Effects of basic fibroblast growth factor on density and morphology of fibroblasts grown on root surfaces with or without conditioning with tetracycline or EDTA.

J Oral Sci. 2007 Sep;49(3):213-20

Authors: Silvério KG, Martinez AE, Rossa C

A study was conducted to evaluate in vitro the effect of root surface conditioning with basic fibroblast growth factor (b-FGF) on morphology and proliferation of fibroblasts. Three experimental groups were used: non-treated, and treated with 50 mug or 125 mug b-FGF/ml. The dentin samples in each group were divided into subgroups according to the chemical treatement received before application of b-FGF: none, or conditioned with tetracycline-HCl or EDTA. After contact with b-FGF for 5 min, the samples were incubated for 24 h with 1 ml of culture medium containing 1 x 10(5) cells/ml plus 1 ml of culture medium alone. The samples were then subjected to routine preparation for SEM, and random fields were photographed. Three calibrated and blind examiners peformed the assessment of morphology and density according to two index systems. Classification and regression trees indicated that the root surfaces treated with 125 mug b-FGF and previously conditioned with tetracycline-HCl or EDTA presented a morphology more suggestive of cellular adhesion and viability (P = 0.004). The density of fibroblasts on samples previously conditioned with EDTA, regardless of treatment with b-FGF, was significantly higher than in the other groups (P < 0.001). The present findings suggest that topical application of b-FGF has a positive influence on both the density and morphology of fibroblasts. (J. Oral Sci. 49, 213-220, 2007).

PMID: 17928728 [PubMed - in process]

Effectiveness of a silicon-based root canal sealer for filling of simulated lateral canals.

Braz Dent J. 2007;18(1):20-3

Authors: Barbizam JV, Souza M, Cecchin D, Dabbel J

The purpose of this in vitro study was to evaluate the ability of a silicon-based root canal sealer, compared to zinc oxide and eugenol and an epoxy resin-based sealers, for filling of simulated lateral canals. Thirty extracted single-rooted human teeth were selected, conventional access was made and the working length was established 1 mm from the apical foramen. Three simulated lateral canals, one in each root third (coronal, middle and apical) were prepared in both the mesial and distal surfaces of each tooth using a size 15 reamer adapted to a low-speed handpiece. Each root canal was instrumented using ProTaper rotary files up to file F3 at the working length, and then irrigated with 2.5% NaOCl followed by EDTA. The teeth were assigned to 3 groups (n=10), according to the root canal sealer: Roeko Seal (Group 1), Sealer 26 (group 2) and Grossman's sealer (Group 3). Gutta-percha cold lateral condensation technique was performed in all groups. Postoperative radiographs were taken and the images were projected for evaluation of the quality of lateral canal filling. Data were submitted to statistical analysis by Kruskal Wallis test at 5% significance level. The results showed that Grossman's sealer filled a larger number of lateral canals than Roeko Seal (p<0.05) and Sealer 26 (p<0.01). It may be concluded that Roeko Seal silicone-based root canal sealer was not as effective as the Grossman's sealer for filling of simulated lateral canals. The lateral canals localized in the apical third of the root were more difficult to be filled.

PMID: 17639195 [PubMed - indexed for MEDLINE]

Radiographic prevalence of root canal ramifications in a sample of root canal treatments in a Brazilian Dental School.

Braz Oral Res. 2007 Apr-Jun;21(2):112-7

Authors: de Quadros I, Zaia AA, Ferraz CC, de Souza Filho FJ, Gomes BP

The aim of this study was to radiographically investigate the presence of root canal ramifications found after endodontic treatment, and to determine any relationship between their presence and the type of the auxiliary chemical substance used. The study evaluated 1,470 endodontic treatments performed by final year undergraduate students at the Dental School of Piracicaba, State University of Campinas (UNICAMP), SP, Brazil, during the period from 1998 to 2000. The X-rays taken during treatment were evaluated in order to establish the presence of ramifications of the root canal system. The initial X-ray did not show the presence of any canal ramifications. After filling, X-rays showed only 3 ramification types: 3.06% of lateral canals, 2.99% of apical deltas, and 0.1% of interradicular canals. The maxillary premolars showed the highest number of lateral canals (n = 13), followed by mandibular premolars (n = 10) and maxillary incisors (n = 10). Apical deltas were mostly found in mandibular molars (n = 14), followed by maxillary incisors (n = 9). Only mandibular molars had interradicular canals. The detection of ramifications increased with the use of EDTA. However, no statistically significant relationship was found between the type of auxiliary chemical substance used and the number of root canal ramifications detected after root canal filling. It was concluded that the frequency of root canal ramifications found radiographically was low in treatments performed by undergraduate students.

PMID: 17589645 [PubMed - in process]

Influence of sodium hypochlorite-based irrigants on the susceptibility of intracanal microbiota to biomechanical preparation.

Braz Dent J. 2006;17(4):310-6

Authors: Soares JA, Pires J&#xFA;nior DR

This study evaluated the microbiological conditions of root canals, using smears and culture from anterior teeth and premolars with necrotic pulps associated with chronic periapical pathologies, before and after biomechanical preparation (BMP). During double-flared instrumentation, 1, 2.5 and 5% sodium hypochlorite (NaOCl)-based irrigants were used in 3 groups: GI (n=39), GII (n=36) and GIII (n=36), respectively. Before BMP, all cultures were positive and the smears showed microbiologically diverse morphotypes, including fusiforms, pleomorphic, rods, cocci and filaments. Quantitatively, 20, 20 and 23 morphotypes were identified in GI, GII and GIII, respectively). After BMP, the percentages of negative cultures in GI, GII and GIII were 74.2%, 86.3% and 93.4% (p>0.05) and the number of morphotypes decreased to 14, 15 and 5, respectively. All teeth with 2 root canals and/or associated fistulas were microbiologically negative after BMP, regardless of irrigant concentration. Gram-negative morphotypes were more susceptible to the action of irrigants. After irrigation with 5% NaOCl, only structural arrangements consisting of Gram-positive cocci and bacilli persisted. Thus, BMP plus 5% NaOCl offered the best antiseptic potential because in the few positive cultures a significant reduction in the number of microbiological morphotypes was also shown (p<0.05).

PMID: 17262145 [PubMed - indexed for MEDLINE]

Smear layer removal and chelated calcium ion quantification of three irrigating solutions.

Braz Dent J. 2006;17(4):306-9

Authors: Marques AA, Marchesan MA, Sousa-Filho CB, Silva-Sousa YT, Sousa-Neto MD, Cruz-Filho AM

The purpose of this study was to evaluate, by scanning electron microscopy (SEM), smear layer removal and quantify, by atomic absorption spectrophotometry, the amount of calcium ion present in the chelating solutions after their use. Sixteen extracted canines were instrumented using the step-back technique and were assigned to 3 groups according to the irrigating solution used: G1: 1 mL 17% EDTAC between each file; G2: 1 mL 17% CDTA; G3: 1 mL 17% EGTA. The solutions were collected after use. The teeth were cleaved longitudinally, evaluated under SEM and assessed for smear layer by blinded examiners and scored from 1 to 4. In order to quantify calcium ion release, the collected solutions were examined by atomic absorption spectrophotometry. Freidman's test was used for statistical analysis of SEM values and showed that canals irrigated with 17% EDTAC and 17% CDTA had significantly less smear layer throughout the canals than 17% EGTA (p<0.01). For analysis of the collected solutions, Tukey's test was used and showed that EDTAC and CDTA had a greater amount of calcium ions (22.8+/-7.54 and 60.6+/-20.67 microg/mL, respectively) compared to EGTA (70.5+/-14.2 microg/mL) (p<0.01). The association both methodologies may contribute to the understanding of how these solutions act in the root canal.

PMID: 17262144 [PubMed - indexed for MEDLINE]

Histological analysis of effects of 24% EDTA gel for nonsurgical treatment of periodontal tissues.

J Oral Sci. 2006 Dec;48(4):207-14

Authors: de Vasconcellos LM, Ricardo LH, Balducci I, de Vasconcellos LG, Carvalho YR

The aim of this study was to investigate, by means of histological and histomorphometric analysis, the effects of 24% ethylenediaminetetraacetic acid (EDTA) gel in periodontal tissue when used in combination with conventional periodontal treatment. Periodontitis was induced in the 2nd upper left permanent molars of 45 male Wistar rats by means of ligature. After 5 weeks, this was removed and debridement was performed. The animals were then randomly divided into 3 groups; group 1: mechanical treatment, group 2: mechanical treatment and EDTA gel application for 2 min, and group 3: mechanical treatment and placebo gel application for 2 min. After the treatment, rinsing was done with 0.9% saline solution for 1 min in all cases, followed by root notching in the deepest part of the pocket. After 4, 10, and 28 days the animals were sacrificed. The averages obtained were evaluated by means of test two-way analysis of variance (ANOVA) and Tukey statistical tests (P < 0.05). The results showed that with respect to the type of treatment employed, there were no statistically significant differences in the vitality of the periodontal tissue. It was concluded that 24% EDTA gel did not interfere with periodontal tissue repair when used in combination with conventional periodontal treatment.

PMID: 17220618 [PubMed - indexed for MEDLINE]

A scanning electron microscopic evaluation of root canal wall cleanliness after calcium hydroxide removal using three irrigation regimens.

