cu(II). All the results demonstrate that BAC@SiO2-EDTA is a potential adsorbent for metal ions in wastewater.

Late calcium EDTA rescues hippocampal CA1 neurons from global ischemia-induced death.

PubMed

Calderone, Agata; Jover, Teresa; Mashiko, Toshihiro; Noh, Kyung-min; Tanaka, Hidenobu; Bennett, Michael V L; Zukin, R Suzanne

2004-11-03

Transient global ischemia induces a delayed rise in intracellular Zn2+, which may be mediated via glutamate receptor 2 (GluR2)-lacking AMPA receptors (AMPARs), and selective, delayed death of hippocampal CA1 neurons. The molecular mechanisms underlying Zn2+ toxicity in vivo are not well delineated. Here we show the striking finding that intraventricular injection of the high-affinity Zn2+ chelator calcium EDTA (CaEDTA) at 30 min before ischemia (early CaEDTA) or at 48-60 hr (late CaEDTA), but not 3-6 hr, after ischemia, afforded robust protection of CA1 neurons in approximately 50% (late CaEDTA) to 75% (early CaEDTA) of animals. We also show that Zn2+ acts via temporally distinct mechanisms to promote neuronal death. Early CaEDTA attenuated ischemia-induced GluR2 mRNA and protein downregulation (and, by inference, formation of Zn2+-permeable AMPARs), the delayed rise in Zn2+, and neuronal death. These findings suggest that Zn2+ acts at step(s) upstream from GluR2 gene downregulation and implicate Zn2+ in transcriptional regulation and/or GluR2 mRNA stability. Early CaEDTA also blocked mitochondrial release of cytochrome c and Smac/DIABLO (second mitochondria-derived activator of caspases/direct inhibitor of apoptosis protein-binding protein with low pI), caspase-3 activity (but not procaspase-3 cleavage), p75NTR induction, and DNA fragmentation. These findings indicate that CaEDTA preserves the functional integrity of the mitochondrial outer membrane and arrests the caspase death cascade. Late injection of CaEDTA at a time when GluR2 is downregulated and caspase is activated inhibited the delayed rise in Zn2+, p75NTR induction, DNA fragmentation, and cell death. The finding of neuroprotection by late CaEDTA administration has striking implications for intervention in the delayed neuronal death associated with global ischemia.

Reactivity of OHrad and e-aq from electron beam irradiation of aqueous solutions of EDTA and aminopolycarboxylic acids

NASA Astrophysics Data System (ADS)

Vel Leitner, N. Karpel; Guilbault, I.; Legube, B.

2003-05-01

Electron beam irradiation of aqueous solutions of EDTA, EDDA NN‧, NTA, IDA and Cu-EDTA was performed in the presence of scavengers for the hydroxyl radicals (methanol) or for the solvated electrons (hydrogen peroxide). Experiments showed that for each molecule, the G-value decreases as the radiation dose increases from 1 to 25 kGy, and for EDTA, when the initial concentration decreases from 10 to 0.1 mmol l-1. At pH 8 and for 5 kGy, the G-values of NTA, IDA, EDTA and EDDA NN‧ removal ascribed to OHrad radicals are, respectively, 0.06, 0.06, 0.15 and 0.20 μmol J-1, whereas for the solvated electrons the G-values were, respectively, 0.01, 0.01, 0.06 and 0.04 μmol J-1. The rate constants of hydroxyl radicals and solvated electrons were determined by comparison with one competitor. For each active species (hydroxyl radical or solvated electron), the reactivity is connected to the number of nitrogen atoms and acetate groups. The rate constants of OHrad radicals are above 1010 and 8.6×109 l mol-1 s-1 for EDDA NN‧ and EDTA, respectively, 2.1×109 l mol-1 s-1 for IDA and 6.1×108 l mol-1 s-1 for NTA. The reactivity of solvated electrons is smaller and the rate constants are in the range 1.9×106-3.7×106 l mol-1 s-1 for NTA, IDA and EDDA NN‧ and equal 1.4×107 l mol-1 s-1 for EDTA. The reactivity of the complex Cu-EDTA towards OHrad does not differ to a large extent from EDTA whereas with e-aq the reactivity of Cu-EDTA is better than EDTA since ke-/Cu-EDTA reaches 2.2×109 l mol-1 s-1. It follows that when both active entities (OHrad and e-aq) are involved in the electron beam irradiation process, the removal of free aminopolycarboxylic acids is mainly due to OHrad radicals. However, the complex Cu-EDTA is concerned by both e-aq and OHrad radicals.

Leaching characteristics of EDTA-enhanced phytoextraction of Cd and Pb by Zea mays L. in different particle-size fractions of soil aggregates exposed to artificial rain.

PubMed

Lu, Yayin; Luo, Dinggui; Lai, An; Liu, Guowei; Liu, Lirong; Long, Jianyou; Zhang, Hongguo; Chen, Yongheng

2017-01-01

Chelator-assisted phytoextraction is an alternative and effective technique for the remediation of heavy metal-contaminated soils, but the potential for heavy metal leaching needs to be assessed. In the present study, a soil column cultivation-leaching experiment was conducted to investigate the Cd and Pb leaching characteristics during assisted phytoextraction of metal-contaminated soils containing different particle-size soil aggregates. The columns were planted with Zea mays "Zhengdan 958" seedlings and treated with combined applications of EDTA and simulated rainfall (pH 4.5 or 6.5). The results were as follows: (1) The greatest uptake of Cd and Pb by Z. mays was observed after treatment with EDTA (2.5 mmol kg -1 soil) and soil aggregates of <1 mm; uptake decreased as the soil aggregate size increased. (2) Simulated rainfall, especially acid rain (pH 4.5), after EDTA applications led to the increasing metal concentrations in the leachate, and EDTA significantly increased the concentrations of both Cd and Pb in the leachate, especially with soil aggregates of <1 mm; metal leachate concentrations decreased as soil particle sizes increased. (3) Concentrations of Cd and Pb decreased with each continuing leachate collection, and data were fit to linear regression models with coefficients of determination (R 2 ) above 0.90 and 0.87 for Cd and Pb, respectively. The highest total amounts of Cd (22.12%) and Pb (19.29%) were observed in the leachate of soils treated with EDTA and artificial acid rain (pH 4.5) with soil aggregates of <1 mm. The application of EDTA during phytoextraction method increased the leaching risk in the following order: EDTA 2.5-1 (pH 4.5) > EDTA 2.5-1 (pH 6.5) > EDTA 2.5-2 (pH 4.5) > EDTA 2.5-4 (pH 4.5) > EDTA 2.5-2 (pH 6.5) > EDTA 2.5-4 (pH 6.5).

Fabrication of a novel NiFe2O4/Zn-Al layered double hydroxide intercalated with EDTA composite and its adsorption behavior for Cr(VI) from aqueous solution

NASA Astrophysics Data System (ADS)

Deng, Lin; Shi, Zhou; Wang, Li; Zhou, Shiqing

2017-05-01

A novel magnetic NiFe2O4/Zn-Al layered double hydroxide intercalated with EDTA composite (NiFe2O4/ZnAl-EDTA LDH) was prepared through modified coprecipitation method and employed for adsorptive removal of Cr(VI) from aqueous solution. The adsorbents were characterized using Brunauer-Emmett-Teller (BET), scanning electron microscopy (SEM), transmission electron microscope (TEM), X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), vibrating sample magnetometer (VSM), and X-ray photoelectron spectroscopy (XPS). Factors affecting the Cr(VI) adsorption, such as initial solution pH, adsorbent dosage, contact time, initial Cr(VI) concentration, temperature and coexisting ions, were studied systematically. Experiments results show that the magnetic NiFe2O4/ZnAl-EDTA LDH exhibits high adsorption efficiency within a wide pH range of 3.0-7.0 (R>80% at Cr(VI) concentration 50 mg L-1, contact time 360 min, and adsorbent dosage 2 g/L) and quick separation property. The adsorption process is fitted well with the Langmuir isotherm and pseudo-second-order kinetic model. The maximum theoretical adsorption capacity is found to be 77.22 mg g-1 at pH 6.0 and 318 K. The positive ΔH value (2.907 kJ mol-1) and negative ΔG value (-4.722 kJ mol-1) at 298-318 K reveals that the adsorption process is feasible, spontaneous and endothermic. Coexisting anions (PO43-, SO42-, CO32-, HCO3-, Cl-, and NO3-) have no significant effect on Cr(VI) removal. The mechanism study indicates that the adsorption of Cr(VI) onto NiFe2O4/ZnAl-EDTA LDH mainly involves electrostatic attraction and ion exchange interaction. It is interesting to note that a proportion of Cr(VI) adsorbed on the adsorbent surface are reduced to Cr(III) during the adsorption process. Results from this study demonstrate the potential utility of the magnetic NiFe2O4/ZnAl-EDTA LDH that could be developed into a viable technology for efficient removal of Cr(VI) from aqueous solution.

Zeta potential study of Sb2S3 nanoparticles synthesized by a facile polyol method in various surfactants

NASA Astrophysics Data System (ADS)

Saxena, Monika; Okram, Gunadhor Singh

2018-05-01

In the present work, we report the successful synthesis of stibnite Sb2S3 nanoparticles (NPs) by a facile polyol method using various surfactant. The structural and optical properties were investigated by X-ray diffraction (XRD), Raman spectroscopy and Zeta potential. Rietveld refinement of XRD data confirms the single phase orthorhombic crystal structure of stibnite Sb2S3. Presence of six obvious Raman modes further confirmed their stoichiometric formation. Effect of different surfactants on the surface charge of Sb2S3 NPs was studied using Zeta potential measurement in deionized water at different pH values. They reveal that these NPs are more stable when it was synthesized in presence of EDTA than that of CTAB or without surfactant samples with high zeta potential. The isoelectronic point was found at pH = 6.4 for pure sample, 3.5 and 7.2 for CTAB and not found for EDTA Sb2S3 samples. This information can be useful for many industrial applications like pharmaceuticals, ceramics, waste water treatment and medicines.

Influence of disodium EDTA on the nucleation and growth of struvite and carbonate apatite

NASA Astrophysics Data System (ADS)

Prywer, Jolanta; Olszynski, Marcin

2013-07-01

The effect of disodium EDTA, as an additive, on the crystallization of struvite and carbonate apatite was studied. The growth of struvite crystals and carbonate apatite occurred in the solution of artificial urine at 37 °C and at the condition emulating real urinary tract infection. The results demonstrate that the addition of disodium EDTA increases the induction time and decreases the growth efficiency compared to the baseline (without disodium EDTA). The struvite crystal mean and median diameters were found to decrease in the presence of disodium EDTA but the crystal morphology and habit remain almost unchanged. Disodium EDTA has demonstrated its potential to be further investigated in the presence of bacteria and in vivo conditions.

Comparison of EDTA and SDS as potential surface impregnation agents for lead adsorption by activated carbon

NASA Astrophysics Data System (ADS)

Chen, Wei-fang; Pan, Ling; Chen, Li-fang; Yu, Zhe; Wang, Qiong; Yan, Chang-cheng

2014-08-01

Ethylene diamine tetraacetic acid (EDTA) and sodium dodecyl sulfate (SDS) were employed to impregnate activated carbons for the purpose of lead removal. The mechanisms of surface impregnation and lead adsorption method of chemical regeneration were investigated. Results showed that the highest impregnation of EDTA and SDS on activated carbon was 0.33 and 0.96 mmol/g, respectively. Adsorption capacities for lead of EDTA and SDS impregnated activated carbons reached 0.29 and 0.24 mmol/g. Rapid small scale column tests of adsorption and regeneration were conducted. Lead adsorption was greatly enhanced by EDTA impregnation. In addition, EDTA impregnated adsorbent was able to be successful regenerated by HNO3 and thus reused.

Reasons for raising the maximum acceptable daily intake of EDTA and the benefits for iron fortification of foods for children 6–24 months of age

PubMed Central

Wreesmann, Carel Theo Jozef

2014-01-01

The current maximum acceptable daily intake (ADI) of ethylenediaminetetraacetic acid (EDTA) of 1.9 mg day−1 per kilogram bodyweight (mg day−1 kgbw−1) limits the daily intake of iron as iron EDTA [ferric sodium EDTA; sodium iron(III) EDTA] to approximately 2–2.5 mg day−1 for children 6–24 months of age. This limit was defined by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) in 1973 based on data from an animal-feed study published in 1963. Other animal studies indicate that this limit can be raised to 4.4 or possibly up to 21.7 mg day−1 kgbw−1, which is 2.3–11.4 times higher than the current value. For nearly 50 years, iron EDTA has been used in France in medicinal syrup for infants 1–6 months of age. The maximum recommended dosage of this drug is 37 times higher than the maximum ADI of EDTA. No adverse health effects have been reported as a result of this medicinal consumption of iron EDTA. Raising the maximum ADI of EDTA to only 4.4 mg day−1 kgbw−1 would enable iron EDTA, an iron fortificant with proven bioavailability in phytate-rich meals, to be added in adequate amounts to cereal-based meals for children 6–24 months of age, who are at risk of iron deficiency. PMID:24521261

Reasons for raising the maximum acceptable daily intake of EDTA and the benefits for iron fortification of foods for children 6-24 months of age.

PubMed

Wreesmann, Carel Theo Jozef

2014-10-01

The current maximum acceptable daily intake (ADI) of ethylenediaminetetraacetic acid (EDTA) of 1.9 mg day(-1) per kilogram bodyweight (mg day(-1)  kgbw(-1) ) limits the daily intake of iron as iron EDTA [ferric sodium EDTA; sodium iron(III) EDTA] to approximately 2-2.5 mg day(-1) for children 6-24 months of age. This limit was defined by the Joint FAO/WHO Expert Committee on Food Additives (JECFA) in 1973 based on data from an animal-feed study published in 1963. Other animal studies indicate that this limit can be raised to 4.4 or possibly up to 21.7 mg day(-1)  kgbw(-1) , which is 2.3-11.4 times higher than the current value. For nearly 50 years, iron EDTA has been used in France in medicinal syrup for infants 1-6 months of age. The maximum recommended dosage of this drug is 37 times higher than the maximum ADI of EDTA. No adverse health effects have been reported as a result of this medicinal consumption of iron EDTA. Raising the maximum ADI of EDTA to only 4.4 mg day(-1)  kgbw(-1) would enable iron EDTA, an iron fortificant with proven bioavailability in phytate-rich meals, to be added in adequate amounts to cereal-based meals for children 6-24 months of age, who are at risk of iron deficiency. © 2014 The Authors. Maternal & Child Nutrition published by John Wiley & Sons Ltd.

Enhanced electrokinetic remediation of lead-contaminated soil by complexing agents and approaching anodes.

PubMed

Zhang, Tao; Zou, Hua; Ji, Minhui; Li, Xiaolin; Li, Liqiao; Tang, Tang

2014-02-01

Optimizing process parameters that affect the remediation time and power consumption can improve the treatment efficiency of the electrokinetic remediation as well as determine the cost of a remediation action. Lab-scale electrokinetic remediation of Pb-contaminated soils was investigated for the effect of complexant ethylenediaminetetraacetic acid (EDTA) and acetic acid and approaching anode on the removal efficiency of Pb. When EDTA was added to the catholyte, EDTA dissolved insoluble Pb in soils to form soluble Pb-EDTA complexes, increasing Pb mobility and accordingly removal efficiency. The removal efficiency was enhanced from 47.8 to 61.5 % when the EDTA concentration was increased from 0.1 to 0.2 M, showing that EDTA played an important role in remediation. And the migration rate of Pb was increased to 72.3 % when both EDTA and acetic acid were used in the catholyte. The "approaching anode electrokinetic remediation" process in the presence of both EDTA and acetic acid had a higher Pb-removal efficiency with an average efficiency of 83.8 %. The efficiency of electrokinetic remediation was closely related to Pb speciation. Exchangeable and carbonate-bounded Pb were likely the forms which could be removed. All results indicate that the approaching anode method in the presence of EDTA and acetic acid is an advisable choice for electrokinetic remediation of Pb-contaminated soil.

Batch affinity adsorption of His-tagged proteins with EDTA-based chitosan.

PubMed

Hua, Weiwei; Lou, Yimin; Xu, Weiyuan; Cheng, Zhixian; Gong, Xingwen; Huang, Jianying

2016-01-01

Affinity adsorption purification of hexahistidine-tagged (His-tagged) proteins using EDTA-chitosan-based adsorption was designed and carried out. Chitosan was elaborated with ethylenediaminetetraacetic acid (EDTA), and the resulting polymer was characterized by FTIR, TGA, and TEM. Different metals including Ni(2+), Cu(2+), and Zn(2+) were immobilized with EDTA-chitosan, and their capability to the specific adsorption of His-tagged proteins were then investigated. The results showed that Ni(2+)-EDTA-chitosan and Zn(2+)-EDTA-chitosan had high affinity toward the His-tagged proteins, thus isolating them from protein mixture. The target fluorescent-labeled hexahistidine protein remained its fluorescent characteristic throughout the purification procedure when Zn(2+)-EDTA-chitosan was used as a sorbent, wherein the real-time monitor was performed to examine the immigration of fluorescent-labeled His-tagged protein. Comparatively, Zn(2+)-EDTA-chitosan showed more specific binding ability for the target protein, but with less binding capacity. It was further proved that this purification system could be recovered and reused at least for 5 times and could run on large scales. The presented M(2+)-EDTA-chitosan system, with the capability to specifically bind His-tagged proteins, make the purification of His-tagged proteins easy to handle, leaving out fussy preliminary treatment, and with the possibility of continuous processing and a reduction in operational cost in relation to the costs of conventional processes.

Cytotoxicity analysis of EDTA and citric acid applied on murine resident macrophages culture.

PubMed

Amaral, K F; Rogero, M M; Fock, R A; Borelli, P; Gavini, G

2007-05-01

To assess the ex vivo cytotoxicity of EDTA and citric acid solutions on macrophages. The cytotoxicity of 17% EDTA and 15% citric acid was evaluated on murine macrophage cultures using MTT-Tetrazolium method [3-(4,5-dimethylthiazol-2-yl)-2,5 diphenyl tetrazolium bromide]. A total of 5 x 10(5) cells were plated in medium culture with 17% EDTA or 15% citric acid. Fresh medium was used as a control. Toxicity values were analysed statistically by anova and Tukey's test (P<0.05) at short (0, 6, 12, 24 h) and medium periods (1, 3, 5, 7 days), using ELISA absorbance. On the short term, both EDTA (0.253 nm) and citric acid (0.260 nm) exhibited cytotoxic effects on macrophage cultures (P<0.05). On the medium term, statistical differences were observed (P<0.05) between the groups. EDTA (0.158 nm) and citric acid (0.219 nm) were cytotoxic when compared with the control group; EDTA-reduced macrophage viability significantly more than citric acid (P<0.05). Both EDTA and citric acid had effects on macrophages cells ex vivo, but citric acid was less toxic in periods from 1 to 7 days of use.

