Polymorphisms of the DNA repair genes XRCC1 and XRCC3 in a Brazilian population

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Structural basis of nuclear import of flap endonuclease 1 (FEN1)

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

Estudos estruturais com a importina-α de mamíferos e peptídeos de sequências de localização nuclear (NLS)

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

Expression of 8-oxoguanine Glycosylase in Human Fetal Membranes

Problem The most common DNA lesion generated by oxidative stress (OS) is 7, 8-dihydro-8-oxoguanine (8-oxoG) whose excision repair is performed by 8-oxoguanine glycosylase (OGG1). We investigated OGG1 expression changes in fetal membranes from spontaneous preterm birth (PTB) and preterm premature rupture of the membranes (pPROM) and its changes in vitro in normal fetal membranes exposed to OS inducer water-soluble cigarette smoke extract (CSE). Method of study DNA damage was determined in amnion cells treated with CSE by comet and FLARE assays. OGG1 mRNA expression and localization in fetal membranes from clinical specimens and in normal term membranes exposed to CSE were examined by QRT-PCR and by immunohistochemistry. Results DNA strand and base damage was seen in amnion cells exposed to CSE. OGG1 expression was 2.5-fold higher in PTB samples compared with pPROM (P=0.045). No significant difference was seen between term and pPROM or PTB and term. CSE treatment showed a nonsignificant decrease in OGG1. OGG1 was localized to both amnion and chorion with less intense staining in pPROM and CSE-treated membranes. Conclusion Increased OS-induced DNA damage predominated by 8-oxoG is likely to persist in fetal cells due to reduced availability of base excision repair enzyme OGG1. This can likely lead to fetal cell senescence associated with some adverse pregnancy outcome.

Functional XPB/RAD25 redundancy in Arabidopsis genome: characterization of AtXPB2 and expression analysis

The xeroderma pigmentosum complementation group B (XPB) protein is involved in both DNA repair and transcription in human cells. It is a component of the transcription factor IIH (TFIIH) and is responsible for DNA helicase activity during nucleotide (nt) excision repair (NER). Its high evolutionary conservation has allowed identification of homologous proteins in different organisms, including plants. In contrast to other organisms, Arabidopsis thaliana harbors a duplication of the XPB orthologue (AtXPB1 and AtXPB2), and the proteins encoded by the duplicated genes are very similar (95% amino acid identity). Complementation assays in yeast rad25 mutant strains suggest the involvement of AtXPB2 in DNA repair, as already shown for AtXPB1, indicating that these proteins may be functionally redundant in the removal of DNA lesions in A. thaliana. Although both genes are expressed in a constitutive manner during the plant life cycle, Northern blot analyses suggest that light modulates the expression level of both XPB copies, and transcript levels increase during early stages of development. Considering the high similarity between AtXPB1 and AtXPB2 and that both of predicted proteins may act in DNA repair, it is possible that this duplication may confer more flexibility and resistance to DNA damaging agents in thale cress. (C) 2004 Elsevier B.V. All rights reserved.

Clinicopathological significance of ERCC1 expression in breast cancer

The excision repair cross-complementation 1 (ERCC1) enzyme plays an essential role in the nucleotide excision repair pathway and is associated with resistance to platinum-based chemotherapy in different types of cancer. The aim of the present study was to evaluate the clinicopathological significance of ERCC1 expression in breast cancer patients. We analyzed the immunohistochemical expression of ERCC1 in a tissue microarray from 135 primary breast carcinomas and correlated the immunohistochemical findings with clinicopathological factors and outcome data. ERCC1 expression analysis was available for 109 cases. In this group, 58 (53.2%) were positive for ERCC1. ERCC1-positive expression was correlated with smaller tumor size (P=0.007) and with positivity for estrogen receptor (P=0.040), but no correlation was found with other clinicopathological features. Although not statistically significant, triple negative breast cancers were more frequently negative for ERCC1 (61.5% of the cases) compared to the non-triple negative breast cancer cases (41.5%). In conclusion, ERCC1 expression correlated significantly with favorable prognostic factors, such as smaller tumor size and ER-positivity, suggesting a possible role for ERCC1 as a predictive and/or prognostic marker in breast cancer. © 2013 Elsevier GmbH.