J Contemp Dent Pract. 2007 Jan 1;8(1):11-8

Authors: Naaman A, Kaloustian H, Ounsi HF, Naaman-Bou Abboud N, Ricci C, Medioni E

AIMS: The aims of this in vitro study were to evaluate debris and smear layer elimination capability effectiveness of 5.25% sodium hypochlorite (NaOCl) after calcium hydroxide removal from the root canal and to determine if ethylenediaminetetraacetic acid (EDTA) and citric acid have any additional effects on debris and smear layer removal. METHODS AND MATERIALS: All irrigation regimens are associated with ultrasonics. Thirty-six pulpless single-rooted teeth were divided into three groups on the basis of the irrigation regimen, namely 5.25% NaOCl alone, 5.25% NaOCl with 17% EDTA pH 7 (NaOCl-EDTA), and 5.25% NaOCl with 50% citric acid (NaOCl-CA). All root canals were instrumented with the Protaper rotary system (Dentsply-Maillefer, Ballaigues, Switzerland) using a crown-down approach. The apical foramen of each tooth was enlarged to a size 30 with a 0.09 taper. After longitudinal sectioning, the dentin walls were microphotographed using scanning electron microscopy (SEM) at a 1500x magnification. Using a grid and a 5-step scale these microphotographs were scored by three investigators in order to evaluate debris and smear layer residuals. Data were statistically evaluated using Kruskal-Wallis and Mann-Whitney tests. RESULTS: In the coronal third the lowest score obtained with NaOCl-CA was similar to NaOCl-EDTA. When using NaOCl-EDTA, mean debris scores were similar to the control group (NaOCl) and lower than NaOCl-CA in the middle and apical thirds. Regarding the mean smear layer score, NaOCl-CA was the lowest in the coronal, middle, and apical thirds with a statistically significant difference (p < 0.001). CONCLUSION: When the total surface of the root was evaluated, NaOCl-EDTA was superior in removing debris, but the association of NaOCl-CA enabled the most effective removal of the smear layer.

PMID: 17211500 [PubMed - indexed for MEDLINE]

A scanning electron microscopic evaluation of different root canal irrigation regimens.

Braz Oral Res. 2006 Jul-Sep;20(3):235-40

Authors: M&#xF4;nika CM, Fröner IC

The purpose of this study was to assess the effectiveness of endodontic irrigants in removing the smear layer from instrumented root canal walls using Scanning Electron Microscopy (SEM). The endodontic irrigants used were: 1% sodium hypochlorite (NaOCl); 1% NaOCl mixed to 17% EDTAC; 2% chlorhexidine gel; and Ricinus communis gel. Photomicrographs of the middle and apical thirds were evaluated with the aid of the Fotoscore - v. 2.0 software. The results indicated that the mixture of sodium hypochlorite and EDTAC completely removed the smear layer from dentinal walls. The other endodontic irrigants were not as efficient in cleansing the root canals.

PMID: 17119706 [PubMed - indexed for MEDLINE]

Scanning electron microscopic analysis of the effect of Carisolv gel on periodontally compromised human root surfaces.

Braz Dent J. 2006;17(2):110-6

Authors: Grisi DC, Theodoro LH, Sampaio JE, Grisi MF, Salvador SL

The aim of this study was to analyze, under scanning electron microscopy (SEM), the morphologic characteristics of root surfaces after application of Carisolv gel in association with scaling and root planing (SRP). Sixty periodontally compromised extracted human teeth were randomly assigned to 6 groups: 1) SRP alone; 2) passive topical application of Carisolv + SRP; 3) active topical application of Carisolv + SRP; 4) multiple applications of Carisolv + SRP; 5) SRP + 24% EDTA; 6) topical application of Carisolv + SRP + 24% EDTA. Carisolv gel was applied to root surfaces for 30 s, followed by scaling and root planing, consisting of 50 strokes with Gracey curettes in an apical-coronal direction, parallel to the long axis of the tooth. The only exception was group 4, in which the roots were instrumented until a smooth, hard and glass-like surface was achieved. All specimens were further analyzed by SEM. The results showed that the treatment with Carisolv caused significant changes in root surface morphology of periodontally compromised teeth only when the chemical agent was actively applied (burnishing technique). Carisolv failed to remove the smear layer completely, especially with a single application, independently of the method of application. Multiple applications of Carisolv were necessary to achieve a smear layer reduction comparable to that obtained with 24% EDTA conditioning.

PMID: 16924336 [PubMed - indexed for MEDLINE]

Effect of rotary instrumentation and of the association of calcium hydroxide and chlorhexidine on the antisepsis of the root canal system in dogs.

Braz Oral Res. 2006 Apr-Jun;20(2):120-6

Authors: Soares JA, Leonardo MR, da Silva LA, Tanomaru Filho M, Ito IY

This study aimed at evaluating the antisepsis of the root canal system (RCS) and periapical region (PR) provided by rotary instrumentation associated with chlorhexidine + calcium hydroxide as intracanal medicament. Chronic periapical lesions were induced in 26 pre-molar roots in two dogs. After microbiological sampling, automatic instrumentation using the Profile system and irrigation with 5.25% sodium hypochlorite solution, with a final rinse of 14.3% EDTA followed by profuse irrigation with physiological saline were carried out in 18 root canals. After drying the canals, a paste based on calcium hydroxide associated with a 2% chlorhexidine digluconate solution was placed inside them. After 21 days, the medication was removed, leaving the root canals empty and coronally sealed. After 96 hours, a final microbiological sample was obtained, followed by histomicrobiological processing by the Brown & Brenn method. Eight untreated root canals represented the control group (C-G). Based on the Mann-Whitney test at a confidence level of 5% (p < 0.05), the procedures of antisepsis used offered significant efficacy (p < 0.05) resulting in 100.0% of the canals free of microorganisms. In the C-G, an elevated incidence of various microbial morphotypes was confirmed in all sites of the RCS, with the presence of microbial colonies in the periapical region. In contrast, the experimental group showed a similar pattern of infection in the RCS, although less intense and a reduced level of periapical infection (p < 0.05). It was concluded that adequate instrumentation followed by the application of calcium hydroxide + chlorhexidine offered significant elimination of microorganisms.

PMID: 16878204 [PubMed - indexed for MEDLINE]

Response of rice (Oryza sativa) with root surface iron plaque under aluminium stress.

Ann Bot (Lond). 2006 Aug;98(2):389-95

Authors: Chen RF, Shen RF, Gu P, Dong XY, DU CW, Ma JF

BACKGROUND AND AIMS: Rice (Oryza sativa) is an aquatic plant with a characteristic of forming iron plaque on its root surfaces. It is considered to be the most Al-tolerant species among the cereal crops. The objective of this study was to determine the effects of root surface iron plaque on Al translocation, accumulation and the change of physiological responses under Al stress in rice in the presence of iron plaque. METHODS: The japonica variety rice, Koshihikari, was used in this study and was grown hydroponically in a growth chamber. Iron plaque was induced by exposing the rice roots to 30 mg L(-1) ferrous iron either as Fe(II)-EDTA in nutrient solution (6 d, Method I) or as FeSO(4) in water solution (12 h, Method II). Organic acid in root exudates was retained in the anion-exchange resin and eluted with 2 m HCl, then analysed by high-performance liquid chromatography (HPLC) after proper pre-treatment. Fe and Al in iron plaque were extracted with DCB (dithionite-citrate-bicarbonate) solution. KEY RESULTS AND CONCLUSIONS: Both methods (I and II) could induce the formation of iron plaque on rice root surfaces. The amounts of DCB-extractable Fe and Al on root surfaces were much higher in the presence of iron plaque than in the absence of iron plaque. Al contents in root tips were significantly decreased with iron plaque; translocation of Al from roots to shoots was significantly reduced with iron plaque. Al-induced secretion of citrate was observed and iron plaque could greatly depress this citrate secretion. These results suggested that iron plaque on rice root surfaces can be a sink to sequester Al onto the root surfaces and Fe ions can pre-saturate Al-binding sites in root tips, which protects the rice root tips from suffering Al stress to a certain extent.

PMID: 16735401 [PubMed - indexed for MEDLINE]

[Root canal therapy of resinified teeth with surgical operating microscope and ultrasonic instruments]

Shanghai Kou Qiang Yi Xue. 2006 Apr;15(2):117-20

Authors: Xu Q, Lin JQ, Mai S, Liu JW

PURPOSE: To evaluate the role of surgical operating microscope and ultrasonic instruments in endodontic treatment of patients who underwent previous resinifying therapy. METHODS: 63 premolars and molars that had been treated with resinifying therapy before were selected for root canal therapy. The coronal portion of resinified root canals were negotiated with ultrasonic instruments under surgical operating microscope, and the apical portion was managed with small size hand files and 15% EDTA. The root canals of all cases were shaped by Nickel-Titanium rotary instruments Hero 642, and obturated with lateral condensation technique. The negotiation of root canal system, instrument fracture, alterations of canal morphology, and operation time were recorded. The efficiency of preparation and obturation was analyzed by radiographs before and after treatment. RESULTS: The root canals of 54 teeth were negotiated, enlarged and obturated, with a success rate of 85.7%. No complications were found, such as vertical fracture, ledge, perforation and instrument separation. Good result of treatment was achieved in the 54 cases. CONCLUSIONS: The use of surgical operating microscope and ultrasonic instruments is proved to be effective in negotiation of coronal portion of the resinified root canals. Nevertheless, the use of ultrasonic instruments in apical or curved portion of root canals is not encouraged.

PMID: 16685346 [PubMed - in process]

The attachment of V79 and human periodontal ligament fibroblasts on periodontally involved root surfaces following treatment with EDTA, citric acid, or tetracycline HCL: an SEM in vitro study.

