Immune regulatory effect of pHSP65 DNA therapy in pulmonary tuberculosis: Activation of CD8+ cells, interferon-γ recovery and reduction of lung injury

A DNA vaccine based on the heat-shock protein 65 Mycobacterium leprae gene (pHSP65) presented a prophylactic and therapeutic effect in an experimental model of tuberculosis. In this paper, we addressed the question of which protective mechanisms are activated in Mycobacterium tuberculosis-infected mice after immune therapy with pHSP65. We evaluated activation of the cellular immune response in the lungs of infected mice 30 days after infection (initiation of immune therapy) and in those of uninfected mice. After 70 days (end of immune therapy), the immune responses of infected untreated mice, infected pHSP65-treated mice and infected pCDNA3-treated mice were also evaluated. Our results show that the most significant effect of pHSP65 was the stimulation of CD8+ lung cell activation, interferon-γ recovery and reduction of lung injury. There was also partial restoration of the production of tumour necrosis factor-α. Treatment with pcDNA3 vector also induced an immune stimulatory effect. However, only infected pHSP65-treated mice were able to produce significant levels of interferon-γ and to restrict the growth of bacilli.

Effect of thermal treatments on tensile strength of commercially cast pure titanium and Ti-6Al-4V alloys

Heating titanium structures is assumed to relieve tensions induced by the casting process as well as possibly optimizing some mechanical properties. The aim of this investigation was to evaluate the effect of thermal treatments on tensile strength of commercially pure titanium (CP Ti) and Ti-6Al-4V alloy. Thirty dumbbell rods, with diameters of 3.0 mm at the central segment and lengths of 42 mm, were cast for each metal using the Rematitan System. CP Ti and Ti-6Al-4V specimens were randomly divided into three groups of ten: a control group that received no thermal treatment and two test groups. One (T1) was heated at 750°C for 2 h and the other (T2) was annealed at 955°C for 1 h and aged at 620°C for 2 h. Tensile strength was measured with a universal testing machine (MTS model 810). Tensile strength means and standard deviations were statistically compared using a Kruskal-Wallis test at a α = 0.05 significance level. No statistically significant differences in tensile strength were observed among CP Ti groups. For the Ti-6Al-4V alloy, the control and T1 groups revealed statistically higher tensile strengths when compared to the T2 group, with no significant difference between the control and T1 groups. © 2005 Springer Science + Business Media, Inc.

Effect of disinfectants on the hardness and roughness of reline acrylic resins

Purpose: Potential effects on hardness and roughness of a necessary and effective disinfecting regimen (1% sodium hypocholorite and 4% chlorhexidine) were investigated for two hard chairside reline resins versus a heat-polymerizing denture base acrylic resin. Materials and Methods: Two standard hard chairside reliners (Kooliner and Duraliner II), one heat-treated chairside reliner (Duraliner II +10 minutes in water at 55°C), and one standard denture base material (Lucitone 550) were exposed to two disinfecting solutions (1% sodium hypochlorite; 4% chlorhexidine gluconate), and tested for two surface properties [Vickers hardness number (VHN, kg/mm2); Roughness (Ra, μm)] for different times and conditions (1 hour after production, after 48 hours at 37 ± 2°C in water, after two disinfection cycles, after 7 days in disinfection solutions, after 7 days in water only). For each experimental condition, eight specimens were made from each material. Data were analyzed by analysis of variance followed by Tukey's test, and Student's t-test (p= 0.05). Results: For Kooliner (from 6.2 ± 0.3 to 6.5 ± 0.5 VHN) and Lucitone 550 (from 16.5 ± 0.4 to 18.4 ± 1.7 VHN), no significant changes in hardness were observed either after the disinfection or after 7 days of immersion, regardless of the disinfectant solution used. For Duraliner II (from 4.0 ± 0.1 to 4.2 ± 0.1 VHN), with and without heat treatment, a small but significant increase in hardness was observed for the specimens immersed in the disinfectant solutions for 7 days (from 4.3 ± 0.2 to 4.8 ± 0.5 VHN). All materials showed no significant change in roughness (Kooliner: from 0.13 ± 0.05 to 0.48 ± 0.24 μm; Duraliner II, with and without heat treatment: from 0.15 ± 0.04 to 0.29 ± 0.07 μm; Lucitone 550: from 0.44 ± 0.19 to 0.49 ± 0.15 μm) after disinfection and after storage in water for 7 days. Conclusions: The disinfectant solutions, 1% sodium hypochlorite and 4% chlorhexidine gluconate, caused no apparent damage on hardness and roughness of the materials evaluated. Copyright © 2006 by The American College of Prosthodontists.

