PURIFICATION AND PROPERTIES OF SHIKIMATE DEHYDROGENASE FROM CUCUMBER (CUCUMIS-SATIVUS L)

Shikimate dehydrogenase (SDH, EC 1.1.1.25) extracted from cucumber pulp (Cucumis sativus L.) was purified 7-fold by precipitation with ammonium sulfate and elution from columns of Sephadex G-25, DEAE-cellulose, and hydroxyapatite. Two activity bands were detected on polyacrylamide gel electrophoresis at the last purification step. pH optimum was 8.7, and molecular weight of 45 000 was estimated on a Sephadex G-100 column. SDH was inhibited competitively by protocatechuic acid with a K(i) value of 2 x 10-4 M. K(m) values of 6 x 10-5 and 1 x 10-5 M were determined for shikimic acid and NADP+, respectively. The enzyme was completely inhibited by HgCl2 and p-(chloromercuri)benzoate (PCMB). NaCl and KCl showed partial protection against inhibition by PCMB. Heat inactivation between 50 and 55-degrees-C was biphasic, and the enzyme was completely inactivated after 10 min at 60-degrees-C. Incubation of SDH with either NADP+ or shikimic acid protected the enzyme against heat inactivation.

Correlation among immune response, morphogenesis of the granulomatous reaction and spleen lymphoid structure in murine experimental paracoccidioidomycosis

We studied the correlation among cellular immune response, the pattern of lung granulomatous lesions and alterations in spleen lymphoid structure in Swiss mice inoculated intravenously with Paracoccidioides brasiliensis strain 18. The animals were evaluated at 24, 48 and 96 h after infection and further studied weekly for 18 weeks by: (i) the macrophage migration inhibition test with phytohemagglutinin (PHA) and P. brasiliensis antigen (PbAg); and (ii) histopathology of the lung and spleen lesions. One group of animals was gamma -irradiated (8 Gy), infected under the same conditions and evaluated for the pattern of lung granulomatous lesions and spleen lymphoid structure at 24, 48 and 96 h after infection. During the first week of infection, the non-irradiated animals presented a positive response to PHA and PbAg, compact granulomas in the lungs and a typical hyperplasia of the spleen white pulp. However, from weeks 2 to 5, a depression of the cell-mediated immunity (CMI) response to PHA and PbAg was observed in association with granulomas presenting only large mononuclear cells and lacking both giant cells and a peripheral halo of small mononuclear cells. This pattern of granuloma formation was similar to that seen in gamma -irradiated animals, whose cells involved in CMI were absent. After week 7, the non-irradiated animals showed granulomas characterized by the presence of giant cells and a peripheral halo of small mononuclear cells. This type of granuloma was formed concomitantly with recovery of the CMI and of the lymphoid structure of the spleen. The results showed a correlation among granulomas composed of large mononuclear cells, hypoplasia of the splenic tissue and impaired CMI. This correlation indicated that although granuloma morphogenesis per se does not depend on the activation of CMI, this response is important at later stages during modulation of the cellular composition of the granulomas.

Acerola's clones of industrial interest

In order to know which clone of acerola is better for acerola industrialization, we studied the pectin methylesterase (PME) specific activity, pectin content and vitamin C content in five different clones of acerola. The pectin yield varied from 1.37 to 2.99% and the highest content of pectin occurred in clones 3 and 5. Ascorbic acid varied significantly from 1157.5 to 1735.5 mg/100 g of pulp in the five clones. The highest content of vitamin C occurred in clone 4. The PME specific activity varied from 0.79 to 2.92 units g(-1)/g of pulp and the highest values occurred in clone 2. We also studied the optimum temperature and the optimum pH of this enzyme. Clones 1, 2, 4 and 5 showed optimum temperature at 90C. Clone 3 showed practically the same specific activity at all temperatures studied. Clones 1 and 4 showed an optimum pH of 9.0 and clone numbers 2, 3 and 5 showed a pH optimum at 8.5.