J Contemp Dent Pract. 2006 Feb 15;7(1):44-59

Authors: Chandra RV, Jagetia GC, Bhat KM

OBJECTIVE: The present in vitro study has been designed to establish and compare the effects of citric acid, EDTA, and tetracycline HCl on human periodontally diseased roots on the structure, attachment, and orientation of V79 (primary Chinese hamster lung fibroblasts) cells and human periodontal ligament fibroblasts (HPDL). MATERIALS AND METHODS: Commercially available V79 cells and HPDL derived from healthy human third molars were used in this study. These fibroblasts were left in solution for seven days in order to attain confluence. Forty single-rooted teeth were obtained from patients diagnosed with periodontitis. The crown part was removed under constant irrigation and the root was split vertically into two equal halves, thus, yielding 80 specimens. Following scaling and root planing, the specimens were washed with phosphate buffered saline (PBS) and kept in 50 microg/ml gentamycin sulphate solution for 24 hours. The root pieces were then treated as follows: citric acid at pH 1, 24% EDTA, or with a 10% solution of tetracycline HCl and were then placed in V79 fibroblast cultures and HPDL cultures. The specimens were harvested after four weeks and were fixed in 2.5% glutaraldehyde in PBS before preparation for scanning electron microscopy (SEM). RESULTS: The behavior of V79 cells was similar to that of human periodontal ligament cells on root conditioned surfaces. V79 and HPDL showed a healthy morphology on root surfaces treated with citric acid and EDTA and a relatively unhealthy appearance on root surfaces treated with tetracycline HCl and distilled water (control group). CONCLUSION: The results suggest the use of citric acid and EDTA as root conditioning agents favorably affects the migration, attachment, and morphology of fibroblasts on human root surfaces, which may play a significant role in periodontal healing and regeneration.

PMID: 16491147 [PubMed - indexed for MEDLINE]

Evaluation of the smear layer removal effectiveness of EDTA using two techniques: an SEM study.

J Contemp Dent Pract. 2006 Feb 15;7(1):9-16

Authors: Tinaz AC, Karadag LS, Ala&#xE7;am T, Mihçioglu T

Obtaining the cleanest canal possible before obturation is one of the goals of endodontic treatment. The purpose of this study was to compare the smear layer removal capability of ethylenediamine tetra-acetic acid (EDTA) application with passive ultrasonic and cotton wrapped on reamer activation. Twelve extracted human teeth with single root canals were used for the study. They were conventionally hand instrumented using reamers and K files. The apical aspect of the canals was enlarged to a #40 file. The teeth were divided into 4 four groups, instrumented and irrigated as follows: Group-A EDTA agitated at the end of preparation with cotton wrapped on a reamer for 1 min; Group-B EDTA applied with ultrasonic agitation for 1 min; Group-C irrigated with EDTA+sodium hypochlorite (NaOCl) (negative control group); and Group-D irrigated with distilled water (positive control group). After scanning electron microscopic study at three different levels, smear layer and dentinal tubules were scored. Means were tested for significance using the Z test. When the techniques were compared, the cotton wrapped on reamer agitation method was as successful as the ultrasonic activation of the files. Although all groups had significantly higher smear layer scores at apical compared to coronal sections, no significant differences were recorded.

PMID: 16491143 [PubMed - indexed for MEDLINE]

Inability of laser and rotary instrumentation to eliminate root canal infection.

J Am Dent Assoc. 2006 Jan;137(1):67-70

Authors: Jha D, Guerrero A, Ngo T, Helfer A, Hasselgren G

BACKGROUND: The authors evaluated the antibacterial effectiveness of laser instrumentation and rotary instrumentation of anterior, single-rooted teeth infected with Enterococcus faecalis. METHODS: The authors divided 35 infected samples into five groups: Group A: inoculation, laser, 17 percent ethylene-diamine-tetra-acetate (EDTA), 2.5 percent sodium hypochlorite (NaOCl) (n=10); Group B: inoculation, laser, 17 percent EDTA, sterile saline (n = 10); Group C: inoculation, rotary, 17 percent EDTA, 2.5 percent NaOCl (n=10); Group D: inoculation, no instrumentation (positive control) (n=5); Group E: no inoculation, no instrumentation (negative control) (n=5). They sampled and incubated dentin shavings from each canal for bacterial growth. RESULTS: In Group A, eight tubes were positive for bacterial growth. In Group B, 10 tubes were positive for bacterial growth. In Group C, six tube were positive for bacterial growth. In Group D, all of the tubes were positive for bacterial growth. In Group E, no tubes showed bacterial growth. The Fisher exact test showed no significant differences among groups A, E and C. CONCLUSION: Neither the laser nor the rotary instrumentation was able to eliminate endodontic infection. CLINICAL IMPLICATIONS: Although lasers have been presented as high-tech tools for disinfecting root canals, the laser was ineffective in this study.

PMID: 16457001 [PubMed - indexed for MEDLINE]

In vitro evaluation of apical sealing in root apex treated with demineralization agents and retrofiled with mineral trioxide aggregate through marginal dye leakage.

Braz Dent J. 2005;16(3):187-91

Authors: Kubo CH, Gomes AP, Mancini MN

The purpose of this study was to evaluate the apical seal in root apex treated with different demineralization agents and retrofilled with mineral trioxide aggregate (MTA) using marginal dye leakage. Fifty-six, human single-rooted teeth were instrumented, filled, resected and had retrofilling cavities prepared with ultrasonic tips. Demineralizing agents were applied before the apical cavities were retrofilled with Pro Root MTA. The specimens were assigned to 4 groups (n=14), as follows: group 1 (no demineralizing agent); group 2 (35% phosphoric acid, for 15 s); group 3 (17% EDTA solution, pH 7, for 3 min); and group 4 (24% EDTA gel, pH 7, for 4 min). The extension of dye (2% rhodamine B, at 37 degrees C, for 24 h) penetration was measured in millimeters using a stereomicroscope. Results were statistically analyzed by ANOVA and Tukey's test at 5% significance level. Among the experimental groups, the least extension of dye penetration was observed in group 1 (1.89 mm), followed by groups 2 (2.18 mm), 4 (2.54 mm) and 3 (2.64 mm). No statistically significant differences (p<0.05) were found in marginal microleakage among groups 1, 2 and 4 and groups 2, 3 and 4. Based on the results obtained in this study, it may be concluded that the application of demineralizing agents cannot be recommended when MTA is used in periradicular surgeries.

PMID: 16429182 [PubMed - in process]

Demineralization effect of EDTA, EGTA, CDTA and citric acid on root dentin: a comparative study.

Braz Oral Res. 2005 Jul-Sep;19(3):188-92

Authors: Sousa SM, Silva TL

The purpose of this study was to biochemically compare the decalcifying effects of 1% EDTA (pH 7.4), 1% EGTA (pH 7.4), 1% CDTA (pH 7.4), 1% citric acid solutions (pH 1.0 and 7.4) and saline solution (control) on root dentin. Forty-eight single-rooted teeth were used in this study. The canals were instrumented by the step-back technique and the roots were randomly divided into six equal experimental groups (n = 8) according to the irrigating agent tested. A total of 30 microL of each solution was pipetted into the root canal and allowed to set undisturbed for 5 minutes. After this time, 15 microL of the solutions were removed from each canal using a Hamilton syringe and placed in a container with 5 mL of deionised water. The microg/mL concentration of calcium ion (Ca2+) extracted from the root canal samples was determined using inductively coupled plasma-atomic emission spectrometry (ICP-AES). Data were analysed by means of the Kruskal-Wallis and Mood's median tests. Citric acid solution at pH 1.0 removed more calcium than at pH 7.4 and than the other chelating solutions tested (p < 0.05). No differences were observed between EDTA and EGTA. Both EDTA and EGTA removed significantly more calcium than CDTA and citric acid at pH 7.4 (p < 0.05). There were no differences between citric acid at pH 7.4 and saline solution, which had the least efficacy for Ca2+ extraction (p > 0.05). These results indicate that citric acid at pH 1.0 is a good alternative as an irrigating solution to remove the smear layer and facilitate the biomechanical procedures.

PMID: 16308606 [PubMed - indexed for MEDLINE]

Blood cell attachment to root surfaces treated with EDTA gel.

Braz Oral Res. 2005 Apr-Jun;19(2):88-92

Authors: Leite FR, Moreira CS, Theodoro LH, Sampaio JE

Root debridement generates a smear layer which contains microorganisms and toxins that could interfere in periodontal healing. For this reason, different substances have been used to remove it and to expose collagen fibers at the tooth surface. Blood element adhesion to demineralized roots and clot stabilization by collagen fibers are extremely important for the success of periodontal surgery. The aim of this study was to evaluate the different patterns of blood element adsorption and adhesion to root surfaces only irrigated with distilled water and after application of a manipulated or an industrialized EDTA gel. Thirty samples were planed, equally divided into three groups and treated with distilled water (control), a manipulated EDTA gel or an industrialized one. Immediately after, samples were exposed to fresh blood and prepared for scanning electron microscopy. Untreated planed dentin presented the best results with blood cells entrapped in a thick web of fibrin. In the manipulated EDTA group, the web of fibrin was thick with sparse blood elements. The worst result was seen with the industrialized EDTA group, in which no blood elements could be seen. Statistical difference was obtained between control and industrialized EDTA groups. Surfaces only irrigated presented the most organized fibrin network and cell entrapment.

PMID: 16292439 [PubMed - indexed for MEDLINE]

Biocompatibility of EDTA, EGTA and citric acid.

