Purification and biochemical characterization of two xylanases produced by Aspergillus caespitosus and their potential for kraft pulp bleaching

Two extracellular xylanases produced by the thermotolerant fungus Aspergillus caespitosus grown in sugar cane bagasse were purified and characterized. Estimated molecular masses were 26.3 and 27 kDa (xyl I); 7.7 and 17.7 kDa (xyl II) for gel filtration and SDS-PAGE, respectively. Optimal temperature for both xylanases was 50-55°C. Optimal pH was 6.5-7.0 for xyl I, and 5.5-6.5 for xyl II. The thermostability (T half) at 55°C was 27.3 min (xyl I) and >90 min (xyl II). Xylanase activity was inhibited by several ions. β-mercaptoethanol activated 59 and 102% xyl I and xyl II activities, respectively. These enzymes preferentially hydrolyzed birchwood xylan, and the K m and V max values were 2.5 mg/ml and 1679 U/mg protein (xyl I), and 3.9 mg/ml and 113 U/mg protein (xyl II). The action of both xylanases mainly that of xyl II, on kraft pulp reduced kappa number and increased pulp viscosity. © 2004 Elsevier Ltd. All rights reserved.

Strong selection against hybrids at a hybrid zone in the Ensatina ring species complex and its evolutionary implications

The analysis of interactions between lineages at varying levels of genetic divergence can provide insights into the process of speciation through the accumulation of incompatible mutations. Ring species, and especially the Ensatina eschscholtzii system exemplify this approach. The plethodontid salamanders E. eschscholtzii xanthoptica and E. eschscholtzii platensis hybridize in the central Sierran foothills of California. We compared the genetic structure across two transects (southern and northern Calaveras Co.), one of which was resampled over 20 years, and examined diagnostic molecular markers (eight allozyme loci and mitochondrial DNA) and a diagnostic quantitative trait (color pattern). Key results across all studies were: (1) cline centers for all markers were coincident and the zones were narrow, with width estimates of 730 m to 2000 m; (2) cline centers at the northern Calaveras transect were coincident between 1981 and 2001, demonstrating repeatability over five generations; (3) there were very few if any putative F1s, but a relatively high number of backcrossed individuals in the central portion of transects: and (4) we found substantial linkage disequilibrium in all three studies and strong heterozygote deficit both in northern Calaveras, in 2001, and southern Calaveras. Both linkage disequilibrium and heterozygote deficit showed maximum values near the center of the zones. Using estimates of cline width and dispersal, we infer strong selection against hybrids. This is sufficient to promote accumulation of differences at loci that are neutral or under divergent selection, but would still allow for introgression of adaptive alleles. The evidence for strong but incomplete isolation across this centrally located contact is consistent with theory suggesting a gradual increase in postzygotic incompatibility between allopatric populations subject to divergent selection and reinforces the value of Ensatina as a system for the study of divergence and speciation at multiple stages. © 2005 The Society for the Study of Evolution. All rights reserved.

Scanning electron microscope analysis of internal adaptation of materials used for pulp protection under composite resin restorations

Purpose: The aim of this study was to evaluate the interfacial microgap with different materials used for pulp protection. The null hypothesis tested was that the combination of calcium hydroxide, resin-modified glass ionomer, and dentin adhesive used as pulp protection in composite restorations would not result in a greater axial gap than that obtained with hybridization only. Materials and Methods: Standardized Class V preparations were performed in buccal and lingual surfaces of 60 caries-free, extracted human third molars. The prepared teeth were randomly assessed in six groups: (1) Single Bond (SB) (3M ESPE, St. Paul, MN, USA); (2) Life (LF) (Kerr Co., Romulus, MI, USA) + SB; (3) LF + Vitrebond (VT) (3M ESPE) + SB; (4) VT + SB; (5) SB + VT; (6) SB + VT + SB. They were restored with microhybrid composite resin Filtek Z250 (3M ESPE), according to the manufacturer's instructions. However, to groups 5 and 6, the dentin bonding adhesive was applied prior to the resin-modified glass ionomer. The specimens were then thermocycled, cross-sectioned through the center of the restoration, fixed, and processed for scanning electron microscopy. The specimens were mounted on stubs and sputter coated. The internal adaptation of the materials to the axial wall was analyzed under SEM with × 1,000 magnification. Results: The data obtained were analyzed with nonparametric tests (Kruskal-Wallis, p ≤ .05). The null hypothesis was rejected. Calcium hydroxide and resin-modified glass ionomer applied alone or in conjunction with each other (p < .001) resulted in statistically wider microgaps than occurred when the dentin was only hybridized prior to the restoration. ©2005 BC Decker Inc.