"Off-the-Shelf" K2-EDTA for Calcific Band Keratopathy.

PubMed

Lee, Marco E; Ouano, Dean P; Shapiro, Brett; Fong, Andrew; Coroneo, Minas T

2018-07-01

To explore the effectiveness of "off-the-shelf" dipotassium-ethylenediaminetetraacetic acid (K2-EDTA) as an alternative to sodium EDTA as chelation therapy in removal of calcific band keratopathy (CBK). This study was a retrospective case series involving 4 patients with CBK who underwent superficial keratectomy and subsequent chelation therapy with K2-EDTA in a single center (Coastal Eye Clinic) by the same surgeon. Visual acuity and symptomatic relief were the main outcomes measured in our study. All 4 participants in this study were female with an average age of 80.3 years. Three of the patients with reasonable baseline visual acuity experienced improved visual acuity at 1 month. The other patient with multiple ocular comorbidities and severely reduced visual potential reported symptomatic pain relief at 1-month follow-up after the intervention. K2-EDTA seems to be an effective alternative to disodium EDTA in its ability to clear calcific plaques and restore visual function. Because of the logistical difficulties associated with acquiring disodium EDTA, and the relative abundance of K2-EDTA in health-care facilities, we believe that our findings warrant further investigation into its use as a more accessible and cost-effective chelating agent in CBK.

Evaluation of ethylenediaminetetraacetic acid (EDTA) solution and gel for smear layer removal.

PubMed

Dotto, Sidney Ricardo; Travassos, Rosana Maria Coelho; de Oliveira, Elias Pandonor Motcy; Machado, Manoel Eduardo de Lima; Martins, José Luiz

2007-08-01

The purpose of this in vitro study was to compare the efficacy of 24% ethylenediaminetetraacetic acid (EDTA) gel and 17% EDTA solution in cleaning dentine walls after root canal instrumentation. Thirty human canine teeth were divided into three groups of 10 teeth each. In Group 1, 1% sodium hypochlorite was used as the irrigating solution; in Group 2, 1% sodium hypochlorite was used with 17% EDTA solution; and in Group 3, 1% sodium hypochlorite was used with 24% EDTA gel. The presence of a smear layer was analysed after instrumentation using scanning electron microscopy. The Kruskal-Wallis test revealed a statistical difference (P < 0.05) between Groups 1 and 2, and also between Groups 1 and 3. No difference was observed between Groups 2 and 3 (P > 0.05). The results indicate that 1% sodium hypochlorite alone does not remove the smear layer and that there was no statistical difference between EDTA gel and EDTA solution in smear layer removal.

Electrochemical Oxidation of EDTA in Nuclear Wastewater Using Platinum Supported on Activated Carbon Fibers

PubMed Central

Zhao, Bo; Zhu, Wenkun; Mu, Tao; Hu, Zuowen; Duan, Tao

2017-01-01

A novel Pt/ACF (Pt supported on activated carbon fibers) electrode was successfully prepared with impregnation and electrodeposition method. Characterization of the electrodes indicated that the Pt/ACF electrode had a larger effective area and more active sites. Electrochemical degradation of ethylenediaminetetra-acetic acid (EDTA) in aqueous solution with Pt/ACF electrodes was investigated. The results showed that the 3% Pt/ACF electrode had a better effect on EDTA removal. The operational parameters influencing the electrochemical degradation of EDTA with 3% Pt/ACF electrode were optimized and the optimal removal of EDTA and chemical oxygen demand (COD) were 94% and 60% after 100 min on condition of the electrolyte concentration, initial concentration of EDTA, current density and initial value of pH were 0.1 mol/L, 300 mg/L, 40 mA/cm2 and 5.0, respectively. The degradation intermediates of EDTA in electrochemical oxidation with 3% Pt/ACF electrode were identified by gas chromatography-mass spectrum (GC-MS). PMID:28754016

Influence of time, temperature and coagulation on the measurement of C3, C3 split products and C4.

PubMed

Sinosich, M J; Teisner, B; Brandslund, I; Fisher, M; Grudzinskas, J G

1982-11-26

Quantitative and qualitative immunoelectrophoretic analyses of circulating C3, C3 split products and C4 were performed in matched EDTA plasma and serum obtained from 5 normal subjects and stored for up to 48 h at room temperature (18 degrees C-22 degrees C) and 4 degrees C. Fluctuations in apparent levels of C3 were greater in serum than plasma stored at room temperature, a fall in levels seen by 24 h being followed by a significant increase. By contrast, levels of C3 did not alter if stored at 4 degrees C. C4 levels in both EDTA plasma and serum remained unchanged for 24 h, a slight decrease being seen at 48 h. Levels of C4 remained constant if samples were stored at 4 degrees C. Crossed immunoelectrophoresis revealed a significant progressive decrease in C3 levels and a simultaneous increase in C3c occurring after 4 h in serum and 8 h in EDTA plasma, stored at room temperature. In studies conducted at 4 degrees C, similar but delayed fluctuations were seen. A progressive and significant increase in C3d levels was seen in both plasma and serum samples stored at room temperature, levels rising to 276% (plasma) and 308% (serum) of levels seen at zero time. At 4 degrees C marginal increases in C3d levels only were observed. These results suggest that in vitro degradation of C3 and C4 are readily facilitated by temperature, time and coagulation, and that conditions of collection and storage of samples must be optimized for the accurate definition of activation of the complement cascade.

The Effect of Four Commonly Used Root Conditioner Agents in Different Time Periods Applied on Periodontally Diseased and Healthy Teeth

PubMed Central

P, Torkzaban; S, Seyedzadeh Sabounchi

2016-01-01

Statement of Problem: Root surface contamination or infection can potentially change the consequences of regenerative periodontal therapies and therefore the modification and disinfection of the contaminated root surfaces are necessary. Objectives: This study aimed to compare the surface characteristics of the extracted human teeth after exposure to four root conditioners in different time periods. Materials and Methods: The study samples were prepared from 40 freshly extracted teeth including 20 affected teeth with periodontal diseases and 20 healthy teeth. After performing root planning, 240 dentinal block samples were prepared and each affected and healthy sample was randomly allocated to receive one of the following root conditioners; Ethylenediaminetetraaceti acid (EDTA), citric acid, doxycycline, and tetracycline or rinsed with normal saline as the control agent. The prepared specimens were evaluated using scanning electron microscope and the inter-group differences and changes in study indices; dentin (%), tubular spaces (%), and diameter of dentinal tubules (μm²) were compared using one-way ANOVA test. Results: In the control group receiving normal saline, the changes in the indicators of dentin, tubular spaces, and diameter of dentinal tubules remained insignificant in all time periods. EDTA, citric acid, and tetracycline had chelating effects on the study indices; however, doxycycline led to gradual decrease of the tubular space and diameter as well as increase in dentin percentage. Conclusions: In different time intervals and when considering healthy or affected tooth surfaces, the effect of conditioning agents could be different. Amongst the four agents used, EDTA and tetracycline consistently increased the diameter of tubules and percentage of patent tubules in both healthy and diseased teeth. PMID:28959749

The effect of biogenic Fe(II) on the stability and sorption of Co(II)EDTA 2- to goethite and a subsurface sediment

NASA Astrophysics Data System (ADS)

Zachara, John M.; Smith, Steven C.; Fredrickson, James K.

2000-04-01

Laboratory experiments were conducted with suspensions of goethite (α-FeOOH) and a subsurface sediment to assess the influence of bacterial iron reduction on the fate of Co(II)EDTA 2-, a representative metal-ligand complex of intermediate stability (log K Co(II)EDTA = 17.97). The goethite was synthetic (ca. 55 m 2/g) and the sediment was a Pleistocene age, Fe(III) oxide-containing material from the Atlantic coastal plain (Milford). Shewanella alga strain BrY, a dissimilatory iron reducing bacterium (DIRB), was used to promote Fe(III) oxide reduction. Sorption isotherms and pH adsorption edges were measured for Co 2+, Fe 2+, Co(II)EDTA 2-, and Fe(II)EDTA 2- on the two sorbents in 0.001 mol/L Ca(ClO 4) 2 to aid in experiment interpretation. Anoxic suspensions of the sorbents in PIPES buffer at pH 6.5-7.0 were spiked with Co(II)EDTA 2- (10 -5 mol/L, 60Co and 14EDTA labeled), inoculated with BrY (1-6 × 10 8 organisms/mL), and the headspace filled with a N 2/H 2 gas mix. The experiments were conducted under non-growth conditions. The medium did not contain PO 43- (with one exception), trace elements, or vitamins. The tubes were incubated under anoxic conditions at 25°C for time periods in excess of 100 d. Replicate tubes were sacrificed and analyzed at desired time periods for pH, Fe(II) TOT, Fe (aq)2+, 60Co, and 14EDTA. Abiotic analogue experiments were conducted where Fe (aq)2+ was added in increasing concentration to Co(II)EDTA 2-/mineral suspensions to simulate the influence of bacterial Fe(II) evolution. The DIRB generated Fe(II) from both goethite and the Milford sediment that was strongly sorbed by mineral surfaces. Aqueous Fe 2+ increased during the experiment as surfaces became saturated; Fe (aq)2+ induced the dissociation of Co(II)EDTA 2- into a mixture of Co 2+, Co(II)EDTA 2-, and Fe(II)EDTA 2- (log K Fe(II)EDTA = 15.98). The extent of dissociation of Co(II)EDTA 2- was greater in the subsurface sediment because it sorbed Fe(II) less strongly than did goethite. The post dissociation sorption behavior of Co 2+ was dependent on pH and the intrinsic sorptivity of the solid phases. Dissociation generally lead to an increase in the sorption (e.g., K d) of Co 2+ relative to EDTA 4- (form unspecified). Sorbed biogenic Fe(II) competed with free Co (aq)2+and reduced its sorption relative to unreduced material. It is concluded that cationic radionuclides such as 60Co or 239/240Pu, which may be mobilized from disposed wastes by complexation with EDTA 4-, may become immobilized in groundwater zones where dissimilatory bacterial iron reduction is operative.

Use of RBC-O and S-MCV parameters of SYSMEX XE-2100 in a patient with RBC cold agglutination.

PubMed

Wang, Hong; Lu, Lin; Zhou, Yun; Liu, Jian; Qian, Min; Tang, Weiming; Jie, Zhang; Pan, Shiyang

2013-01-01

Sometimes EDTA blood of erythrocyte agglutination cannot be well resolved by incubation at 37 degrees C. In this case report, however, such a specimen was detected from a lymphoma patient at room temperature by using RBC-O and S-MCV parameters of the SYSMEX XE-2100 hematology analyzer. The specimen was diluted with 0.9% NaCL solution at 1:1 before measurement. HCT, MCV, and MCHC, corrected by RBC-O, HGB and S-MCV, were all in their normal ranges. This case indicates that RBC-O and S-MCV parameters of XE-2100 can be used in the routine blood examination of erythrocyte agglutination specimen at room temperature.

The role of EDTA in phytoextraction of hexavalent and trivalent chromium by two willow trees.

PubMed

Yu, Xiao-Zhang; Gu, Ji-Dong

2008-04-01

Effects of the synthetic chelator ethylenediamine tetraacetate (EDTA) on uptake and internal translocation of hexavalent and trivalent chromium by plants were investigated. Two different concentrations of EDTA were studied for enhancing the uptake and translocation of Cr from the hydroponic solution spiked with K(2)CrO(4) or CrCl(3) maintained at 24.0 +/- 1 degrees C. Faster removal of Cr(3+) than Cr(6+) by hybrid willows (Salix matsudana Koidz x Salix alba L.) from the plant growth media was observed. Negligible effect of EDTA on the uptake of Cr(6+) was found, but significant decrease of the Cr concentration in roots was measured. Although the translocation of Cr(6+) within plant materials was detected in response to EDTA concentration, the amount of Cr(6+) translocated to the lower stems was considerably small. EDTA in the nutrient media showed a negative effect on the uptake of Cr(3+ )by hybrid willows; the removal rates of Cr(3+ )were significantly decreased. Translocation of Cr(3+) into the stems and leaves was undetectable, but roots were the exclusive sink for Cr(3+) accumulation. Weeping willows (Salix babylonica L.) showed lower removal rates for both chemical forms of Cr than hybrid willows. Although EDTA had a minor effect on Cr(6+ )uptake by weeping willows, positive effect on Cr(6+ )translocation within plant materials was observed. It was also determined that EDTA in plant growth media significantly decreased the amount of Cr(3+) taken up by plants, but significantly increased Cr(3+) mobilization from roots to stems. Results indicated that EDTA was unable to increase the uptake of Cr(6+) by both plant species, but translocation of Cr(6+)-EDTA within plant materials was possible. Addition of EDTA in the nutrient media showed a strong influence on the uptake and translocation of Cr(3+) in both willows. Cr(3+)-EDTA in tissues of weeping willows was more mobile than that in hybrid willows. The information has important implications for the use of metal chelator in plant nutritional research.

Nitric oxide removal by combined urea and FeIIEDTA reaction systems.

PubMed

He, Feiqiang; Deng, Xianhe; Chen, Min

2017-02-01

(NH 2 ) 2 CO as well as Fe II EDTA is an absorbent for simultaneous desulfurization and denitrification. However, they have their own drawbacks, like the oxidation of Fe II EDTA and the low solubility of NO in urea solution. To overcome these defects, A mixed absorbent containing both (NH 2 ) 2 CO and Fe II EDTA was employed. The effects of various operating parameters (urea and Fe II EDTA concentration, temperature, inlet oxygen concentration, pH value) on NO removal were examined in the packed tower. The results indicated that the NO removal efficiency increased with the decrease of oxygen concentration as well as the increase of Fe II EDTA concentration. The NO removal efficiency had little change with a range of 25-45 °C, and sharply decreased at the temperature of above 55 °C. The NO removal efficiency initially increases up to the maximum value and then decreases with the increase of pH value as well as the raise of urea concentration. In addition, the synergistic mechanism of (NH 2 ) 2 CO and Fe II EDTA on NO removal was investigated. Results showed that urea could react with Fe II EDTA-NO to produce Fe II EDTA, N 2 , and CO 2 , and hinder oxidation of Fe II EDTA. Finally, to evaluate the effect of SO 3 2- on NO removal, a mixed absorbent containing Fe II EDTA, urea, and Na 2 SO 3 was employed to absorb NO. The mixed absorbent could maintain more than 78% for 80 min at 25 °C, pH = 7.0, (NH 2 ) 2 CO concentration of 5 wt%, Fe II EDTA concentration of 0.02 M, O 2 concentration of 7% (v/v), and Na 2 SO 3 concentration of 0.2 M. Copyright © 2016 Elsevier Ltd. All rights reserved.

Chelate-Assisted Heavy Metal Movement Through the Root Zone

NASA Astrophysics Data System (ADS)

Kirkham, M.; Madrid, F.; Liphadzi, M. S.

2001-12-01

Chelating agents are added to soil as a means to mobilize heavy metals for plant uptake during phytoremediation. Yet almost no studies follow the displacement of heavy metals through the vadose zone following solubilization with chelating agents. The objective of this work was to determine the movement of heavy metals through the soil profile and their absorption by barley (Hordeum vulgare L.) in a soil amended with biosolids and in the presence of a chelating agent (EDTA). Twelve columns 75 cm in height and 17 in diameter were packed with a Haynie very fine sandy loam (coarse-silty, mixed, calcareous, mesic Mollic Udifluvents) and watered with liquid biosolids applied at the surface at a rate of 120 kg N/ha. Three weeks after plants germinated, soil was irrigated with a solution of the disodium salt of EDTA added at a rate of 0.5 g/kg soil. Four treatments were imposed: columns with no plants and no EDTA; columns with no plants plus EDTA; columns with plants and no EDTA; and columns with plants and EDTA. Columns were watered intensively for 35 days until two pore volumes of water had been added, and the leachates were collected daily. With or without plants, columns with EDTA had lower total concentrations of Cu, Zn, Cd, Ni, and Pb in the surface 20 cm than columns without EDTA. Concentrations of the heavy metals in this layer were not afffected by the presence of roots. Iron in leachate was followed as an indicator metal for movement to groundwater. No iron appeared in the leachate without EDTA, either in the columns with plants or without plants. The peak concentration of iron in the leachate occurred three days earlier in the columns without plants and EDTA compared to the columns with plants and EDTA. The results indicated the importance of vegetation on retarding heavy metal leaching to groundwater during chelate-facilitated phytoremediation.

Mobility of arsenic and its compounds in soil and soil solution: the effect of soil pretreatment and extraction methods.

PubMed

Száková, J; Tlustos, P; Goessler, W; Frková, Z; Najmanová, J

2009-12-30

The effect of soil extraction procedures and/or sample pretreatment (drying, freezing of the soil sample) on the extractability of arsenic and its compounds was tested. In the first part, five extraction procedures were compared with following order of extractable arsenic portions: 2M HNO(3)>0.43 M CH(3)COOH>or=0.05 M EDTA>or=Mehlich III (0.2M CH(3)COOH+0.25 M NH(4)NO(3)+0.013 M HNO(3)+0.015 M NH(4)F+0.001 M EDTA) extraction>water). Additionally, two methods of soil solution sampling were compared, centrifugation of saturated soil and the use of suction cups. The results showed that different sample pretreatments including soil solution sampling could lead to different absolute values of mobile arsenic content in soils. However, the interpretation of the data can lead to similar conclusions as apparent from the comparison of the soil solution sampling methods (r=0.79). For determination of arsenic compounds mild extraction procedures (0.05 M (NH(4))(2)SO(4), 0.01 M CaCl(2), and water) and soil solution sampling using suction cups were compared. Regarding the real soil conditions the extraction of fresh samples and/or in situ collection of soil solution are preferred among the sample pretreatments and/or soil extraction procedures. However, chemical stabilization of the solutions should be allowed and included in the analytical procedures for determination of individual arsenic compounds.

Organic acids on the growth, anatomical structure, biochemical parameters and heavy metal accumulation of Iris lactea var. chinensis seedling growing in Pb mine tailings.

