LipL53, a temperature regulated protein from Leptospira interrogans that binds to extracellular matrix molecules

The regulation of gene expression by environmental signals, such as temperature and osmolarity, has been correlated with virulence. In this study, we characterize the protein LipL53 from Leptospira interrogans, previously shown to react with serum sample of individual diagnosed with leptospirosis and to be up-regulated by shift to physiological osmolarity. The recombinant protein was expressed in Escherichia coli system, in insoluble form, recovered by urea solubilization and further refolded by decreasing the denaturing agent concentration during the purification procedure. The secondary structure content of the recombinant LipL53, as assessed by circular dichroism, showed a mixture of beta-strands and alpha-helix. The presence of LipL53 transcript at 28 degrees C was only detected within the virulent strains. However, upon shifted of attenuated cultures of pathogenic strains from 28 degrees C to 37 degrees C and to 39 degrees C, this transcript could also be observed. LipL53 binds laminin, collagen IV, cellular and plasma fibronectin in dose-dependent and saturable manner. Animal challenge studies showed that LipL53, although immunogenic, elicited only partial protection in hamsters. LipL53 is probably surface exposed as seen through immunofluorescence confocal microscopy. Our results suggest that LipL53 is a novel temperature regulated adhesin of L. interrogans that may be relevant in the leptospiral pathogenesis. (C) 2009 Elsevier Masson SAS. All rights reserved.

Influence of Photoperiod and Temperature on the Larval Behavioral Diapause of Amblyomma cajennense (Acari: Ixodidae)

Larval behavioral diapause was shown to be the major factor controlling the 1-yr generation pattern of Amblyomma cajennense (F.) (Acari: Ixodidae) in Brazil. During fieldwork, this behavior was shown to coincide with long daylength (>12 h) and high mean ground temperature (approximate to 25 degrees C), which prevail during spring-summer in Brazil. The current study evaluated biological parameters of engorged females, their eggs, and the resultant larvae inside plastic pots planted with the grass Brachiaria decumbens Stapf. held in incubators set with different combinations of temperature and photoperiod. Both the long daylength (photoperiod 14:10 [L:D]h) and high temperature (25 degrees C) during larval hatching induced larval behavioral diapause, characterized by the confinement of hatched larvae on the ground below the vegetation for many weeks. When long daylength was present during hatching, but temperature was low (15 degrees C), larvae did not enter diapause. Similarly, when short daylength (10:14 or 12:12) was present during larval hatching, larvae did not enter diapause regardless whether temperature was high (25 degrees C). Termination of diapause was induced by shifting photoperiod from 14:10 to 12:12 or the temperature from 25 to 15 degrees C. When applied to field conditions, the present results indicate that both high ground mean temperature (approximate to 25 degrees C) and long daylength (>12 h) during spring-summer (October-March) are responsible for the induction and maintenance of A. cajennense larval behavioral diapause in the field. Furthermore, both the low ground mean temperature (-20 degrees C) and the short daylength (<12h) during autumn (April-May) are responsible for termination of larval behavioral diapause in the field.

Genotype effects on body temperature in dairy cows under grazing conditions in a hot climate including evidence for heterosis

We compared diurnal patterns of vaginal temperature in lactating cows under grazing conditions to evaluate genotype effects on body temperature regulation. Genotypes evaluated were Holstein, Jersey, Jersey x Holstein and Swedish Red x Holstein. The comparison of Holstein and Jersey versus Jersey x Holstein provided a test of whether heterosis effects body temperature regulation. Cows were fitted with intravaginal temperature recording devices that measured vaginal temperature every 15 min for 7 days. Vaginal temperature was affected by time of day (P < 0.0001) and genotype x time (P < 0.0001) regardless of whether days in milk and milk yield were used as covariates. Additional analyses indicated that the Swedish Red x Holstein had a different pattern of vaginal temperatures than the other three genotypes (Swedish Red x Holstein vs others x time; P < 0.0001) and that Holstein and Jersey had a different pattern than Jersey x Holstein [(Holstein + Jersey vs Jersey x Holstein) x time, P < 0.0001]. However, Holstein had a similar pattern to Jersey [(Holstein vs Jersey) x time, P > 0.10]. These genotype x time interactions reflect two effects. First, Swedish Red x Holstein had higher vaginal temperatures than the other genotypes in the late morning and afternoon but not after the evening milking. Secondly, Jersey x Holstein had lower vaginal temperatures than other genotypes in the late morning and afternoon and again in the late night and early morning. Results point out that there are effects of specific genotypes and evidence for heterosis on regulation of body temperature of lactating cows maintained under grazing conditions and suggest that genetic improvement for thermotolerance through breed choice or genetic selection is possible.