Attractiveness of citrus pulp and orange albedo extracts to Atta sexdens rubropilosa (Hymenoptera: Formicidae)

The present study aimed to compare the attractiveness of industrial citrus pulp with the handmade orange albedo to the workers of Atta sexdens rubropilosa. For this, filter paper fragments were impregnated with organic extracts obtained through chemical extraction and sequential fractioning with hexane and dichloromethane and offered to different field nests. It was verified that the industrial citrus pulp extract is as good as the handmade orange albedo extract. This preference is discussed keeping in mind the chemical, behavioral and nutritional factors.

Sorption isotherm, glass transitions and state diagram for freeze-dried plum skin and pulp

Differential scanning calorimetry (DSC) was used to determine phase transitions of freeze-dried plums. Samples at low and intermediate moisture contents, were conditioned by adsorption at various water activities (0.11≤a w≤0.90) at 25°C, whereas in the high moisture content region (a w>0.90) samples were obtained by direct water addition, with the resulting sorption isotherm being well described by the Guggenheim-Anderson-deBoer (GAB) model. Freeze-dried samples of separated plum skin and pulp were also analysed. At a w≤0.75, two glass transitions were visible, with the glass transition temperature (T g) decreasing with increasing a w due to the water plasticising effect. The first T g was attributed to the matrix formed by sugars and water. The second one, less visible and less plasticised by water, was probably due to macromolecules of the fruit pulp. The Gordon-Taylor model represented satisfactorily the matrix glass transition curve for a w≤0.90. In the higher moisture content range T g remained practically constant around T g′ (-57.5°C). Analysis of the glass transition curve and the sorption isotherm indicated that stability at a temperature of 25°C, would be attained by freeze dried plum at a water activity of 0.04, corresponding to a moisture content of 12.9% (dry basis). © 2006 SAGE Publications.

Effects of hemostatic agents on the histomorphologic response of human dental pulp capped with calcium hydroxide

Objective: To evaluate the response of human pulps capped with a calcium hydroxide [Ca(OH)2] cement after bleeding control with 2 hemostatic agents. Method and Materials: Pulps were exposed on the occlusal floor, and the bleeding was controlled either with saline solution (SS) or 2.5% sodium hypochlorite (NaOCI) (SH). After that, the pulp was capped with Ca(OH) 2 cement and restored with resin composite. After 30 (groups SS30 and SH30) and 60 (groups SS60 and SH60) days, the teeth were extracted and processed with hematoxylin-eosin and categorized in a histologic score system. The data were subjected to Kruskal-Wallis and Mann-Whitney tests (α = .05). Results: Regarding dentin bridge formation, an inferior response of SH60 group was observed when compared to SS60 (P < .05). The response of the SH30 group generally was similar to that of the groups treated with saline solution. However, after 60 days, 2.5% NaOCl showed a trend toward having an inferior response. Conclusion: Using saline solution as a hemostatic agent before pulp capping with Ca(OH)2 resulted in a significantly better histomorphologic response than using 2.5% NaOCl as a hemostatic agent before capping with Ca(OH)2.

In vitro assessment of pulp chamber temperature of different teeth submitted to dental bleaching associated with LED/laser and halogen lamp appliances

This study sought to assess the pulp chamber temperature in different groups of human teeth that had been bleached using hydrogen peroxide gel activated with halogen lamps or hybrid LED/laser appliances. Four groups of ten teeth (maxillary central incisors, mandibular incisors, mandibular canines, and maxillary canines) were used. A digital thermometer with a K-type thermocouple was placed inside pulp chambers that had been filled with thermal paste. A 35% hydrogen peroxide-based red bleaching gel was applied to all teeth and photocured for a total of three minutes and 20 seconds (five activations of 40 seconds each), using light from an LED/laser device and a halogen lamp. The temperatures were gauged every 40 seconds and the data were analyzed by three-way ANOVA, followed by Tukey's test. Regardless of the light source, statistically significant differences were observed between the groups of teeth. The mean temperature values (±SD) were highest for maxillary central incisors and lowest for mandibular canines. The halogen lamp appliance produced more pulp chamber heating than the LED/laser appliance. The increase in irradiation time led to a significant increase in temperature.