PubMed

Han, Yu-Lin; Huang, Su-Zhen; Yuan, Hai-Yan; Zhao, Jiu-Zhou; Gu, Ji-Guang

2013-08-01

The effect of citric acid (CA) and ethylene diamine tetraacetic acid (EDTA) on the growth, anatomical structure, physiological responses and lead (Pb) accumulation of Iris lactea var. chinensis seedling growing in Pb mine tailings for 30 days were studied. Results showed that the dry weights (DW) of roots decreased significantly under both levels of CA. The DWs of leaves and roots treated with 2 mmol/kg EDTA decreased significantly and were 23 and 54 %, respectively, lower than those of the control. The tolerant indexes of I. lactea var. chinensis under all treatments of organic acids were lower than control. The root tip anatomical structure was little affected under the treatments of 2 mmol/kg CA and 2 mmol/kg EDTA compared with control. However, the formation of photosynthesizing cells was inhibited by the treatment of 2 mmol/kg EDTA. The concentrations of chlorophyll a, chlorophyll b and total carotenoids in the leaves treated with 2 mmol/kg EDTA significantly decreased. Higher CA level and lower EDTA level could trigger the synthesis of ascorbic acid and higher level of EDTA could trigger the synthesis of glutathione. CA and EDTA could promote Pb accumulation of I. lactea var. chinensis and Pb concentration in the leaves and roots at 2 mmol/kg EDTA treatment increased significantly and reached to 160.44 and 936.08 μg/g DW, respectively, and 1.8 and 1.6 times higher than those of the control. The results indicated that I. lactea var. chinensis could be used to remediate Pb tailing and the role of EDTA in promoting Pb accumulation was better than CA did.

Effect of Different Protocols in Preconditioning With EDTA in Sclerotic Dentin and Enamel Before Universal Adhesives Applied in Self-etch Mode.

PubMed

Martini, E C; Parreiras, S O; Gutierrez, M F; Loguercio, A D; Reis, A

The aim of this study was to investigate the effect of different protocols of 17% ethylene diamine tetra-acetic acid (EDTA) conditioning on the etching pattern and immediate bond strength of universal adhesives to enamel and sclerotic dentin. Forty bovine teeth with sclerotic dentin and 20 human third molars were randomly divided into eight groups resulting from the combination of the main factors surface treatment (none, two-minute EDTA conditioning manual application, 30-second EDTA manual application, 30-second EDTA sonic application) and adhesives systems (Scotchbond Universal Adhesive [SBU] and Prime & Bond Elect [PBE]). Resin-dentin and enamel-dentin bond specimens were prepared and tested under the microtensile bond strength (μTBS) and microshear bond strength (μSBS) tests, respectively. The etching pattern produced on the unground enamel and the sclerotic dentin surfaces under the different protocols and adhesive systems was evaluated under scanning electron microscopy. For enamel, only the main factor adhesive was significant (p<0.0001), with SBU showing the highest μSBS. In sclerotic dentin, the lowest mean was observed for the group without EDTA application and the highest mean in the group with EDTA application with the sonic device for 30 seconds. Regardless of the EDTA protocol, the highest means of μTBS were observed for SBU (p<0.05). EDTA conditioning improves the bonding performance of universal adhesives in the self-etch mode on sclerotic dentin, mainly when applied for 30 seconds with the aid of a sonic device. EDTA pretreatment also improves the retentive etching pattern of enamel, but it does not result in higher enamel bond strength.

How do low doses of desferrioxamine B and EDTA affect the phytoextraction of metals in sunflower?

PubMed

Cornu, J Y; Dépernet, C; Garnier, C; Lenoble, V; Braud, A; Lebeau, T

2017-08-15

The aim of this study was to compare the efficiency of siderophore desferrioxamine B (DFOB) and EDTA in increasing the phytoextraction of metals in sunflower. A 28-day pot experiment was conducted in a metal-contaminated soil supplied with 200μmolkg -1 of DFOB or EDTA. Pore water was collected and pseudo-polarographic analyses were conducted to assess the impact of the two chelators on the mobility and speciation of metals in the liquid phase. Our results showed that DFOB is not an efficient mobilizing agent of divalent metals in soil. Adding DFOB selectively increased the mobility of trivalent metals while the supply of EDTA simultaneously increased the mobility of both trivalent and divalent metals. EDTA significantly reduced the labile fractions of Cd, Cu, (Pb) and Zn measured in the porewater. The labile concentration of Cd and Zn measured in presence of EDTA was even less than that measured in the control. As expected from the pore water analysis, the addition of DFOB did not affect the phytoextraction of any divalent metals. In contrast, the addition of EDTA enhanced Cu and Ni phytoextraction in sunflower 2.0 to 2.8 fold for Cu and 1.3 to 2.3 fold for Ni, depending on the cultivar. This result supports different hypotheses regarding the forms and the related pathways in which metals are taken up in presence of EDTA. Based on the results obtained for Ni, whose uptake is rate limited by its internalization across the cell membrane, the direct uptake of metal-EDTA complexes via the non-selective apoplastic pathway is hypothesized to contribute the most to the overall uptake of metals in presence of EDTA, even added at "low" concentrations. Copyright © 2017 Elsevier B.V. All rights reserved.

Enhanced phytoextraction of Pb and other metals from artificially contaminated soils through the combined application of EDTA and EDDS.

PubMed

Luo, Chunling; Shen, Zhenguo; Li, Xiangdong; Baker, Alan J M

2006-06-01

Chemically enhanced phytoextraction is achieved by the application of chelates to soils. Using pot experiments, the effect of the combined application of EDTA and EDDS on the uptake of Cu, Pb, Zn and Cd by Zea mays L. was studied. Among the tested application ratios of 1:1, 1:2, and 2:1 (EDTA/EDDS), 2:1 of EDTA:EDDS was the most efficient ratio for increasing the concentrations of Cu, Pb, Zn and Cd in the shoots. The combined application of 3.33 mmol kg(-1) soil of EDTA+1.67 mmol kg(-1) soil of EDDS produced 650 mg kg(-1) of Pb in the shoots, which was 2.4 and 5.9 times the concentration of Pb in the shoots treated with 5 mmol kg(-1) of EDTA and EDDS alone, respectively. The total phytoextraction of Pb reached 1710 microg kg(-1) soil, which was 2.1 and 6.1 times the total Pb from 5 mmol kg(-1) EDTA and EDDS alone, respectively. The combined application of EDTA and EDDS also significantly increased the translocation of Pb from the roots to the shoots. The mechanism of enhancing the phytoextraction of Pb by the combined application of EDTA+EDDS did not involve a change in the pH of the soil. The increase in the phytoextraction of Pb by the shoots of Z. mays L. was more pronounced than the increase of Pb in the soil solution with the combined application of EDTA and EDDS. It was thought that the major role of EDDS might be to increase the uptake and translocation of Pb from the roots to the shoots of plants.

Interaction between chlorhexidine digluconate and EDTA.

PubMed

Rasimick, Brian J; Nekich, Michelle; Hladek, Megan M; Musikant, Barry L; Deutsch, Allan S

2008-12-01

The combination of chlorhexidine and EDTA produces a white precipitate. The aim of this study was to determine if the precipitate involves the chemical degradation of chlorhexidine. The precipitate was produced and redissolved in a known amount of dilute trifluoroacetic acid. The amount of chlorhexidine and EDTA present in the dissolved precipitate was determined by reverse-phase high performance liquid chromatography (HPLC) with ultraviolet detection at 288 nm. More than 90% of the precipitate's mass was found to be EDTA or chlorhexidine. The remainder is suspected to be water, gluconate, and sodium. Parachloroaniline, a potentially carcinogenic decomposition product of chlorhexidine, was not detected in the precipitate (the limit of detection was 1%). The molar ratio of chlorhexidine to EDTA in the precipitate was about 1.6 to 1. Based on the results, chlorhexidine forms a salt with EDTA rather than undergoing a chemical reaction.

Hydroponic phytoremediation of Cd, Cr, Ni, As, and Fe: can Helianthus annuus hyperaccumulate multiple heavy metals?

PubMed

January, Mary C; Cutright, Teresa J; Van Keulen, Harry; Wei, Robert

2008-01-01

Sundance sunflowers were subjected to contaminated solutions containing 3, 4, or 5 heavy metals, with and without EDTA. The sunflowers exhibited a metal uptake preference of Cd=Cr>Ni, Cr>Cd>Ni>As and Fe>As>Cd>Ni>Cr without EDTA and Cr>Cd>Ni, Fe>As>Cd>Cr>Ni with EDTA. As uptake was not affected by other metals, but it decreased Cd and Ni concentration in the stems. The presence of Fe improved the translocation of the other metals regardless of whether EDTA was present. In general, EDTA served as a hindrance to metal uptake. For the experiment with all five heavy metals, EDTA decreased Cd in the roots and stems from 2.11 to 1.36 and from 2.83 to 2.3 2mg g(-1) biomass, respectively. For the same conditions, Ni in the stems decreased from 1.98 to 0.94 mg g(-1) total metal uptake decreased from 14.95 mg to 13.89 mg, and total biomass decreased from 2.38 g to 1.99 g. These results showed an overall negative effect in addition of EDTA. However it is unknown whether the negative effect was due to toxicity posed by EDTA or the breaking of phytochelatin-metal bonds. The most important finding was the ability of Sundance sunflowers to achieve hyperaccumulator status for both As and Cd under all conditions studied. Ni hyperaccumulator status was only achieved in the presence of three metals without EDTA.

Optimal antimicrobial catheter lock solution, using different combinations of minocycline, EDTA, and 25-percent ethanol, rapidly eradicates organisms embedded in biofilm.

PubMed

Raad, Issam; Hanna, Hend; Dvorak, Tanya; Chaiban, Gassan; Hachem, Ray

2007-01-01

Antimicrobial lock solutions may be needed to salvage indwelling catheters in patients requiring continuous intravenous therapy. We determined the activity of minocycline, EDTA, and 25% ethanol, alone or in combination, against methicillin-resistant Staphylococcus aureus and Candida parapsilosis catheter-related bloodstream infection strains in two established models of biofilm colonization. Biofilm-colonized catheter segments from a modified Robbins device and a silicone disk biofilm colonization model were exposed to these antimicrobial agents for 15 or 60 min, respectively. After exposure, segments were sonicated and cultured. To determine regrowth after incubation at 37 degrees C, following the brief exposure to the antimicrobial agents, an equal number of segments were washed, reincubated for 24 h, and then sonicated and cultured. The triple combination of minocycline-EDTA (M-EDTA) in 25% ethanol was the only antimicrobial lock solution that completely eradicated S. aureus and C. parapsilosis in biofilm of all segments tested in the two models, and it completely prevented regrowth. In addition, M-EDTA in 25% ethanol was significantly more effective in rapidly eradicating the growth or regrowth of methicillin-resistant S. aureus and C. parapsilosis biofilm colonization in the two models than the other solutions--minocycline, EDTA, M-EDTA, 25% ethanol, and EDTA in ethanol. We conclude that M-EDTA in 25% ethanol is highly effective at rapidly eradicating S. aureus and C. parapsilosis embedded in biofilm adhering to catheter segments.

Influence of matrix metalloproteinase synthetic inhibitors on dentin microtensile bond strength of resin cements.

PubMed

Stape, T H S; Menezes, M S; Barreto, B C F; Aguiar, F H B; Martins, L R; Quagliatto, P S

2012-01-01

This study evaluated the effect of dentin pretreatment with 2% chlorhexidine (CHX) or 24% ethylenediamine tetra-acetic acid gel (EDTA) on the dentin microtensile bond strength (μTBS) of resin cements. Composite blocks were luted to superficial noncarious human dentin (n=10) using two resin cements (RelyX ARC [ARC] and RelyX U100 [U100]) and three dentin pretreatments (without pretreatment-control, CHX, and EDTA). CHX was applied for 60 seconds on the acid-etched dentin in the ARC/CHX group, and for the same time on smear layer-covered dentin in the U100/CHX group. EDTA was applied for 45 seconds on smear-covered dentin in the U100/EDTA group, and it replaced phosphoric acid conditioning in the ARC/EDTA group for 60 seconds. After storage in water for 24 hours, specimens were prepared for microtensile bond strength testing. The results were submitted to two-way analysis of variance (ANOVA) followed by Tukey test. ARC produced significantly higher μTBS (p<0.05) compared to the U100, except when EDTA was used. For ARC, no pretreatment and CHX produced higher μTBS than EDTA. For U100, EDTA produced higher μTBS; no statistical difference occurred between CHX pretreatment and when no pretreatment was performed. While CHX did not affect immediate dentin bond strength of both cements, EDTA improved bond strength of U100, but it reduced dentin bond strength of ARC.

Lead poisoning of horses in the vicinity of a battery recycling plant.

PubMed

Palacios, H; Hibarren, I; Olalla, M J; Cala, V

2002-05-06

The diagnosis of lead poisoning in horses living on farmland in the vicinity of a battery recycling plant was based on clinical signs as well as on laboratory findings. Chemical analysis of six surface soils (0-15 cm) and herbage samples taken at different distances to the recycling plant showed very high total lead levels in the closest sites to the facility and a clear decrease with distance. Total lead levels in soil samples ranged from 127 to 5657 mg kg(-1), with more than 70% of lead extractable by EDTA in the most polluted soils. Lead levels in the aerial part of herbage samples were in the range of 113-4741 mg kg(-1). A water washing pre-treatment of the vegetal samples considerably diminished the concentration of lead, suggesting that airborne lead particles from the facility emissions were fixed on the shoots. The analysis of samples taken from six dead horses showed lead concentrations, expressed as mg kg(-1) (d.w.), as follows: blood: 0.20-0.89; liver: 2.5-15; kidney: 1.70-6.75. Lead intake levels, estimated according to the ingestion rate of Grammineae forage, illustrates that the apported lead through the ingestion of vegetation growing in the closest sites to the recycling plant was approximately 99.5 mg Pb/kg body weight/day surpassing the fatal dosage for horses of 2.4 mg Pb/kg body weight/day reported by Hammond and Aronson, Ann NY Acad Sci, 1964; 111: 595-611.

EDTA analysis on the Roche MODULAR analyser.

PubMed

Davidson, D F

2007-05-01

Patient specimens can be subject to subtle interference from cross contamination by liquid-based, potassium-containing EDTA anticoagulant, leading to misinterpretation of results. A rapid method for EDTA analysis to detect such contamination is described. An in-house EDTA assay on the Roche MODULAR analyser was assessed for accuracy and precision by comparison with an adjusted calcium difference measurement (atomic absorption and o-cresolphthalein complexone colorimetry). EDTA method versus adjusted calcium difference showed: slope = 1.038 (95% confidence interval [CI] 0.949-1.131); intercept = 0.073 (95% CI 0.018-0.132) mmol/L; r = 0.914; n = 94. However, inter-assay precision of the calcium difference method was estimated to be poorer (coefficient of variation 24.8% versus 3.4% for the automated colorimetric method at an EDTA concentration of 0.25 mmol/L). Unequivocal contamination was observed at an EDTA concentration of > or =0.2 mmol/L. The automated method showed positive interference from haemolysis and negative interference from oxalate. The method was unaffected by lipaemia (triglycerides <20 mmol/L), icterus (bilirubin <500 micromol/L), glucose (<100 mmol/L), iron (<100 micromol/L), and citrate, phosphate or fluoride (all <2.5 mmol/L). The automated colorimetric assay described is an accurate, precise and rapid (3 min) means of detecting EDTA contamination of unhaemolysed biochemistry specimens.

Frequency of dog erythrocyte antigen 1.1 in 4 breeds native to different areas in Turkey.

PubMed

Ergul Ekiz, Elif; Arslan, Murat; Ozcan, Mukaddes; Gultekin, Guldal Inal; Gulay, Ozlem Yildiz; Kirmizibayrak, Turgut; Giger, Urs

2011-12-01

Dog erythrocyte antigen (DEA) 1.1 is the most important RBC antigen clinically, as it is highly immunogenic and causes acute hemolytic transfusion reactions (HTR) in sensitized dogs. The aims of this study were to determine the frequency of DEA 1.1 expression in 4 Turkish dog breeds, and to estimate the potential risk of HTR when blood from a DEA 1.1-positive donor is administered to a DEA 1.1-negative recipient following sensitization by a prior mismatched transfusion. EDTA blood samples (n = 178) were typed for DEA 1.1 using a commercial gel-column agglutination test (ID-Gel-Test Canine DEA 1.1). Probabilities of sensitization and risk of an HTR were calculated. The frequency of positivity for DEA 1.1 among Kars (n = 59), Kangal (n = 53), Akbash (n = 50), and Catalburun (n = 16) breeds was 71.2%, 67.9%, 60.0%, and 50.0%, respectively. Potential risk for occurrence of an HTR after administration of blood from a dog of the same breed ranged from 12.5% to 14.8%, whereas HTR induced by blood of a dog from a different breed ranged from 7.2% to 25.3%. The frequency of DEA 1.1-positive dogs among 4 Turkish breeds is high compared with that of most other breeds previously surveyed. The predicted risk of both sensitization and occurrence of DEA 1.1-related HTR following transfusion between dogs of either the same or different Turkish breeds was considerable. Although few dogs are transfused ≥4 days after the first transfusion, we recommend that (1) all donors and recipients be typed for DEA 1.1, (2) DEA 1.1-negative recipients receive only DEA 1.1-negative blood, and (3) blood be cross-matched prior to transfusing any dog ≥4 days after the first transfusion. These guidelines are also applicable to other breeds and countries. © 2011 American Society for Veterinary Clinical Pathology.

Effect of 5% Chlorine Dioxide Irrigant on Micro Push Out Bond Strength of Resin Sealer to Radicular Dentin: An In Vitro Study

PubMed Central

Devarasanahalli, Swapna V; Aswathanarayana, Ranjini M; Rashmi, K; Gowda, Yashwanth; Nadig, Roopa R

2017-01-01

Introduction Chlorine dioxide (ClO2) has been recently investigated as a possible root canal irrigant due to its broad spectrum of antimicrobial action, tissue dissolution and smear layer removal properties. Literature is scarce on the effect of chlorine dioxide irrigation on the resin sealer dentin bond strength. Aim To compare 5% chlorine dioxide (ClO2) with or without Ethylene Diamine Tetra Acetic acid (EDTA) with 3% Sodium hypochlorite (NaOCl) and EDTA combination as endodontic irrigants on the adhesion of AH Plus sealer to radicular dentin using micro- Push out Bond Strength (µPBS) test. Materials and Methods Forty freshly extracted central incisors were decoronated and randomly divided into four groups based on the different irrigation regimes followed during irrigation: Group I - 3% NaOCl + 17% EDTA, Group II - 5% ClO2 + 17% EDTA, Group III - 5% ClO2 and Group IV – Saline, and canal enlarged till Protaper F3. All the samples were obturated with F3 gutta-percha cones using AH Plus sealer and sectioned perpendicular to long axis to obtain 1mm thick slices from the middle and coronal portions for µPBS measurement in universal testing machine followed by assessment of failure pattern under stereomicroscope. Data was analysed using One-way analysis of variance (ANOVA), Bonferroni and t-test. Results Bond strength values were in the following order: Group I>Group II>Group III>Group IV, with no statistically significant difference amongst experimental groups on intergroup comparison, except with saline. The µPBS values were more in coronal third than middle third in all specimens, with no statistical significant difference. Mode of failure showed mixed patterns in all experimental groups except saline. Conclusion In the present study, the bond strength values of ClO2 were comparable with conventional NaOCl and EDTA combination and hence, ClO2 can be considered as an effective alternative endodontic irrigant. PMID:28658907

Preservation of water samples for arsenic(III/V) determinations: An evaluation of the literature and new analytical results

USGS Publications Warehouse

McCleskey, R. Blaine; Nordstrom, D. Kirk; Maest, A.S.

