Decalcification dynamic of dog mineralized tissue by microwaves

The aim of this article is to present the decalcification process dynamic of mineralized tissue in dogs, teeth and jaw, comparing the traditional decalcification method, immersion, and microwave, immersion followed by irradiation using a domestic microwave oven, accompanying the liberation of calcium through spectrophotometer of atomic absorption. It was used as decalcified agent, EDTA solution or nitric acid. The results showed that with the use of nitric acid (5%), after 15 days, the irradiated fragments could be processed for histological analysis, otherwise the tooth not irradiated need to be submerged for 65 days. The EDTA decalcified action was slower than the nitric acid. The histological observations of the irradiated samples showed an excellent preservation of the morphological characteristics, independently of the decalcified agent used. © 2007 Sociedad Chilena de Anatomía.

Production and partial characterization of keratinase produced by a microorganism isolated from poultry processing plant wastewater

A Streptomyces was isolated from poultry plant wastewater, showed high keratinolytic activity when cultured on feather meal medium. Optimum keratinolytic activity was observed at 40°C and pH 8.0. The enzyme also showed to be stable between 40 and 60°C. The keratinolytic activity was not inhibited by EDTA, DMSO and Tween 80. On the other hand, CaCl2, ZnCl2, and BaCl2 slightly inhibited the keratinolytic activity. The Streptomyces isolated might be useful in leather, keratin waste treatment, animal feeding industry, and also cosmetic industry. © 2008 Academic Journals.

Myotoxic phospholipases A2 isolated from Bothrops brazili snake venom and synthetic peptides derived from their C-terminal region: Cytotoxic effect on microorganism and tumor cells

This paper reports the purification and biochemical/pharmacological characterization of two myotoxic phospholipases A2 (PLA2s) from Bothrops brazili venom, a native snake from Brazil. Both myotoxins (MTX-I and II) were purified by a single chromatographic step on a CM-Sepharose ion-exchange column up to a high purity level, showing Mr ∼ 14,000 for the monomer and 28,000 Da for the dimer. The N-terminal and internal peptide amino acid sequences showed similarity with other myotoxic PLA2s from snake venoms, MTX-I belonging to Asp49 PLA2 class, enzymatically active, and MTX-II to Lys49 PLA2s, catalytically inactive. Treatment of MTX-I with BPB and EDTA reduced drastically its PLA2 and anticoagulant activities, corroborating the importance of residue His48 and Ca2+ ions for the enzymatic catalysis. Both PLA2s induced myotoxic activity and dose-time dependent edema similar to other isolated snake venom toxins from Bothrops and Crotalus genus. The results also demonstrated that MTXs and cationic synthetic peptides derived from their 115-129 C-terminal region displayed cytotoxic activity on human T-cell leukemia (JURKAT) lines and microbicidal effects against Escherichia coli, Candida albicans and Leishmania sp. Thus, these PLA2 proteins and C-terminal synthetic peptides present multifunctional properties that might be of interest in the development of therapeutic strategies against parasites, bacteria and cancer. © 2008 Elsevier Inc. All rights reserved.

Efeito de um quelante de cálcio, um detergente e da lecitina de soja sobre a qualidade do sêmen caprino congelado-descongelado

The aim of this paper was verifying the effect of the Equex STM Paste and EDTA addition to a Tris-egg yolk extender, on the postthaw goat sperm viability. Nine semen samples of two adult goats were collected by artificial vagina and cryopreserved. It was also objective of this study, to evaluate the utilization of a soybean lecithin based commercial extender (Bioexcell® - IMV, L'Aigle, French) for the goat semen freezing. They were formed five experimental groups: TRIS; TRIS+EDTA; TRIS+EQUEX; TRIS+EDTA+EQUEX e Bioexcell. After evaluation, the semen was diluted in the five extenders and packed in 0.25mL straws with 100 million of motile spermatozoa. The samples were cooled at 0,46°C/min to 5°C, submitted at 75min of equilibration time and frozen in liquid nitrogen vapour. The thawing was accomplished in 37°C water bath for 50s. There were no differences (P>0,05) on the means of post-thaw total and progressive sperm motility among the groups TRIS, TRIS+EQUEX and TRIS+EQUEX+EDTA. The Bioexcell group obtained the least (P<0,05) percentage of post-thawing total and progressive sperm motility. After the thermotolerance test, it was observed the greatest (P<0,05) rates of total and progressive sperm motility in the Equex STM groups (TRIS+EQUEX and TRIS+EQUEX+EDTA). Thus, it can be affirmed that the Equex addition promotes better maintenance rates in the pos-thaw sperm viability, when compared with the extenders that did not contain it.

