Laminin-binding epitope on gp43 from Paracoccidioides brasiliensis is recognized by a monoclonal antibody raised against Staphylococcus aureus laminin receptor

Adhesion is regarded as an important step in the pathogenesis of several microorganisms. Thus, the ability to recognize extracellular matrix proteins, such as laminin or fibronectin, has been correlated with invasiveness. Studying the already characterized laminin-binding protein of Paracoccidioides brasiliensis, the 43 kDa glycoprotein (gp43), we evaluated whether MAb 1.H12, raised against the laminin-binding protein from Staphylococcus aureus, cross-reacts with that fungal protein. By immunoblot analysis we show that MAb 1.H12 recognizes gp43. This interaction is able to inhibit the laminin-mediated adhesion to epithelial cells as well as the P. brasiliensis infection in vivo. Moreover, through immunoenzymatic assays, we show that MAb 1.H12 recognizes gp43 in solid phase and that this interaction is partially inhibited by the addition of anti-gp43 MAbs. These results show that MAb 1.H12 recognizes the gp43, suggesting the presence of an epitope similar to those found in the other laminin-binding proteins from phylogenetically very distant cells. These findings reinforce the possibility of evolutionary conservation of such epitopes.

Three-dimensional reconstruction of fracture surfaces: Area matching algorithms for automatic parallax measurements

This paper describes two solutions for systematic measurement of surface elevation that can be used for both profile and surface reconstructions for quantitative fractography case studies. The first one is developed under Khoros graphical interface environment. It consists of an adaption of the almost classical area matching algorithm, that is based on cross-correlation operations, to the well-known method of parallax measurements from stereo pairs. A normalization function was created to avoid false cross-correlation peaks, driving to the true window best matching solution at each region analyzed on both stereo projections. Some limitations to the use of scanning electron microscopy and the types of surface patterns are also discussed. The second algorithm is based on a spatial correlation function. This solution is implemented under the NIH Image macro programming, combining a good representation for low contrast regions and many improvements on overall user interface and performance. Its advantages and limitations are also presented.

Tracheobronchial consequences of the use of heat and moisture exchangers in dogs

Purpose: To determine the effect of heat and moisture exchange (HME) on the tracheobronchial tree (TBT) using a unidirectional anesthesic circuit with or without CO2 absorber and high or low fresh gas flow (FGF), in dogs. Methods: Thirty-two dogs were randomly allocated to four groups: G1 (n = 8) valvular circuit without CO2 absorber and high FGF (5 L·min-1); G2 (n = 8) as G1 with HME; G3 (n = 8) circuit with CO2 absorber with a low FGF (1 L·min-1); G4 (n = 8) as G3 with HME. Anesthesia was induced and maintained with pentobarbital. Tympanic temperature (TT), inhaled gas temperature (IGT), relative (RH) and absolute humidity (AH) of inhaled gas were measured at 15 (control), 60, 120 and 180 min of controlled ventilation. Dogs were euthanized and biopsies in the areas of TBT were performed by scanning electron microscopy. Results: The G2 and G4 groups showed the highest AH (>20 mgH2O·L-1) and G1 the lowest (< 10 mgH2O·L-1) and G3 was intermediate (<20 mgH2O·L-1) (P < 0.01). There was no difference of TT and IGT among groups. Alterations of the mucociliary system were greatest in G1, least in G2 and G4, and intermediate in G3. Conclusion: In dogs, introduction of HME to a unidirectional anesthetic circuit with/without CO2 absorber and high or low FGF preserved humidity of inspired gases. HME attenuated but did not prevent alterations of the mucociliary system of the TBT.