Braz Dent J. 2005;16(1):3-8

Authors: Sousa SM, Bramante CM, Taga EM

This in vivo study evaluated, through the physicochemical assay method for quantification of enhanced vascular permeability, the irritating potential of EDTA, EGTA, citric acid and saline. Thirty-two male Wister rats were anesthetized and four experimental sites were demarcated on their backs. Injections of 2% Evans blue (20 mg/kg) were administered intravenously into the lateral caudal vein. The test solutions were immediately injected intradermally (0.01 mL) into the experimental sites. The animals were killed 30 min, 1, 3 and 6 h after injection of the solutions and each piece of skin was submerged in formamide and incubated at 45 masculineC for 72 h. After filtration, the optical density was measured in a spectrophotometer and the total amount of dye extracted from the samples was calculated by means of a standard calibration curve. Data were analyzed statistically by two-way ANOVA and Tukey's HSD test. Compared to control, EDTA had the greatest volume of dye followed by EGTA and citric acid, for all time periods. There were statistically significant differences between all solutions (p<0.01). Considering the periods assessed, a significant difference was observed between the 3- and 6-h groups (p<0.05), but not between the 30-min and 1-h groups. Among the organic acids evaluated in this study, citric acid yielded the lowest amount of extracted dye. This indicates that the citric acid was the least irritating solution.

PMID: 16113926 [PubMed - indexed for MEDLINE]

[Comparison of the effects of 5 irragant combinations on cleaning of anterior straight root canal walls]

Shanghai Kou Qiang Yi Xue. 2005 Jun;14(3):293-7

Authors: Li C, DU R, Zhu YQ

PURPOSE: To evaluate the effect of 5 kinds of irrigant combinations on cleaning of anterior straight root canal walls. METHODS: 25 extracted human maxillary anterior teeth, with a single straight root were randomly divided into 5 groups. They were instrumented with K-files and conventional preparation technique. The irrigations were as follow: in group 1,the canals were irrigated with 1% NaOCl and 3% H2O2 in sequence during and after instrumentation.In group 2,the canals were irrigated with 1% NaOCl during instrumentation and after instrumentation being irrigated with 17% EDTA. In group 3,irrigation during and after instrumentation was accomplished using 17% EDTA and 1% NaOCl in sequence. In group 4,irrigation during instrumentation was accomplished using 17% EDTA and 1% NaOCl in sequence, and after instrumentation being irrigated with 17% EDTA.In group 5,the canals were irrigated with 17% EDTA, 1% Triton X-100 and 1% NaOCl in sequence during instrumentation and irrigated with 17% EDTA after instrumentation. The total amount of irrigant was 22ml and the irrigation time was 7 minutes in each root canal. The root was split longitudinally and 3 specimens in each group were evaluated regarding to the smear layer, debris and erosion of dentinal tubules with SEM at the coronal, middle and apical thirds. RESULTS: In group 1, typical amorphous smear layer and abundance of debris were seen on the wall of canals. In group 2, the smear layer was removed partly in coronal and middle thirds, but the apical surface of the canal walls were covered with a typical amorphous smear layer. Although irrigants of group 3 could remove smear layer effectively, it resulted in moderate erosion of dentinal tubules. In group 5, part of the smear layer was removed on the surface of coronal and middle thirds but abundance of debris were present on the surface at the apical levels. Severe erosion was noted on the root canal surfaces in this group. Irrigation of group 4 had the best result: it not only removed smear layer effectively ,but also had no erosion on dentinal tubules. CONCLUSION: Using the combination of 17% EDTA and 1% NaOCl can remove smear layer effectively and has no erosion on the dentinal tubules under the condition of strict time control and irrigation sequence. Supported by Phosphor Plan (No.01QB14025) of Science and Technology of Shanghai Municipality.

PMID: 15995780 [PubMed - in process]

Changes of root morphology and Pb uptake by two species of Elsholtzia under Pb toxicity.

J Zhejiang Univ Sci B. 2005 Jun;6(6):546-52

Authors: Peng HY, Tian SK, Yang XE

Elsholtzia argyi and Elsholtzia splendens, which are Chinese endemic Pb/Zn mined and Cu mined ecotype respectively, were investigated in the aspect of their response to Pb toxicity in the presence or absence of EDTA addition. After 8 d's Pb treatment, root length, root surface area and root volume of E. splendens decreased much more than those of E. argyi, and reduced considerably with increase of Pb, while no marked change was noted for root average diameter. Compared to E. argyi, length of root with diameter (D)<0.2 mm was significantly reduced for E. splendens as Pb increasing. Root with cross-sectional area of D<0.1 mm for E. splendens was at Pb> or =10 mg/L, while for E. argyi, it was at Pb> or =25 mg/L. DW of E. splendens decreased much more than that of E. argyi with increase of Pb. E. argyi exhibited much more tolerance to Pb toxicity than E. splendens. Treatment with 100 mg/L Pb plus 50 mmol/L EDTA significantly decreased the length and surface area of D< or =0.2 mm root, increased the length and surface area of 0.2< or =D< or =0.8 mm root for the case of E. argyi, while for E. splendens, length and surface area of D<0.6 mm root reduced, as compared to 100 mg/L Pb treatment, alone. At 100 mg/L Pb, shoot Pb accumulation in E. splendens and E. argyi were 27.9 and 89.0 microg/plant DW respectively, and much more Pb was uptaken by the root and translocated to the stem of E. argyi as compared to E. splendens. Treatment of the plant with 100 mg/L Pb plus 50 mmol/L EDTA increase leaf Pb accumulation from 16.8 to 84.9 g/plant for E.splendens and from 18.8 to 52.5 g/plant for E. argyi, while both root and stem Pb pronouncedly reduced for both Elsholtzia species. The increased translocation of Pb to the leaf of E. splendens than that of E. argyi at the treatment of 100 mg/L Pb plus 50 mmol/L EDTA should be further investigated.

PMID: 15909342 [PubMed - indexed for MEDLINE]

Influence of Er: YAG laser irradiation on apical sealing of four different sealers.

Braz Dent J. 2004;15(3):190-3

Authors: Mello I, Robazza CR, Antoniazzi JH

The sealing of the root canal system is of fundamental importance for successful endodontic treatment. To obtain an adequate apical seal, many factors must be considered such as the presence of smear layer and the sealer applied. After canal preparation, this layer must be removed because it prevents close contact between the dentinal walls and the sealing material. The goal of this study was to evaluate the sealing ability of four different sealers after smear layer removal with either 17% EDTA-T irrigation or Er:YAG laser irradiation of 46 teeth. The canals were sealed with four different sealers: Sealapex, Ketac Endo, AH Plus and N-Rickert. The method for smear layer removal did not influence apical sealing. AH Plus and N-Rickert allowed less dye leakage when compared to Sealapex and Ketac Endo.

PMID: 15798821 [PubMed - in process]

Short- and longer-term effects of the willow root system on metal extractability in contaminated dredged sediment.

J Environ Qual. 2004 May-Jun;33(3):976-83

Authors: Vervaeke P, Tack FM, Lust N, Verloo M

Willow (Salix spp.) stands are often proposed as vegetation covers for the restoration and stabilization of contaminated and derelict land. Planting willows on dredged sediment disposal sites for biomass production can be an alternative to traditional capping techniques. However, with the introduction of willow stands on dredged sediment disposal sites, the possibility of increased contaminant availability in the root zone must be acknowledged as it can increase the risk of leaching. Two trials investigated the availability of Cd, Zn, Cu, and Pb in the root zones of willows grown on contaminated sediment. To assess the effects of willow root growth on metal extractability and mobility, bulk and rhizosphere sediment samples were extracted with deionized water, ammonium acetate at pH 7, and ammonium acetate-EDTA at pH 4.65. A rhizobox experiment was used to investigate the short-term effect of willow roots on metal availability in oxic and anoxic sediment. Longer-term effects were assessed in a field trial. The rhizobox trial showed that Cd, Zn, and Cu extractability in the rhizosphere increased while the opposite was observed for Pb. This was attributed to the increased willow-induced oxidation rate in the root zone as a result of aeration and evapotranspiration, which masked the direct chemical and biological influences of the willow roots. The field trial showed that Cu and Pb, but not Cd, were more available in the root zone after water and ammonium acetate (pH 7) extraction compared with the bulk sediment. Sediment in the root zone was better structured and aggregated and thus more permeable for downward water flows, causing leaching of a fraction of the metals and significantly lower total contents of Cd, Cu, and Pb. These findings indicate that a vegetation cover strategy to stabilize sediments can increase metal availability in the root zone and that potential metal losses to the environment should be considered.

PMID: 15224934 [PubMed - indexed for MEDLINE]

Transmembrane electron transport in sealed and NAD(P)H-loaded right-side-out plasma membrane vesicles isolated from maize (Zea mays L.) roots.

J Exp Bot. 2004 Jun;55(401):1343-9

Authors: Menckhoff M, L&#xFC;thje S

Electron transport across plasma membranes has been observed in vivo in several plant species and tissues after the application of ferricyanide (hexacyanoferrate III, HCF III). In the present work, a transmembrane electron flow was demonstrated in sealed and NAD(P)H-loaded right-side-out (apoplastic-side-out) plasma membrane vesicles isolated from maize (Zea mays L.) roots. HCF III was reduced at a rate of up to 126 nmol min(-1) mg(-1) protein by NADPH-loaded vesicles, while reduction rates with NADH-loaded vesicles were several-fold lower. Coincident with the reduction of HCF III, NAD(P)H oxidation was observed inside the vesicles. The dependence of reduction on K+ indicated an electrogenic transmembrane electron flow. Application of 100 microM calcium decreased HCF III reduction up to 66%, while pre-incubation with 200 microM warfarin or diphenylene iodonium inhibited transmembrane electron transport only weakly. Fe(3+)-EDTA was not reduced significantly by NADPH-loaded plasma membrane vesicles, whereas XTT was reduced at a rate of 765 pmol min(-1) mg(-1) protein. The results suggested a major function for NADPH in transmembrane electron flow and were discussed in conjunction with in vivo experiments.

PMID: 15155782 [PubMed - indexed for MEDLINE]

The effects of water and acetone-based dentin adhesives on apical microleakage.