Comparative analysis of root surface smear layer removal by different etching modalities or erbium:yttrium-aluminum-garnet laser irradiation. A scanning electron microscopy study

The purpose of this study was to evaluate the effect of erbium:yttrium-aluminum-garnet (Er:YAG) laser (2.94 mu m) irradiation on the removal of root surface smear layer of extracted human teeth and to compare its efficacy with that of citric acid, ethylenediamine tetra-acetic acid (EDTA), or a gel containing a mixture of tetracycline hydrochloride (HCl) and citric acid, using scanning electron microscopy (SEM). Thirty human dentin specimens were randomly divided into six groups: G1 (control group), irrigated with 10 ml of physiologic saline solution; G2, conditioned with 24% citric acid gel; G3, conditioned with 24% EDTA gel; G4, conditioned with a 50% citric acid and tetracycline gel; G5, irradiated with Er:YAG laser (47 mJ/10 Hz/5.8 J/cm(2)/pulse); G6, irradiated with Er:YAG laser (83 mJ/10 Hz/10.3 J/cm(2)/pulse). Electron micrographs were obtained and analyzed according to a rating system. Statistical analysis was conducted with Kruskal-Wallis and Mann-Whitney tests (P < 0.05). G1 was statistically different from all the other groups; no statistically significant differences were observed between the Er:YAG laser groups and those undergoing the other treatment modalities. When the two Er:YAG laser groups were compared, the fluency of G6 was statistically more effective in smear layer removal than the one used in G5 (Mann-Whitney test, P < 0.01). Root surfaces irradiated by Er:YAG laser had more irregular contours than those treated by chemical agents. It can be concluded that all treatment modalities were effective in smear layer removal. The results of our study suggest that the Er:YAG laser can be safely used to condition diseased root surfaces effectively. Furthermore, the effect of Er:YAG laser irradiation on root surfaces should be evaluated in vivo so that its potential to enhance the healing of periodontal tissues can be assessed.

Influence of Final Rinse Technique on Ability of Ethylenediaminetetraacetic Acid of Removing Smear Layer

Introduction: There is ongoing debate regarding the ideal sequence, volume, and concentration of irrigants, length of time for irrigation, and irrigation technique to achieve debridement of the root canal system. The aim of this study was to verify the impact of the final rinse technique on smear layer removal ability of 17% ethylenediaminetetraacetic acid (EDTA). Methods: Sixteen single-rooted human teeth were instrumented and divided into 2 groups at the final rinse step according to the following final rinse techniques used: continuous rinse group, continuous rinse with EDTA during 3 minutes, and rinse and soaking group, rinse with 1 mL of EDTA, soaking of the canal for 2 minutes and 30 seconds, and rinse completion with the remaining 4 mL for 30 seconds. The specimens were split lengthwise and observed under scanning electron microscope. Results: Data were analyzed with Kruskal-Wallis and Dunn tests. The continuous rinse group presented more debris-free surfaces when compared with the rinse and soaking group (P <. 01). When the root canal areas were compared within the groups, no statistical differences were found (P > .05). Conclusions: It can be concluded that a continuous rinse with 5 mL of EDTA for 3 minutes can more efficiently remove the smear layer from root canal walls. (J Endod 2010;36:512-514)

Qualitative Analysis of the Removal of the Smear Layer in the Apical Third of Curved Roots: Conventional Irrigation versus Activation Systems