Tissue response to an epoxy resin-based root canal sealer

The objective of the present study was to evaluate two different types of root canal sealers: AH Plus (an epoxy resin-based sealer) and Fill Canal (a zinc oxide-eugenol based sealer). A total of 34 root canals with vital pulp from dogs' premolars were used. After instrumentation, the root canals were filled with gutta-percha and AH Plus or gutta-percha and Fill Canal sealers using a classical technique of lateral condensation. After histological processing, the sections were stained with hematoxylineosin or Mallory's trichrome stain. Inflammatory cells or areas of necrosis were not associated with AH Plus. Hard tissue formation apically to the material was observed in 14 specimens. The Fill Canal sealer presented an inflammatory response of moderate intensity in the periapical region, mainly adjacent to the material.

Reactionary dentinogenesis after applying restorative materials and bioactive dentin matrix molecules as liners in deep cavities prepared in nonhuman primate teeth

Objective: the aim of this in vivo study was to evaluate the response of the pulp-dentin complex following application of resin-modified glass-ionomer cement, calcium hydroxide hard-setting cement and EDTA-soluble preparation of dentine matrix proteins (ESDP) in deep cavities prepared in non-human primate teeth. Methods: Eighteen deep Class V buccal cavities were prepared in premolars of four capuccin monkeys. In Groups 1 and 2, the cavity floor was lined with ESDP or a resin-modified glass-ionomer cement (Vitrebond - 3M ESPE), respectively. In Group 3 (control), the cavity was lined with a hard setting calcium hydroxide cement (Dycal - Dentsply). The cavities were subsequently filled with amalgam. After 6 months, the animals were sacrificed and the teeth were prepared for microscopic assessment. Six-micron thick serial sections were stained with H/E, Masson's trichrome and Brown & Brenn techniques. Results: No inflammatory pulpal response was observed for all experimental and control Groups. However, the amount of reactionary dentin deposition differed between groups in the rank order ESDP (Group 1) > calcium hydroxide (Group 3) > resin-modified glass-ionomer (Group 2). These differences were statistically significant. Conclusions: All materials were biocompatible when applied in deep cavities. ESDP stimulated higher deposition of reactionary dentin matrix than Vitrebond and Dycal.

Pulpal temperature increases with Er : YAG laser and high-speed handpieces

Statement of problem. During tooth preparation, both high-speed handpieces and lasers generate heat, which, if not controlled, can cause pulpal necrosis.Purpose. The aim of this study was to compare temperature increases produced by a high-speed dental handpiece with those produced by a relatively new instrument, the Er:YAG (erbium: yttrium-aluminum-garnet) laser.Material and methods. Thirty bovine mandibular incisors were reduced to an enamel/dentin thickness of 2.5 mm. Class V preparations were completed to a depth of 2.0 mm, measured with a caliper or by a mark oil the burs. A thermocouple was placed inside the pulp chamber to determine temperature increases (degreesC). Analysis was performed on the following groups (n = 10): Group 1, high-speed handpiece without water cooling, Group 11, high-speed handpiece with water cooling (30 mL/min), and Group III, the noncontact Er:YAG laser (2.94 mum at 350 mJ/10 Hz) with water cooling (4.5 mL/min). The temperature increases were recorded by a computer linked to the thermocouples. The data were analyzed using the Kruskal-Wallis test. The Dunn multiple comparison test was used as post hoc test (alpha = .05).Results. The average temperature rises were: 11.64degreesC (+/-4.35) for Group 1, 0.96degreesC (+/-0.71) for Group 11, and 2.69degreesC (+/-1.12) for Group III. There were no statistical differences between Groups 11 and III, both 11 and III differed from Group I significantly (P = .000 and P = .002, respectively).Conclusion. The preparations made with the high-speed and the laser instrument generated similar heat increases under water cooling. Water cooling was essential to avoid destructive temperature increases when using both the high-speed handpiece and laser.