2004-01-01

Published literature on preservation procedures for stabilizing aqueous inorganic As(III/V) redox species contains discrepancies. This study critically evaluates published reports on As redox preservation and explains discrepancies in the literature. Synthetic laboratory preservation experiments and time stability experiments were conducted for natural water samples from several field sites. Any field collection procedure that filters out microorganisms, adds a reagent that prevents dissolved Fe and Mn oxidation and precipitation, and isolates the sample from solar radiation will preserve the As(III/V) ratio. Reagents that prevent Fe and Mn oxidation and precipitation include HCl, H 2SO4, and EDTA, although extremely high concentrations of EDTA are necessary for some water samples high in Fe. Photo-catalyzed Fe(III) reduction causes As(III) oxidation; however, storing the sample in the dark prevents photochemical reactions. Furthermore, the presence of Fe(II) or SO 4 inhibits the oxidation of As(III) by Fe(III) because of complexation reactions and competing reactions with free radicals. Consequently, fast abiotic As(III) oxidation reactions observed in the laboratory are not observed in natural water samples for one or more of the following reasons: (1) the As redox species have already stabilized, (2) most natural waters contain very low dissolved Fe(III) concentrations, (3) the As(III) oxidation caused by Fe(III) photoreduction is inhibited by Fe(II) or SO4.

Comparison of efficacy of pulverization and sterile paper point techniques for sampling root canals.

PubMed

Tran, Kenny T; Torabinejad, Mahmoud; Shabahang, Shahrokh; Retamozo, Bonnie; Aprecio, Raydolfo M; Chen, Jung-Wei

2013-08-01

The purpose of this study was to compare the efficacy of the pulverization and sterile paper point techniques for sampling root canals using 5.25% NaOCl/17% EDTA and 1.3% NaOCl/MTAD (Dentsply, Tulsa, OK) as irrigation regimens. Single-canal extracted human teeth were decoronated and infected with Enterococcus faecalis. Roots were randomly assigned to 2 irrigation regimens: group A with 5.25% NaOCl/17% EDTA (n = 30) and group B with 1.3% NaOCl/MTAD (n = 30). After chemomechanical debridement, bacterial samplings were taken using sterile paper points and pulverized powder of the apical 5 mm root ends. The sterile paper point technique did not show growth in any samples. The pulverization technique showed growth in 24 of the 60 samples. The Fisher exact test showed significant differences between sampling techniques (P < .001). The sterile paper point technique showed no difference between irrigation regimens. However, 17 of the 30 roots in group A and 7 of the 30 roots in group B resulted in growth as detected by pulverization technique. Data showed a significant difference between irrigation regimens (P = .03) in pulverization technique. The pulverization technique was more efficacious in detecting viable bacteria. Furthermore, this technique showed that 1.3% NaOCl/MTAD regimen was more effective in disinfecting root canals. Published by Elsevier Inc.

Biodegradation of Metal-EDTA Complexes by an Enriched Microbial Population

PubMed Central

Thomas, Russell A. P.; Lawlor, Kirsten; Bailey, Mark; Macaskie, Lynne E.

1998-01-01

A mixed culture utilizing EDTA as the sole carbon source was isolated from a mixed inoculum of water from the River Mersey (United Kingdom) and sludge from an industrial effluent treatment plant. Fourteen component organisms were isolated from the culture, including representatives of the genera Methylobacterium, Variovorax, Enterobacter, Aureobacterium, and Bacillus. The mixed culture biodegraded metal-EDTA complexes slowly; the biodegradability was in the order Fe>Cu>Co>Ni>Cd. By incorporation of inorganic phosphate into the medium as a precipitant ligand, heavy metals were removed in parallel to EDTA degradation. The mixed culture also utilized a number of possible EDTA degradation intermediates as carbon sources. PMID:9546167

Thumbnail Sketches: EDTA-Type Chelating Agents in Everyday Consumer Products: Some Medicinal and Personal Care Products.

ERIC Educational Resources Information Center

Hart, J. Roger

1984-01-01

Discusses various ethylenediaminetetraacetate (EDTA)-type chelating agents found in ophthalmic products, personal care products, and disinfectants. Also discusses the properties and action of these EDTA agents. (JN)

EDTA: a synthetic draw solute for forward osmosis.

PubMed

Lutchmiah, Kerusha; Post, Jan W; Rietveld, Luuk C; Cornelissen, Emile R

2014-01-01

The draw solution is the driving force of the forward osmosis (FO) process; however, the solute loss of the draw solute to the feed side is a general, financial limitation for most applications. The anthropogenic amino acid ethylenediaminetetraacetic acid (EDTA) was investigated as a draw solution for FO. At concentrations of approximately 1.0 osmol/kg, EDTA demonstrated comparable water fluxes (Jv = 5.29 L/m(2) h) to the commonly used salt, NaCl (Jv = 4.86 L/m(2) h), and both produced better water fluxes than glucose (Jv = 3.46 L/m(2) h). EDTA showed the lowest solute loss with Js (reverse solute loss or solute leakage) = 0.54 g/m(2) h. The molecular weight, degree of ionisation and charge of EDTA played a major role in this efficiency and EDTA was therefore well rejected by the membrane, showing a low Js/Jv ratio of 0.10 g/L. Owing to the low solute loss of EDTA and its resistance to biodegradation, this compound has the potential to be used as a draw solute for FO during long periods without requiring much replenishment.

Comparative in vitro studies on disodium EDTA effect with and without Proteus mirabilis on the crystallization of carbonate apatite and struvite

NASA Astrophysics Data System (ADS)

Prywer, Jolanta; Olszynski, Marcin; Torzewska, Agnieszka; Mielniczek-Brzóska, Ewa

2014-06-01

Effect of disodium EDTA (salt of ethylenediamine tetraacetic acid) on the crystallization of struvite and carbonate apatite was studied. To evaluate such an effect we performed an experiment of struvite and carbonate apatite growth from artificial urine. The crystallization process was induced by Proteus mirabilis to mimic the real urinary tract infection, which usually leads to urinary stone formation. The results demonstrate that disodium EDTA exhibits the effect against P. mirabilis retarding the activity of urease - an enzyme produced by these microorganisms. The spectrophotometric results demonstrate that, with and without P. mirabilis, the addition of disodium EDTA increases the induction time and decreases the growth efficiency compared to the baseline (without disodium EDTA). These results are discussed from the standpoint of speciation of complexes formed in the solution of artificial urine in the presence of disodium EDTA. The size of struvite crystals was found to decrease in the presence of disodium EDTA. However, struvite crystals are larger in the presence of bacteria while the crystal morphology and habit remain unchanged.

Simulation of the influence of EDTA on the sorption of heavy metals by humic acids

NASA Astrophysics Data System (ADS)

Kropacheva, T. N.; Didik, M. V.; Kornev, V. I.

2015-04-01

The results of mathematical simulation of sorption equilibria with the participation of divalent cations of heavy metals (HMs), chelant (EDTA), and insoluble forms of humic acids (HAs) are discussed. It is shown that the formation of chelates of metals with EDTA in solutions results in the decreasing sorption of the metals by humic acids. We also analyzed the effect of the acidity of the medium and the HM: EDTA: HA ratio (in a wide range) on the desorption of metals. The desorbing effect of EDTA on the metals is the highest at pH 3-5 and increases with an increase in the concentration of EDTA and a decrease in the concentration of HAs. With respect to the remobilization of metals under the impact of EDTA, the metal cations can be arranged into the following sequence: Cu(II) > Ni(II) > Pb(II) ≫ Cd(II) > Co(II) > Zn(II). The obtained data have been used to analyze the remobilization / extraction of HMs from soils with a high content of humic substances.

Comparison between B·R·A·H·M·S PCT direct, a new sensitive point-of-care testing device for rapid quantification of procalcitonin in emergency department patients and established reference methods - a prospective multinational trial.

PubMed

Kutz, Alexander; Hausfater, Pierre; Oppert, Michael; Alan, Murat; Grolimund, Eva; Gast, Claire; Alonso, Christine; Wissmann, Christoph; Kuehn, Christian; Bernard, Maguy; Huber, Andreas; Mueller, Beat; Schuetz, Philipp

2016-04-01

Procalcitonin (PCT) is increasingly being used for the diagnostic and prognostic work up of patients with suspected infections in the emergency department (ED). Recently, B·R·A·H·M·S PCT direct, the first high sensitive point-of-care test (POCT), has been developed for fast PCT measurement on capillary or venous blood samples. This is a prospective, international comparison study conducted in three European EDs. Consecutive patients with suspicion of bacterial infection were included. Duplicate determination of PCT was performed in capillary (fingertip) and venous whole blood (EDTA), and compared to the reference method. The diagnostic accuracy was evaluated by correlation and concordance analyses. Three hundred and three patients were included over a 6-month period (60.4% male, median age 65.2 years). The correlation between capillary or venous whole blood and the reference method was excellent: r2=0.96 and 0.97, sensitivity 88.1% and 93.0%, specificity 96.5% and 96.8%, concordance 93% and 95%, respectively at a 0.25 μg/L threshold. No significant bias was observed (-0.04 and -0.02 for capillary and venous whole blood) although there were 6.8% and 5.1% outliers, respectively. B·R·A·H·M·S PCT direct had a shorter time to result as compared to the reference method (25 vs. 144 min, difference 119 min, 95% CI 110-134 min, p<0.0001). This study found a high diagnostic accuracy and a faster time to result of B·R·A·H·M·S PCT direct in the ED setting, allowing shortening time to therapy and a more wide-spread use of PCT.

Evaluation of the Aptima HIV-1 Quant Dx Assay Using Plasma and Dried Blood Spots

PubMed Central

Sahoo, Malaya K.; Varghese, Vici; White, Elizabeth; Winslow, Meg; Katzenstein, David A.; Shafer, Robert W.

2016-01-01

HIV-1 RNA quantitation in plasma, or virus load testing, is the primary method by which the response to antiretroviral therapy is monitored. Here we describe evaluation of the Aptima HIV-1 Quant Dx assay (Aptima) performed on the automated Panther system. The clinical performance of Aptima was compared to that of the Cobas AmpliPrep/Cobas TaqMan HIV-1 Test v2.0 (CAP/CTM) using 162 EDTA plasma samples collected from patients undergoing HIV-1 monitoring. Overall agreement was 84.0% (136/162), with a kappa statistic of 0.723 (standard error, 0.047; 95% confidence interval [CI], 0.630 to 0.815), indicating substantial agreement. Using the 86 clinical samples quantifiable by both methods, Passing-Bablok regression revealed a regression line of Y = (1.069 × X) − 0.346 (95% CI of the slope [1.003 to 1.139] and intercept [−0.666 to −0.074]), and Bland-Altman analysis demonstrated a mean difference (Aptima-CAP/CTM) of −0.075 log10 copies/ml (95% limits of agreement of −0.624 to 0.475), consistent with negative bias. Comparison of Aptima results for paired dried blood spot (DBS) and plasma specimens archived from participants in the Peninsula AIDS Research Cohort Study (PARC) demonstrated an overall agreement of 94.7% (90/95) when 1,000 copies/ml was used as the threshold. In conclusion, the Aptima HIV-1 Quant Dx assay provides a suitable alternative for HIV-1 monitoring in plasma and DBS. PMID:27535684

Evaluation of the Aptima HIV-1 Quant Dx Assay Using Plasma and Dried Blood Spots.

PubMed

Sahoo, Malaya K; Varghese, Vici; White, Elizabeth; Winslow, Meg; Katzenstein, David A; Shafer, Robert W; Pinsky, Benjamin A

2016-10-01

HIV-1 RNA quantitation in plasma, or virus load testing, is the primary method by which the response to antiretroviral therapy is monitored. Here we describe evaluation of the Aptima HIV-1 Quant Dx assay (Aptima) performed on the automated Panther system. The clinical performance of Aptima was compared to that of the Cobas AmpliPrep/Cobas TaqMan HIV-1 Test v2.0 (CAP/CTM) using 162 EDTA plasma samples collected from patients undergoing HIV-1 monitoring. Overall agreement was 84.0% (136/162), with a kappa statistic of 0.723 (standard error, 0.047; 95% confidence interval [CI], 0.630 to 0.815), indicating substantial agreement. Using the 86 clinical samples quantifiable by both methods, Passing-Bablok regression revealed a regression line of Y = (1.069 × X) - 0.346 (95% CI of the slope [1.003 to 1.139] and intercept [-0.666 to -0.074]), and Bland-Altman analysis demonstrated a mean difference (Aptima-CAP/CTM) of -0.075 log10 copies/ml (95% limits of agreement of -0.624 to 0.475), consistent with negative bias. Comparison of Aptima results for paired dried blood spot (DBS) and plasma specimens archived from participants in the Peninsula AIDS Research Cohort Study (PARC) demonstrated an overall agreement of 94.7% (90/95) when 1,000 copies/ml was used as the threshold. In conclusion, the Aptima HIV-1 Quant Dx assay provides a suitable alternative for HIV-1 monitoring in plasma and DBS. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

Lead toxicosis of captive vultures: case description and responses to chelation therapy.

PubMed

Pikula, Jiri; Hajkova, Pavlina; Bandouchova, Hana; Bednarova, Ivana; Adam, Vojtech; Beklova, Miroslava; Kral, Jiri; Ondracek, Karel; Osickova, Jitka; Pohanka, Miroslav; Sedlackova, Jana; Skochova, Hana; Sobotka, Jakub; Treml, Frantisek; Kizek, Rene

2013-01-16

Lead, a serious threat for raptors, can hamper the success of their conservation. This study reports on experience with accidental lead intoxication and responses to chelation therapy in captive Cinereous (Aegypius monachus) and Egyptian (Neophron percnopterus) Vultures. Soil contamination by lead-based paint sanded off the steel aviary resulted in poisoning of eight Cinereous and two Egyptian Vultures. A male Egyptian Vulture developed signs of apathy, polydipsia, polyuria, regurgitation, and stupor, and died on the next day. Liver, kidney and blood lead concentrations were 12.2, 8.16 and 2.66 μg/g, respectively. Laboratory analyses confirmed severe liver and kidney damage and anaemia. Blood Pb levels of Pb-exposed Cinereous Vultures were 1.571 ± 0.510 μg/g shortly after intoxication, decreased to 0.530 ± 0.165 μg/g without any therapy in a month and to 0.254 ± 0.097 μg/g one month after CaNa(2)EDTA administration. Eight months later, blood lead levels decreased to close to the background of the control group. Blood parameters of healthy Pb-non-exposed Cinereous Vultures were compared with those of the exposed group prior to and after chelation therapy. Iron levels in the lead-exposed pre-treatment birds significantly decreased after chelation. Haematocrit levels in Pb-exposed birds were significantly lower than those of the controls and improved one month after chelation. Creatine kinase was higher in pre-treatment birds than in the controls but normalised after therapy. Alkaline phosphatase increased after chelation. A marked increase in the level of lipid peroxidation measured as thiobarbituric acid reactive species was demonstrated in birds both prior to and after chelation. The ferric reducing antioxidant power was significantly lower in pre-treatment vultures and returned to normal following chelation therapy. Blood metallothionein levels in lead-exposed birds were higher than in controls. Reduced glutathione dropped after CaNa(2)EDTA therapy, while oxidised glutathione was significantly lower in both pre- and post-treatment birds. A chick in an egg produced by a Cinereous Vulture female two months after lead toxicosis died on day 40 of artificial incubation. Lead concentrations in foetal tissues were consistent with levels causing avian mortality. The reported blood parameters and reproduction impairment in captive birds may have implications for professionals dealing with lead exposure in wild birds.

Accuracy and reproducibility of a new contrast clearance method for the determination of glomerular filtration rate.

PubMed Central

O'Reilly, P H; Brooman, P J; Martin, P J; Pollard, A J; Farah, N B; Mason, G C

1986-01-01

A new method for determining the glomerular filtration rate was analysed prospectively. The method uses an x ray fluorescence technique to measure disappearance from the plasma of injected non-ionic iodinated contrast media. Eighty seven patients were studied. Fifty four had an intravenous dose of 100 ml iohexol (Omnipaque) and 33 had 50 ml iohexol. Clearances of chromium-51 labelled edetic acid (51Cr-EDTA) were measured simultaneously. In the patients given 100 ml iohexol there was excellent correlation with 51Cr-EDTA clearance (r = 0.90). The correlation using 50 ml iohexol was also good (r = 0.85). Correlation between creatinine clearance and clearance of 51Cr-EDTA in 33 patients was less satisfactory (r = 0.69). There were no adverse reactions to the contrast media. The equipment used for measuring contrast clearance was robust and simple to operate. Freezing plasma samples in 10 studies and re-examining them weekly for six weeks showed no significant variation in results; hence reproducibility was good. This new and accurate method for determining the glomerular filtration rate merits further study and might find a useful place in routine clinical practice. Images FIG 1 PMID:3089467

Antioxidant activity of oregano, parsley, and olive mill wastewaters in bulk oils and oil-in-water emulsions enriched in fish oil.

PubMed

Jimenez-Alvarez, D; Giuffrida, F; Golay, P A; Cotting, C; Lardeau, A; Keely, Brendan J

2008-08-27

The antioxidant activity of oregano, parsley, olive mill wastewaters (OMWW), Trolox, and ethylenediaminetetraacetic acid (EDTA) was evaluated in bulk oils and oil-in-water (o/w) emulsions enriched with 5% tuna oil by monitoring the formation of hydroperoxides, hexanal, and t-t-2,4-heptadienal in samples stored at 37 degrees C for 14 days. In bulk oil, the order of antioxidant activity was, in decreasing order (p < 0.05), OMWW > oregano > parsley > EDTA > Trolox. The antioxidant activity in o/w emulsion followed the same order except that EDTA was as efficient an antioxidant as OMWW. In addition, the total phenolic content, the radical scavenging properties, the reducing capacity, and the iron chelating activity of OMWW, parsley, and oregano extracts were determined by the Folin-Ciocalteau, oxygen radical absorbance capacity, ferric reducing antioxidant power, and iron(II) chelating activity assays, respectively. The antioxidant activity of OMWW, parsley, and oregano in food systems was related to their total phenolic content and radical scavenging capacity but not to their ability to chelate iron in vitro. OMWW was identified as a promising source of antioxidants to retard lipid oxidation in fish oil-enriched food products.