J Contemp Dent Pract. 2004 May 15;5(2):93-101

Authors: Karadag LS, Bala O, Türköz E, Mihçioğlu T

In this study, the aim was to assess the in vitro apical microleakage of a resin-based sealer used with two different adhesives. Thirty nine freshly extracted maxillary incisors were used. The teeth were decoronated at the cemento-enamel junction with a water-cooled fissure bur. Chemo-mechanical debridement of the root canals was accomplished with the step-back technique. The smear layer was removed by 19% ethylenediamine tetra acetic acid (EDTA). The roots were then divided into three experimental groups of thirteen teeth in each. Specimens in group 1 were filled with gutta-percha, AH Plus sealer, and water-based adhesive system (Syntac Single Component). Group 2 specimens were filled with gutta-percha, AH Plus sealer, and acetone-based dentin adhesive (Prime & Bond NT ). Specimens of group 3 were filled with only gutta-percha and AH Plus sealer (no adhesive was applied). The teeth were immersed into 2% methylene blue solution. Apical sealing qualities were assessed by measuring the linear dye penetration with a stereomicroscope. Dentin tubule penetration was observed under scanning electron microscopy (SEM). Results showed no statistically significant difference between the materials used, however, the leakage in group 2 was less than group 1 and 3.

PMID: 15150637 [PubMed - indexed for MEDLINE]

Smear layer removal capacity of disinfectant solutions used with and without EDTA for the irrigation of canals: a SEM study.

Pesqui Odontol Bras. 2003 Oct-Dec;17(4):349-55

Authors: Menezes AC, Zanet CG, Valera MC

The purpose of this study was to carry out a scanning electron microscopic (SEM) analysis of the cleaning qualities and smear layer removal from root canal walls, instrumented and irrigated with 2.5% NaOCl, 2.0% chlorhexidine and saline solutions. Fifty extracted teeth were used in this study. All teeth were radiographed to determine the existence of a single canal. The crowns were cut at the cervical limit and the root canals were instrumented with K-type files up to size 45. During root canal preparation, irrigations were made with the different solutions being evaluated: Group 1: 2.5% NaOCl (10 roots); Group 2: 2.5% NaOCl and 17% EDTA for 2 minute (10 roots); Group 3: 2.0% chlorhexidine (10 roots); Group 4: 2.0% chlorhexidine and 17% EDTA for 2 minutes (10 roots); Group 5: saline solution (5 roots); Group 6: saline solution and 17% EDTA for 2 minutes (5 roots). After instrumentation, the canals were irrigated with each one of the solutions and the roots were cut in the buccolingual direction for SEM analysis, at the cervical, middle and apical thirds, to ascertain the presence or absence of smear layer and debris. SEM analysis was performed by three calibrated examiners and scores were submitted to Kruskal-Wallis test at the significance level of p = 5%. Results showed that the use of 17% EDTA decreased the smear layer significantly (p < 0.05) for all evaluated solutions in all thirds. When EDTA was not used, a significantly higher quantity of smear layer on the apical third was observed only in the NaOCl groups. The use of 17% EDTA was significant for debris removal except for the chlorhexidine groups. The following conclusion could be drawn: the use of 17% EDTA was necessary to enhance cleanness of the root canals.

PMID: 15107918 [PubMed - indexed for MEDLINE]

Effectiveness of EDTA and EDTA-T brushing on the removal of root surface smear layer.

Pesqui Odontol Bras. 2003 Oct-Dec;17(4):319-25

Authors: Sampaio JE, Rached RS, Pilatti GL, Theodoro LH, Batista LH

The purpose of this study was to compare the removal of root surface smear layer following active application of EDTA gel and EDTA-T (texapon) gel in different concentrations (5%, 10%, 15%, 20% and 24%), using scanning electron microscopy. A total of 220 dentin blocks obtained from the root surfaces of extracted teeth were divided into 3 groups: Group I - (control) application of saline solution (n = 20); Group II - EDTA gel (pH 7.0) was applied in the following concentrations: 5%, 10%, 15%, 20% and 24% (n = 100); Group III - EDTA-T gel (pH 7.0) applied in the same concentrations described above (n = 100). The photomicrographs were evaluated by one calibrated examiner using a smear layer removal index and following statistical analysis (Kruskal-Wallis test). The results demonstrated that the specimens treated with EDTA and EDTA-T gel presented a better smear layer removal than the control group (p < 0.01); no statistically significant differences were observed between the EDTA and EDTA-T groups and between the concentrations tested (Mann-Whitney, p > 0.05). Within the limits of this study, it can be concluded that all treatment modalities effectively removed the smear layer from the root surface. The addition of texapon into the EDTA gel formulation did not increase its effectiveness.

PMID: 15107913 [PubMed - indexed for MEDLINE]

[Experimental research on preparation of small root canals with EDTA and cirtric acid]

Shanghai Kou Qiang Yi Xue. 1998 Jun;7(2):67-9

Authors: Chen YZ, Weng YL

OBJECTIVE:To study the effects of EDTA and citric acid on small root canals so as to provide experimental basis for clinical applications.METHODS:17% EDTA and 50% citric acid are applied to Group A and Group B respectively for three days, each group having 14 extracted teeth with small root canals. The data obtained during the three days' experiment are analyzed with statistic method.RESULTS:The 14 small root canals applied with 17% EDTA solution for three days are all effective, the overall effectiveness rate being 100% and 11 small root canals show an expansion level of 4mm or over, 9 of which reachthe length required for treatment, 11 out of the 14 small root canals applied with 50% citric acid solution for three days are effective (three are not effective), the overall effectiveness rate being 78.57%. 8 small root canals show an expansion level of over 4 mm and all of them reach the length required for treatment.Based upon statistic treatment, the expansion level of small root canals of both Group A and Group B has an obvious difference (P<0.01) after the application.CONCLUSION:EDTA should be the recommended medicine for chemical preparation of small root canals.

PMID: 15071668 [PubMed - as supplied by publisher]

[The effect of EDTA with different density on adhesive power of casting post and its scanning electromicroscopy (SEM) observation]

Shanghai Kou Qiang Yi Xue. 1999 Mar;8(1):7-10

Authors: Li YL, Liao HB

OBJECTIVE:To evaluate the effect of EDTA with different density on adhesive power of casting post.METHODS:Four different density of EDTA(5%,10%,15%,17%)were used to rinse the smear layer of root canal for 60 extracted teeth before they were cemented with ZP;on the other,a SEM observation was made on the rinse of root canal and the combination of adhesive and root canal.Meanwhile,a comparison was also made, after the rinse of EDTA,of tensile strength of casting post after their cement.RESULTS:EDTA with different density have different effect in the rinse of smear layer of root canal,it was proved by both SEM observation and the records of tensile strength of casting post. CONCLUSION:EDTA with density 10% is more effective in both the rinse of smear layer of root canal and the enhancement of adhesive power of casting post.

PMID: 15048303 [PubMed - as supplied by publisher]

Influence of passive ultrasonic activation on the penetration depth of different sealers.

J Contemp Dent Pract. 2004 Feb 15;5(1):115-25

Authors: Karadag LS, Tinaz AC, Mih&#xE7;ioğlu T

Correct use of instruments and irrigating solutions are major factors during chemo-mechanical preparation of root canals. Ultrasonically activated files and solutions are used for debridement of canals. In this study 42 single rooted anterior human teeth were used. The roots were divided into two groups. One group received an application of 17% ethylenediamine tetra acetic acid (EDTA) of 1 ml for 1 min using an ultrasonically energized file. The other group received an application of 17% EDTA of 1 ml for 0.5 min using an ultrasonically energized file. The groups were divided again into two subgroups. The roots were filled with a resin based sealer, AH26 and a glass ionomer sealer--Endion with lateral condensation technique. The roots were separated into two halves. The specimens were viewed through a scanning electron microscope (SEM). Statistical differences between the groups and regions were analyzed with Variance and Duncan tests. Under the conditions of this study, it can be said that ultrasonically activated irrigation did not reduce the smear layer effectively at 1 min and 0.5 min time intervals. According to this result, no difference was observed between the penetration of sealers.

PMID: 14973565 [PubMed - indexed for MEDLINE]

Modeling organic carbon dynamics and cadmium fate in long-term sludge-amended soil.

J Environ Qual. 2004 Jan-Feb;33(1):181-91

Authors: Bergkvist P, Jarvis N

A model is described that may help to resolve uncertainty and controversy over the long-term consequences of sludge applications to arable land, especially with regard to the effects of sludge adsorption characteristics on trace metal solubility and bioavailability (e.g., the sludge "time bomb" or sludge "protection" hypotheses). Mass balances of organic and inorganic material derived from sludge and crop residues are simulated. Each pool has a potentially different adsorption affinity for trace metals, and this leads to changes in the adsorption capacity of sludge-amended soil that influence leaching and crop uptake. Model simulations were compared with measured changes in organic carbon and ethylenediaminetetraacetic acid (EDTA)-extractable cadmium contents in a clay loam soil following 41 years of sludge applications. The model adequately reproduced the data, although discrepancies in the vertical distribution of Cd were attributed to the effects of macropore transport and root-uptake driven recirculation. A Monte Carlo sensitivity analysis demonstrated that the most important parameters affecting leaching and crop uptake were the Cd loading and parameters controlling adsorption, especially the partition coefficient for sludge-derived inorganic material and the exponent regulating the effect of pH on sorption. Scenario simulations show that no general conclusions can be drawn with respect to the validity of the sludge "time bomb" and sludge "protection" hypotheses. Either may occur, or neither, depending on three key system parameters: the ratio of sludge adsorption capacity to the initial adsorption capacity of the soil, the proportion of the sludge adsorption capacity contributed by the inorganic fraction, and the sludge Cd loading.