Sealing evaluation of the cervical base in intracoronal bleaching

Discoloration of non-vital teeth is an esthetic deficiency frequently requiring bleaching treatment. The purpose of this study was to evaluate in vitro the cervical base efficacy in order to prevent or to minimize the leakage along the root canal filling and into the dentinal tubules. Thirty-eight extracted single-root human teeth were used, which were biomechanically prepared, filled, and divided into three experimental groups: G1, a cervical base was applied (3 mm of thickness) below the cemento-enamel junction, with resin-modified glass-ionomer cement (Vitremer); G2, the base was done with glass-ionomer cement (Vidrion R); and G3 (Control), did not receive any material as base. A mixture of sodium perborate and hydrogen peroxide 30% was placed inside the pulp chamber for 3 days, and the access opening was sealed with Cimpat. This procedure was repeated thrice. Soon after this, a paste of calcium hydroxide was inserted into the pulp chamber for 14 days. All teeth were covered with two layers of sticky wax, except the access opening, and immersed in blue India Ink for 5 days. The results did not show statistically significant differences between the three groups concerning the leakage inside the dentinal tubules. Regarding the apical direction, a statistical difference (ANOVA P < 0.05) was observed among the experimental group G1 and control group G3. No statistically significant difference was observed between G2 and G3 groups. Therefore, the placement of a cervical base before internal bleaching procedures is still recommended.

Effect of irrigating solution and calcium hydroxide root canal dressing on the repair of apical and periapical tissues of teeth with periapical lesion

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Prevalence of different types of accessory canals in the furcation area of third molars

Background: Although there are several studies that show the prevalence and diameter of accessory root canals in the furcation area, there is a scarceness of studies that observe the trajectory and different types of cavo-interradicular canals. The aim of this study was to verify the prevalence of the different morphologic types of accessory canals in the furcation region in an attempt to show their trajectories.Methods: Forty submerged mandibular third molars were used, which were extracted and decalcified so that the microtomy procedure in the mesio-distal axial plane could be performed, obtaining semiserial sections with thicknesses of 5 mu m. The sections were stained with hematoxylin and eosin and observed under optical microscopy.Results: All of the morphologic types were found, whereas the proper accessory canals, type-A canals, were present in 10% of the specimens. The most prevalent canals were the sealed ones (type D), with a prevalence of 87.5%, followed by the blind ones (type B), with a prevalence of 75%. The loop accessory canals (type C), observed in only 5% of the teeth, were the least prevalent ones.Conclusions: Different morphologic types of accessory canals were found in the furcation area of submerged mandibular molars. The histologic method was effective to show the canal trajectories.

Rheological properties of frozen concentrated orange juice (FCOJ) as a function of concentration and subzero temperatures

Data on flow properties of Frozen Concentrated Orange Juice (FCOJ) produced from oranges cv. Pera-Rio (65.04 Brix, 8.8% w/w pulp content, 2.5% w/w pectin, 3.84% citric acid, 1.293 g cm(-3)) from -18 to 0 degrees C were fitted with appropriate predictive models. The power law model was found to be the most appropriate to fit the flow curves obtained for FCOJ between 46.56 and 65.04 degrees Brix. In higher concentrations, thixotropy was observed and showed more temperature dependence. A single equation combining Arrhenius and exponential relationships was applied to describe the temperature effect and shear rate on the quantity of breakdown of FCOJ.

Evaluation of subsurface dentin microhardness after Er : yttrium-aluminum-garnet and Nd : yttrium-aluminum-garnet laser irradiation

Background and objectives: To assess the microhardness of dentin subsurface after Er:yttrium-aluminum-garnet (YAG) and Nd:YAG laser irradiation. Study design/materials and methods: Twenty-four bovine incisors, without pulp, were used. The vestibular surface was worn out until the dentin was reached and divided in mesial and distal regions. The samples were divided into two groups: GI-distal, irradiated by Er: YAG laser, and GII-distal, irradiated by Nd: YAG laser. The mesial area was protected so as to not receive the laser irradiation. The measurements were made on Vickers digital microhardmeter. Results: For GI-there was no significant statistical difference, Cl(-4.59 to 0.78), between the values of irradiated (55.61 +/- 4.38) and unirradiated (57.51 +/- 4.00) areas. For GII-the values were higher for the irradiated (62.21 +/- 6.48) compared to the unirradiated (57.82 +/- 5.42) area, CI(1.65 +/- to 7.13). Conclusions: There was an increase of dentin microhardness when the Nd: YAG was used, but the Er: YAG did not cause significant alterations in dentin microhardness. (c) 2007 Laser Institute of America.