Mechanism of permeability-enhancing effect of EDTA and boric acid on the corneal penetration of 4-[1-hydroxy-1-methylethyl]-2-propyl-1-[4-[2-[tetrazole-5-yl]phenyl]phenyl] methylimidazole-5-carboxylic acid monohydrate (CS-088).

PubMed

Kikuchi, Takayuki; Suzuki, Masahiko; Kusai, Akira; Iseki, Ken; Sasaki, Hitoshi; Nakashima, Kenichiro

2005-08-11

This study was conducted to clarify the penetration properties of 4-[1-hydroxy-1-methylethyl]-2-propyl-1-[4-[2-[tetrazole-5-yl]phenyl]phenyl]methylimidazole-5-carboxylic acid monohydrate (CS-088), an ophthalmic agent, and the mechanism of the permeability-enhancing effect of EDTA and boric acid (EDTA/boric acid) on the corneal penetration of CS-088. In the absence of additives, corneal permeability decreased with increasing concentration of CS-088 as CS-088 monomers self-associate to form dimers. Presence of EDTA/boric acid caused no significant changes in the physicochemical properties of CS-088, the apparent partition coefficient or the mean particle size of CS-088. EDTA/boric acid induced only a slight change in the zeta potential of liposomes used as a model of the biological membrane. On the other hand, EDTA/boric acid significantly increased membrane fluidity of liposomes, whereas other buffering agents tested did not. This effect was synergistic and concentration-dependent for both EDTA and boric acid as was observed in in vitro corneal penetration of CS-088. In accordance with the result, the rate of CS-088 permeation into the liposomes significantly increased by the addition of EDTA/boric acid. Therefore, it was demonstrated that EDTA/boric acid promotes corneal penetration of CS-088 through the transcellular pathway by increasing membrane fluidity. Conversely, other buffering agents decreased corneal permeability of CS-088 by inducing further self-association of CS-088 aggregates.

Experimental inflammation following dural application of complete Freund's adjuvant or inflammatory soup does not alter brain and trigeminal microvascular passage.

PubMed

Lundblad, Cornelia; Haanes, Kristian A; Grände, Gustaf; Edvinsson, Lars

2015-01-01

Migraine is a paroxysmal, disabling primary headache that affects 16 % of the adult population. In spite of decades of intense research, the origin and the pathophysiology mechanisms involved are still not fully known. Although triptans and gepants provide effective relief from acute migraine for many patients, their site of action remains unidentified. It has been suggested that during migraine attacks the leakiness of the blood-brain barrier (BBB) is altered, increasing the passage of anti-migraine drugs. This study aimed to investigate the effect of experimental inflammation, following dural application of complete Freund's adjuvant (CFA) or inflammatory soup (IS) on brain and trigeminal microvascular passage. In order to address this issue, we induced local inflammation in male Sprague-Dawley-rats dura mater by the addition of CFA or IS directly on the dural surface. Following 2, 24 or 48 h of inflammation we calculated permeability-surface area product (PS) for [(51)Cr]-EDTA in the trigeminal ganglion (TG), spinal trigeminal nucleus, cortex, periaqueductal grey and cerebellum. We observed that [(51)Cr]-EDTA did not pass into the central nervous system (CNS) in a major way. However, [(51)Cr]-EDTA readily passed the TG by >30 times compared to the CNS. Application of CFA or IS did not show altered transfer constants. With these experiments we show that dural IS/CFA triggered TG inflammation, did not increase the BBB passage, and that the TG is readily exposed to circulating molecules. The TG could provide a site of anti-migraine drug interaction with effect on the trigeminal system.

Role of neutrophilic elastase in ethanol induced injury to the gastric mucosa

DOE Office of Scientific and Technical Information (OSTI.GOV)

Kvietys, P.R.; Carter, P.R.

1990-02-26

Intragastric administration of ethanol (at concentrations likely to be encountered by the mucosa during acute intoxication) produces gastritis. Recent studies have implicated neutrophils in the gastric mucosal injury induced by luminal ethanol. The objective of the present study was to assess whether neutrophilic elastase contributes to the ethanol-induced gastric mucosal injury. Sprague-Dawley rats were instrumented for perfusion of the gastric lumen with saline or ethanol. Mucosal injury was quantitated by continuously measuring the blood-to-lumen clearance of {sup 51}Cr-EDTA. The experimental protocol consisted of a 40 minute control period (saline perfusion) followed by three successive 40 minute experimental periods (ethanol perfusion).more » During the three experimental periods the concentration of ethanol was progressively increased to 10, 20, and 30%. The experiments were performed in untreated animals and in animals pretreated with either Eglin c (an inhibitor of elastase and cathepsin G activity) or L 658 (a specific inhibitor of elastase activity). The effects of ethanol on EDTA clearance (x control) in untreated (n = 9) and L658 treated (n = 5) animals are shown in the Table below. Pretreatment with L 658 significantly attenuated the ethanol-induced increases in EDTA clearance. Pretreatment with Eglin c (n = 6) also provided some protection against ethanol-induced injury, but not to the extent as that provided by L658. The results of the authors studies suggest that neutrophilic elastase contributes to a gastric mucosal injury induced by luminal perfusion of the stomach with physiologically relevant concentrations of ethanol.« less

21 CFR 573.360 - Disodium EDTA.

Code of Federal Regulations, 2010 CFR

2010-04-01

..., FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.360 Disodium EDTA. The food additive disodium EDTA (disodium ethylenediaminetetraace- tate... food additive contains a minimum of 99 percent disodium ethylenediaminetetraacetate dihydrate...

21 CFR 573.360 - Disodium EDTA.

Code of Federal Regulations, 2014 CFR

2014-04-01

..., FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.360 Disodium EDTA. The food additive disodium EDTA (disodium ethylenediaminetetraace- tate... food additive contains a minimum of 99 percent disodium ethylenediaminetetraacetate dihydrate...

21 CFR 573.360 - Disodium EDTA.

Code of Federal Regulations, 2013 CFR

2013-04-01

..., FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.360 Disodium EDTA. The food additive disodium EDTA (disodium ethylenediaminetetraace- tate... food additive contains a minimum of 99 percent disodium ethylenediaminetetraacetate dihydrate...

21 CFR 573.360 - Disodium EDTA.

Code of Federal Regulations, 2012 CFR

2012-04-01

..., FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.360 Disodium EDTA. The food additive disodium EDTA (disodium ethylenediaminetetraace- tate... food additive contains a minimum of 99 percent disodium ethylenediaminetetraacetate dihydrate...

21 CFR 573.360 - Disodium EDTA.

Code of Federal Regulations, 2011 CFR

2011-04-01

..., FEEDS, AND RELATED PRODUCTS FOOD ADDITIVES PERMITTED IN FEED AND DRINKING WATER OF ANIMALS Food Additive Listing § 573.360 Disodium EDTA. The food additive disodium EDTA (disodium ethylenediaminetetraace- tate... food additive contains a minimum of 99 percent disodium ethylenediaminetetraacetate dihydrate...

Effect of EDTA conditioning upon the retention of fibre posts luted with resin cements.

PubMed

Rasimick, B J; Shah, R P; Musikant, B L; Deutsch, A S

2008-12-01

To determine if a final rinse with EDTA improves the short-term in-tooth retentive strength of adhesively luted fibre-reinforced composite posts. Six different commercial fibre post systems were tested. Post spaces in extracted human anterior teeth were irrigated with either 6% NaOCl or 6% NaOCl followed by 17% EDTA. Posts were luted using the manufacturer's recommended bonding agent and luting cement. Two different sizes of post were tested for each post system. After 24 h of storage, the retentive strength of the post to the tooth (14 per group, 336 total) was tested using a pull-out technique. Three-way anova and the Student-Newman-Kuels test were used to test the effects of irrigation regimen, post size and post brand on the retentive strength of the fibre post. Almost every sample failed at the interface between dentine and luting cement. Irrigation method did not have any significant effect (P > 0.14) for any group other than ParaPost Fibre White (P < 0.001). The overall retention of a post appeared to be correlated to the amount of surface texture on the post; the smooth ICEPost was the least retentive brand, the textured FibreKor and the ParaPost Fibre White posts were moderately retained, and the threaded Flexi-Post Fibre and the Flexi-Flange Fibre posts were the most retentive brands tested. For all brands, larger diameter posts were more retentive than smaller diameter posts (P < 0.008). A final rinse of EDTA did not improve the short-term retention of fibre posts except for ParaPost Fibre White.

Improvement effects of cytokinin on EDTA assisted phytoremediation and the associated environmental risks.

PubMed

Luo, Jie; Cai, Limei; Qi, Shihua; Wu, Jian; Gu, X W Sophie

2017-10-01

Soil samples containing excess Cd (0.82 mg kg -1 ), Pb (92.7 mg kg -1 ) and Cu (72.7 mg kg -1 ) relative to their corresponding safe thresholds (0.3, 80 and 50 mg kg -1 , respectively) from a notorious e-waste disposing and recycling place in southern China were phytoremediated with EDTA addition to evaluate the promotion effects of cytokinin on the remediation efficiency of Eucalyptus globulus. Biomass production of the plant, evapotranspiration amount of the soil, metals accumulation in plant organs and the volume of leachate under various treatments were compared. Relative to the planting control, EDTA application shortened the time required for Cd, Pb and Cu decontamination by 1.7-5.5 times but led to significantly more leachate (996 vs 1256 mL), indicating the negative influence of the chelate treatment on the species and the surrounding environment. The foliar application of cytokinin can expand the advantage and alleviate the adverse impact of individual EDTA application simultaneously as manifested by the increased biomass yield, less time consumption for purification and decreased leachate volume. Cytokinin accelerated the transpiration rate of the plant proved by the least volume of leachate in individual cytokinin treatment. The major factors for effective phytoremediation were the resistance of species to high concentrations of contaminants and less environmental risks generation during the remediation processes. Therefore, synergistic use of such components provides more efficient decontamination of metals and more security for the environment. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of EDTA and citric acid on absorption of heavy metals and growth of Moso bamboo.

PubMed

Zhang, Xiaowei; Zhong, Bin; Shafi, Mohammad; Guo, Jia; Liu, Chen; Guo, Hua; Peng, Danli; Wang, Ying; Liu, Dan

2018-04-30

The effect of EDTA and citric acid on accumulation, toxicity of heavy metals (Cu, Zn, Cd, and Pb), and growth of Moso bamboo was investigated in current experiment. The availability of heavy metals in soil and its uptake by plants has indicated toxicity. The results revealed that EDTA and citric acid has reduced biomass of Moso bamboo but non-significant difference in biomass was observed compared with control. Application of EDTA (10 mmol kg -1 ) has significantly improved copper (Cu) by 56.5 and 84.9% in roots and above ground parts of plants. Application of EDTA (10 mmol kg -1 ) has significantly enhanced lead (Pb) by 51.8 and 210.8% in roots and above ground parts of Moso bamboo. Furthermore, treatment of EDTA has significantly improved activities of water-soluble Cd, Cu, and Pb in soil by 98.9, 70.1, and 73.1 times compared with control. In case of contents of diethylenetriaminepentaacetic acid (DTPA)-extractable metals, the treatment of EDTA (10 mmol kg -1 ) has produced maximum increase of 244.5 mg kg -1 Zn and 157.9 mg kg -1 Pb, respectively. It is concluded that effect of EDTA was superior compared with citric acid for improvement of phytoremediation potential of Moso bamboo.

A blood-testis barrier restricting passage from blood into rete testis fluid but not into lymph

PubMed Central

Setchell, B. P.; Voglmayr, J. K.; Waites, G. M. H.

1969-01-01

1. A permeability barrier in or around the seminiferous tubules of rams has been demonstrated by studying the rate of passage of a variety of substances from blood plasma into fluid collected from the rete testis and into testicular lymph. 2. All substances studied passed readily into testicular lymph. 3. Tritiated water, urea, ethanol and bicarbonate in rete testis fluid equilibrated with blood plasma within 3 hr; Na+, K+, Rb+, Cl-, I-, CNS-, creatinine and galactose entered slowly and p-aminohippurate (PAH), glutamate, iodinated albumin, inulin and [51Cr]EDTA did not appear in rete testis fluid at all. 4. Rubidium was excluded relative to iodoantipyrine from the testes of control and hypophysectomized rats and from rat testes heated to 37, 40, 43 and 45° C; no such exclusion was seen in testes of rats which had been given cadmium chloride 5 months earlier so as to destroy the seminiferous tubules. 5. It is suggested that this permeability barrier will regulate the access to the seminiferous epithelium of some constituents of blood plasma, isolate the germinal cells immunologically and help to maintain the concentration differences between rete testis fluid and lymph or blood plasma. PMID:4973530

Effect of oral calcium disodium EDTA on iron absorption in a human model of iron overdose.

PubMed

Matteucci, Michael J; Habibe, Michael; Robson, Kristie; Baldassano, Accursia A; Riffenburgh, Robert H; Tanen, David A

2006-01-01

Anecdotal case reports and animal models have suggested that the administration of CaNa2EDTA (EDTA) may be effective in reducing the absorption of iron after an oral iron overdose. We designed this study to determine the effect of orally administrated EDTA with or without activated charcoal (AC) on iron absorption after a mild iron ingestion in healthy human volunteers. A randomized, crossover study was conducted in eight healthy human volunteers. All subjects ingested 5 mg/kg of elemental iron in the form of ferrous sulfate. One hour post ingestion, subjects were randomized to receive 35 mg/kg EDTA, EDTA plus 50 grams of AC, or water. Serial iron levels were obtained at baseline and every hour for the first 6 hours, then at 8, 12, and 24 hours. A 2-week washout was used between study arms. The Kruskal-Wallis test was used for the following comparisons between treatment groups: baseline serum iron levels, area under time-concentration curves (AUCs) from baseline to 12 hours and baseline to 24 hours, and peak iron levels. Baseline serum iron levels did not differ among the three treatment groups (p = 0.844). AUCs were not different among groups (p = 0.746 for 12 hr, p = 0.925 for 24 hr). AUC medians (with 95% binomial confidence bounds) for control, EDTA, EDTA + AC groups, respectively, for 12 hr were: 2813 (2298, 3561), 2570 (1669, 3476), and 2654 (2125, 3600); and for 24 hr were: 4083 (3488,5314), 4139 (2666, 5547), and 4274 (3336, 5577). Peak serum iron levels did not differ among treatment groups (p = 0.481). Peak iron level medians in microg/dL (with 95% binomial confidence bounds) were for control: 329 (253, 382), for EDTA: 271 (184, 375), and for EDTA + AC: 285 (229, 352). Orally administered EDTA did not significantly reduce iron absorption when administered 1 hour post iron ingestion during the 12 or 24-hour period following the ingestion of 5 mg/kg of elemental iron in healthy human volunteers.

Gel Electrophoresis--The Easy Way for Students

ERIC Educational Resources Information Center

VanRooy, Wilhelmina; Sultana, Khalida

2010-01-01

This article describes a simple, inexpensive, easy to conduct gel-electrophoresis activity using food dyes. It is an alternative to the more expensive counterparts which require agarose gel, DNA samples, purchased chamber and Tris-borate-EDTA buffer. We suggest some learning activities for senior biology students along with comments on several…

Cytotoxic and genotoxic potential of Cr(VI), Cr(III)-nitrate and Cr(III)-EDTA complex in human hepatoma (HepG2) cells.

PubMed

Novotnik, Breda; Ščančar, Janez; Milačič, Radmila; Filipič, Metka; Žegura, Bojana

2016-07-01

Chromium (Cr) and ethylenediaminetetraacetate (EDTA) are common environmental pollutants and can be present in high concentrations in surface waters at the same time. Therefore, chelation of Cr with EDTA can occur and thereby stable Cr(III)-EDTA complex is formed. Since there are no literature data on Cr(III)-EDTA toxicity, the aim of our work was to evaluate and compare Cr(III)-EDTA cytotoxic and genotoxic activity with those of Cr(VI) and Cr(III)-nitrate in human hepatoma (HepG2) cell line. First the effect of Cr(VI), Cr(III)-nitrate and Cr(III)-EDTA on cell viability was studied in the concentration range from 0.04 μg mL(-1) to 25 μg mL(-1) after 24 h exposure. Further the influence of non-cytotoxic concentrations of Cr(VI), Cr(III)-nitrate and Cr(III)-EDTA on DNA damage and genomic stability was determined with the comet assay and cytokinesis block micronucleus cytome assay, respectively. Cell viability was decreased only by Cr(VI) at concentrations above 1.0 μg mL(-1). Cr(VI) at ≥0.2 μg mL(-1) and Cr(III) at ≥1.0 μg mL(-1) induced DNA damage, while after Cr(III)-EDTA exposure no formation DNA strand breaks was determined. Statistically significant formation of micronuclei was induced only by Cr(VI) at ≥0.2 μg mL(-1), while no influence on the frequency of nuclear buds nor nucleoplasmic bridges was observed at any exposure. This study provides the first evidence that Cr(III)-EDTA did not induce DNA damage and had no influence on the genomic stability of HepG2 cells. Copyright © 2016 Elsevier Ltd. All rights reserved.

Synergetic effects of DA-6/GA₃ with EDTA on plant growth, extraction and detoxification of Cd by Lolium perenne.