Polimorfismo nos genes metilenotetrahidrofolato redutase e cistationina-beta-sintase e a sua relação com eventos vaso-oclusivos na doença falciforme

Sickle cell disease is an inflammatory condition with a pathophysiology that involves vaso-occlusive episodes. Mutations of the methylenetetrahydrofolate reductase (MTHFR) and cystathionine beta-synthase (CBS) genes are risk factors for vascular disease. Due to the importance of identifying risk factors for vaso-occlusive events in sickle cell patients, we investigated the frequencies of the C677T and 844ins68 mutations of the MTHFR and CBS genes, respectively. Three hundred patients with Hb SS, HB SC and HbS/Beta thalassemia, from Brasília, Goiânia, Rio de Janeiro, São Jose do Rio Preto and São Paulo were evaluated. Samples of 5 mL of venous blood were collected in EDTA after informed consent was received from patients. Classical diagnostic methods were used to confirm the hemoglobin phenotypes. The hemoglobin genotypes and polymorphisms studied were evaluated by Restriction Fragment Length Polymorphism and Allele Specific amplification. The results showed that 93 patients (31.00%) were heterozygous and 13 (4.33%) homozygous for the C677T mutation and 90 were heterozygotes (30.00%) and 8 homozygous (2.66%) for the 844ins68 mutation, both with significant differences for genotype frequency between the localities. The allelic frequencies are in Hardy-Weinberg equilibrium for both polymorphisms. The frequency of mutations was significant and the presence of related vaso-occlusive events was more common in patients with Hb SS (p = 0007). The 844ins68 mutation was approximately three times more frequent in patients with vaso-occlusive complications (p = 0011). The C677T mutation did not prove to be associated with risk of vaso-occlusive events (p = 0.193). A C677T-844ins68 interaction occurred in 12.08% of the patients, doubling the risk of vaso-occlusive manifestations. The frequencies of the polymorphisms are consistent with those expected in the Brazilian population. The presence of the 844ins68 mutation of the CBS gene proved to be a potential risk factor for vaso-occlusive events in sickle cell patients.

In vitro evaluation of the effect of dietary acids and toothbrushing on human root dentin permeability

OBJECTIVES: The purpose of this in vitro study was to quantify the alterations in human root dentin permeability after exposure to dietary acids and to evaluate the effect of toothbrushing after acid application. METHOD AND MATERIALS: Extracted human third molars had their crowns sectioned above the CEJ, pulp tissue removed, and cervical root dentin exposed using a high-speed bur (approximately 1 mm in depth of substance loss). From each root fragment, one specimen was prepared. A total of 25 specimens were used and distributed randomly into five groups. The specimens were attached to a hydraulic pressure apparatus to evaluate the alterations of root dentin permeability after exposure to different acids. Dentin permeability was measured after the following sequential steps: (1) treatment with EDTA for 3 minutes to obtain the maximum permeability; (2) root planing to create a smear layer; (3) exposure to different acidic substances for 5 minutes (vinegar, cola drink, lemon juice, white wine, and orange juice); and (4) brushing for 3 minutes. RESULTS: All acidic substances increased dentin permeability after root planing. Lemon juice produced higher values for permeability when compared to the other substances (P = .009); moreover, orange juice showed similar results (P < .02) except when compared to vinegar (P = .12). Brushing right after acid exposure significantly reduced dentin permeability except in the vinegar group (P = .07). CONCLUSION: Under the experimental conditions, dietary acids increased root dentin permeability, and immediate brushing reduced permeability levels.

Aplicação da metodologia de superfície de resposta na remediação de um solo arenoso contaminado artificialmente com cobre

Soil flushing is an alternative remediation technology for soils contaminated with heavy metals, which the main contaminant removing process consists in percolating an extraction solution. This work aimed to use the response surface methodology to point out combinations among the parameters of the extraction solution (Na2EDTA concentration, volume e pH) in order to reduce the concentration of copper in a sandy soil to risk levels lower than the intervention levels for exposure scenarios adopted by the Environmental Company of Sao Paulo State. Thus, a series of tests in leaching columns were carried out using a Fluvisol artificially contaminated (1257,3 mg kg-1). The tests were conducted in triplicate and setup a central composite rotatable design with 15 different parameters combinations of the extraction solution and one replicate in the center point. Using 5% significance level, the adjusted model (R2 = 0,98) indicated combinations of Na2EDTA concentration, pH and volume of the extraction solution which allow reduction of copper concentration below levels reported by environmental agency of Sao Paulo State for industrial, residential, agricultural or maximum exposure scenarios.