Response of human dental pulp capped with MTA and calcium hydroxide powder

Objectives: To compare the response of human dental pulp capped with a mineral trioxide aggregate (MTA) and Ca(OH) 2 powder. Methods and Material: Pulp exposures were performed on the occlusal floor of 40 permanent premolars. The pulp was then capped with either Ca(OH) 2 powder (CH) or MTA and restored with resin composite. After 30 days (groups CH30 and MTA30) and 60 days (groups CH60 and MTA60), the teeth were extracted and processed for HE and categorized in a histological score system. The data were subjected to Kruskal-Wallis and Conover tests (α=0.05). Results: In regard to dentin bridge formation, CH30 showed a tendency towards superior performance compared to MTA30 (p>0.05), although the products showed comparable results at day 60. In the item Inflammation and General State of the Pulp (p>0.05), CH showed a tendency towards presenting a higher inflammatory response. In the item Other Pulpal Findings, MTA and Ca(OH) 2 showed equal and excellent performance after 30 and 60 days (p>0.05). Conclusion: After 30 days, Ca(OH) 2 powder covered with calcium hydroxide cement showed faster hard tissue bridge formation compared to MTA. After 60 days, Ca(OH) 2 powder or MTA materials showed a similar and excellent histological response with the formation of a hard tissue bridge in almost all cases with low inflammatory infiltrate. © Operative Dentistry, 2008.

Evaluation of two mineral trioxide aggregate compounds as pulp-capping agents in human teeth

Aim: The present randomized, controlled prospective study evaluated the histomorphological response of human dental pulps capped with two grey mineral trioxide aggregate (MTA) compounds. Methodology: Pulp exposures were performed on the occlusal floor of 40 human permanent pre-molars. The pulp was capped either with ProRoot (Dentsply) or MTA-Angelus (Angelus) and restored with zinc oxide eugenol cement. After 30 and 60 days, teeth were extracted and processed for histological examination and the effects on the pulp were scored. The data were subjected to Kruskal-Wallis and Conover tests (α = 0.05). Results: In five out of the 40 teeth bacteria were present in pulp tissue. No significant difference was observed between the two materials (P > 0.05) in terms of overall histological features (hard tissue bridge, inflammatory response, giant cells and particles of capping materials). Overall, 94% and 88% of the specimens capped with MTA-Angelus and ProRoot, respectively, showed either total or partial hard tissue bridge formation (P > 0.05). Conclusions: Both commercial materials ProRoot (Dentsply) and MTA-Angelus (Angelus) produced similar responses in the pulp when used for pulp capping in intact, caries-free teeth. © 2009 International Endodontic Journal.

Influence of the coloring agent concentration on bleaching gel and pulp chamber temperatures during dental bleaching

This study evaluated the Influence of the coloring agent concentration on the temperature of the gel layer and pulp chamber during dental bleaching with an LED/laser light source. Ten human incisors and a digital thermometer with K-type thermocouples were used. Using a high-speed spherical diamond bur, endodontic access was gained through openings on the lingual faces until pulp chamber was exposed. One end of the thermocouple was placed on the labial surface (immersed in bleaching gel) and the other end in the pulp chamber. The same 10 specimens were used in the 12 groups, according to the type and concentration of bleaching gel. Each bleaching gel was used in four different concentrations: manipulated without coloring, with normal quantity recommended by the manufacturer, with double the recommended amount of coloring, and with triple the recommended amount of coloring. The temperature rise was measured every 30 seconds for three minutes with a K-type thermocouple. The data were analyzed by ANOVA to examine the concentration and type of bleaching gel. This test was followed by Tukey's test, which was performed Independently for the gel at the labial surface and the pulp chamber (a = 5%). For both surfaces, values of p = 0.00 were obtained for all factors and for the Interaction between them. The varying concentrations of coloring agent produced statistically significant differences in terms of temperature increase for both the gel layer and the pulp chamber during activation.

Evaluation of the dentin-pulp complex after cavity preparation with ultrasonic diamond tip

The aim of this paper was to compare the dentin-pulp complex response to cavity preparation in human teeth using ultrasonic chemical vapor deposition (CVD) diamond tip and high-speed diamond bur. Class V buccal cavities were randomly prepared in 40 premolars from 14 patients aged 11 to 15 years. The cutting time was recorded and the cavities had the axial walls protected with gutta-percha and were filled with glass ionomer cement. The teeth were extracted at intervals of 0, 5, 10 and 20 days, and were decalcified, sectioned and stained by Hematoxylin & Eosin, Masson's Trichrome and Brown & Brenn techniques. The inflammatory response and cell disorganization were blindly evaluated by two examiners. The remaining dentin thickness (RDT) was measured by a linear scale using computer software. Statistical analysis by one-way ANOVA showed no statistically significant difference (P≤0.05) among the cavities prepared with either type of instrument, with mean RDT of 1132.50 mm. Cutting time and the pulp-dentin complex responses were analyzed statistically by Kruskal-Wallis and Dunn tests (P≤0.05). The ultrasonic CVD diamond tip took 5 times longer to prepare the cavities and there were no typical inflammatory pulp responses in cavities prepared with either type of cutting instrument, only mild to moderate cell disorganization was present. Even taking longer to cut the dental substrate, the ultrasonic CVD diamond tip produced similar pulp response compared to the conventional high-speed diamond bur.