Regression of the lateral oviducts during the larval-adult transformation of the reproductive system of Melipona quadrifasciata and Frieseomelitta varia

Toward the end of the larval phase (pre-pupa), the reproductive systems of Melipona quadrifasciata and Frieseomelitta varia workers are anatomically similar. Scanning electron microscopy showed that during this developmental phase the right and left ovaries are fused and form a heart-shaped structure located above the midgut. Each ovary is connected to the genital chamber by a long and slender lateral oviduct. During pupal development, the lateral oviducts of workers from both species become extremely reduced due to a drastic process of cell death, as shown by transmission electron microscopy. During the lateral oviduct shortening, their simple columnar epithelial cells show some signs of apoptosis in addition to necrosis. Cell death was characterized by cytoplasmic vesiculation, peculiar accumulation of glycogen, and dilation of cytoplasmic organelles such as mitochondria and rough endoplasmic reticulum. The nuclei, at first irregularly contoured, became swollen, with chromatin flocculation and various areas of condensed chromatin next to the nuclear envelope. At the end of the pupal phase, deep recesses marked the nuclei. At emergence, worker and queen reproductive systems showed marked differences, although reduction in the lateral oviducts was an event occurring in both castes. However, in queens the ovarioles increased in length and the spermatheca was larger than that of workers. At the external anatomical level, the reproductive system of workers and queens could be distinguished in the white- and pink-eyed pupal phase. The metamorphic function of the death of lateral oviduct cells, with consequent oviduct shortening, is discussed in terms of the anatomical reorganization of the reproductive system and of the ventrolateral positioning of adult worker bee ovaries. (C) 2000 Wiley-Liss, Inc.

Use of one-bottle adhesive as an intermediate bonding layer to reduce sealant microleakage on saliva-contaminated enamel

Purpose: To evaluate the influence of three different adhesives, each used as an intermediary layer, on microleakage of sealants applied under condition of salivary contamination. Materials and Methods: Six different experimental conditions were compared, 3 with adhesives and 3 without. After prophylaxis and acid etching of enamel, salivary contamination was placed for 10 s. In Group SC the sealant was applied after saliva without bonding agent and then light-cured. In Group SCA, after saliva, the surface was air dried, and then the sealant was applied and cured. In Groups ScB, SB and PB, a bonding agent (Scotchbond Dual Cure/3M, Single Bond/3M and Prime & Bond 2.1/Dentsply, respectively) was applied after the saliva and prior to the sealant application and curing. After storage in distilled water at 37°C for 24 hrs, the teeth were submitted to 500 thermal cycles (5°C and 55°C), and silver nitrate was used as a leakage tracer. Leakage data were collected on cross sections as percentage of total enamel-sealant interface length. Representative samples were evaluated under SEM. Results: Sealants placed on contaminated enamel with no bonding agent showed extensive microleakage (94.27% in SC; 42.65% in SCA). The SEM revealed gaps as wide as 20 μm in areas where silver nitrate leakage could be visualized. In contrast, all bonding agent groups showed leakage less than 6.9%. Placement of sealant with a dentin-bonding agent on contaminated enamel significantly reduced microleakage (P< 0.0001). The use of a bonding agent as an intermediary layer between enamel and sealant significantly reduced saliva's effect on sealant microleakage.

Morphology of the ventral lobe of the prostate and seminal vesicles in an ethanol-drinking strain of rats (UChA and UChB)

Chronic alcoholism alters reproduction and therefore may be responsible for alterations of prostate and seminal vesicles, which are the subject of this analysis in UCh ethanol-drinking rats. The prostate and seminal vesicles of 20 animals were submitted to macroscopic, light microscopy, electron microscopy and morphometric analysis. The UCh rats showed atrophy of the epithelium and reduction of the weight of the prostate and seminal vesicle, liver hypertrophy and fat infiltration and alterations of the hypothalamus-pituitary axis. Ethanol induces changes in the weight and in the epithelium of prostate and seminal vesicles and hypothalamus-pituitary axis of UCh rats.

Morphology and histochemistry of the hyolingual apparatus in chameleons

We reexamined the morphological and functional properties of the hyoid, the tongue pad, and hyolingual musculature in chameleons. Dissections and histological sections indicated the presence of five distinctly individualized pairs of intrinsic tongue muscles. An analysis of the histochemical properties of the system revealed only two fiber types in the hyolingual muscles: fast glycolytic and fast oxidative glycolytic fibers. In accordance with this observation, motor-endplate staining showed that all endplates are of the en-plaque type. All muscles show relatively short fibers and large numbers of motor endplates, indicating a large potential for fine muscular control. The connective tissue sheet surrounding the entoglossal process contains elastin fibers at its periphery, allowing for elastic recoil of the hyolingual system after prey capture. The connective tissue sheets surrounding the m. accelerator and m. hyoglossus were examined under polarized light. The collagen fibers in the accelerator epimysium are configured in a crossed helical array that will facilitate limited muscle elongation. The microstructure of the tongue pad as revealed by SEM showed decreased adhesive properties, indicating a change in the prey prehension mechanics in chameleons compared to agamid or iguanid lizards. These findings provide the basis for further experimental analysis of the hyolingual system. © 2001 Wiley-Liss, Inc.

