Surveillance for zoonotic vector-borne infections using sick dogs from southeastern Brazil

For many vector-borne organisms, dogs can be used as sentinels to estimate the risk of human infection. The objective of this study was to use dogs as sentinels for multiple vector-borne organisms in order to evaluate the potential for human infection with these agents in southeastern Brazil. Blood from 198 sick dogs with clinicopathological abnormalities consistent with tick-borne infections were selected at the São Paulo State University Veterinary Teaching Hospital in Botucatu and tested for DNA and/or antibodies against specific vector-borne pathogens. At least one organism was detected in 88% of the dogs, and Ehrlichia canis DNA was amplified from 78% of the blood samples. Bartonella spp. seroreactivity was found in 3.6%. Leishmania chagasi antibodies were detected in 1% of the dogs. There was no serological or polymerase chain reaction evidence of infection with Anaplasma phagocytophilum, Borrelia burgdorferi, Ehrlichia chaffeensis, Ehrlichia ewingii, and Rickettsia rickettsii. The full E. canis 16S rRNA gene sequence of one of the Brazilian strains obtained in this study was identical to the causative agent of human ehrlichiosis in Venezuela. Ehrlichia canis may pose a human health hazard and may be undiagnosed in southeastern Brazil, whereas exposure to the other organisms examined in this study is presumably infrequent.

Effects of chemical disinfectants on the transverse strength of denture base acrylic resins

The disinfection of dental prostheses by immersion in a chemical solution should be capable of rapid inactivation of pathogenic microorganisms, without causing any adverse effect on the denture base resins. This study evaluated the effect of disinfection immersion on the transverse strength of two heat-cured resins. The denture base resins (Lucitone 550 and QC 20) were polymerized according to the manufacturers' instructions. After polymerization, the specimens were polished, and then stored in water at 37 degreesC for 50 +/- 2 h prior immersion in one of the following solutions for 10 min: 4% chlorhexidine, 1% sodium hypochlorite and 3.78% sodium perborate. The specimens were submitted to disinfection twice, simulating when dentures come from the patient and before being returned to the patient. Ten specimens were made for each group. The transverse strength was evaluated by a 3-point bend test. The flexural strength of the two denture base acrylic resins evaluated remained unaffected after immersion in the three solutions evaluated. In general, the QC 20 resin specimens exhibited lower transverse strength than the Lucitone 550 resin specimens, regardless of immersion solutions.

Hardness and compressive strength of indirect composite resins: effects of immersion in distilled water

The aim of this study was to evaluate the effect of ageing in distilled water on the hardness and compressive strength of a direct composite resin Z100, a feldspatic porcelain (Noritake) and three indirect composites (Artglass, Solidex and Targis). For the Vickers hardness tests, five disk-shaped specimens (2 x 4 mm) of each material were prepared according to the manufacturers' instructions. The hardness tests were conducted using a Vickers diamond indentor. Compressive strength measurements were recorded on cylindrical specimens with a diameter of 6 mm and a length of 12 mm. The compression tests were carried out with a constant cross-head speed of 0.5 mm min(-1) on a mechanical test machine. For each material, 10 specimens were tested after 7 days of dry storage at 37 +/- 1 degreesC and 10 specimens were tested after water storage at 37 +/- 1 degreesC for 180 days. Noritake porcelain specimens showed higher hardness values than the composites. Among the composite materials, Z100 promoted the highest VHN values, regardless of the ageing periods. The results showed that Solidex and Z100 had the highest compressive strength values. Ageing in water reduced the hardness for all composites, but had no long-term effect on the compressive strength.

Comparison between wound healing in induced diabetic and nondiabetic rats after low-level laser therapy

Objective: the aim of this work was to compare the effect of low-level laser therapy (LLLT) on the wound healing process in nondiabetic and diabetic rats. Background Data: Among the clinical symptoms caused by diabetes mellitus, a delay in wound healing is a potential risk for patients. It is suggested that LLLT can improve wound healing. Methods: the tissue used for this study was extracted from animals suffering from diabetes, which was induced by Streptozotocin (R), and from nondiabetic rats. Animals were assembled into two groups of 25 rats each (treated and control) and further subdivided into two groups: diabetic (n = 15) and nondiabetic (n = 10). A full-thickness skin wound was made on the dorsum. area, with a round 8-mm hole-punch. The treated group was irradiated by a HeNe laser at 632.8 nm, with the following parameters: 15 mW, exposition time of 17 sec, 0.025 cm(2) irradiated area, and energy density of 10 J/cm(2). Square full-thickness skin samples (18 mm each side, including both injured and noninjured tissues) were obtained at 4, 7, and 15 days after surgery and analyzed by qualitative and quantitative histological methods. Results: Quantitative histopathological analysis confirmed the results of the qualitative analysis through histological microscope slides. When comparing tissue components (inflammatory cells, vessels and fibroblast/area), we found that treated animals had a less intense inflammatory process than controls. Conclusion: Results obtained by both qualitative and quantitative analyses suggested that irradiation of rats with HeNe (632.8 nm), at the tested dose, promoted efficient wound healing in both nondiabetic and diabetic rats as, compared to the control group.