Pulp tissue dissolution when the use of sodium hypochlorite and EDTA alone or associated

Purpose: The aim of the present study was to evaluate the tissue dissolving capacity of various concentrations of sodium hypochlorite either alone or in combination with 17% EDTA. Methods: Eighty bovine pulp fragments were prepared, and their weight was determined using a precision balance. Each pulp fragment was immersed for 2 hours in a solution/mixture that was based on the following groups: G1-saline solution; G2-0.5% NaOCl; G3-1.0% NaOCl; G4-2.5% NaOCl; G5-17% EDTA; G6-0.5% NaOCl+17% EDTA; G7-1.0% NaOCl+ 17% EDTA; and G8-2.5% NaOCl+17% EDTA. The final weight was measured, and the weight loss was calculated. A statistical analysis was performed using either the Student's t-test for paired samples or an ANOVA and Tukey tests (P<0.05 was considered to be significant). Results: We measured a significant difference between the sample weight before and after treatment for each of the tested groups (P<0.05). The 2.5% sodium hypochlorite solution (G4) completely dissolved the pulp tissue within the test period. NaOCl+EDTA was less effective than sodium hypochlorite alone at dissolving the pulp tissue (P<0.05), and EDTA alone (G5) did not markedly dissolve the pulp tissue. Conclusion: Using EDTA together with NaOCl reduced the tissue dissolving properties compared with NaOCl alone, regardless of the concentration of NaOCl that was used. © 2011 Só et al.; licensee EDIPUCRS.

Shelf Life of Fresh and Pasteurized Organic Passion Fruit (Passiflora Edulis F. Flavicarpa Deg.) Pulp

The shelf life of the organic passion fruit pulp, both fresh and pasteurized at 70C and 90C and stored under refrigeration, was evaluated. The heat treatment did not affect the sensory and physicochemical characteristics of the pasteurized pulps when compared with the fresh pulp, except for the ascorbic acid content. The pulps were also microbiologically safe. The pulps pasteurized at 70 and 90C were suitable for consumption for a minimum shelf-life period of 207 days of storage under refrigeration and for the fresh pulp it was attributed a shelf-life period of 60 to 90 days. The pulp pasteurized at 70C showed higher acceptance scores for all the attributes and purchase intention scores, suggesting a more stable behavior and higher sensory quality. Practical Applications: This work intended to evaluate the influence of the minimum pasteurization on the sensory acceptance and microbiological and physicochemical characteristics of the organic passion fruit pulp stored under refrigeration, with the aim to identify the shelf life. Heat treatment is one of the processes used for food preservation. Lower pasteurization temperature than that used by the Brazilian industries and storage under refrigeration showed to be appropriate for passion fruit pulp quality. In this way, the microbiological, physicochemical and sensory features of passion fruit were preserved. This work can be used as a reference for passion fruit pasteurization, which is able to increase the shelf life of this fruit while preserving its desirable original features. Journal Compilation © 2012 Wiley Periodicals, Inc.

A hydrogel scaffold that maintains viability and supports differentiation of dental pulp stem cells

Objectives: The clinical translation of stem cell-based Regenerative Endodontics demands further development of suitable injectable scaffolds. Puramatrix™ is a defined, self-assembling peptide hydrogel which instantaneously polymerizes under normal physiological conditions. Here, we assessed the compatibility of Puramatrix™ with dental pulp stem cell (DPSC) growth and differentiation. Methods: DPSC cells were grown in 0.05-0.25% Puramatrix™. Cell viability was measured colorimetrically using the WST-1 assay. Cell morphology was observed in 3D modeling using confocal microscopy. In addition, we used the human tooth slice model with Puramatrix™ to verify DPSC differentiation into odontoblast-like cells, as measured by expression of DSPP and DMP-1. Results: DPSC survived and proliferated in Puramatrix™ for at least three weeks in culture. Confocal microscopy revealed that cells seeded in Puramatrix™ presented morphological features of healthy cells, and some cells exhibited cytoplasmic elongations. Notably, after 21 days in tooth slices containing Puramatrix™, DPSC cells expressed DMP-1 and DSPP, putative markers of odontoblastic differentiation. Significance: Collectively, these data suggest that self-assembling peptide hydrogels might be useful injectable scaffolds for stem cell-based Regenerative Endodontics. © 2012 Academy of Dental Materials.