Root instrumentation with an erbium:yttrium-aluminum-garnet laser: Effect on the morphology of fibroblasts

Objective: The purpose of this study was to evaluate the effect of erbium:yttrium-aluminum-garnet laser instrumentation of root surfaces on the morphology of fibroblasts from continuous lineage. Method and materials: Dentinal slices with 4 mm2 of surface area were obtained from teeth extracted for severe periodontal involvement. Specimens were assigned to one of three treatment groups: group 1, application of the laser with an energy level of 250 mJ at 103 pulses per second; group 2, application of the laser with an energy level of 80 mJ at 166 pulses per second; and group 3, similar to group 2, but with concomitant water irrigation of the device. The specimens were incubated in multiwell plates containing cell culture media. After 24 hours, the specimens were submitted to routine preparation for scanning electron microscopy. Three independent and blind examiners used photomicrographs to evaluate the morphology of the fibroblasts: 0 = without cells; 1 = flat cells; 2 = round cells; and 3 = combination of round and flat cells. Results: Statistical analysis indicated that there were significant differences among treatment groups and that group 3 was significantly different from groups 1 and 2. Conclusion: There was no difference between groups 1 and 2 in the morphology of fibroblasts. Laser instrumentation with concomitant irrigation impaired the adhesion of fibroblasts to dentinal surfaces.

EM evaluation of bacterial biofilm and microorganisms on the apical external root surface of human teeth

The aim of this study was to evaluate the presence of bacterial biofilm on the external surface of the root apex in teeth with pulp necrosis, with and without radiographically visible periapical lesions, and in teeth with a vital pulp. Twenty-one teeth were extracted, eight with pulp necrosis and periapical lesions, eight with pulp necrosis without radiographically visible periapical lesions, and five with a vital pulp. The roots were sectioned, and the root apexes (+/- 3 mm) were processed for scanning electron microscope evaluation. The surface of the apical root was evaluated for the presence of microorganisms, root resorption, and biofilm. There were no microorganisms on the apical root surface of either teeth with pulp vitality or with pulp necrosis with no radiographically visible periapical lesions. Microorganisms were always present in teeth with pulp necrosis and radiographically visible periapical lesions. These included cocci, bacilli, and filaments and the presence of an apical biofilm. Apical biofilm is clinically important because microbial biofilms are inherently resistant to antimicrobial agents and cannot be removed by biomechanical preparation alone. This may cause failure of endodontic treatment as a consequence of persistent infection.

Structure of the pelvic and penile urethra - Relationship with the ducts of the sex accessory glands of the Mongolian gerbil (Meriones unguiculatus)

The urethra is the main place of entry for sexually transmitted pathogens. However, there is little literature on the morphology of the urogenital system, principally the urethra and ducts of the sex accessory glands. The Mongolian gerbil is an insectivorous, herbivorous and monogamous rodent with nocturnal habits; it has been used successfully as a laboratory animal since the 1960s. Therefore, the objective of the present paper was to describe the structure and ultrastructure of the urethra and its relations to the ducts of the accessory sex glands of the Mongolian gerbil (Meriones unguiculatus), contributing to the understanding of the reproductive biology of the rodent and aiming to provide data for future experimental studies. Conventional techniques of light and scanning electron microscopy were utilized. The urethra and ducts of the accessory sex glands are similar to those of the albino rat and the mouse. However, there is variation in drainage type among accessory sex glands for the inner urethra. The ducts of the seminal vesicle, the ductus deferens, drain their contents independently into the ampullary duct that opens in the urethra. The ducts of the prostate, coagulating and bulbourethral glands drain their contents independently into the urethra.