Introduction: The aim of this study was to evaluate the effectiveness of different irrigant agitation techniques on smear layer removal in curved root canals. Methods: Mesiobuccal canals of 62 extracted lower molars with a curvature of 33 degrees were used and instrumented up to Pro Taper F2. The samples were divided into 3 experimental groups according to the final irrigation: conventional irrigation, ultrasonic irrigation, and sonic irrigation by using the Endo Activator system. The control group was composed of 2 specimens without any final irrigation. In all of the experimental groups, 5 mL of 17% ethylenediaminetetraacetic acid was used for 1 minute, and 5 mL of 2.5% NaOCl was used for 30 seconds. The analysis of the apical region was performed via scanning electron microscopy by 3 examiners. The data were submitted to the Kruskal-Wallis and Dunn tests (P<.05). Results: The activation systems removed significantly more smear layer than did conventional irrigation. Conclusions: Sonic and ultrasonic irrigation resulted in better removal of the smear layer in the apical third of curved root canals than did conventional irrigation. (J Endod 2011;37:1268-1271)

Does a final rinse to remove smear layer interfere on dentin permeability of root canals?

Objective. The objective of this study was to assess the influence of different final irrigating solutions on dentin permeability and smear layer removal using the same specimens and relate the results obtained. Study design. Forty anterior human teeth were instrumented and divided into 4 groups (n = 10) at the final rinse step, according to the irrigant used: G I (control) - 1% NaOCl; G II - 17% EDTA; G III - 17% EDTAT; and G IV - Biopure MTAD. The canals were filled with 0.5% methylene blue and maintained in bottles for 48 hours. The roots were transversally split in coronal, middle, and apical fragments. The specimens were photographed and analyzed regarding dye penetration. The fragments were then axially split and prepared for SEM. The photomicrographs were analyzed and qualified by scores. Results. Only the EDTA-T group exhibited statistical difference in which the apical third had less dentin permeability (P < .05). When a decalcifying agent was used, smear layer was removed, which did not happen in the NaOCl group. Regarding smear layer removal, differences were found only in the EDTA group in which the apical third presented more smear layer (P < .05). No correlation was found for both studies (r = 0.4207). Conclusions. There was not an even relationship between the results from both studies, which inferes that higher or lower dentinal permeability does not necessarily correspond to a higher or lower amount of smear layer. The analysis of dentin permeability and smear layer removal was shown to be a feasible procedure using the same specimens. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2009; 107: e47-e51)

Effects of Nd:YAG Laser Irradiation on the Hybrid Layer of Different Adhesive Systems

Purpose: The aim of this in vitro study was to evaluate the microtensile bond strength (mu TBS) and hybrid layer morphology of different adhesive systems, either followed by treatment with Nd:YAG laser irradiation or not. Previous studies have shown the effects of Nd:YAG laser irradiation on the dentin surface at restoration margins, but there are few reports about the significance of the irradiation on the hybrid layer. Materials and Methods: The flattened coronal and root dentin samples of 24 bovine teeth were randomly divided into 8 groups, according to the adhesive system used - Scotchbond Multi Purpose (SBMP) or Clearfil SE Bond (CSEB) - and were either irradiated with Nd:YAG or not, with different parameters: 0.8 W/10 Hz, 0.8 W/20 Hz, 1.2 W/10 Hz, 1.2 W/20 Hz. The left sides of specimens were the control groups, and right sides were irradiated. A composite crown was built over bonded surfaces and stored in water (24 h at 37 degrees C). Specimens were sectioned vertically into slabs that were subjected to mu TBS testing and observed by SEM. Results: Control groups (27.81 +/- 1.38) showed statistically higher values than lased groups (21.37 +/- 0.99), and CSEB control group values (31.26 +/- 15.71) were statistically higher than those of SBMP (24.3 +/- 10.66). There were no significant differences between CSEB (20.34 +/- 10.01) and SBMP (22.43 +/- 9.82) lased groups. Among parameters tested, 0.8 W/10 Hz showed the highest value (25.54 +/- 11.74). Nd:YAG laser irradiation caused dentin to melt under the adhesive layer of both adhesive systems tested. Conclusion: With the parameters used in this study, Nd:YAG laser irradiation of the hybrid layer promoted morphological changes in dentin and negatively influenced the bond strength of both adhesive systems.