Leaf spray fertilization of boron and zinc on productivity and fruit quality of Japanese pear tree

The cultivation of fruit plants from temperate climate in tropical or subtropical regions can be a good income alternative for the producer. However, due to the little existent information about cultivation of those fruit plants, the producers use imported techniques of other producing areas, or even an association of practices used for other fruit plants, pointing out the leaf spray fertilization of micronutrients without appropriate scientific base. In this context, the objective of this study was to verify the effect of the leaf spray fertilization of B and Zn on productivity and fruit quality of Japanese pear tree. The experiment was conducted from 2004 to 2005, in Ilha Solteira, in northwestern São Paulo State-Brazil. The climate is, according to the Köpppen Classification, tropical wet and dry (Aw). The 'Okusankichi' cultivar, grafted on Pyrus communis L. rootstock was used as well as doses of 110 g.ha-1 of B and 250 g.ha-1 of Zn in each application. The treatments were: T1. water, T2. boric acid, T3. zinc sulfate, T4. T2 + T3, T5. boric acid + urea + citric acid + EDTA, T6. zinc sulfate + urea + citric acid + EDTA, T7. T5 + T6, T8. boric acid + urea + citric acid + EDTA + sodium molibdate + sulfur + calcium chloride, T9. zinc sulfate + urea + citric acid + EDTA + Fe sulfate + Mn sulfate + Mg sulfate and, T10. T8+T9. A randomized blocks design was used and the averages were compared by Tukey test. In the first crop the mixture of boric acid with quelating agents were efficient to supply B to the plants and zinc sulfate plus quelating agents were efficient to increase Zn leaf content. However, the productivity and the fruit quality were not influenced by the leaf spray of B and Zn. In the second crop the leaf content of B and Zn and the productivity were not influenced by the leaf spray; the boric acid and the zinc sulfate with or without quelating agents increased the contents of total soluble solids and, the boric acid with or without quelating agents increased the contents of total titratable acidity.

Calcium hydroxide intracanal dressing removal with different rotary instruments and irrigating solutions: A scanning electron microscopy study

This study evaluated the efficacy of 2 types of rotary instruments employed in association with sodium hypochlorite (NaOCl) or EDTA in removing calcium hydroxide (CH) residues from root canals dentin walls. Forty-two mandibular human incisors were instrumented with the ProTaper System up to F2 instrument, irrigated with 2.5% NaOCl followed by 17% EDTA and filled with a CH intracanal dressing. After 7 days, the CH dressing was removed using 4 techniques: NiTi rotary instrument size 25, 0.06 taper (K3 Endo) and irrigation with 17% EDTA (Group 1), NiTi rotary F1 instrument (ProTaper) and irrigation with 17% EDTA (Group 2), NiTi rotary instrument size 25, 0.06 taper and irrigation with 2.5% NaOCl (Group 3) and NiTi rotary F1 instrument and irrigation with 2.5% NaOCl (Group 4). Two roots without intracanal dressing were used as negative controls. Teeth were evaluated by scanning electron microscopy, in the cervical and apical canal thirds. None of the techniques removed the CH dressing completely. In the apical and cervical thirds, F1 instrument was better than instrument size 25, 0.06 taper in removing CH residues (p<0.05), regardless of the final irrigating solution. No difference was found between the irrigating solutions in the groups of F1 instrument and of instrument size 25, 0.06 taper (p>0.05). The negative controls had no CH residues on the dentin walls. In conclusion, the ProTaper F1 instrument was better than K3 Endo instrument size 25, 0.06 taper in the removal of CH intracanal medication, regardless of irrigating solution used.