Effect of the association of nystatin with a tissue conditioner on its ultimate tensile strength

Purpose: This study evaluated the ultimate tensile strength of a tissue conditioner without nystatin incorporation (GI - control group) and the same tissue conditioner modified by the addition of nystatin in two concentrations: GII - 500,000 International Units (U) and GIII - 1,000,000 U, in which each milligram of the medicament corresponded to 6079 U. Materials and Methods: Dumbbell-shaped specimens (N = 7) with a central cross-sectional area of 33 × 6 × 3 mm were produced for the three experimental groups. After polymerization following manufacturer's instructions, specimens were immersed in distilled water at 37°C for either 24 hours or 7 days and then tested in tension in the MTS 810 at 40 mm/minute. Data were analyzed by two-way ANOVA followed by Tukey's test, at 95% level of confidence. Results: The means (force-grams (gf) ± standard deviation) of the ultimate tensile strength were: GI - 634.29 ± 122.80; GII - 561.92 ± 133.56; and GIII - 547.30 ± 73.47 for 24-hour storage, and GI - 536.68 ± 54.71; GII - 467.50 ± 143.51; and GIII - 500.62 ± 159.76 for 7-day storage. There were no statistically significant differences among the three experimental groups (p > 0.05). The ultimate tensile strength means of all experimental groups after 7 days were significantly lower than those observed after 24 hours (p = 0.04). Conclusions: The results of this study suggest that the addition of nystatin into the tissue conditioner investigated in concentrations below 1,000,000 U did not affect its ultimate tensile strength. Copyright © 2006 by The American College of Prosthodontists.

Effect of microwave disinfection procedures on torsional bond strengths of two hard chairside denture reline materials

Purpose: This study evaluated the potential effects of denture base resin water storage time and an effective denture disinfection method (microwave irradiation at 650 W for 6 minutes) on the torsional bond strength between two hard chairside reline resins (GC Reline and New Truliner) and one heat-polymerizing denture base acrylic resin (Lucitone 199). Materials and Methods: Cylindrical (30 x 3.9 mm) denture base specimens (n = 160) were stored in water at 37°C (2 or 30 days) before bonding. A section (3.0 mm) was removed from the center of the specimens, surfaces prepared, and the reline materials packed into the space. After polymerization, specimens were divided into four groups (n = 10): Group 1 (G1) - tests performed after bonding; Group 2 (G2) - specimens immersed in water (200 ml) and irradiated twice (650 W for 6 minutes); Group 3 (G3) - specimens irradiated daily until seven cycles of disinfection; Group 4 (G4) - specimens immersed in water (37°C) for 7 days. Specimens were submitted to a torsional test (0.1 Nm/min), and the torsional strengths (MPa) and the mode of failure were recorded. Data from each reline material were analyzed by a two-way analysis of variance, followed by Neuman-Keuls test (p = 0.05). Results: For both Lucitone 199 water storage periods, before bonding to GC Reline resin, the mean torsional strengths of G2 (2 days - 138 MPa; 30 days - 132 MPa), G3 (2 days - 126 MPa; 30 days - 130 MPa), and G4 (2 days - 130 MPa; 30 days - 137 MPa) were significantly higher (p < 0.05) than G1 (2 days - 108 MPa; 30 days - 115 MPa). Similar results were found for Lucitone 199 specimens bonded to New Truliner resin, with G1 specimens (2 days - 73 MPa; 30 days - 71 MPa) exhibiting significantly lower mean torsional bond strength (p < 0.05) than G2 (2 day - 86 MPa; 30 days - 90 MPa), G3 (2 days - 82 MPa; 30 days - 82 MPa), and G4 specimens (2 days - 78 MPa; 30 days - 79 MPa). The adhesion of both materials was not affected by water storage time of Lucitone 199 (p > 0.05). GC reline showed a mixed mode of failure (adhesive/cohesive) and New Truliner failed adhesively. Conclusions: Up to seven microwave disinfection cycles did not decrease the torsional bond strengths between the hard reline resins, GC Reline and New Truliner to the denture base resin Lucitone 199. The effect of additional disinfection cycles on reline material may be clinically significant and requires further study. Copyright © 2006 by The American College of Prosthodontists.