PMID: 14964373 [PubMed - indexed for MEDLINE]

In vitro evaluation of the effects of the interaction between irrigating solutions, intracanal medication and Er:YAG laser in dentin permeability of the endodontic system.

Pesqui Odontol Bras. 2003 Jul-Sep;17(3):278-85

Authors: Rald DP, Lage-Marques JL

The purpose of this study was to evaluate in vitro the effects of different associations between irrigating solutions (EDTA-T and citric acid), intracanal medicament (NDP), and Er:YAG laser irradiation on dentin permeability. Fifty-one extracted single-rooted teeth were instrumented and divided into seven groups. Groups GI and GII had final irrigation with a demineralizing solution only (EDTA-T and citric acid, respectively). Groups GIII and GIV had final irrigation with EDTA-T and citric acid, respectively, plus an association of irrigating solution and Er:YAG laser. Groups GV and GVI had final irrigation with EDTA-T and citric acid, respectively, plus an association of intracanal medication and Er:YAG laser. Group GVII (control group) had final irrigation with distilled water. All root canals were filled with NDP associated with rhodamine B dye. After the experimental period, the samples were transversely cut into six 2.0 mm thick slices for subsequent reading using the ImageLab software. Analysis of the results allowed us to conclude that there were statistically significant differences (p < 0.05) between the groups as to the penetration of the dye-intracanal medication solution. Groups III and IV presented smaller values of dentinal permeability when compared to the other groups. The best results were obtained with the interaction between a demineralizing irrigating solution and the association of intracanal medicament and laser Er:YAG (groups V and VI). In these groups the observed penetration of the intracanal medicament plus dye solution in the apical third was, on average, 29% greater than in the other groups.

PMID: 14762508 [PubMed - indexed for MEDLINE]

Biogeochemistry of fluoride in a plant-solution system.

J Environ Qual. 2003 Nov-Dec;32(6):2230-7

Authors: Mackowiak CL, Grossl PR, Bugbee BG

Fluoride (F-) pollutants can harm plants and the animals feeding on them. However, it is largely unknown how complexing and chelating agents affect F bioavailability. Two studies were conducted that measured F- bioavailability and uptake by rice (Oryza sativa L.). In the first study, rice was grown in solution culture (pH 5.0) with 0, 2, or 4 mM F- as KF to compare the interaction of F- with humic acid (HA) and with a conventional chelating agent, N-hydroxyethylenthylenediaminetriacetic acid (HEDTA). In the second study, F was supplied at 0, 0.5, 1.0, and 2.0 mM KF with an additional 2 mM F- treatment containing solution Ca at 2x (2 mM Ca) the level used in the first study, to test the effect added Ca had on F- availability and uptake. Total biomass was greatest with HEDTA and F- < 1 mM. Leaf and stem F concentrations increased exponentially as solution F- increased linearly, with nearly no F partitioning into the seed. Results suggest that F was taken up as HF0 while F- uptake was likely restricted. Additionally, F- competed with HA for Ca, thus preventing the formation of Ca-HA flocculents. The addition of soluble Ca resulted in the precipitation of CaF2 solids on the root surface, as determined by tissue analysis and energy dispersive X-ray spectroscopy.

PMID: 14674546 [PubMed - indexed for MEDLINE]

[A comparative study of root smear layer removal using different pH EDTA salts]

Shanghai Kou Qiang Yi Xue. 2003 Jun;12(3):197-9

Authors: Ma R, Liu Z, Zhu CL

OBJECTIVE: To evaluate the effect of different pH EDTA salts on removing root canal smear layers. METHODS: Sixty human teeth with single root were instrumented using step-back technique, then were irrigated with several irrigating solutions including A: 0.9% saline; B: 5.25% NaOCl +3% H2O2; C: 15% EDTA (pH = 6.5); D: 15% EDTA (pH = 13); E: 15% EDTA (pH = 6.5) 25% NaOCl +3% H2O2; F: 15% EDTA (pH = 13) 25% NaOCl +3% H2O2. After the teeth were split, the root canal walls were examined with scanning electron microscopy at the coronal, middle and apical thirds for smear layer removal. RESULTS: Except A and B group, all the groups were effective on removing smear layer at the coronal, middle thirds of the root canal, group C had a stronger effect to remove smear layer than group D (P < 0.05), group E was the most effective among these groups. However, these groups were all ineffective on removing smear layer at the apical thirds of root canal. CONCLUSIONS: 15% EDTA (pH = 6.5) 25% NaOCl +3% H2O2 was the most effective irrigation on removing smear layer.

PMID: 14661329 [PubMed - indexed for MEDLINE]

[Intratubular penetration of root canal sealers]

Pesqui Odontol Bras. 2002 Oct-Dec;16(4):332-6

Authors: de Deus G, Gurgel Filho ED, Ferreira CM, Coutinho Filho T

The purpose of this study was to evaluate the capacity of penetration of four endodontic sealers (Endo Fill, Sealapex, AH Plus and Pulp Canal Sealer) into dentinal tubules. Seventy-two extracted human maxillary anterior teeth were utilized in this study. The teeth were cleaned and shaped by means of the balanced-forces technique. The work length was established at 1 mm beyond the apex. Copious irrigation with 10 ml of 5.25% sodium hypochlorite was carried out. The teeth were divided in 8 groups - 4 had the smear layer maintained, and 4 had it removed. The smear layer was removed with a commercial solution of 17% EDTA, and the root canal system was flushed for 3 min. Finally, the roots were irrigated with 3 ml of 5.25% sodium hypochlorite. All teeth were sealed by means of the technique of the condensation wave with a medium nonstandardized cone. After filling, the roots were grooved, longitudinally split and examined under a scanning electron microscope (SEM). The focus of observation was the interface between the dentin and the sealing material. The Rickert sealer (Pulp Canal) presented the maximum penetration depths into the dentinal tubules, and Sealapex, the minimum. The Spearman test was used to determine whether there were significant differences between the groups. The removal of smear layer allowed significant penetration of the sealers (p <= 0.01).

PMID: 12612772 [PubMed - indexed for MEDLINE]

Nitric oxide improves internal iron availability in plants.

Plant Physiol. 2002 Dec;130(4):1852-9

Authors: Graziano M, Beligni MV, Lamattina L

Iron deficiency impairs chlorophyll biosynthesis and chloroplast development. In leaves, most of the iron must cross several biological membranes to reach the chloroplast. The components involved in the complex internal iron transport are largely unknown. Nitric oxide (NO), a bioactive free radical, can react with transition metals to form metal-nitrosyl complexes. Sodium nitroprusside, an NO donor, completely prevented leaf interveinal chlorosis in maize (Zea mays) plants growing with an iron concentration as low as 10 microM Fe-EDTA in the nutrient solution. S-Nitroso-N-acetylpenicillamine, another NO donor, as well as gaseous NO supply in a translucent chamber were also able to revert the iron deficiency symptoms. A specific NO scavenger, 2-(4-carboxy-phenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, blocked the effect of the NO donors. The effect of NO treatment on the photosynthetic apparatus of iron-deficient plants was also studied. Electron micrographs of mesophyll cells from iron-deficient maize plants revealed plastids with few photosynthetic lamellae and rudimentary grana. In contrast, in NO-treated maize plants, mesophyll chloroplast appeared completely developed. NO treatment did not increase iron content in plant organs, when expressed in a fresh matter basis, suggesting that root iron uptake was not enhanced. NO scavengers 2-(4-carboxy-phenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide and methylene blue promoted interveinal chlorosis in iron-replete maize plants (growing in 250 microM Fe-EDTA). Even though results support a role for endogenous NO in iron nutrition, experiments did not establish an essential role. NO was also able to revert the chlorotic phenotype of the iron-inefficient maize mutants yellow stripe1 and yellow stripe3, both impaired in the iron uptake mechanisms. All together, these results support a biological action of NO on the availability and/or delivery of metabolically active iron within the plant.

PMID: 12481068 [PubMed - indexed for MEDLINE]

Changes in the rhizosphere of metal-accumulating plants evidenced by chemical extractants.

J Environ Qual. 2002 Sep-Oct;31(5):1561-9

Authors: Hammer D, Keller C

The plants Salix viminalis L. (common osier) and Thlaspi caerulescens J. Presl & C. Presl have been studied often because of their high potential to extract heavy metals from soils. The soil properties favoring this phytoextraction are not yet fully known. In this study we compared three frequently used single-extracting agents (NaNO3, diethylenetriaminepentaacetic acid [DTPA], and ethylenediaminetetra-acetic acid [EDTA]) with a sequential extraction procedure to describe changes in the different Cd, Cu, and Zn pools in the rhizosphere of S. viminalis and T. caerulescens grown on calcareous and acidic Swiss soils in a pot experiment. The sequential extraction was used to assess the chemical affinities of these heavy metals (HM) in the soil whereas the single extractants were used for estimating the bioavailable HM pools in the soils. Cadmium depletion in several pools was most apparent in the acidic soil, with a significant decrease observed in the NaNO3-, DTPA-, and EDTA-extractable fractions following T. caerulescens growth compared with control pots. The sequential extraction showed that most Cd extracted by the plant from the acidic soil originated from the organic pool, which implies that heavy metals bound to organic matter may constitute a significant part of the bioavailable Cd pool in soils. In the calcareous soil only a small amount of Cd was taken up by T. caerulescens, and this came mainly from the carbonate-bound fraction. This study shows that T. caerulescens, and to a lesser extent S. viminalis, can alter the heavy metal distribution in different soil pools within 90 d.

PMID: 12371173 [PubMed - indexed for MEDLINE]

The effect of EDTA on the attachment and growth of cultured human gingival fibroblasts in periodontitis-affected root surface.