Influence of dental exposure to oral environment on smear layer removal and collagen exhibition after using different conditioning agents

Although in vitro studies have shown encouraging results for root surface conditioning with demineralizing agents, in vivo studies have failed to show its benefits in periodontal healing. This can be attributed to several factors, among which, the hypermineralization of dental surface. Therefore, this in vitro study compared, using scanning electron microscopy (SEM), the effect of root surface conditioning with different conditioners (1% and 25% citric acid, 24% EDTA and 50 mg/mL tetracycline hydrochloride) in impacted teeth and in teeth that had their roots exposed to the oral environment. One trained examiner assessed the SEM micrographs using a root surface modification index. There was a tendency of more root surface modification in the group of impacted teeth, suggesting that the degree of root mineralization influences its chemical demineralization.

Pharmacokinetics of dexmedetomidine administered intravenously in isofluraneanesthetized cats

Objective-To determine the pharmacokinetics of dexmedetomidine administered as a short-duration IV infusion in isoflurane-anesthetized cats. Animals-6 healthy adult domestic female cats. Procedures-Dexmedetomidine hydrochloride was injected IV (10 μg/kg over 5 minutes [rate, 2 μg/kg/min]) in isoflurane-anesthetized cats. Blood samples were obtained immediately prior to and at 1, 2, 5, 6, 7, 10, 15, 30, 60, 90, 120, 240, and 480 minutes following the start of the IV infusion. Collected blood samples were transferred to tubes containing EDTA, immediately placed on ice, and then centrifuged at 3,901 X g for 10 minutes at 4°C. The plasma was harvested and stored at -20°C until analyzed. Plasma dexmedetomidine concentrations were determined by means of liquid chromatography-mass spectrometry. Dexmedetomidine plasma concentration-time data were fitted to compartmental models. Results-A 2-compartment model with input in and elimination from the central compartment best described the disposition of dexmedetomidine administered via short-duration IV infusion in isoflurane-anesthetized cats. Weighted mean ± SEM apparent volume of distribution of the central compartment and apparent volume of distribution at steady-state were 402 ± 47 mL/kg and 1,701 ± 200 mL/kg, respectively; clearance and terminal half-life (harmonic mean ± jackknife pseudo-SD) were 6.3 ± 2.8 mL/min/kg and 198 ± 75 minutes, respectively. The area under the plasma concentration curve and maximal plasma concentration were 1,061 ± 292 min·ng/mL and 17.6 ± 1.8 ng/mL, respectively. Conclusions and Clinical Relevance-Disposition of dexmedetomidine administered via short-duration IV infusion in isoflurane-anesthetized cats was characterized by a moderate clearance and a long terminal half-life.

Blood cells attachment after root conditioning and PRP application: An in vitro study

Aim: Root conditioning is aimed at smear layer removal and at dental matrix collagen exposure, which may promote periodontal regeneration. This in vitro study assessed smear layer removal, collagen fiber exposure and the influence of PRP (platelet-rich plasma) application on adhesion of blood cells to the root surface using scanning electron microscopy (SEM). Materials and methods: Scaled root samples (n = 160) were set in five groups and conditioned with: group I - control group (saline solution); group II (EDTA 24%); group III (citric acid 25%); group IV (tetracycline hydrochloride 50 mg/ml); group V (sodium citrate 30%). Eighty samples were assessed using the root surface modification index (RSMI). The other eighty samples were set in two groups. The first group (n = 40) received PRP gel application with a soft brush and the second group (n = 40) received PRP application and then a blood drop. The fibrin clot formation was assessed in the first group and the blood cells adhesion was assessed in the second group using the BEAI (blood elements adhesion index). A previously trained, calibrated, and blind examiner evaluated photomicrographs. Statistical analysis was performed using the Kruskal-Wallis's and Dunn's tests. Results: Group III attained the best results for RSMI and BEAI. Moreover, it was the only group showing fibrin clot formation. Conclusion: Citric acid was the most efficient conditioner for smear layer removal, collagen fiber exposure and blood cell adhesion. Moreover, it was the only group showing fibrin clot formation after PRP application. Clinical significance: This study demonstrated that root conditioning followed by PRP application may favor blood cell adhesion on root surface which may optimize periodontal healing.