Effect of environmental mycobacteria (Mycobacterium avium) on immunity induced by a DNA vaccine (DNAhsp65) against tuberculosis

The efficacy of BCG vaccine (attenuated Mycobacterium bovis) against pulmonary tuberculosis varies enormously among different populations. The prevailing hypothesis attributes this variation to interactions between the vaccine and mycobacteria common in the environment. Studies have revealed that most protective antigens expressed by the antituberculous vaccine are conserved in M. avium, supporting the hypothesis that exposure to environmental mycobacteria generates a cross-reactive immune response that interferes with BCG efficacy. In this study we investigated the effect of a prior exposure to heat-killed M. avium on the immune response and the protective efficacy induced by a genetic vaccine pVAXhsp65 (hsp65 gene from M. leprae inserted in pVAX vector) against experimental tuberculosis. To evaluate the effect on the immune response, female BALB/c mice were initially injected with distinct doses (0.08×106, 4×106, and 200×10 6) of heat-killed M. avium by subcutaneous route. Three weeks later, the animals were immunized with 3 doses of DNAhsp65 by intramuscular route (100μg/15 days apart). Control groups received only M. avium, vaccine (pVAXhsp65), vector (pVAX) or saline solution. Cytokine production and antibody levels were determined by ELISA. To evaluate the effect on the protective efficacy, animals were initially sensitized with 200×106 heat-killed CFU of M. avium by subcutaneous route and then immunized with 3 doses of pVAXhsp65 (100μg/15 days apart) by intramuscular route. Control groups were injected with saline, pVAX (4 doses), pVAXhsp65 (4 doses), M. avium or M. avium plus pVAX (3 doses). Fifteen days after last DNA dose, the animals were infected with 1×104 viable CFU of H37Rv M. tuberculosis by intratracheal route. Thirty days after challenge, the animals were sacrificed and the bacterial burden was determined by counting the number of CFU in the lungs. Lung histological sections were also analyzed. Splenic cells from primed animals produced more IL-5 but less IFN-gamma than non-primed ones. Also, prior contact with M. avium determined higher production of IgG1 and IgG2a anti-hsp65 antibodies in comparison to control groups. However, this higher immune response did not decrease the bacterial burden in the lungs. In addition, prior sensitization with M. avium decreased the parenchyma preservation observed in the group immunized only with pVaxhsp65. These results indicate that environmental mycobacteria can interfere with immunity and protective efficacy induced by DNAhsp65.

Vickers hardness of cast commercially pure titanium and Ti-6Al-4V alloy submitted to heat treatments

The purpose of this study was to evaluate the effect of heat treatments on the Vickers hardness of commercially pure titanium and Ti-6Al-4V cast alloys. Six-millimeter-diameter cylindrical specimens were cast in a Rematitan System. Commercially pure titanium and Ti-6Al-4V alloy specimens were randomly assigned to 3 groups (n=10) that received the following heat treatments: control (no heat treatment); treatment 1 (T1): heating at 750°C for 2 h; and treatment 2 (T2): annealing at 955°C for 1 h and aging at 620°C for 2 h. After heat treatments, the specimens were embedded in acrylic resin and their surface was ground and polished and hardness was measured. Vickers hardness means (VHN) and standard deviations were analyzed statistically by Kruskal-Wallis test at 5% significance level. For commercially pure titanium, Vickers hardness means of group T2 (259.90 VHN) was significantly higher than those of the other groups (control - 200.26 VHN and T1 - 202.23 VHN), which presented similar hardness means to each other (p>0.05). For Ti-6Al-4V alloy, statistically significant differences were observed among the three groups: T2 (369.08 VHN), T1 (351.94 VHN) and control (340.51 VHN) (p<0.05). The results demonstrated different hardness of CP Ti and Ti-6Al-4V when different heat treatments were used. For CP Ti, VHN means of T2 group was remarkably higher than those of control and T1 group, which showed similar VHN means to each other. For Ti-6Al-4V alloy, however, VHN means recorded for each group may be presented as follows: T2>T1>control.

Effect of thermal treatment under oxygen atmosphere on the superconducting properties of RuSr2GdCu2O8

The discovery of the spatial uniform coexistence of superconductivity and ferromagnetism in rutheno-cuprates, RuSr2GdCu2O8 (Ru-1212), has spurred an extraordinary development in the study of the competition between magnetism and superconductivity. However, several points of their preparation process and characterization that determine their superconductive behavior are still obscure. The improvement of sample preparation conditions involves some thermal treatments in inert atmosphere. The first treatment results in the immediate formation of Sr2GdRuO 6. Using the CuO composition as a precursor, we produced Ru-1212. To turn it metallic and superconductor, besides the previous treatment, a final sinterization is carried out in oxygen flow for several days. Three Ru-1212 samples were produced by varying the last sinterization time (two, four, and six days under oxygen flow). Through measurements of x-ray diffraction, scanning electron microscopy, differential thermal analysis, magnetic susceptibility and mechanical spectroscopy, it was studied the influence of the treatments under oxygen atmosphere on the structural and superconducting properties of the material.