PubMed

He, Shanying; Wu, Qiuling; He, Zhenli

2014-12-01

Research is needed to improve efficiency of phytoextraction of heavy metals from contaminated soils. A pot experiment was carried out to study the effects of plant growth regulators (PGRs) (diethyl aminoethyl hexanoate (C18H33NO8, DA-6) and gibberellic acid 3 (C19H22O6, GA3)) and/or EDTA on Cd extraction, subcellular distribution and chemical forms in Lolium perenne. The addition of EDTA or PGRs significantly enhanced Cd extraction efficiency (P<0.05), with the decreasing order of: 1 μM DA-6>10 μM DA-6>10 μM GA3>2.5 mmol kg(-1) EDTA>other treatments of PGR alone. PGRs+EDTA resulted in a further increase in Cd extraction efficiency, with EDTA+1 μM DA-6 being the most efficient. At the subcellular level, about 44-57% of Cd was soluble fraction, 18-44% in cell walls, and 12-25% in cellular organelles fraction. Chemical speciation analysis showed that 40-54% of Cd was NaCl extractable, 7-23% HAc extractable, followed by other fractions. EDTA increased the proportions of Cd in soluble and cellular organelles fraction, as well as the metal migration in shoot; therefore, the toxicity to plant increased and plant growth was inhibited. Conversely, PGRs fixed more Cd in cell walls and reduced Cd migration in shoot; thus, metal toxicity was reduced. In addition, PGRs promoted plant biomass growth significantly (P<0.05), with 1 μM DA-6 being the most effective. A combination of DA-6/GA3 with EDTA can alleviate the adverse effect of EDTA on plant growth, and the treatment of EDTA+1 μM DA-6 appears to be optimal for improving the remediation efficiency of L. perenne for Cd contaminated soil. Copyright © 2014 Elsevier Ltd. All rights reserved.

[Synergistic effects of lysozyme with EDTA-2Na on antibacterial activity].

PubMed

Li, Xiao-man; Wang, Xiao-yan; Gao, Xue-jun

2015-02-18

To evaluate the synergistic antibacterial effects of lysozyme with ethylenediaminetetraacetic acid disodium salt (EDTA-2Na) on Enterococcus faecalis (E. faecalis) and Porphyromonas endodontalis (P. endodontalis). E. faecalis and P. endodontalis were cultured and adjusted to 10(8) CFU/mL. Then 0.3, 0.5, 1, 2, 5, 10, 50, 100, 150 and 300 g/L of lysozyme were prepared with deionized water; and the lysozyme solutions were mixed with 0.5, 1.0, 2.0 g/L of EDTA-2Na, respectively. The bacteria and lysosome with/without EDTA-2Na interacted for 15 min, then water-soluble tetrazolium (WST) working solution was added and the activity of the bacteria was calculated by measuring optical densities at 450 nm and 630 nm with microplate spectrophotometer. Regarding the pure lysozyme from 0.5 g/L to 150 g/L, more E. faecalis and P. endodontalis were inhibited when the concentration of lysozyme was higher, especially for E. faecalis. There was synergistic effect of lysozyme with EDTA-2Na on antibacterial activity, which was related to the concentration of lysozyme. On E. faecalis, the antibacterial activity of lysozyme with EDTA-2Na was 1.2-3.7 folds than the pure lysozyme when the concentration of lysozyme was 0.5-50 g/L (P<0.05), and on P. endodontalis, the antibacterial activity of lysozyme with EDTA-2Na was 1.3-3.5 folds than the pure lysozyme when the concentration of lysozyme was 0.5-10 g/L (P<0.05). When the concentration of lysozyme was higher than 100 g/L, EDTA-2Na did not show synergistic effect on the antibacterial activity (P>0.05). For E. faecalis and P. endodontalis, a low concentration of lysozyme with EDTA-2Na showed significant synergistic antibacterial activity, while a high concentration of lysozyme with EDTA-2Na did not.

A Comparative Study of [CaEDTA](2-) and [MgEDTA](2-): Structural and Dynamical Insights from Quantum Mechanical Charge Field Molecular Dynamics.

PubMed

Tirler, Andreas O; Hofer, Thomas S

2015-07-09

Structure and dynamics of [MgEDTA](2-) and [CaEDTA](2-) complexes in aqueous solution have been investigated via quantum mechanical/molecular mechanical (QM/MM) simulations. While for the first a 6-fold octahedral complex has been observed, the presence of an additional coordinating water ligand has been observed in the latter case. Because of rapidly exchanging water molecules, this 7-fold coordination complex was found to form pentagonal bipyramidal as well as capped trigonal prismatic configurations along the simulation interchanging on the picosecond time scale. Also in the case of [MgEDTA](2-) a trigonal prismatic configuration has been observed for a very short time period of approximately 1 ps. This work reports for the first time the presence of trigonal prismatic structures observed in the coordination sphere of [MgEDTA](2-) and [CaEDTA](2-) complexes in aqueous solution. In addition to the detailed characterization of structure and dynamics of the systems, the prediction of the associated infrared spectra indicates that the ion-water vibrational mode found at approximately 250 cm(-1) provides a distinctive measure to experimentally detect the presence of the coordinating water molecule via low-frequency IR setups.

Oxidative degradation stability and hydrogen sulfide removal performance of dual-ligand iron chelate of Fe-EDTA/CA.

PubMed

Miao, Xinmei; Ma, Yiwen; Chen, Zezhi; Gong, Huijuan

2017-09-05

Catalytic oxidation desulfurization using chelated iron catalyst is an effective method to remove H 2 S from various gas streams including biogas. However, the ligand of ethylenediaminetetraacetic acid (EDTA), which is usually adopted to prepare chelated iron catalyst, is liable to be oxidative degraded, and leads to the loss of desulfurization performance. In order to improve the degradation stability of the iron chelate, a series of iron chelates composed of two ligands including citric acid (CA) and EDTA were prepared and the oxidative degradation stability as well as desulfurization performance of these chelated iron catalysts were studied. Results show that the iron chelate of Fe-CA is more stable than Fe-EDTA, while for the desulfurization performance, the situation is converse. For the dual-ligand iron chelates of Fe-EDTA/CA, with the increase of mol ratio of CA to EDTA in the iron chelate solution, the oxidative degradation stability increased while the desulfurization performance decreased. The results of this work showed that Fe-EDTA/CA with a mol ratio of CA:EDTA = 1:1 presents a relative high oxidative degradation stability and an acceptable desulfurization performance with over 90% of H 2 S removal efficiency.

Formation of hydroxyl radicals and Co3+ in the reaction of Co(2+)-EDTA with hydrogen peroxide. Catalytic effect of Fe3+.

PubMed

Eberhardt, M K; Santos, C; Soto, M A

1993-05-07

Co2+ ions (Co(NO3)2.6H2O) react with H2O2 only in presence of EDTA to yield OH radicals and Co3+. This reaction was carried out in unbuffered aqueous solutions (pH = 2.6). The formation of Co3+ was confirmed by spectroscopy. The Co(3+)-EDTA complex shows two typical absorptions at 382 nm and 532 nm. The Co(3+)-EDTA complex can be prepared by a number of oxidizing agents, like Fe3+, Fe(3+)-EDTA, Ag+, Ag2+, Ce4+, and hydroxyl radicals. Since Fe3+ oxidizes Co(2+)-EDTA to Co(3+)-EDTA and Fe2+ we initiate a chain reaction for .OH formation. Our results show that there are two modes for H2O2 decomposition: (1) One electron transfer to give OH radicals and (2) Decomposition of H2O2 to H2O and O2 without intermediate .OH formation. This reaction depends strongly on the pH of the buffer. The H2O2 decomposition increases with increasing pH and increasing Co2+ concentration.

Crystal and molecular structure of Sr{sub 2}(Edta) . 5H{sub 2}O, Sr{sub 2}(H{sub 2}Edta)(HCO{sub 3}){sub 2} . 4H{sub 2}O, and Sr{sub 2}(H{sub 2}Edta)Cl{sub 2} . 5H{sub 2}O strontium ethylenediaminetetraacetates

DOE Office of Scientific and Technical Information (OSTI.GOV)

Polyakova, I. N., E-mail: polyakova@igic.ras.ru; Poznyak, A. L.; Sergienko, V. S.

2009-03-15

Three Sr{sup 2+} compounds with the Edta{sup 4-} and H{sub 2}Edta{sup 2-} ligands-Sr{sub 2}(Edta) . 5H{sub 2}O (I), Sr{sub 2}(H{sub 2}Edta)(HCO{sub 3}){sub 2} . 4H{sub 2}O (II), and Sr{sub 2}(H{sub 2}Edta)Cl{sub 2} . 5H{sub 2}O (III)-are synthesized, and their crystal structures are studied. In I, the Sr(1) atom is coordinated by the hexadentate Edta{sup 4-} ligand following the 2N + 4O pattern and by two O atoms of the neighboring ligands, which affords the formation of zigzag chains. The Sr(2) atom forms bonds with O atoms of five water molecules and attaches itself to a chain via bonds with threemore » O atoms of the Edta{sup 4-} ligands. The Sr(1)-O and Sr(2)-O bond lengths fall in the ranges 2.520(2)-2.656(3) and 2.527(3)-2.683(2) A, respectively. The Sr(1)-N bonds are 2.702(3) and 2.743(3) A long. In II and III, the H{sub 2}Edta{sup 2-} anions have a centrosymmetric structure with the trans configuration of the planar ethylenediamine fragment. The N atoms are blocked by acid protons. In II, the environment of the Sr atom is formed by six O atoms of three H{sub 2}Edta ligands, two O atoms of water molecules, and an O atom of the bicarbonate ion, which is disordered over two positions. In III, the environment of the Sr atom includes six O atoms of four H{sub 2}Edta{sup 2-} ligands and three O atoms of water molecules. The coordination number of the Sr atoms is equal to 8 + 1. In II and III, the main bonds fall in the ranges 2.534(3)-2.732(2) and 2.482(2)-2.746(3) A, whereas the ninth bond is elongated to 2.937(3) and 3.055(3) A, respectively. In II, all the structural elements are linked into wavy layers. The O-H-O interactions contribute to the stabilization of the layer and link neighboring layers. In III, hydrated Sr{sup 2+} cations and H{sub 2}Edta{sup -} anions form a three-dimensional [Sr{sub 2}(H{sub 2}Edta)(H{sub 2}O){sub 3}]{sub n}{sup 2n+} framework. The Cl{sup -} anions are fixed in channels of the framework by hydrogen bonds with four water molecules. In II and III, the N-H groups form four-center N-H-O{sub 3} hydrogen bonds, which include one intermolecular and two intramolecular components. PACS numbers: 61.66.Hq.« less

An in vitro evaluation of passive ultrasonic agitation of different irrigants on smear layer removal after post space preparation: a scanning electron microscopic study.

PubMed

Srirekha, A; Rashmi, K; Hegde, Jayshree; Lekha, S; Rupali, K; Reshmi, George

2013-09-01

This study evaluated the removal of debris and smear layer after post space preparation using different irrigations and passive ultrasonic agitation. Sixty human premolars were decoronated and post space prepared after endodontic therapy. The samples were then randomly divided into three experimental groups (Groups A, B, C) and one control group (Group D) with fifteen samples in each group. Groups A and B samples were treated with 10 % citric acid and 17 % ethylenediamintetraacetic acid (EDTA), respectively and passive ultrasonic agitation was done, rinsed with sodium hypochlorite and finally flushed with saline. Group C samples were conditioned with 36 % phosphoric acid and then rinsed with saline. The control group was treated with 3 % sodium hypochlorite, passive ultrasonic agitation done and flushed with saline. The samples were sectioned and evaluated for debris and smear layer removal under scanning electron microscope. 10 % citric acid showed the best removal of smear layer when compared with 17 % EDTA and 36 % phosphoric acid, but was not statistically significant (p > 0.05). The difference in scoring for debris and smear layer removal in the coronal, middle and apical third of post space of experimental groups in comparison with control group was statistically significant (p < 0.001).

Phosphorus Elimination at Sodium Silicate from Quartz Sand Roasted with Complexation using Chitosan-EDTA

NASA Astrophysics Data System (ADS)

Wahyuningsih, S.; Ramelan, A. H.; Suharty, N. S.; Handayani, M.; Firdiyono, F.; Sulistiyono, E.; Munawaroh, H.; Sari, P. P.; Kristiawan, Y. R.

2018-03-01

A phosphorus elimination from sodium silicate solution has been studied. Phosphorus elimination was performed by adding chitosan-EDTA to remove cation phosphorus. Characterization of chitosan-EDTA material was performed using FT-IR, while the decreasing level of phosphorus content was analyzed by quantitative analysis using spectrophotometer UV-Vis refers to SNI 06-6989-2004. The results showed that the content of the sodium silicate can be reduced up to 67.1% through Chitosan-EDTA complexation with phosphorus.

Ethylenediaminetetraacetic acid in endodontics

PubMed Central

Mohammadi, Zahed; Shalavi, Sousan; Jafarzadeh, Hamid

2013-01-01

Ethylenediaminetetraacetic acid (EDTA) is a chelating agent can bind to metals via four carboxylate and two amine groups. It is a polyamino carboxylic acid and a colorless, water-soluble solid, which is widely used to dissolve lime scale. It is produced as several salts, notably disodium EDTA and calcium disodium EDTA. EDTA reacts with the calcium ions in dentine and forms soluble calcium chelates. A review of the literature and a discussion of the different indications and considerations for its usage are presented. PMID:24966721

Manufacture and Incorporation of Liposome-Entrapped Ethylenediaminetetraacetic Acid into Model Miniature Gouda-Type Cheese and Subsequent Effect on Starter Viability, pH, and Moisture Content.

PubMed

McAuliffe, Lisa N; Kilcawley, Kieran N; Sheehan, Jeremiah J; McSweeney, Paul L H

2016-10-25

Liposome-encapsulated ethylenediaminetetraacetic acid (EDTA) was incorporated into a model miniature Gouda-type cheese (20 g) in order to assess its effect on rennet gelation, starter viability, pH, and moisture content. EDTA was encapsulated within 2 different food-grade proliposome preparations, Pro-Lipo Duo and Pro-Lipo C (50% and 40% unsaturated soybean phospholipids and 50% and 60% aqueous medium, respectively), using the following high-shear technologies: Ultra-Turrax (5000 rpm), 2-stage homogenization (345 bar), or microfluidization (690 bar). Liposome size distribution was affected by the high-shear technology employed with the proportion of large vesicles (>100 nm) decreasing in the order microfluidization < 2-stage homogenization < Ultra-Turrax. All EDTA-containing liposomes were stable during 28 d refrigerated storage, with no significant (P ≤ 0.05) change in size distribution or EDTA entrapment efficiency (%EE). Liposome composition affected the entrapment of EDTA, with Pro-Lipo C having a significantly greater %EE than Pro-Lipo Duo, 63% and 54%, respectively. For this reason, Pro-Lipo C EDTA liposomes, with and without EDTA, were incorporated into model miniature Gouda-type cheese. Addition of liposome-encapsulated EDTA to milk during cheese making did not impact pH or rennet gel formation. No differences in composition or pH were evident in liposome-treated cheeses. The results of this study show that the incorporation of liposome-encapsulated EDTA into milk during cheese manufacture did not affect milk fermentation, moisture content, or pH, suggesting that this approach may be suitable for studying the effects of calcium equilibrium on the texture of brine-salted cheeses. © 2016 Institute of Food Technologists®.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Yan, Minyan; Shen, Yi, E-mail: shenyiemail@163.com; Zhao, Li

Graphical abstract: The MoO{sub 3} powder, which exhibits highly dispersed floral aggregated-like structure built up by superimposed and staggered nanoflakes with a diameter of 1-1.5 {mu}m and a thickness of 0.1-0.2 {mu}m, has been successfully obtained when the molar ratio of EDTA/Mo{sup 6+} is 0.05:1. The EDTA inducer obviously enlarges the surface area and apparently enhances the reactivity of MoO{sub 3} powders, making it show greater absorptive capacity to the excitation light and better photochromic properties than the pure MoO{sub 3} powder. Highlights: {yields} EDTA as organic inducing agent. {yields} EDTA inducer at EDTA/Mo{sup 6+} molar ratio of 0.05:1 enablesmore » growth of flower-like microspheres. {yields} The formation of flower-like MoO{sub 3} makes its photochromic properties strongly enhanced. -- Abstract: In this study, the photochromic MoO{sub 3} powder with novel morphology has been synthesized via hydrothermal method, using ethylene diamine tetraacetice acid (EDTA) as organic inducing agent. The influence of EDTA on the morphology, structure and photochromic properties of MoO{sub 3} powder has been characterized by X-ray diffraction (XRD), scanning electron microscopy (SEM), transmission electron microscopy (TEM), as well as ultraviolet and visible spectroscopy (UV-vis) and color difference meter. When the molar ratio of EDTA/Mo{sup 6+} is 0.05:1, the EDTA-induced MoO{sub 3} powder is found to have 3D flower-like morphologies and excellent photochromic properties. Furthermore, the possible growth mechanism of the flower-like structure and the photochromic mechanism of MoO{sub 3} powder are also discussed in detail.« less

A new Pb{sup II}(ethylenediaminetetraacetate) coordination polymer with a two-dimensional layer structure

DOE Office of Scientific and Technical Information (OSTI.GOV)

Zhao, D., E-mail: iamzd@hpu.edu.cn; Zhang, R. H.; Li, F. F.

2016-12-15

A new Pb{sup II}−edta{sup 4–} coordination polymer, Pb{sub 2}(edta)(H{sub 2}O){sub 0.76} (edta{sup 4–} = ethylenediaminetetraacetate) was synthesized under hydrothermal condition. Single crystal X-ray analysis reveals that it represents a novel two-dimensional (2D) Pb{sup 2+}–edta{sup 4–} layer structure with a (4,8{sup 2})-topology. Each edta{sup 4–} ligand employs its four carboxylate O and two N atoms to chelate one Pb{sup II} atom (hexa-coordinated) and connects five Pb{sup II} atoms (ennea-coordinated) via its four carboxylate groups to form 2D layer framework. Adjacent layers are packed into the overall structure through vander Waals interactions.

Characterization of Insulin Degrading Enzyme and other Aβ Degrading Proteases in Human Serum: a Role in Alzheimer’s disease?

PubMed Central

Liu, Zhiheng; Zhu, Haihao; Fang, Guang Guang; Walsh, Kathryn; Mwamburi, Maya; Wolozin, Benjamin; Abdul-Hay, Same O.; Ikezu, Tsuneya; Lessring, Malcolm A.; Qiu, Wei Qiao

2013-01-01

Sporadic Alzheimer’s disease (AD) patients have low amyloid-β peptide (Aβ) clearance in the central nervous system (CNS). The peripheral Aβ clearance may also be important but its role in AD remains unclear. We aimed to study the Aβ degrading proteases including insulin degrading enzyme (IDE), angiotensin converting enzyme (ACE) and others in blood. Using the fluorogenic substrate V—a substrate of IDE and other metalloproteases, we showed that human serum degraded the substrate V, and the activity was inhibited by adding increasing dose of Aβ. The existence of IDE activity was demonstrated by the inhibition of insulin, amylin or EDTA, and further confirmed by immunocapture of IDE using monoclonal antibodies. The involvement of ACE was indicated by the ability of the ACE inhibitor, lisinopril, to inhibit the substrate V degradation. To test the variations of substrate V degradation in humans, we used serum samples from a homebound elderly population with cognitive diagnoses. Compared with the elderly who had normal cognition, those with probable AD and amnestic mild cognitive impairment (amnestic MCI) had lower peptidase activities. Probable AD or amnestic MCI as an outcome remained negatively associated with serum substrate V degradation activity after adjusting for the confounders. The elderly with probable AD had lower serum substrate V degradation activity compared with those who had vascular dementia. The blood proteases mediating Aβ degradation may be important for the AD pathogenesis. More studies are needed to specify each Aβ degrading protease in blood as a useful biomarker and a possible treatment target for AD. PMID:22232014

Evaluation of the tratment of metal-EDTA complexes using Ti0{sub 2} photocatalysis

DOE Office of Scientific and Technical Information (OSTI.GOV)

Madden, T.; Datyte, A.K.; Prairie, M.R.