Influence of different volumes of EDTA for final rinse on smear layer removal

Objective. The aim of this study was to analyze the influence of different volumes of 17% EDTA for final rinse on smear layer removal on the different areas of the root canal. Study design. Forty single-rooted teeth were instrumented using rotary instruments. The teeth were divided into 3 test groups according to the EDTA volume for final rinse (5 mL, 10 mL, 15 mL) and 1 control group (10 mL of 1% sodium hypochlorite). The roots were axially split into halves, and the smear layer removal from the canals was determined under scanning electron microscope. The data were analyzed using Kruskal Wallis and Dunn tests. Results. The 3 experimental groups showed no statistical difference (P > .05); however, when the test groups were compared to the control group statistically significant differences were found (P < .01). The root canal wall surfaces of teeth in the control group showed the presence of heavy smear layer through the entire length of the root canals. The other groups showed mainly smear layer-free surfaces or a small amount of debris. When coronal, middle and apical thirds were compared, there was no statistically significant difference (P > .05). Conclusions. Based on the results, it appears that a final rinse with 5 mL of EDTA per canal provides good smear layer removal, with root canal walls free of debris and mostly open dentinal tubules in all areas. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2008;106:e40-e43)

Low-fluence CO(2) laser irradiation decreases enamel solubility

This study investigated whether subablative-pulsed CO(2) laser (10.6 mu m) irradiation, using fluences lower than 1 J/cm(2), was capable of reducing enamel acid solubility. Fifty-one samples of bovine dental enamel were divided into three groups: control group, which was not irradiated (CG); group laser A (LA) irradiated with 0.3 J/cm ; and group laser B (LB) irradiated with 0.7 J/cm(2). After irradiation, the samples were subjected to demineralization in an acetate buffer solution and were then analyzed by SEM. A finite-element model was used to calculate the temperature increase. The calcium and phosphorous content in the demineralization solution were measured with an ICP-OES. ANOVA and the t-test pairwise comparison (p < 0.016) revealed that LB showed significantly lower mean Ca and P content values in the demineralization solution than other groups. A reduction in the enamel solubility can be obtained with pulsed CO(2) laser irradiation (0.7 J/cm(2), 135 mJ/pulse, 74 Hz, 100 mu s) without any surface photomodification and a less than 2 degrees C temperature increase at a 3-mm depth from the surface.

Can the durability of one-step self-etch adhesives be improved by double application or by an extra layer of hydrophobic resin?

Objectives: This study evaluated the immediate and 6-month resin-dentin mu-bond strength (mu TBS) of one-step self-etch systems (Adper Prompt L-Pop [AD] 3M ESPE; Xeno III [XE] Dentsply De Trey; iBond [iB] Heraeus Kulzer) under different application modes. Materials and methods: Dentin oclusal surfaces were exposed by grinding with 600-grit SiC paper. The adhesives were applied according to the manufacturer`s directions [MD], or with double application of the adhesive layer [DA] or following the manufacturer`s directions plus a hydrophobic resin layer coating [HL]. After applying the adhesive resins, composite crowns were built up incrementally. After 24-h water storage, the specimens were serially sectioned in ""x"" and ""y"" directions to obtain bonded sticks of about 0.8 mm 2 to be tested immediately [IM] or after 6 months of water storage [6M] at a crosshead speed of 0.5 mm/min. The data from each adhesive was analyzed by a two-way repeated measures ANOVA (mode of application vs. storage time) and Tukey`s test (alpha = 0.05). Results: The adhesives performed differently according to the application mode. The DA and HL either improved the immediate performance of the adhesive or did not differ from the MD. The resin-dentin bond strength values observed after 6 months were higher when a hydrophobic resin coat was used than compared to those values observed under the manufacturer`s directions. Conclusions: The double application of one-step self-etch system can be safety performed however the application of an additional hydrophobic resin layer can improve the immediate resin-dentin bonds and reduce the degradation of resin bonds over time. (c) 2008 Elsevier Ltd. All rights reserved.