Effect of milk treatment on acidification, physicochemical characteristics, and probiotic cell counts in low fat yogurt

The effect of milk treatment (heat, high hydrostatic pressure - HHP, or combined heat and HHP) on acidification, physicochemical characteristics, and probiotic cell counts in low fat yogurt was studied. All samples were analyzed for fermentation time, pH, titratable acidity, total solids, water-holding capacity, syneresis, Hunter L*, a*, and b* values, Streptococcus thermophilus, Lactobacillus delbrueckii ssp bulgaricus, L. acidophilus, and Bifidobacterium longum. The application of HHP combined with thermal treatment resulted in yogurt gels with attractive physicochemical characteristics and high water-holding capacity. In addition to this, the milk treatment did not affect the probiotic bacteria growth. The balance of strains in the starter culture and level of inoculation influenced the yogurt fermentation and properties. The use of combined heat and HHP to treat milk before yogurt fermentation could be an alternative process for obtaining high quality, additive-free healthy products.

The effect of water immersion on the shear bond strength between chairside reline and denture base acrylic resins: Basic science research

Purpose: The effect of water immersion on the shear bond strength (SBS) between 1 heat-polymerizing acrylic resin (Lucitone 550-L) and 4 autopolymerizing reline resins (Kooliner-K, New Truliner-N, Tokuso Rebase Fast-T, Ufi Gel Hard-U) was investigated. Specimens relined with resin L were also evaluated. Materials and Methods: One hundred sixty cylinders (20 × 20 mm) of L denture base resin were processed, and the reline resins were packed on the prepared bonding surfaces using a split-mold (3.5 × 5.0 mm). Shear tests (0.5 mm/min) were performed on the specimens (n = 8) after polymerization (control), and after immersion in water at 37°C for 7, 90, and 180 days. All fractured surfaces were examined by scanning electron microscopy (SEM) to calculate the percentage of cohesive fracture (PCF). Shear data were analyzed with 2-way ANOVA and Tukey's test; Kruskall-Wallis test was used to analyze PCF data (α = 0.05). Results: After 90 days water immersion, an increase in the mean SBS was observed for U (11.13 to 16.53 MPa; p < 0.001) and T (9.08 to 13.24 MPa, p = 0.035), whereas resin L showed a decrease (21.74 MPa to 14.96 MPa; p < 0.001). The SBS of resins K (8.44 MPa) and N (7.98 MPa) remained unaffected. The mean PCF was lower than 32.6% for K, N, and T, and higher than 65.6% for U and L. Conclusions: Long-term water immersion did not adversely affect the bond of materials K, N, T, and U and decreased the values of resin L. Materials L and U failed cohesively, and K, N, and T failed adhesively. © 2007 by The American College of Prosthodontists.

Effect of a post-polymerization treatments on the flexural strength and vickers hardness of reline and acrylic denture base resins

This study evaluated the effect of water-bath and microwave post-polymerization treatments on the flexural strength and Vickers hardness of four autopolymerizing reline resins (Duraliner II-D, Kooliner-K, Tokuso Rebase Fast-TR and Ufi Gel Hard C-UGH) and one heat-polymerized acrylic resin (Lucitone 550-L), processed using two polymerization cycles (short cycle - 90 minutes at 73°C and 100°C for 30 minutes; and long cycle - 9 hours at 71°C). For each material, thirty specimens (64 x 10 x 3.3 mm) were made and divided into 3 groups (n=10). Specimens were tested after: processing (control group); water-bath at 55°C for 10 minutes (reline materials) or 60 minutes (L); and microwave irradiation. Flexural strength tests were performed at a crosshead speed of 5 mm/min using a three-point bending device with a span of 50 mm. The flexural strengths values were calculated in MPa. One fragment of each specimen was submitted to Vickers hardness test. Data were analyzed by 2-way ANOVA followed by Tukey's HSD test (α=0.05). L microwaved specimens (short cycle) exhibited significantly higher flexural strength means than its respective control group (p<0.05). Water-bath promoted a significant increase (p<0.05) in flexural strength of K and L (long cycle). The hardness of the tested materials was not influenced by the post-polymerization treatments. Post-polymerization treatments could be used to improve the flexural strength of some materials tested.