1996-03-01

This study has demonstrated the feasibility of TiO{sub 2} photocatalysis to treat EDTA and several metal-EDTA complexes that can be found in industrial wastewaters. For the EDTA complexes of metals capable of photodeposition, such as Cu and Pb, certain reaction conditions were shown to facilitate the simultaneous complex degradation and photodeposition of these metals onto the catalyst. With metals that do not easily photodeposit, such as Ni and Cd, it is shown that the complex degradation is still facilitated, and can enhance other metals removal processes after photocatalytic treatment. Because the treatment of these metal-EDTA complexes typically requires special measures,more » there may exist situations where TiO{sub 2} photocatalysis could actually be the preferred method of treatment. However, its use should be compared economically to other more established advanced oxidation technologies. This necessity is demonstrated in the economic comparison to ozone treatment for EDTA degradation alone, where ozone treatment appears to be the clear choice in this application.« less

Calcium EDTA toxicity: renal excretion of endogenous trace metals and the effect of repletion on collagen degradation in the rat.

PubMed

Braide, V B

1984-01-01

Studies on total hydroxyproline concentrations in urine of rats infused with toxic doses of CaEDTA at 6 mmol/kg per 24 hr for 48 hr or injected i.p. with the chelate at 4.8 mmol/kg/day for 10 days, indicate a two- to six-fold increase in urine excretion of the imino acid. This is due to increased degradation of collagen induced by CaEDTA. CaEDTA infusion was also shown to enhance urine excretion of some trace metals (Zn, Mn, Cu and Fe). Rats infused with CaEDTA for 36 hr showed a gradual fall in concentration of hydroxyproline in the urine, following cessation of chelate infusion. The decline in hydroxyproline concentrations was faster in rats receiving trace metal (Zn, Co, Mn or Ni) treatment during the post-CaEDTA infusion period; suggesting that the metals may affect collage, making the protein less susceptible to degradation in the body.

Risks of using EDTA as an agent for trace metals dosing in anaerobic digestion of olive mill solid waste.

PubMed

Serrano, A; Pinto-Ibieta, F; Braga, A F M; Jeison, D; Borja, R; Fermoso, F G

2017-12-01

Low concentrations of trace elements in many organic wastes recommend their supplementation in order to avoid potential limitations. Different chelating agents have been used to ensure an adequate trace metal pool in the soluble fraction, by forming dissolved complexes. Ethylenediaminetetraacetic acid (EDTA) is probably the most common, although several negative effects could be associated with its usage. Biomethane potential tests were performed using Olive Mill Solid Waste as the substrate, supplementing different combinations of Fe, Co, Ni, Ba, always under the presence of EDTA. Results show that Ni and Co slightly recovered biodegradability. However, Ba supplementation resulted in worsening the methane yield coefficient in all cases. High concentration of EDTA led to decrease in the activity of anaerobic digestion. High availability of EDTA induces the capture of trace metals like Co or Ni, key trace metals for anaerobic biomass activity. While supplementing trace metals, the addition of Ba and/or EDTA must be carefully considered.

Anti-biofilm properties of the antimicrobial peptide temporin 1Tb and its ability, in combination with EDTA, to eradicate Staphylococcus epidermidis biofilms on silicone catheters.

PubMed

Maisetta, Giuseppantonio; Grassi, Lucia; Di Luca, Mariagrazia; Bombardelli, Silvia; Medici, Chiara; Brancatisano, Franca Lisa; Esin, Semih; Batoni, Giovanna

2016-08-01

In search of new antimicrobials with anti-biofilm potential, in the present study activity of the frog-skin derived antimicrobial peptide temporin 1Tb (TB) against Staphylococcus epidermidis biofilms was investigated. A striking ability of TB to kill both forming and mature S. epidermidis biofilms was observed, especially when the peptide was combined with cysteine or EDTA, respectively. Kinetics studies demonstrated that the combination TB/EDTA was active against mature biofilms already after 2-4-h exposure. A double 4-h exposure of biofilms to TB/EDTA further increased the therapeutic potential of the same combination. Of note, TB/EDTA was able to eradicate S. epidermidis biofilms formed in vitro on silicone catheters. At eradicating concentrations, TB/EDTA did not cause hemolysis of human erythrocytes. The results shed light on the anti-biofilm properties of TB and suggest a possible application of the peptide in the lock therapy of catheters infected with S. epidermidis.

Highly concentrated EDTA gel improves cleaning efficiency of root canal preparation in vitro.

PubMed

Putzer, P; Hoy, L; Günay, H

2008-12-01

Debris and smear layer, as a product of mechanical root canal instrumentation, reduce the effectiveness of pharmacological substances to prevent post-treatment diseases and impair direct contact of filling materials with a clean dentinal surface. The aim of this in vitro study was to investigate the presence and localization of debris and smear layer via scanning electron microscope analysis after standardized root canal preparation with different chelating agents. Dentin surfaces received treatment with: (1) 15% ethylenediaminetetraacetic acid (EDTA), (2) 18.6% EDTA (3) and 24% EDTA or without any demineralizing chemicals as control. Forty vertically split human premolars were sputtered and divided into coronal, middle, and apical sections, followed by a randomized, blinded score evaluation using five scores. Pairwise comparisons of all treatment groups against a control group have been performed by Mann-Whitney U test and the Kruskal-Wallis test. Debris grades showed no significant difference between the three regions of the root canals, except for 18.6% EDTA in the central third. Smear layer and smear plug removal was concentration-dependent. Removal of the smear layer in the three areas showed that there was a statistically significant difference between all parts when using 18.6% and 24% EDTA concentrations compared with the control. The best smear layer removal in the apical region was observed using a 24% EDTA gel as chelating agent and lubricant. The usage of EDTA gel >/=18.6% presented a better cleaning regime when compared to the control group.

The effect of EDTA on Helianthus annuus uptake, selectivity, and translocation of heavy metals when grown in Ohio, New Mexico and Colombia soils.

PubMed

Turgut, Cafer; Pepe, M Katie; Cutright, Teresa J

2005-02-01

The use of two EDTA concentrations for enhancing the bioavailability of cadmium, chromium, and nickel in three natural soils (Ohio, New Mexico and Colombia) was investigated. The resulting uptake, translocation and selectivity with Helianthus annuus after mobilization were also examined. In general, plants grown in the sandy-loam Ohio soil had a higher uptake that resulted in a selectivity and total metal content of Cd>Cr>Ni and 0.73 mg and Cr>Cd>Ni and 0.32 mg for 0.1 and 0.3 g kg-1 EDTA, respectively. With the silty-loam New Mexico soil, although the total metal uptake was not statistically different the EDTA level did alter the selectivity; Cd>Cr>Ni (0.1 g kg-1 EDTA) and Cd>Cr>Ni (0.3 g kg-1 EDTA). Conversely, with the Colombian (sandy clay loam) soil increasing the EDTA level resulted in a higher total metal uptake (0.62 mg) than the 0.1 g kg-1 (0.59 mg) treatment. For all three soils, the translocation of Cd was limited. Evaluating the mobile metal fraction with and without EDTA determined that the chelator was capable of overcoming mass transfer limitations associated with the expandable clay fraction in the soils. Root wash results and root biomass concentrations indicated that Cd sorption was occurring. Therefore limited Cd translocation was attributed to insufficient phytochelatin levels.

The Molecular Prevalence of Viral Infections in Transplant Candidates with Bone Marrow Suppression, Shiraz, Southern Iran, 2010

PubMed Central

Mohammadi, B.; Yaghobi, R.; Dehghani, M.; Behzad Behbahani, A.

2013-01-01

Background: Transient bone marrow suppression, characterized by acute inability of the bone marrow to produce circulating blood cells, may strongly relate to the pathogenesis of some viral infections. Objective: To study the prevalence of some DNA and RNA viruses in patients with transient bone marrow suppression. Methods: EDTA-treated blood samples were collected from 27 patients with clinically- and laboratory-confirmed transient bone marrow suppression. The genomic DNA of hepatitis B virus, adenovirus, polyomavirus BK, and parvovirus B19, and genomic RNA of hepatitis C and G viruses were extracted and amplified by sensitive and specific in-house simple and nested PCR and RT-PCR protocols, respectively. The risk factors that might be related to the studied viral infections were analyzed. Results: Hepatitis B virus infection was diagnosed in 9 (33%) of 27 patients; adenovirus infection in 2 (7%); and parvovirus B19 infection in 7 (26%) of 27 patients. The genomic DNA of polyomovirus BK was not detected in any patients. Both hepatitis C and G viruses were found in 3 (11%) of 27 patients. Conclusion: Diagnosis of the high prevalence of hepatitis B virus, and parvovirus B19 in patients with transient bone marrow suppression, reflects the importance of these viral infections in introducing bone marrow suppression. This hypothesis should be confirmed in further studies. PMID:25013658

Immunization of Aged Pigs with Attenuated Pseudorabies Virus Vaccine Combined with CpG Oligodeoxynucleotide Restores Defective Th1 Immune Responses

PubMed Central

Chu, Pinpin; Ma, Miaopeng; Shi, Juqing; Cai, Haiming; Huang, Chaoyuan; Li, Huazhou; Jiang, Zhenggu; Wang, Houguang; Wang, Weifang; Zhang, Shuiqing; Zhang, Linghua

2013-01-01

Background and Aims Attempts to immunize aged subjects often result in the failure to elicit a protective immune response. Murine model studies have shown that oligonucleotides containing CpG motifs (CpG ODN) can stimulate immune system in aged mice as effectively as in young mice. Since many physiological and pathophysiological data of pigs can be transferred to humans, research in pigs is important to confirm murine data. Here we investigated whether immunization of aged pig model with attenuated pseudorabies virus vaccine (PRV vaccine) formulated with CpG ODN could promote a successful development of immune responses that were comparable to those induced in young pigs in a similar manner. Methodology Young and aged pigs were immunized IM with PRV vaccine alone, or in combination with CpG ODN respectively. At days 3, 7, 14 post immunization sera were assayed by ELISA for IgG titres, at day 7 for IgG1 and IgG2 subtypes titres. All blood samples collected in evacuated test tubes with K-EDTA at day 7 were analyzed for flow cytometer assay. Blood samples at day 7 collected in evacuated test tubes with heparin were analysed for antigen-specific cytokines production and peripheral blood mononuclear cells (PBMCs) proliferative responses. Results CpG ODN could enhance Th1 responses (PRV-specific IgG2/IgG1 ratio, proliferative responses, Th1 cytokines production) when used as an adjuvant for the vaccination of aged pigs, which were correlated with enhanced CD4+ T cells percentage, decreased CD4+CD8+CD45RO+ T cells percentage and improved PRV-specific CD4+ T cells activation. Conclusions Our results demonstrate a utility for CpG ODN, as a safe vaccine adjuvant for promoting effective systemic immune responses in aged pig model. This agent could have important clinical uses in overcoming some of age-associated depressions in immune function that occur in response to vaccination. PMID:23785433

New evidence that a large proportion of human blood plasma cell-free DNA is localized in exosomes

PubMed Central

Jiang, Chao; Krzyzanowski, Gary D.; Ryan, Wayne L.

2017-01-01

Cell-free DNA (cfDNA) in blood is used as a source of genetic material for noninvasive prenatal and cancer diagnostic assays in clinical practice. Recently we have started a project for new biomarker discovery with a view to developing new noninvasive diagnostic assays. While reviewing literature, it was found that exosomes may be a rich source of biomarkers, because exosomes play an important role in human health and disease. While characterizing exosomes found in human blood plasma, we observed the presence of cfDNA in plasma exosomes. Plasma was obtained from blood drawn into K3EDTA tubes. Exosomes were isolated from cell-free plasma using a commercially available kit. Sizing and enumeration of exosomes were done using electron microscopy and NanoSight particle counter. NanoSight and confocal microscopy was used to demonstrate the association between dsDNA and exosomes. DNA extracted from plasma and exosomes was measured by a fluorometric method and a droplet digital PCR (ddPCR) method. Size of extracellular vesicles isolated from plasma was heterogeneous and showed a mean value of 92.6 nm and a mode 39.7 nm. A large proportion of extracellular vesicles isolated from plasma were identified as exosomes using a fluorescence probe specific for exosomes and three protein markers, Hsp70, CD9 and CD63, that are commonly used to identify exosome fraction. Fluorescence dye that stain dsDNA showed the association between exosomes and dsDNA. Plasma cfDNA concentration analysis showed more than 93% of amplifiable cfDNA in plasma is located in plasma exosomes. Storage of a blood sample showed significant increases in exosome count and exosome DNA concentration. This study provide evidence that a large proportion of plasma cfDNA is localized in exosomes. Exosome release from cells is a metabolic energy dependent process, thus suggesting active release of cfDNA from cells as a source of cfDNA in plasma. PMID:28850588

New evidence that a large proportion of human blood plasma cell-free DNA is localized in exosomes.

PubMed

Fernando, M Rohan; Jiang, Chao; Krzyzanowski, Gary D; Ryan, Wayne L

2017-01-01

Cell-free DNA (cfDNA) in blood is used as a source of genetic material for noninvasive prenatal and cancer diagnostic assays in clinical practice. Recently we have started a project for new biomarker discovery with a view to developing new noninvasive diagnostic assays. While reviewing literature, it was found that exosomes may be a rich source of biomarkers, because exosomes play an important role in human health and disease. While characterizing exosomes found in human blood plasma, we observed the presence of cfDNA in plasma exosomes. Plasma was obtained from blood drawn into K3EDTA tubes. Exosomes were isolated from cell-free plasma using a commercially available kit. Sizing and enumeration of exosomes were done using electron microscopy and NanoSight particle counter. NanoSight and confocal microscopy was used to demonstrate the association between dsDNA and exosomes. DNA extracted from plasma and exosomes was measured by a fluorometric method and a droplet digital PCR (ddPCR) method. Size of extracellular vesicles isolated from plasma was heterogeneous and showed a mean value of 92.6 nm and a mode 39.7 nm. A large proportion of extracellular vesicles isolated from plasma were identified as exosomes using a fluorescence probe specific for exosomes and three protein markers, Hsp70, CD9 and CD63, that are commonly used to identify exosome fraction. Fluorescence dye that stain dsDNA showed the association between exosomes and dsDNA. Plasma cfDNA concentration analysis showed more than 93% of amplifiable cfDNA in plasma is located in plasma exosomes. Storage of a blood sample showed significant increases in exosome count and exosome DNA concentration. This study provide evidence that a large proportion of plasma cfDNA is localized in exosomes. Exosome release from cells is a metabolic energy dependent process, thus suggesting active release of cfDNA from cells as a source of cfDNA in plasma.

21 CFR 73.2120 - Disodium EDTA-copper.

Code of Federal Regulations, 2011 CFR

2011-04-01

... 21 Food and Drugs 1 2011-04-01 2011-04-01 false Disodium EDTA-copper. 73.2120 Section 73.2120 Food and Drugs FOOD AND DRUG ADMINISTRATION, DEPARTMENT OF HEALTH AND HUMAN SERVICES GENERAL LISTING OF COLOR ADDITIVES EXEMPT FROM CERTIFICATION Cosmetics § 73.2120 Disodium EDTA-copper. (a) Identity. The...

Heat stress increases the efficiency of EDTA in phytoextraction of heavy metals.

PubMed

Chen, Ya-Hua; Mao, Ying; He, Shi-Bin; Guo, Peng; Xu, Ke

2007-04-01

Solution culture and pot experiments were carried out to investigate the effects of root damage on phytoextraction of heavy metals. In hydroponics, roots of corn (Zea mays L.) seedlings were pretreated with heating stress, and then were exposed to 250 microM Pb+250 microM EDTA solutions for 7d. The results showed that the preheating treatment significantly increased Pb transportation from roots to shoots. In pot experiments, the effect of hot EDTA solution (95 degrees C) on the accumulation of heavy metal in the shoot of corn and pea (Pisum sativum L.) was also examined. Compared to normal EDTA (25 degrees C) treatment, application of hot EDTA solution to the soil surface increased the total removal of Pb in shoots of corn and pea by about 8- and 12-fold, respectively, in an artificially multimetal-contaminated soil. In addition, hot EDTA solution increased the shoot Cu removal by about 6-fold for corn and 9-fold for pea, respectively, in a naturally Cu-contaminated soil. These results suggested that exposure of roots to high temperature could increase the efficiency of EDTA on the accumulation of heavy metals in shoots. This new approach can help to minimize the amount of chelate applied in the field and reduce the potential risk of heavy metals' leaching.

Degradation of Ethylenediaminetetraacetic Acid by Microbial Populations from an Aerated Lagoon

PubMed Central

Belly, R. T.; Lauff, J. J.; Goodhue, C. T.

1975-01-01

The ferric chelate of ethylenediaminetetraacetic acid (EDTA) was biologically degraded by a mixed population of microorganisms present in an aerated lagoon receiving this chemical in its feed. As determined radiorespirometrically, 28% of the acetate-2-C and 30% of the ethylene position of the ammonium ferric chelate of [14C]EDTA was recovered as 14CO2 after 5 days. In a separate experiment using gas liquid chromatography and the sodium ferric chelate, as much as 89% disappearance of EDTA (0.1% wt/vol) was observed during a similar time period. Optimum 14CO2 evolution was observed at a pH value between 7 and 8 and at room temperature. Degradation of NH4Fe-[2-14C]EDTA was stimulated by the addition of either unlabeled NaFe-EDTA, nitrilotriacetic acid or ethylenediamine, and inhibited by the addition of a variety of different sugars and amino acids. Consistent with the biological nature of this degradation, little or no 14CO2 evolution was observed after heat treatment of the microorganisms at 100 C for 10 min, or after the addition of formalin or antibiotics to the incubation mixtures. Gas-liquid chromatography and mass spectral analyses were performed to demonstrate EDTA disappearance and to identify various possible intermediates of EDTA degradation. PMID:239630

The effect of EDTA, cations, and various buffers on the morphology of erythrocyte membranes: an electron-microscopic study.