Effect of air-powder system on titanium surface on fibroblast adhesion and morphology.

PURPOSE: To evaluate the number and morphology of fibroblasts grown on machined titanium healing abutments treated with an airpowder system. MATERIALS AND METHODS: Twenty-six abutments were assigned to two experimental groups: control (no treatment) and treated (exposed to the Prophy-Jet for 30 seconds). The specimens were incubated for 24 hours with fibroblastic cells in multiwell plates, followed by routine laboratory processing for scanning electron microscope analysis. The specimens were photographed at x 350, and the cell number was counted on an area of approximately 200 um2. RESULTS: No significant differences were found on morphology between the groups (P > 0.05); however, the control group presented a significantly greater amount of cells (71.44 +/- 31.93, mean +/- SD) in comparison with treated group (35.31 +/- 28.14), as indicated by a nonpaired t test (P = 0.001). CONCLUSION: The use of an air-abrasive prophylaxis system on the surface of titanium healing abutments reduced the cells proliferation but did not influence cell morphology.

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

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.

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

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.

Effect of 2% chlorhexidine on microtensile bond strength of composite to dentin

Purpose: To evaluate the effect of 2% chlorhexidine on the microtensile bond strength of composite resin to dentin treated with three dentin bonding systems. Materials and Methods: Flat dentinal surfaces were prepared in 24 extracted human third molars. Teeth were randomly divided into 8 distinct experimental groups according to the adhesive applied (Prime & Bond NT, Single Bond and Clearfil SE Bond), the application (yes/no) of chlorhexidine, and the time point at which it was applied (before or after acid etching the dentin). Composite resin blocks were built up over treated surfaces, and teeth were then stored in water at 37°C for 24 h. Samples were thermocycled, stored under the same conditions, and then vertically sectioned, thus obtaining specimens with 1.0 ± 0.1 mm2 cross-sectional area. Specimens were stressed in tension at 0.5 mm/min crosshead speed. Bond strength results were evaluated using a one-way ANOVA (p < 0.05). The modes of failures were verified using optical microscopy. Dentin disks were obtained from 3 additional teeth treated in the same manner for observation under SEM. The most representative samples of fractured specimens were also observed under SEM. Results: No statistically significant differences of bond strength values were found between any groups. Failures occurred mainly within the bond; exclusively adhesive fractures (adhesive-dentin) were not observed. Conclusion: The 2% chlorhexidine solution, applied before or after acid etching of the dentin, did not interfere with the microtensile bond strength of composite resin to the dentin treated with Prime & Bond NT, Single Bond, or Clearfil SE Bond bonding systems.

Effectiveness of Microwave Sterilization on Three Hard Chairside Reline Resins

Purpose: The aim of this study was to evaluate the effectiveness of microwave irradiation sterilization on hard chairside reline resins. Materials and Methods: Specimens of three reline resins (Kooliner, Tokuso Rebase, and Ufi Gel Hard) were fabricated and subjected to ethylene oxide sterilization. The specimens were then individually inoculated (107 cfu/mL) with Tryptic Soy Broth media containing one of the tested microorganisms (C albicans, S aureus, B subtilis, and P aeruginosa). After 48 hours at 37°C, the samples were vortexed for 1 minute and allowed to stand for 9 minutes, followed by a short vortex to resuspend any organisms present. After inoculation, 40 specimens of each material were immersed in 200 mL of water and subjected to microwave irradiation at 650 W for 6 minutes. Forty non-irradiated specimens were used as positive controls. Replicate specimens (25 μL) of suspension were plated at dilutions of 10-3 to 10-6 on plates of selective media appropriate for each organism. All plates were incubated at 37°C for 48 hours. After incubation, colonies were counted, and the data were statistically analyzed by the Kruskal-Wallis test. Twelve specimens of each material were prepared for SEM. Results: All immersed specimens showed consistent sterilization of all the individual organisms after microwave irradiation. SEM examination indicated an alteration in cell morphology after microwave irradiation. Conclusion: Microwave sterilization for 6 minutes at 650 W proved to be effective for the sterilization of hard chairside reline resins.