Cytotaxonomic analysis of Brazilian species of the genus Amazona (Psittacidae, Aves) and confirmation of the genus Salvatoria (Ribeiro, 1920)

The karyotypes of 12 species of Psittacidae of the genus Amazona were studied: A. aestiva, A. amazonica, A. brasiliensis, A. autumnalis, A. farinosa, A, festiva, A. kawalli, A. ochrocephala, A. pretrei, A. rhodocorytha, A. vinacea and A. xanthops. The metaphases were obtained using a short term culture of leather pulp. Eleven of the twelve analyzed species were karyotypically homogeneous, with only a few divergences in chromosomes 2 and 3. The species A. xanthops showed large karyotypic differences compared to the genus Amazona. Consequently, the genus Salvatoria (Ribeiro, Rev. Mus. Paul. 12: 1-82, 1920) was confirmed, and A. xanthops renamed Salvatoria xanthops. The study showed the chromosomic conservation of the genus Amazona and the need for further taxonomic studies of the karyotype of the Psittacidae.

Effect of a pilot washing system on dicofol levels in orange matrix

An efficient analytical method is described for the analysis of dicofol residues in pulp and orange peel. Samples are mixed with Celite and transferred to chromatographic columns prepacked with silica gel. Dicofol is eluted with ethyl acetate, and the extracts are analyzed by gas chromatography with electron capture detection. Mean recoveries for dicofol at levels of 0.5, 2.0, 5.0, and 10 mg/kg ranged from 87 to 95% with relative standard deviation values between 2.6 and 9.0%. To investigate the effect of a pilot washing system on dicofol residues in oranges, the analytical procedure was applied to samples submitted to different treatments with commercial formulations under field and laboratory conditions. The orange samples with and without washing were analyzed in duplicate, and the results indicated that washing under the described conditions did not allow a complete removal of dicofol residues from orange peel.

Pectinolytic enzyme production by Bacillus species and their potential application on juice extraction

Five Bacillus strains isolated from decaying vegetable material were cultivated on wheat bran and endo-polygalacturonases, exo-polygalacturonase and pectin lyase activities in the crude enzymatic solution obtained were determined. Highest activity was observed for all enzymes when fermentation was carried out at 28 degreesC, the highest activity values were obtained after 120 h of cultivation for exo-PG and after 48 h for endo-PG and PL. The use of the enzymatic solution for treatment of fruits and vegetable mash afforded a high juice extraction and a pulp with good pressing characteristics.

Consequences of crown fractures with pulpal exposure: histopathological evaluation in dogs

When a crown fracture involving pulpal exposure is produced, the therapeutic treatment to be applied depends to a great extent on the general histopathological condition of the exposed pulp. Hence, the objective of this study was to evaluate histopathological and bacteriological changes occurring in dental tissue and periradicular tissue of crown-fractured teeth with pulpal exposure. Twenty-four anterior teeth (central and lateral incisors) from the maxillary teeth of four young, adult Mongrel dogs were used. At 48 and 72 h after performing the crown fractures, the animals were sacrificed and the results evaluated. Both observation periods revealed the existence of an area of superficial inflammation with the formation of hyperplastic tissue towards the external surface. Intense neutrophilic infiltrate was observed below it. Mean depth of inflammation was greater at 48 h (4633.33 mum) than at 72 h (3933.33 mum), perhaps coinciding with the bigger pulp chamber opening (x1332.14 mum at 48 h vs. x479.52 mum at 72 h). Upon approaching the cervical portion, the inflammation became less. Bacterial contamination was constant in all the cases evaluated, worsening the histopathological findings with exposure time. This study demonstrates that when a crown fracture with pulpal exposure is produced, the success in treating it depends partly on how quickly therapeutic treatment is administered.