Adhesion of real seal to human root dentin treated with different solutions

The aim of this study was to evaluate the effects of different irrigants on sealer-dentin bond strength when using Real Seal. Thirty single-rooted teeth were divided into 3 groups. In one group, the teeth were irrigated with 3 mL of 2.5% NaOCl after each file change, flushed with 17% EDTA for 3 min and finally rinsed with 3 mL of 2.5% NaOCl. In the other two groups, rinse with NaOCl was replaced with 2% chlorhexidine gluconate (CHX) and 0.9% saline, respectively. Each root was sectioned transversally into apical, middle and coronal thirds to obtain 2-mm-thick slices. Each slice was filled with Real Seal and Resilon. Push-out test was used to analyze bond strength and failure modes were classified as adhesive, cohesive or mixed, according to SEM observations. The push-out test did not reveal any statistically significant difference (p>0.05) between the irrigants. However, the groups exhibited significantly different (p<0.05) bond strengths in terms of the root canal third. Higher bond strength was observed at the apical third when compared with coronal third, while middle third presented intermediary values. Fifteen specimens were analyzed by SEM (5 per group). Eleven specimens exhibited adhesive failures (5 in saline, 4 in NaOCl and 2 in CHX group); 2 cohesive failures were observed in the CHX group, and 1 mixed failure each was observed in the CHX and NaOCl groups. The tested irrigants did not influence the bond strength of Resilon and Real Seal to dentin. The apical third exhibited higher mean bond strengths and adhesive failures were predominant.

Persistence of epoxy-based sealer residues in dentin treated with different chemical removal protocols

The presence of residual endodontic sealer in the pulp chamber may cause discoloration of the dental crown and interfere with the adhesion of restorative materials. The aim of this study was to compare the efficacy of different solvents in removing residues of an epoxy resin-based sealer (AH Plus) from the dentin walls of the pulp chamber, by scanning electron microscopy (SEM). Forty-four bovine incisor dental crown fragments were treated with 17% EDTA and 2.5% NaOCl. Specimens received a coating of AH Plus and were left undisturbed for 5 min. Then, specimens were divided in four groups (n = 10) and cleaned with one of the following solutions: isopropyl alcohol, 95% ethanol, acetone solution, or amyl acetate solution. Negative controls (n = 2) did not receive AH Plus, while in positive controls (n = 2) the sealer was not removed. AH Plus removal was evaluated by SEM, and a score system was applied. Data were analyzed by Kruskal-Wallis and Dunn tests. None of the solutions tested was able to completely remove AH Plus from the dentin of the pulp chamber. Amyl acetate performed better than 95% ethanol and isopropyl alcohol (p < 0.05), but not better than acetone (p > 0.05) in removing the sealer from dentin. No significant differences were observed between acetone, 95% ethanol, and isopropyl alcohol (p > 0.05). It was concluded that amyl acetate and acetone may be good options for cleaning the pulp chamber after obturation with AH Plus. SCANNING 35:17-21, 2013. © 2012 Wiley Periodicals, Inc. © Wiley Periodicals, Inc.

Wound healing of dehiscence defects following different root conditioning modalities: An experimental study in dogs

Objective: The purpose of this study was to investigate the periodontal healing pattern of dehiscence-type defects following different chemical root conditioning modalities. Materials and methods: Buccal osseous dehiscence defects were created on six teeth of seven dogs. After dental plaque accumulation, defects were treated with sterile saline solution (control group) or one chemical conditioning modality: citric acid (CA group), ethylenediaminetetraacetic acid (EDTA group), tetracycline (TTC group), citric acid + tetracycline (CA + TTC group), or tetracycline + citric acid (TTC + CA group). After 3 months of healing, clinical parameters were evaluated, and the animals were killed. Histological sections were processed, and a computer-assisted histometric analysis was used to evaluate the formation of new cementum, new bone, and epithelial apical migration. Results: All treatments yielded significant improvements in terms of probing depth decrease and clinical attachment level gain compared to baseline values; however, without significant differences among the groups (p > 0.05; one-way ANOVA). The highest amount of new cementum was noted in the EDTA group (3.72 ± 0.83 mm, 77.6 %), while the lowest amount of new bone was observed in the TTC group (0.7 ± 0.94 mm, 14.3 %). However, no statistically significant differences could be observed among the groups regarding epithelial apical migration, new cementum, and alveolar bone formation (p > 0.05). Conclusion: Chemical root surface conditioning did not promote any significant improvement in periodontal healing pattern of dehiscence-type defects in dogs. Clinical Relevance: Chemical root surface conditioning after surgical debridement did not promote positive or negative effects on periodontal healing pattern of dehiscence-type defects. © 2012 Springer-Verlag Berlin Heidelberg.