950 resultados para Human identification protocols


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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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Objective In studies on Candida albicans adhesion to surfaces, diverse protocols have been used for collection and preparation of saliva samples. Thus, this study investigated whether variations in the centrifugation parameters and number of donors of saliva would influence the adhesion of C. albicans to a denture base resin. Methods Resin acrylic samples (n = 72) were made and then divided into four groups: (a) control – specimens were left without preconditioning in saliva; (b) three experimental groups, in which the specimens were preconditioned with saliva collected from 15 volunteers and centrifuged at 12 000 g for 5 min (G1); from 15 volunteers and centrifuged at 18 000 g for 30 min (G2); and from one volunteer and centrifuged at 12 000 g for 5 min (G3). Candida adhesion was evaluated by both the 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-[(phenylamino) carbonyl]-2H-tetrazolium hydroxide (XTT) reduction method and crystal violet staining. Data were analyzed by one-way analyses of variance (P = 0.05). Results For XTT reduction assay, groups G2, G3, and control were not significantly different, whereas group G1 showed significantly higher absorbance value than control. For crystal violet staining there were no significant differences among all groups. Conclusion Variations in the centrifugation parameters and number of donors of saliva may influence C. albicans adhesion to denture base resins.

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The intense valuation of an esthetic pleasant smile guided the dentistry to bleached tooth due the popularity of whitening treatments. The consequence of it is an increasing interest in searching the effect of peroxides in hard dental tissues. The aim of this work was to analyze qualitatively in vitro the human enamel after three different bleaching treatments: Opalescence PF 10%, White Class 7.5% and Opalescence Xtra Boost 38%, correlating the structural changes in the surface of the enamel with its respective pH. A total of 40 sound human pre-molars were randomly divided into four groups of 10 elements, which had been immersed in artificial saliva during all the experiment. Bleaching protocols followed the recommendations of the respective manufacturers. Each bleached sample and control group were submitted to a scanning electronic microscopy analysis and compared with one another. Bleaching agents used in this experiment had modified the morphologic aspect of the surface of the dental enamel; however, it did not have correlation between the degrees of severity of the alterations and pH. There is a correlation between hydrogen peroxide concentration and changes in the enamel, where G4 showed more severe alterations, followed for G3 and G2.

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Objective: The aim of this study to investigate the effects of different polymerization protocols on the cuspal movement in class II composite restorations. Materials and methods: Human premolar teeth were prepared with class II cavities and then restored with composite and three-step and two-step etch-and-rinse adhesive systems under different curing techniques (n = 10). It was used a lightemittingdiode curing unit and the mode of polymerization were: standard (exposure for 40 seconds at 700 mW/cm2), pulse-delay (initial exposure for 6 seconds at 350 mW/cm2 followed by a resting period of 3 minutes and a final exposure of 37 seconds at 700 mW/cm2) and soft-start curing (exposure 10 seconds at 350 mW/cm2 and 35 seconds at 700 mW/cm2). The cuspal distance (µm) was measured before and after the restorative procedure and the difference was recorded as cuspal movement. The data were submitted to two-way ANOVA and Bonferroni test (p < 0.05). Results: The type of adhesive system did not influenced the cuspal movement for all the curing methods. Standard protocol showed the highest values of cuspal movement and was statistically different from the pulse-delay and soft-start curing modes. Conclusion: Although the cuspal displacement was not completely avoided, alternative methods of photocuring should be considered to minimize the clinical consequences of composites contraction stress.

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Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

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The taeniasis-cysticercosis complex is a zoonosis of great medical and economic importance where humans play an important role as the carrier of adult stage of Taenia solium and Taenia saginata. This paper describes PCR standardization that can be applied in human fecal samples for taeniasis diagnosis. DNA extraction was achieved with DNAzol reagent, after egg disruption with glass beads. DNA prepared from fecal specimens was first purified and PCR amplified generating fragments of 170 and 600 bp. The assay described herein provides an important tool for T. saginata identification in human fecal samples.

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Since bleaching has become a popular procedure, the effect of peroxides on dental hard tissues is of great interest in research. Purpose: The aim of this in vitro study was to perform a qualitative analysis of the human enamel after the application of in-office bleaching agents, using Scanning Electron Microscopy (SEM). Materials and Methods: Twenty intact human third molars extracted for orthodontic reasons were randomly divided into four groups (n=5) treated as follows: G1- storage in artificial saliva (control group); G2- four 30-minute applications of 35% carbamide peroxide (total exposure: 2h); G3- four 2-hour exposures to 35% carbamide peroxide (total exposure: 8h); G4- two applications of 35% hydrogen peroxide, which was light-activated with halogen lamp at 700mW/cm2 during 7min and remained in contact with enamel for 20min (total exposure: 40min). All bleaching treatments adopted in this study followed the application protocols advised by manufacturers. Evaluation of groups submitted to 35% carbamide peroxide was carried out after two time intervals (30 minutes and 2 hours per session), following the extreme situations recommended by the manufacturer. Specimens were prepared for SEM analysis performing gold sputter coating under vacuum and were examined using 15kV at 500x and 2000x magnification. Results: Morphological alterations on the enamel surface were similarly detected after bleaching with either 35% carbamide peroxide or 35% hydrogen peroxide. Surface porosities were characteristic of an erosive process that took place on human enamel. Depression areas, including the formation of craters, and exposure of enamel rods could also be detected. Conclusion: Bleaching effects on enamel morphology were randomly distributed throughout enamel surface and various degrees of enamel damage could be noticed. Clinical significance: In-office bleaching materials may adversely affect enamel morphology and therefore should be used with caution.