J Contemp Dent Pract. 2001 Feb 15;2(1):13-23

Authors: Babay N

The purpose of the present study was to analyze the effects of 5% and 24% EDTA on the attachment of gingival fibroblasts to periodontally diseased root surfaces. A flat root surface was created on human teeth that were extracted due to severe periodontitis. The teeth were etched with the following concentrations of etylediaminetetraacetic acid (EDTA) for two minutes: 5% (group I) and 24% (group II). Group III was soaked in saline and served as a control. The specimens and fibroblasts were incubated in a culture medium for 24 hours each day for one and two weeks and photographed using scanning electron microscopy. Each specimen was examined for the migration of cells into the etched and non-etched root surface. No fibroblasts could be detected on the saline groups. More fibroblasts could attach to the surface treated with 24% EDTA than with 5% EDTA. It was concluded that supersaturated EDTA at 24% enhances the attachment of gingival fibroblasts to the root surface.

PMID: 12167940 [PubMed - indexed for MEDLINE]

Solution structure of the SL1 RNA of the M1 double-stranded RNA virus of Saccharomyces cerevisiae.

Biophys J. 2001 Apr;80(4):1957-66

Authors: Yoo JS, Cheong HK, Lee BJ, Kim YB, Cheong C

The 20-nucleotide SL1 VBS RNA, 5'-GGAGACGC[GAUUC]GCGCUCC (bulged A underlined and loop bases in brackets), plays a crucial role in viral particle binding to the plus strand and packaging of the RNA. Its structure was determined by NMR spectroscopy. Structure calculations gave a precisely defined structure, with an average pairwise root mean square deviation (RMSD) of 1.28 A for the entire molecule, 0.57 A for the loop region (C8-G14), and 0.46 A for the bulge region (G4-G7, C15-C17). Base stacking continues for three nucleotides on the 5' side of the loop. The final structure contains a single hydrogen bond involving the guanine imino proton and the carbonyl O(2) of the cytosine between the nucleotides on the 5' and 3' ends of the loop, although they do not form a Watson-Crick base pair. All three pyrimidine bases in the loop point toward the major groove, which implies that Cap-Pol protein may recognize the major groove of the SL1 loop region. The bulged A5 residue is stacked in the stem, but nuclear Overhauser enhancements (NOEs) suggest that A5 spends part of the time in the bulged-out conformation. The rigid conformation of the upper stem and loop regions may allow the SL1 VBS RNA to interact with Cap-Pol protein without drastically changing its own conformation.

PMID: 11259308 [PubMed - indexed for MEDLINE]

Zinc deficiency up-regulates expression of high-affinity phosphate transporter genes in both phosphate-sufficient and -deficient barley roots.

Plant Physiol. 2000 Sep;124(1):415-22

Authors: Huang C, Barker SJ, Langridge P, Smith FW, Graham RD

Phosphate (P) is taken up by plants through high-affinity P transporter proteins embedded in the plasma membrane of certain cell types in plant roots. Expression of the genes that encode these transporters responds to the P status of the plants, and their transcription is normally tightly controlled. However, this tight control of P uptake is lost under Zn deficiency, leading to very high accumulation of P in plants. We examined the effect of plant Zn status on the expression of the genes encoding the HVPT1 and HVPT2 high-affinity P transporters in barley (Hordeum vulgare L. cv Weeah) roots. The results show that the expression of these genes is intimately linked to the Zn status of the plants. Zn deficiency induced the expression of genes encoding these P transporters in plants grown in either P-sufficient or -deficient conditions. Moreover, the role of Zn in the regulation of these genes is specific in that it cannot be replaced by manganese (a divalent cation similar to Zn). It appears that Zn plays a specific role in the signal transduction pathway responsible for the regulation of genes encoding high-affinity P transporters in plant roots. The significance of Zn involvement in the regulation of genes involved in P uptake is discussed.

PMID: 10982454 [PubMed - indexed for MEDLINE]

Nitrate assimilation in chicory roots (Cichorium intybus L.) which acquire radial growth.

J Exp Bot. 2000 Mar;51(344):539-46

Authors: Druart N, Goupil P, Dewaele E, Boutin JP, Rambour S

Nitrate assimilation was analysed in chicory plants (Cichorium intybus L. cv. Turbo) during the early vegetative growth. Nitrate reductase (NR, EC 1.6.6.1) activity (NRA) was measured in roots and leaves at different developmental stages. During phase I, which corresponds to the structural growth (21-42 DAS), nitrate reduction mainly occurred in the roots. At the onset of the tuber formation (phase II), which is characterized by the formation of a cambium inducing a radial growth (42-63 DAS), NRA rapidly decreased in roots and developed in leaves. A tight correlation was found between the nitrate content, the amino acid level and NRA in roots and leaves. Northern blot and ELISA analysis showed that both levels of NR mRNA and NR protein were not modified during the time-course of the experiment suggesting that modification of nitrate assimilation was not controlled at a transcriptional level. In vitro NRA assayed in presence of either Mg2+ ions or EDTA showed that NR was influenced at least in part by a reversible phosphorylation/dephosphorylation reaction. Okadaic acid, a serine-threonine protein phosphatases inhibitor, strongly decreased NRA. Conversely, staurosporine, a serine-threonine protein kinases inhibitor, did not significantly change NRA in roots or leaves. Therefore, NRA was regulated at a post-translational level during the early vegetative growth by modifying the phosphorylation balance of the NR protein in chicory.

PMID: 10938810 [PubMed - indexed for MEDLINE]

Blood circulation as source for osteopontin in acellular extrinsic fiber cementum and other mineralizing tissues.

J Dent Res. 1999 Nov;78(11):1688-95

Authors: VandenBos T, Bronckers AL, Goldberg HA, Beertsen W

Osteopontin (OPN) is one of the major non-collagenous proteins in root cementum and other mineralized tissues. Although most of this mineral-seeking protein is thought to be produced by local tissue cells, some of it might enter the mineralizing matrix from the blood. To test this hypothesis, we followed the distribution of a single dose of purified porcine or rat 125I-labeled OPN injected i.v. in rats, in mineralizing and non-mineralizing tissues and in subcutaneously implanted collagenous implants. The animals were killed 30 or 48 hrs after injection. Tissues (calvaria, tibia, lower and upper jaws) were harvested and processed for radioautography and biochemical analysis. Tissues as well as calcifying collagenous implants proved to have taken up radiolabel. In EDTA extracts of long bones, the majority of the radiolabel was demonstrated to be associated with intact OPN. The iodinated protein was also found in the acellular extrinsic fiber cementum (acellular cementum) layer investing the continuously growing incisors, in laminae limitantes, cement lines, and in forming bone near the mineralization front. Further, the label was present in the circumpulpal dentin of the incisors, and some of it appeared to have been incorporated into developing enamel. It is concluded that OPN in acellular cementum and other mineralizing tissues may-at least partially-originate from sources outside the direct environment following its transportation via serum.

PMID: 10576164 [PubMed - indexed for MEDLINE]

Soybean root nodule acid phosphatase.

Plant Physiol. 1997 Jun;114(2):597-604

Authors: Penheiter AR, Duff SM, Sarath G

Acid phosphatases are ubiquitous enzymes that exhibit activity against a variety of substrates in vitro, although little is known about their intracellular function. In this study, we report the isolation, characterization, and partial sequence of the major acid phosphatase from soybean (Glycine max L.) root nodules. The phosphatase was purified predominantly as a heterodimer with subunits of 28 and 31 kD; homodimers of both subunits were also observed and exhibited phosphatase activity. In addition to the general phosphatase substrate, p-nitrophenyl phosphate, the heterodimeric form of the enzyme readily hydrolyzed 5'-nucleotides, flavin mononucleotide, and O-phospho-L-Tyr. Low or negligible activity was observed with ATP or polyphosphate. Purified nodule acid phosphatase was stimulated by magnesium, inhibited by calcium and EDTA, and competitively inhibited by cGMP and cAMP with apparent Ki values of 7 and 12 microM, respectively. Partial N-terminal and internal sequencing of the nodule acid phosphatase revealed homology to the soybean vegetative storage proteins. There was a 17-fold increase in enzyme activity and a noticeable increase in protein levels detected by immunoblotting methods during nodule development. Both of these parameters were low in young nodules and reached a peak in mature, functional nodules, suggesting that this enzyme is important for efficient nodule metabolism.

PMID: 9193092 [PubMed - indexed for MEDLINE]

The pH Requirement for in Vivo Activity of the Iron-Deficiency-Induced "Turbo" Ferric Chelate Reductase (A Comparison of the Iron-Deficiency-Induced Iron Reductase Activities of Intact Plants and Isolated Plasma Membrane Fractions in Sugar Beet).

Plant Physiol. 1996 Jan;110(1):111-123

Authors: Susin S, Abadia A, Gonzalez-Reyes JA, Lucena JJ, Abadia J

The characteristics of the Fe reduction mechanisms induced by Fe deficiency have been studied in intact plants of Beta vulgaris and in purified plasma membrane vesicles from the same plants. In Fe-deficient plants the in vivo Fe(III)-ethylenediaminetetraacetic complex [Fe(III)-EDTA] reductase activity increased over the control values 10 to 20 times when assayed at a pH of 6.0 or below ("turbo" reductase) but increased only 2 to 4 times when assayed at a pH of 6.5 or above. The Fe(III)-EDTA reductase activity of root plasma membrane preparations increased 2 and 3.5 times over the controls, irrespective of the assay pH. The Km for Fe(III)-EDTA of the in vivo ferric chelate reductase in Fe-deficient plants was approximately 510 and 240 [mu]M in the pH ranges 4.5 to 6.0 and 6.5 to 8.0, respectively. The Km for Fe(III)-EDTA of the ferric chelate reductase in intact control plants and in plasma membrane preparations isolated from Fe-deficient and control plants was approximately 200 to 240 [mu]M. Therefore, the turbo ferric chelate reductase activity of Fe-deficient plants at low pH appears to be different from the constitutive ferric chelate reductase.