Cryopreservation of dog spermatozoa: Effect of bovine serum albumin on acrosomal integrity and pregnancy rates after artificial insemination

The objective of the present study was to compare the in vitro and in vivo profile of frozen dog semen with Tris-bovine serum albumin (TB) and Tris-egg yolk (TE) extenders. Twenty dogs were used as donors. Each dog was stimulated by penile massage and only the sperm-rich fraction was collected weekly until 40 ejaculates were obtained. After macroscopic and microscopic analysis, equal parts of each ejaculate were diluted with TB and TE by the one-step method at 37 °C. The semen was added to 0.5-mL French straws which presented normal characteristics before freezing and after thawing. Acrosomal integrity was evaluated by double Trypan blue-Giemsa staining, in which alive intact (LI), alive reacted (LR), dead intact (DI) and dead reacted (DR) spermatozoa, were identified by the time of thawing and up to 4 h of incubation at 39 °C, the TE being significantly superior to TB (P<0,01) in the LI and LR variables. The TB being significantly superior to TE (P<0,01) in the DR variable. Female dogs in natural heat were submitted to artificial insemination, 20 receiving TE-semen and 20 receiving TB-semen with the Osiris probe (IMV, L'Aigle, France) and the numbers indicate that TE was significantly better than TB (P<0,01) to pregnancy rate and number of puppies/delivery. We concluded from this study, that TE was better than TB, because this, induced an eady acrossome reaction in dog's sperm.

Effects of atrazine on female Wistar rats: Morphological alterations in ovarian follicles and immunocytochemical labeling of 90 kDa heat shock protein

Fertility in female mammals may be affected by a variety of endocrine disrupters present in the environment. Herbicide atrazine is an example of endocrine disrupter employed in agriculture, which disrupts estrous cyclicity in rats. Aiming to characterize morphologically the effect of low and sublethal doses of atrazine on the ovaries of Wistar rats, in an effort to determine the possible intrafollicular target site through which this herbicide acts adult females were submitted to both subacute and subchronic treatments. Additionally, immunocytochemical labeling of 90 kDa heat shock protein (HSP90) was performed in order to evaluate the role played by this protein in the ovary, under stressed conditions induced by herbicide exposure. The results indicated that atrazine induced impaired folliculogenesis, increased follicular atresia and HSP90 depletion in female rats submitted to subacute treatment, while the subchronic treatment with low dose of atrazine could compromise the reproductive capacity reflected by the presence of multioocytic follicle and stress-inducible HSP90. © 2007 Elsevier Ltd. All rights reserved.

Influence of storage period and effect of different brands of acrylic resin on the dimensional accuracy of the maxillary denture base

The aim of this study was to evaluate the dimensional changes of denture bases made from different resins after different storage periods. For this purpose, 25 sets of plaster models/resin bases were prepared using 4 acrylic resins submitted to two types of polymerization: 1- QC-20 submitted to polymerization by microwave energy; 2- QC-20 submitted to polymerization by water hot bath; 3- Vipi Cril submitted to polymerization by water hot bath; 4- Vipi Wave submitted to polymerization by microwave energy; and 5- Onda Cryl submitted to polymerization by microwave energy. After polymerization, the specimens were sectioned for accuracy readings using a comparison microscope. Readings were taken at 3 points: the crests of the right (A) and left (B) ridges, and the median region of the palate, in 4 different periods. The data obtained were submitted to two-way ANOVA and Tukey's test at 5% significance level. The greatest distortions were found in the posterior palatal region of the base (M), with statistically significant difference (p<0.05) for the studied resins. All acrylic resins presented dimensional changes and the storage period influenced these alterations.

Heat transfer in a packed sugar cane bagasse bed

Heat transfer in a packed bed of sugar cane bagasse, which is a potential biofuel used in cars and industries, percolated with air flow was studied. The fibers were washed, sieved, oven dried, and afterwards moisture content was adjusted to 4 and 47%. The relative humidity of the air, packing bed technique, and the initial moisture content of the porous media did not have a significant effect on the outlet temperature of the bed. Air flow rate influenced the averaged radial temperature profile, but not the temperature measured at the nearest position to the tube wall. At the end of the experiments, moisture segregation was observed, the lower bed depths being drier than the higher ones. This is an abstract of a paper presented at the 18th International Congress of Chemical Process Engineering (Praque, Czech Republic 8/24-28/2008).