K-bi-Lipschitz equivalence of real function-germs

In this paper we prove that the set of equivalence classes of germs of real polynomials of degree less than or equal to k, with respect to K-bi-Lipschitz equivalence, is finite.

Effects of Nd : YAG laser irradiation on root canal dentin wall: A scanning electron microscopic study

Objective: the purpose of this study was to evaluate, by scanning electron microscopy (SEM), the effects of Nd:YAG laser irradiation applied perpendicular or parallel to the root canal dentin wall. Methods: Thirty human teeth were divided into two groups: Group A (20 roots), laser application with circular movements, parallel to the dentin root surface; and Group B (10 roots), roots cut longitudinally and laser applied perpendicular to the root surface. Group A was subdivided into A1 (10 roots), laser application with 100 mJ, 15 Hz and 1.5 W; and A2 (10 roots) with 160 mJ, 15 Hz, and 2.4 W. Group B was subdivided into B1 (10 hemisections) and B2 (10 hemi-sections) with parameters similar to A I and A2. Four applications of 7-sec duration were performed, with a total exposure of 28 sec. SEM evaluations were made in the cervical, middle, and apical thirds, with 500X and 2000X magnifications. Morphological changes scores were attributed, and the results were submitted to Kruskal Wallis statistical test (5%). Results: Significant statistical differences were found between groups A and B (p = 0.001). In groups A1 and A2, few areas of dentin melting were observed. In groups B1 and B2, areas of melting dentin covering dentin surface were observed. Conclusions: It was concluded that intracanal laser application with circular movements (parallel to the surface) produces limited morphological changes in root canal dentin wall.

Granulocyte colony-stimulating factor expression from transduced vascular smooth muscle cells provides sustained neutrophil increases in rats

Granulocyte colony-stimulating factor (G-CSF) regulates granulocyte precursor cell proliferation, neutrophil survival, and activation. Cyclic hematopoiesis, a disease that occurs both in humans and grey collie dogs is characterized by cyclical variations in blood neutrophils. Although the underlying molecular defect is not known, long-term daily administration of recombinant G-CSF eliminates the severe recurrent neutropenia, indicating that expression of G-CSF by gene therapy would be beneficial. As a prelude to preclinical studies in affected collie dogs, we monitored hematopoiesis in rats receiving vascular smooth muscle cells transduced to express G-CSF. Cells transduced with LrGSN, a retrovirus expressing rat G-CSF, were implanted in the carotid artery and control animals received cells transduced with LASN, a retrovirus expressing human adenosine deaminase (ADA). Test animals showed significant increases in neutrophil counts for at least 7 weeks, with mean values of 3,670 +/- 740 cells/mu l in comparison to 1,870 +/- 460 cells/mu l in controls (p < 0.001). Thus, in rats G-CSF gene transfer targeted at vascular smooth muscle cells initiated sustained production of 1,800 neutrophils/mu l, a cell number that would provide clinical benefit to patients. Lymphocytes, red cells and platelets were not different between control and test animals (p > 0.05). These studies indicate that retrovirally transduced vascular smooth muscle cells can provide sustained clinically useful levels of neutrophils in vivo.

Microleakage and nanoleakage: Influence of laser in cavity preparation and dentin pretreatment