PMID: 12226175 [PubMed - as supplied by publisher]

Calcium/Calmodulin Activation of Soybean Glutamate Decarboxylase.

Plant Physiol. 1995 Jun;108(2):543-549

Authors: Snedden WA, Arazi T, Fromm H, Shelp BJ

Recently, we provided preliminary evidence for calcium (Ca2+)/calmodulin (CaM) stimulation of plant glutamate decarboxylase (GAD; EC 4.1.1.15). In the present study, a detailed characterization of the phenomenon is described. GAD was partially purified from various soybean (Glycine max L. Merr.) tissues (developing seed coat and cotyledons, leaf, and root) in the presence of EDTA by a combination of ammonium sulfate precipitation and anion-exchange fast protein liquid chromatography. GAD activity showed a sharp optimum at pH 5.8, with about 12% of maximal activity at pH 7. It was stimulated 2- to 8-fold (depending on the tissue source) in the presence of Ca2+/CaM at pH 7 but not at pH 5.8. Furthermore, when the protease inhibitor phenylmethylsulfonyl fluoride was omitted from the purification procedure, GAD activity was insensitive to Ca2+/CaM but was similar in magnitude to CaM-stimulated activity. The stimulation by Ca2+/CaM was fully inhibited by the CaM antagonists N-(6-aminohexyl)-5-chloro-1-naphthalenesulfon-amide and trifluoperazine. With saturating CaM or Ca2+, the concentrations of Ca2+ and CaM required for half-maximal stimulation were about 7 to 11 [mu]M and 25 nM, respectively. The effect of Ca2+ and CaM appeared to be through a 2.4-fold stimulation of Vmax and a 55% reduction in Km. The results suggested that GAD is activated via Ca2+ signal transduction.

PMID: 12228492 [PubMed - as supplied by publisher]

Tensile properties of mineralized and demineralized human and bovine dentin.

J Dent Res. 1994 Jun;73(6):1205-11

Authors: Sano H, Ciucchi B, Matthews WG, Pashley DH

The relative contribution of the matrix of dentin to the physical properties of dentin is unknown but thought to be small. The objective of this study was to test the hypothesis that the demineralized matrix of dentin contributes little to the strength of dentin by measuring and comparing the ultimate tensile strength and modulus of elasticity of mineralized and demineralized dentin. Small slabs (4 x 0.5 x 0.5 mm) of bovine and human dentin were tested in a microtensile testing device in vitro. Human coronal mineralized dentin gave a mean ultimate tensile strength (UTS) of 104 MPa. Bovine incisor coronal dentin exhibited a UTS of 91 MPa, and bovine root dentin failed at 129 MPa. The modulus of elasticity of mineralized bovine and human dentin varied from 13 to 15 MPa. When dentin specimens were demineralized in EDTA, the UTS and modulus of elasticity fell to 26-32 MPa and 0.25 GPa, respectively, depending on dentin species. The results indicate that collagen contributes about 30% of the UTS of mineralized dentin, which is higher than was expected.

PMID: 8046110 [PubMed - indexed for MEDLINE]

Bradyrhizobium japonicum delta-aminolevulinic acid dehydratase is essential for symbiosis with soybean and contains a novel metal-binding domain.

J Bacteriol. 1993 Nov;175(22):7222-7

Authors: Chauhan S, O'Brian MR

The Bradyrhizobium japonicum hemA gene product delta-aminolevulinic acid (ALA) synthase is not required for symbiosis of that bacterium with soybean. Hence, the essentiality of the subsequent heme synthesis enzyme, ALA dehydratase, was examined. The B. japonicum ALA dehydratase gene, termed hemB, was isolated and identified on the basis of its ability to confer hemin prototrophy and enzyme activity on an Escherichia coli hemB mutant, and it encoded a protein that was highly homologous to ALA dehydratases from diverse organisms. A novel metal-binding domain in the B. japonicum ALA dehydratase was identified that is a structural composite of the Mg(2+)-binding domain found in plant ALA dehydratases and the Zn(2+)-binding region of nonplant ALA dehydratases. Enzyme activity in dialyzed extracts of cells that overexpressed the hemB gene was reconstituted by the addition of Mg2+ but not by addition of Zn2+, indicating that the B. japonicum ALA dehydratase is similar to the plant enzymes with respect to its metal requirement. Unlike the B. japonicum hemA mutant, the hemB mutant strain KP32 elicited undeveloped nodules on soybean, indicated by the lack of nitrogen fixation activity and plant hemoglobin. We conclude that the hemB gene is required for nodule development and propose that B. japonicum ALA dehydratase is the first essential bacterial enzyme for B. japonicum heme synthesis in soybean root nodules. In addition, we postulate that ALA is the only heme intermediate that can be translocated from the plant to the endosymbiont to support bacterial heme synthesis in nodules.

PMID: 8226669 [PubMed - indexed for MEDLINE]

Effect of Fe-Catalyzed Photooxidation of EDTA on Root Growth in Plant Culture Media.

Plant Physiol. 1991 Jul;96(3):843-847

Authors: Hangarter RP, Stasinopoulos TC

Light from fluorescent lamps can induce formaldehyde production and iron deficiency in plant nutrient culture medium. Formaldehyde is produced from EDTA when it is oxidized by the photochemical reduction of ferric iron and it can accumulate to inhibitory levels. The medium becomes iron deficient because iron becomes unchelated and forms an unavailable precipitate as EDTA is oxidized. The combination of light-induced formaldehyde production and loss of available iron reduces the ability of the culture media to support growth of Arabidopsis thaliana roots. Removing ultraviolet and blue wavelengths with a yellow acrylic filter is a simple and effective means of preventing Fecatalyzed photooxidation of EDTA in plant culture media.

PMID: 16668263 [PubMed - as supplied by publisher]

Does Iron Deficiency in Pisum sativum Enhance the Activity of the Root Plasmalemma Iron Transport Protein?

Plant Physiol. 1990 Nov;94(3):1353-1357

Authors: Grusak MA, Welch RM, Kochian LV

Roots of Fe-sufficient and Fe-Deficient pea (Pisum sativum L.) were studied to determine the effect of Fe-deficiency on the activity of the root-cell plasmalemma Fe(2+) transport protein. Rates of Fe(III) reduction and short-term Fe(2+) influx were sequentially determined in excised primary lateral roots using Fe(III)-ethylene-diaminetetraacetic acid (Fe[III]-EDTA). Since the extracellular Fe(2+) for membrane transport was generated by root Fe(III) reduction, rates of Fe(2+) influx for each root system were normalized on the basis of Fe(III) reducing activity. Ratios of Fe(2+) influx to Fe(III) reduction (micromole Fe(2+) absorbed/micromole Fe[III] reduced) revealed no enhanced Fe(2+) transport capacity in roots of Fe-deficient peas (from the parental genotype, Sparkle) or the functional Fe-deficiency pea mutant, E107 (derived from Sparkle), relative to roots of Fe-sufficient Sparkle plants. Data from studies using 30 to 100 micromolar Fe(III)-EDTA indicated a linear relationship between Fe(2+) influx and Fe(III) reduction (Fe(2+) generation), while Fe(2+) influx saturated at higher concentrations of Fe(III)-EDTA. Estimations based on current data suggest the Fe(2+) transport protein may saturate in the range of 10(-4.8) to 10(-4) molar Fe(2+). These results imply that for peas, the physiological rate limitation to Fe acquisition in most well-aerated soils would be the root system's ability to reduce soluble Fe(III)-compounds.

PMID: 16667840 [PubMed - as supplied by publisher]

Purification and characterization of cytosolic aldolase from carrot storage root.

Biochem J. 1990 Jul 1;269(1):133-9

Authors: Moorhead GB, Plaxton WC

A single fructose-1,6-bisphosphate (FBP) aldolase has been detected in extracts from carrot storage roots (Daucus carota L.). The enzyme was purified 850-fold to electrophoretic homogeneity and a final specific activity of 26.3 mumols of FBP utilized/min per mg of protein. SDS/PAGE of the final preparation revealed a single protein-staining band of 40 kDa. The native molecular mass was determined by analytical gel filtration to be 159 kDa, indicating that the enzyme is a homotetramer. Denaturing isoelectric focusing revealed two predominant protein-staining bands, with pI values of 5.6 and 5.7. The enzyme is a class I aldolase, since EDTA or metal ions had no effect on its activity. The enzyme was relatively heat-stable, had an activation energy (Ea) of 68.3 kJ.mol-1, and had an absorption coefficient of 8.08 x 10(4) M-1.cm-1 at 280 nm. Km values for FBP and sedoheptulose 1,7-bisphosphate (SBP) were both determined to be 6 microM (pH optima 7.4). The specificity constant with FBP was 2.6 times that obtained with SBP. Ribose 5-phosphate, 6-phosphogluconate, MgAMP, glucose 1-phosphate and phosphoenolpyruvate (PEP) were inhibitors. PEP was a mixed-type inhibitor with respect to FBP (Ki = 3.2 mM, K'i = 5.1 mM). No activators were found. Rabbit anti-(carrot aldolase) polyclonal antibodies immunoprecipitated the activity of both carrot root aldolase and spinach leaf cytosolic aldolase, but not that of spinach leaf plastid aldolase. Western-blot analysis also revealed cross-reactivity with cytosolic, but not plastid, spinach leaf aldolase, indicating that the single carrot root aldolase is cytosolic.

PMID: 2198022 [PubMed - indexed for MEDLINE]