Effect of temperature and heating rate on the sintering of leucite-based dental porcelains

The aim of this work was to determine the effect of temperature and heating rate on the densification of four leucite-based dental porcelains: two low-fusion (Dentsply Ceramco and Ivoclar) and two high-fusion commercial porcelains (Dentsply Ceramco). Porcelain powders were characterized by differential thermal analysis (DTA), X-ray diffraction (XRD), particle size distribution, helium picnometry, and by scanning electron microscopy. Test specimens were sintered from 600 to 1050 degrees C, with heating rates of 55 degrees C/min and 10 degrees C/min. The bulk density of the specimens was measured by the Archimedes method in water, and microstructures of fracture surfaces were analyzed by scanning electron microscopy (SEM). The results showed that densification of specimens increased with the increase in temperature. The increase in the heating rate had no effect on the densification of the porcelains studied. Both high-fusion materials and one of the low-fusing porcelains reached the maximum densification at a temperature that was 50 degrees C lower than that recommended by the manufactures. (C) 2011 Elsevier Ltd and Techna Group S.r.l. All rights reserved.

Effect of ion-exchange temperature on mechanical properties of a dental porcelain

The objective of this study was to determine the influence of different ion-exchange temperatures on the biaxial flexural strength (sigma(f)), hardness (HV) and indentation fracture resistance (K(IF)) of a dental porcelain. Disk-shaped specimens were divided into five groups (n = 10) and submitted to an ion-exchange procedure using KNO(3) paste for 15 min in the following temperatures (degrees C); (I) 430; (II) 450; (III) 470; (IV) 490; (V) 510; and control (no ion exchange). The value of sigma(f) was determined in artificial saliva at 37 degrees C. The values of HV and K(IF) were obtained using 3 Vickers indentations in each specimen (19.6 N). Results showed that ion exchange increases significantly the properties of the material as compared to the control and no significant differences were found among the temperatures tested for any of the properties studied. (C) 2010 Elsevier Ltd and Techna Group S.r.l. All rights reserved.

Microshear Bond Strength of Self-etching Systems Associated with a Hydrophobic Resin Layer

Purpose: To evaluate in vitro the microshear bond strength of adhesive systems applied to dentin according to manufacturers` instructions, associated or not with a hydrophobic layer of unfilled resin. Materials and Methods: Six self-etching adhesives (Clearfil SE Bond, Kuraray Medical; AdheSE, lvoclar Vivadent; Xeno III, Dentsply; I Bond, Heraeus-Kulzer; Bond Force, Tokuyama; Futurabond DC, Voco) were tested. The labial dentin of sixty bovine incisors was exposed, and the teeth were divided into two groups according to the application or not of an extra hydrophobic resin layer (Scotchbond Multi Purpose Plus, bottle 3). Six composite cylinders (Filtek Z250, 3M ESPE) were built up on each treated surface. Specimens were stored in distilled water at 37 C for 24 h and then subjected to the microshear bond strength test in a universal testing machine at a crosshead speed of 0.5 mm/min. Microshear bond strength values were analyzed by 2-way ANOVA and Tukey`s post-hoc test. Failure mode was determined using a stereomicroscope under 20X magnification. Results: The application of the hydrophobic resin layer did not affect bond strength, except for AdheSE. However, the bond strengths with the hydrophobic layer were similar among the six tested systems (Clearfil: 17.1 +/- 7.9; AdheSE: 14.5 +/- 7.1; Xeno III: 12.8 +/- 7.7; I Bond: 9.5 +/- 5.8; Bond Force: 17.5 +/- 4.1; Futurabond: 7.7 +/- 2.3). When used as recommended by the manufacturers, Bond Force presented statistically higher bond strength than AdheSE and I Bond (p < 0.05) (Clearfil 10.4 +/- 4.9; AdheSE 1.6 +/- 1.6; Xeno III: 9.0 +/- 3.8; I Bond: 3.0 +/- 1.5; Bond Force: 14 +/- 3.9; Futurabond: 8.8 +/- 3.8). Failure mode was predominantly adhesive. Conclusion: The bond strength of the self-etching systems tested was not significantly affected by the application of a hydrophobic layer, but a significant improvement was observed in AdheSE.