Objective: the purpose of this study was to verify if the application of the Nd:YAG laser following pretreatment of dentin with adhesive systems that were not light cured in class V cavities and were prepared with Er:YAG laser would promote better sealing of the gingival margins when compared to cavities prepared the conventional way. Background Data: Previous studies had shown that the pretreatment of dentin with laser irradiation after the application of an adhesive system is efficient in achieving higher shear bond and tensile bond strength. Materials and Methods: Er:YAG laser (Kavo-Key, Germany) with 350 mJ, 4 Hz, and 116.7 J/cm(2) was used for cavity preparation. The conventional preparation was made with diamond bur mounted in high-speed turbine. Dentin treatment was accomplished using an Nd:YAG laser (Pulse Master 1000, ADT. USA) at 60 mJ, 10 Hz, and 74.65/cm(2) following application of the adhesive system. The cavities were stored with Single Bond/Z100 and Prime & Bond NT/TPH. Eighty bovine incisors were used, and class V preparations were done at buccal and lingual surfaces divided into eight groups: (1) Er:YAG preparation + Prime & Bond NT + TPH; (2) Er:YAG preparation + Single Bond + Z100; (3) Er:YAG preparation + Single Bond + Nd:YAG + Z100; (4) Er:YAG preparation + Prime & Bond NT + Nd:YAG + TPH; (5) conventional preparation + Prime & Bond NT + TPH; (6) conventional preparation + Single Bond + Z100; (7) conventional preparation + Single Bond + Nd:YAG + Z100; (8) conventional preparation + Prime & Bond NT + Nd:YAG + TPH. All specimens were thermocycled for 300 full cycles between 5 degreesC +/- 2 degreesC and 55 degreesC +/- 2 degreesC (dwell time of 30 sec), and stored in 50% silver nitrate solution for 24 h soaked in photodeveloping solution and exposed to fluorescent light for 6 h. After this procedure, the specimens were sectioned longitudinally in 3 portions and the extension of microleakage at the gingival wall was determined following a criteria ranging from 0 to 4 using scanning electron microscopy (SEM). The medium portion sectioned of each specimen was polished and prepared for nanoleakage avaliation by SEM. Results: Kruskall-Wallis and Miller statistical tests determined that group 3 presented less microleakage and nanoleakage. Conclusion: Application of the Nd:YAG laser following pretreatment of dentin with adhesive Single Bond non-photocured Single Bond adhesive in cavities prepared with Er:YAG promote better sealing of the gingival margins.

Nitric oxide production in blowfly hemolymph after yeast inoculation

Although insects lack the adaptive immune response of the mammalians, they manifest effective innate immune responses that include both cellular and humoral components. Cellular responses are mediated by hemocytes and Immoral responses include the activation of proteolytic cascades that initiate many events, including NO production. In this work, we determined NO production in Chrysomya megaccphala hemolymph and hemocytes after yeast inoculation. Assays were performed with non-infected controls (NIL), saline-injected larvae (SIL) or larvae injected with Saccharomyces cerevisiae (YIL). The hemolymph of injected groups was collected 0.5, 1, 2, 4, 12, 24 or 48 h post-injection. NO levels in SIL were comparable to those measured in NIL until 12 h, which might be considered the basal production, increasing at 24 and 48 h post-injection, probably in response to the increased larval fragility after cuticle rupture. YIL exhibited significantly higher levels of NO than were found in other groups, peaking at 24 h. L-NAME and EDTA caused a significant reduction of NO production in YIL at this time, suggesting the activity of a Ca2+ -dependent NOS. Plasmatocytes and granular cells phagocytosed the yeasts. Plasmatocytes initiated the nodule formation and granular cells were the only hemocyte type to produce NO. These results permit us to conclude that yeasts induced augmented NO production in C. megacephala hemolymph and granular cells are the hemocyte type involved with the generation of this molecule. (c) 2005 Elsevier B.V. All rights reserved.

Secretion of metalloendopeptidase 24.15 (EC 3.4.24.15)

The metalloendopeptidase EP24.15 (EC3.4.24.15) is a neuropeptide-metabolizing enzyme present in neural and endocrine tissues, presumably functioning extracellularly, Because the majority of the EP24.15 activity is identified in the soluble fraction of cellular homogenates, suggesting that the enzyme is primarily an intracellular protein, we addressed the issue of how EP24.15 arrives in the extracellular environment, We utilized a model system of neuroendocrine secretion, the AtT20 cell, According to both enzymatic activity and immunologic assays, EP24.15 was synthesized in and released from AtT20 cells. Under basal conditions and after stimulation by corticotropin-releasing hormone or the calcium ionophore A23187, EP24.15 activity accumulated in the culture medium. This secretion was not attributable to cell damage, as judged by the absence of release of cytosolic enzyme markers and the ability to exclude trypan blue dye. Pulse-chase analysis and subcellular fractionation of AtT20 cell extracts suggested that the mechanism of EP24.15 secretion is not solely via classical secretory pathways, Additionally, drugs which disrupt the classical secretory pathway, such as Brefeldin A and nocodazole, blocked A23187-stimulated EP24.15 release yet had no effect on basal EP24.15 release, suggesting differences in the basal and stimulated pathways of secretion for EP24.15. In summary, EP24.15 appears to be secreted from AtT20 pituitary cells into the extracellular milieu, where the enzyme can participate in the physiologic metabolism of neuropeptides.