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Dermatophytes are adapted to infect skin, hair and nails by their ability to utilize keratin as a nutrient source. Trichophyton rubrum is an anthropophilic fungus, causing up to 90% of chronic cases of dermatophytosis. The understanding of the complex interactions between the fungus and its host should include the identification of genes expressed during infection. To identify the genes involved in the infection process, representational difference analysis (RDA) was applied to two cDNA populations from T. rubrum, one transcribed from the RNA of fungus cultured in the presence of keratin and the other from RNA generated during fungal growth in minimal medium. The analysis identified differentially expressed transcripts. Genes related to signal transduction, membrane protein, oxidative stress response, and some putative virulence factors were up-regulated during the contact of the fungus with keratin. The expression patterns of these genes were also verified by real-time PCR, in conidia of T. rubrum infecting primarily cultured human keratinocytes in vitro, revealing their potential role in the infective process. A better understanding of this interaction will contribute significantly to our knowledge of the process of dermatophyte infection.

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To evaluate the transdentinal cytotoxicity of resin-based luting cements (RBLCs), with no HEMA in their composition, to odontoblast-like cells. Human dentine discs 0.3 mm thick were adapted to artificial pulp chambers (APCs) and placed in wells of 24-well plates containing 1 mL of culture medium (DMEM). Two categories of HEMA-free RBLCs were evaluated: group 1, self-adhesive Rely X Unicem (RU; 3M ESPE), applied directly to the dentine substrate; and group 2, Rely X ARC (RARC; 3M ESPE), applied to dentine previously acid-etched and treated with a bonding agent. In group 3 (control), considered as representing 100% cell metabolic activity, no treatment was performed on dentine. The APC/disc sets were incubated for 24 h or 7 days at 37 °C and 5% CO2 . Then, the extracts (DMEM + dental materials components that diffused through dentine) were applied to cultured odontoblast-like MDPC-23 cells for 24 h. After that, the cell viability (MTT assay), cell morphology (SEM), total protein production (TP) and alkaline phosphatase (ALP) activity were assessed. Data from MTT assay and TP production were analysed by Kruskal-Wallis and Mann-Whitney tests (α = 5%). Data from ALP activity were analysed by one-way anova and Tukey's test (α = 5%). In group 1, a slight reduction in cell viability (11.6% and 16.8% for 24-h and 7-day periods, respectively) and ALP activity (13.5% and 17.9% for 24-h and 7-day periods, respectively) was observed, with no significant difference from group 3 (control) (P > 0.05). In group 2, a significant reduction in cell viability, TP production and ALP activity compared with group 3 (control) occurred (P < 0.05), regardless of incubation time. Alteration in MDPC-23 cell morphology was observed only in group 2. HEMA-free Rely X ARC cement caused greater toxicity to odontoblast-like MDPC-23 cells than did Rely X Unicem cement when both resin-based luting materials were applied to dentine as recommended by the manufacturer.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

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Fusobacterium nucleatum is one of the most common anaerobic bacteria present in the oral cavity and is often isolated from infections involving other body sites. To characterise F. nucleatum strains from patients attending a teaching hospital in Nigeria in order to provide information on the methods for accurate identification of anaerobes in clinical specimen. Fusobacterium nucleatum specie from 50 patients presenting with oro-facial infections were studied by culture on Fusobacterium selective agar and fastidious anaerobe agar. The isolates were characterised based on colonial morphology, microscopy, lipase production, susceptibility to kanamycin and colistin and resistance to vancomycin. Biochemical tests were performed using a commercial test kit. The identity of the isolates was confirmed based on molecular characterization performed using polymerase chain reaction (PCR) analysis. Forty-eight (96%) F. nucleatum isolates were obtained from the 50 patients by culture and all the isolates were identified by colonial appearance and microscopy based on their unique spindle shape with tapered ends. Only 26 (54.2%) of the 48 isolates were identified by commercial API 20A test kit while PCR confirmed the identity of all the isolates. Anaerobes are involved in human infections and their study is quite cumbersome due to tedious nature and high cost of the techniques involved. Cultural method is reliable in the isolation and identification of F. nucleatum species. PCR is a rapid and simple method that can complement the phenotypic identification of anaerobes and would assist in their full identification.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)