PubMed

Pinteric, L; Manery, J F; Chaudry, I H; Madapallimattam, G

1975-05-01

Membranes of human erythrocytes were prepared by stepwise osmotic hemolysis in Ca2+-free solutions. Examination with the electron microscope after negative staining showed some short, conelike protuberances on the surface of about 20 percent of the ghosts, while 80 percent were round, intact spheres. After Ca2+ treatment, all membranes were round and intact. After exposure to ethylenediaminetetraacetic acid (EDTA) (1.0 mM, pH 7.4), the entire ghost surface was covered with long, thin extrusions called stromalytic forms (about 460 per cell). Their sizes, shapes, and fine structure are described. Exposure to ionic calcium (1.4 times 10-minus 4M) abolished the EDTA-induced stromalytic forms. A second exposure to EDTA reversed this Ca2+ effect. ATP, like EDTA, produced stromalytic forms. EDTA-induced stromalytic forms were also abolished by Zn2+, La3+, and Nd3+ at concentrations of 1-5 times 10-minus 4 M. Mg2+ at 10-minus 2 M was ineffective. Ghosts were prepared by graded lysis in various buffers. Those prepared in phosphate were the most stable and provided consistent EDTA effects and Ca2+ reversal. Ghosts in Tris-HCl showed breakdown unless salt was added. Moderately satisfactory ghosts were also obtained in Hepes-NaOH buffer and salt.

Impact of extra virgin olive oil and ethylenediaminetetraacetic acid (EDTA) on the oxidative stability of fish oil emulsions and spray-dried microcapsules stabilized by sugar beet pectin.

PubMed

Polavarapu, Sudheera; Oliver, Christine M; Ajlouni, Said; Augustin, Mary Ann

2012-01-11

The influence of EDTA on lipid oxidation in sugar beet pectin-stabilized oil-in-water emulsions (pH 6, 15% oil, wet basis), prepared from fish oil (FO) and fish oil-extra virgin olive oil (FO-EVOO) (1:1 w/w), as well as the spray-dried microcapsules (50% oil, dry basis) prepared from these emulsions, was investigated. Under accelerated conditions (80 °C, 5 bar oxygen pressure) the oxidative stability was significantly (P < 0.05) higher for FO and FO-EVOO formulated with EDTA, in comparison to corresponding emulsions and spray-dried microcapsules formulated without EDTA. The EDTA effect was greater in emulsions than in spray-dried microcapsules, with the greatest protective effect obtained in FO-EVOO emulsions. EDTA enhanced the oxidative stability of the spray-dried microcapsules during ambient storage (~25 °C, a(w) = 0.5), as demonstrated by their lower concentration of headspace volatile oxidation products, propanal and hexanal. These results show that the addition of EDTA is an effective strategy to maximize the oxidative stability of both FO emulsions and spray-dried microcapsules in which sugar beet pectin is used as the encapsulant material.

Reduction in heat-induced gastrointestinal hyperpermeability in rats by bovine colostrum and goat milk powders.

PubMed

Prosser, C; Stelwagen, K; Cummins, R; Guerin, P; Gill, N; Milne, C

2004-02-01

Male Sprague-Dawley rats were assigned to one of three dietary groups [standard diet (Cont; n = 8), standard diet plus bovine colostrum powder (BColost 1.7 g/kg; n = 8), or goat milk powder (GMilk 1.7 g/kg; n = 8)] to determine the ability of these supplements to reduce gastrointestinal hyperpermeability induced by heat. Raising core body temperature of rats to 41.5 degrees C increased transfer of (51)Cr-EDTA from gut into blood 34-fold relative to the ambient temperature value (P < 0.05) in the Cont group of rats, indicative of increased gastrointestinal permeability. Significantly less (P < 0.01) (51)Cr-EDTA was transferred into the blood of rats in either the BColost (27% of Cont) or GMilk group (10% of Cont) after heating, showing that prior supplementation with either bovine colostrum or goat milk powder significantly reduced the impact of heat stress on gastrointestinal permeability. The changes in the BColost group were not significantly different than those of the GMilk group. The potential mechanism of the protective effect of bovine colostrum and goat milk powders may involve modulation of tight junction permeability, because both powders were able to maintain transepithelial resistance in Madin Darby canine kidney cells challenged with EGTA compared with cells maintained in media only. The results show that bovine colostrum powder can partially alleviate the effects of hyperthermia on gastrointestinal permeability in the intact animal. Moreover, goat milk powder was equally as effective as bovine colostrum powder, and both may be of benefit in other situations where gastrointestinal barrier function is compromised.

A PET Tracer for Renal Organic Cation Transporters, ¹¹C-Metformin: Radiosynthesis and Preclinical Proof-of-Concept Studies.

PubMed

Jakobsen, Steen; Busk, Morten; Jensen, Jonas Brorson; Munk, Ole Lajord; Zois, Nora Elisabeth; Alstrup, Aage K O; Jessen, Niels; Frøkiær, Jørgen

2016-04-01

Organic cation transporters (OCTs) in the kidney proximal tubule (PT) participate in renal excretion of drugs and endogenous compounds. PT function is commonly impaired in kidney diseases, and consequently quantitative measurement of OCT function may provide an important estimate of kidney function. Metformin is a widely used drug and targets OCT type 2 located in the PT. Thus, we hypothesized that (11)C-labeled metformin would be a suitable PET tracer for quantification of renal function. (11)C-metformin was prepared by (11)C-methylation of 1-methylbiguanide. In vitro cell uptake of (11)C-metformin was studied in LLC-PK1 cells in the presence of increasing doses of unlabeled metformin. In vivo small-animal PET studies in Sprague-Dawley rats were performed at baseline and after treatment with OCT inhibitors to evaluate renal uptake of (11)C-metformin. Kidney and liver pharmacokinetics of (11)C-metformin was investigated in vivo by dynamic (11)C-metformin PET/CT in 6 anesthetized pigs, and renal clearance of (11)C-metformin was compared with renal clearance of (51)Cr-ethylenediaminetetraacetic acid (EDTA). Formation of (11)C metabolites was investigated by analysis of blood and urine samples. The radiochemical yield of (11)C-metformin was 15% ± 3% (n= 40, decay-corrected), and up to 1.5 GBq of tracer were produced with a radiochemical purity greater than 95% in less than 30 min. Dose-dependent uptake of (11)C-metformin in LLC-PK1 cells was rapid. Rat small-animal PET images showed (11)C-metformin uptake in the kidney and liver, the kinetics of which were changed after challenging animals with OCT inhibitors. In pigs, 80% of the injected metformin dose was rapidly present in the kidney, and a high dose of metformin caused a delayed renal uptake and clearance compared with baseline consistent with transporter-mediated competition. Renal clearance of (11)C-metformin was approximately 3 times the renal clearance of (51)Cr-EDTA. We successfully synthesized an (11)C-metformin tracer, and PET studies in rats and pigs showed a rapid kidney uptake from the blood and excretion into the bladder similar to other radiopharmaceuticals developed for γ-camera renography. © 2016 by the Society of Nuclear Medicine and Molecular Imaging, Inc.

Development and optimization of an in vitro cultivation protocol allows for isolation of Borrelia miyamotoi from patients with hard tick-borne relapsing fever.

PubMed

Koetsveld, J; Kolyasnikova, N M; Wagemakers, A; Toporkova, M G; Sarksyan, D S; Oei, A; Platonov, A E; Hovius, J W

2017-07-01

Borrelia miyamotoi has been shown to infect humans in Eurasia and North America causing hard tick-borne relapsing fever (HTBRF). In vitro cultivation of B. miyamotoi was described recently; but clinical isolation of relapsing fever Borrelia is cumbersome. Our aim was to develop a straightforward protocol enabling B. miyamotoi isolation directly from the blood of patients. Modified Kelly-Pettenkorfer (MKP-F) medium, with or without anticoagulants, or blood from healthy human volunteers, was spiked with B. miyamotoi spirochaetes in vitro. Subsequently, either media or plasma was used for cultivation directly, or after an additional centrifugation step. This isolation protocol was tested in a clinical setting on patients suspected of HTBRF. Dipotassium-EDTA, trisodium citrate and lithium heparin inhibited growth of B. miyamotoi at concentrations ≥250 μg/mL, 2.5 mM and 1 IU/mL, respectively. However, when plasma originating from human blood containing B. miyamotoi spirochaetes was subjected to an additional centrifugation step at 8000 g, suspended and inoculated into fresh MKP-F media, positive cultures were observed within 2 weeks. Of importance, this straightforward protocol allowed for isolation of B. miyamotoi from six out of nine patients with confirmed HTBRF. Direct culture from K 2 -EDTA, trisodium citrate and lithium heparin plasma containing B. miyamotoi is hampered due to anticoagulants. Using a simple centrifugation protocol we were able to circumvent this detrimental effect, allowing for the first clinical isolation of B. miyamotoi. This will be of value for future research on the pathogenesis, genetics, diagnosis, therapy and epidemiology of HTBRF and other tick-borne relapsing fevers. Copyright © 2017 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Total plasma clearance versus urinary plasma clearance of (51)Cr-EDTA in patients with cirrhosis with and without fluid retention.

PubMed

Henriksen, Ulrik L; Hansen, Hanne B; Ring-Larsen, Helmer; Bendtsen, Flemming; Henriksen, Jens H

2015-01-01

In patients with fluid retention, the total plasma clearance of (51)Cr-EDTA (ClP) may overestimate the glomerular filtration rate (GFR). The present study was therefore undertaken in order to compare ClP with the urinary plasma clearance of (51)Cr-EDTA (ClU) in patients with cirrhosis with and without fluid retention. A total of 136 patients with cirrhosis (24 without fluid retention, 112 with ascites) received a quantitative intravenous injection of (51)Cr-EDTA followed by plasma and quantitative urinary samples for 5 hours. ClP was determined from the injected dose relative to the plasma concentration-time area, extrapolated to infinity. ClU was determined as urinary excretion relative to the plasma concentration-time area up to voiding. In patients without fluid retention, the difference between ClP and ClU (ClP - ClU = ClΔ) was mean 4.5 mL/min/1.73 m(2). In patients with ascites, ClΔ was significantly higher (17.6 mL/min/1.73 m(2), p < 0.0001). ClΔ increased with lower values of GFR (r = - 0.458, p < 0.001). Repeated measurements of ClU in a subgroup of patients with fluid retention (n = 25) gave almost identical values. Different types of corrections of one-pool clearance were almost identical with ClP, except for higher clearance values, which were somewhat underestimated by the former. In patients with fluid retention and ascites ClP and corrected one-pool clearance overestimates GFR substantially. Although ClU may underestimate GFR slightly, patients with ascites should collect urine quantitatively in order to obtain a reliable measurement of GFR.

Effect of irrigation on surface roughness and fatigue resistance of controlled memory wire nickel-titanium instruments.

PubMed

Cai, J-J; Tang, X-N; Ge, J-Y

2017-07-01

To investigate the effect of irrigation on the surface roughness and fatigue resistance of HyFlex and M3 controlled memory (CM) wire nickel-titanium instruments. Two new files of each brand were analysed by atomic force microscopy (AFM). Then, the instruments were dynamically immersed in either 5.25% sodium hypochlorite (NaOCl) or 17% ethylene diamine tetraacetic acid (EDTA) solution for 10 min, followed by AFM analysis. The roughness average (Ra) and root mean square (RMS) values were analysed statistically using an independent sample t-test. Then, 36 files of each brand were randomly assigned to three groups (n = 12). Group 1 (the control group) was composed of new instruments. Groups 2 and 3 were dynamically immersed in 5.25% NaOCl and 17% EDTA solutions for 10 min, respectively. The number of rotations to failure for various groups was analysed using the one-way analysis of variance software. For M3 files, the Ra and RMS values significantly increased (P < 0.05) after the immersion. For the HyFlex file, the Ra and RMS values significantly increased (P < 0.05) only in EDTA, but not (P > 0.05) NaOCl. The resistance to cyclic fatigue of both HyFlex and M3 files did not significantly decrease (P > 0.05) by immersing in 5.25% NaOCl and 17% EDTA solutions. Except the HyFlex files immersed in NaOCl, the surface roughness of other files exposed to irrigants increased. However, a change in the surface tomography of CM wire instruments caused by contact with irrigants for 10 min did not trigger a decrease in cyclic fatigue resistance. © 2016 International Endodontic Journal. Published by John Wiley & Sons Ltd.

Streptomycin and EDTA decrease the number of desmin-negative fibers following stretch injury.

PubMed

Willems, Mark E T; Stauber, William T

2005-09-01

Streptomycin and ethylene diamine tetraacetic acid (EDTA) were used to examine the role of extracellular calcium in stretch-induced muscle injury. Streptomycin was injected in one group of rats, three times daily for 8 days (S, 300 mg.kg(-1).day(-1) intraperitoneally). In another group, EDTA was administered (150 mg.kg(-1) IP) 20 min before and 24 h after the injury protocol. Untreated rats (C) served as controls. Muscle injury was produced by 40 stretches of active dorsiflexor muscles by ankle rotation from 80 degrees to 130 degrees (velocity 1.75 rad.s(-1)). Ten minutes after the injury protocols, all animals lost the same amount of isometric force at both low and high stimulation frequencies (20 HZ; S, 56 +/- 6%; EDTA, 47 +/- 7%; C, 55 +/- 4%) and 120 HZ; S, 11 +/- 3%, EDTA, 13 +/- 3%; C, 11 +/- 3%). Tibialis anterior (TA) muscles were removed after 48 h for morphometric analysis. In both streptomycin-and EDTA-treated rats, the percent of injured (i.e., desmin-negative) myofibers in TA was reduced compared to untreated, injured muscles (S, 0.35 +/- 0.08%; EDTA, 0.64 +/- 0.19%; C, 1.81 +/- 0.43%). Thus, streptomycin and EDTA treatment did not alter the development of muscle weakness (i.e., isometric force deficit), but almost abolished the histopathologic changes. This study shows that the mechanisms for muscle weakness and histopathologic changes (inflammation) following repeated muscle strains can largely be dissociated from each other and helps explain why there is no correlation between isometric force deficits and the number of pathologic cells.

EDTA chelation effects on urinary losses of cadmium, calcium, chromium, cobalt, copper, lead, magnesium, and zinc.

PubMed

Waters, R S; Bryden, N A; Patterson, K Y; Veillon, C; Anderson, R A

2001-12-01

The efficacy of a chelating agent in binding a given metal in a biological system depends on the binding constants of the chelator for the particular metals in the system, the concentration of the metals, and the presence and concentrations of other ligands competing for the metals in question. In this study, we make a comparison of the in vitro binding constants for the chelator, ethylenediaminetetraacetic acid, with the quantitative urinary excretion of the metals measured before and after EDTA infusion in 16 patients. There were significant increases in lead, zinc, cadmium, and calcium, and these increases roughly corresponded to the expected relative increases predicted by the EDTA-metal-binding constants as measured in vitro. There were no significant increases in urinary cobalt, chromium, or copper as a result of EDTA infusion. The actual increase in cobalt could be entirely attributed to the cobalt content of the cyanocobalamin that was added to the infusion. Although copper did increase in the post-EDTA specimens, the increase was not statistically significant. In the case of magnesium, there was a net retention of approximately 85% following chelation. These data demonstrate that EDTA chelation therapy results in significantly increased urinary losses of lead, zinc, cadmium, and calcium following EDTA chelation therapy. There were no significant changes in cobalt, chromium, or copper and a retention of magnesium. These effects are likely to have significant effects on nutrient concentrations and interactions and partially explain the clinical improvements seen in patients undergoing EDTA chelation therapy.

Effect of Decellularization Protocol on the Mechanical Behavior of Porcine Descending Aorta

PubMed Central

Fitzpatrick, John C.; Clark, Peter M.; Capaldi, Franco M.

2010-01-01

Enzymatic-detergent decellularization treatments may use a combination of chemical reagents to reduce vascular tissue to sterilized scaffolds, which may be seeded with endothelial cells and implanted with a low risk of rejection. However, these chemicals may alter the mechanical properties of the native tissue and contribute to graft compliance mismatch. Uniaxial tensile data obtained from native and decellularized longitudinal aortic tissue samples was analyzed in terms of engineering stress and fit to a modified form of the Yeoh rubber model. One decellularization protocol used SDS, while the other two used TritonX-100, RNase-A, and DNase-I in combination with EDTA or sodium-deoxycholate. Statistical significance of Yeoh model parameters was determined by paired t-test analysis. The TritonX-100/EDTA and 0.075% SDS treatments resulted in relatively variable mechanical changes and did not effectively lyse VSMCs in aortic tissue. The TritonX-100/sodium-deoxycholate treatment effectively lysed VSMCs and was characterized by less variability in mechanical behavior. The data suggests a TritonX-100/sodium-deoxycholate treatment is a more effective option than TritonX-100/EDTA and SDS treatments for the preparation of aortic xenografts and allografts because it effectively lyses VSMCs and is the least likely treatment, among those considered, to promote a decrease in mechanical compliance. PMID:20689621

ON THE EFFECT OF ETHYLENEDIAMINETETRAACETATE (EDTA) ON THE ACCUMULATION COEFFICIENT OF DIFFERENT RADIOISOTOPES FROM AQUEOUS SOLUTION BY FRESH-WATER PLANTS (in Russian)

DOE Office of Scientific and Technical Information (OSTI.GOV)

Timofeeva-Resovskaya, E.A.; Timofeev-Resovskii, N.V.

1960-01-01

The effects of ethylenediaminetetraacetate on the accumulation coefficients of Ce/sup 144/, Co/sup 60/, Cs/sup 137/, Fe/sup 59/, Nb /sup 95/, Ru/ sup 106/, S/sup 35/, Sr/sup 90/, Y/sup 91/, Zn/sup 65/, and Zr/sup 95/ from aqueous solutions were studied. Eight grams of elodea (Elodea canadinsis Rich,), aquatic plant (Ceratophyllum demersum L), duckweed (Lemna minor L.), and chara (Chara fragilis Desw.) were placed in three liters of water containirg 400 mg of EDTA per liter. Tabulated data show 4 groups of isotopic reactions to EDTA. The first group includes strontium and cesium (whose uptake increases in the presence of EDTA), themore » second is represented hy sulfur (which does not react to EDTA), the third group consists of zirconium, niobium, and ruthenium (whose uptake is reduced 2 to 3 fold in the presence of EDTA), and the fourth group contains iron, cobalt, zinc, yttrium, and cerium (whose uptake is reduced 10 to 100 fold). (R.V.J.)« less