Effects of β-glucan extracted from Agaricus blazei on the expression of ERCC5, CASP9, and CYP1A1 genes and metabolic profile in HepG2 cells

The polysaccharide β-glucan has biological properties that stimulate the immune system and can prevent chronic pathologies, including cancer. It has been shown to prevent damage to DNA caused by the chemical and physical agents to which humans are exposed. However, the mechanism of β-glucan remains poorly understood. The objective of the present study was to verify the protective effect of β-glucan on the expression of the genes ERCC5 (involved in excision repair of DNA damage), CASP9 (involved in apoptosis), and CYP1A1 (involved in the metabolism of xenobiotics) using real-time polymerase chain reaction and perform metabolic profile measurements on the HepG2 cells. Cells were exposed to only benzo[a]pyrene (B[a]P), β-glucan, or a combination of B[a]P with β-glucan. The results demonstrated that 50 μg/mL β-glucan significantly repressed the expression of the ERCC5 gene when compared with the untreated control cells in these conditions. No change was found in the CASP9 transcript level. However, the CYP1A1 gene expression was also induced by HepG2 cells exposed to B[a]P only or in association with β-glucan, showing its effective protector against damage caused by B[a]P, while HepG2 cells exposed to only β-glucan did not show CYP1A1 modulation. The metabolic profiles showed moderate bioenergetic metabolism with an increase in the metabolites involved in bioenergetic metabolism (alanine, glutamate, creatine and phosphocholine) in cells treated with β-glucan and to a lesser extent treated with B[a]P. Thus, these results demonstrate that the chemopreventive activity of β-glucan may modulate bioenergetic metabolism and gene expression. © 2013 The Author(s).

Desenvolvimento e validação de uma técnica quantitativa de análise de imagem para avaliação do teste do cometa corado pela prata

The comet assay is a method of DNA damage analysis widely used to quantify oxidative damage, crosslinks of DNA, apoptosis and genotoxicity of chemicals substances as chemical, pharmaceuticals, agrochemicals products, among others. This technique is suitable to detect DNA strand breaks, alkali-labile sites and incomplete excision repair sites and is based on the migration of DNA fragments by microeletroforesis, DNA migrates for the anode forming a “tail”, and the formed image has the appearance of a comet. The slides can be stained with fluorescence or silver, having differences in the microscopy type used for the analysis and the possibility of storage of the slides, moreover, the first one is a stained-method with more difficulties of accomplishment. The image analysis can be performed by a visual way, however, there is a disadvantage as the subjectivity on the results, that can be minimized by an automated method of digital analysis. This process was studied in this report with the aim to perceive the validation of the digital analysis turning it a quantitative method with larger reproductibility, minimizing the variability and imprecision due to the subjective analysis. For this validation we selected 50 comets photographed in a standardized way and printed, afterwards, pictures were submitted to three experienced appraisers, who quantified them manually. Later, the images were processed by free software ImageJ 1.38x, printed and quantified manually by the same appraisers. The intraclass correlation was higher to comet measures after image processing. Following, an algorithm of automated digital analysis from the measures of the comet was developed; the values obtained were compared with those 12 estimated manually after the processing resulting high correlation among the measures. The use of image analysis systems increases ...(Complete abstract click electronic access below)

Effect of different repair techniques on the accuracy of repositioning the fractured denture base

Objective:This study investigated the efficacy of different techniques for the union of fragments of a denture before repair and on the accuracy of the reposition.Materials and methods:For this study, 20 maxillary dentures made with Lucitone 550 heat-cured resin were used. Points were determined with a scanner on the cusp of the teeth, as a measurement of the segments. After digitisation, each model was exported to the AUTOCAD R 14 program and two-dimensional measurements of the distances between the marked points were made. After the initial analysis, the dentures were fractured into two segments using an impact test machine. For the repair, maxillary dentures were divided into two groups; in the first, the repair was carried out using Kerr's sticky wax and in the second group, Super Bonder was used to join the fragments, with subsequent inclusion of DENTSPLY((R)) Repair Material resin. After the repair, the points of the maxillary dentures were measured again. The numerical values obtained were tabulated to compare the measurements before fracture and after the repair. For statistical analysis, analysis of variance was employed, using a single factor and double factor, followed by the Tukey test with a reliability of 95%.Results:The results demonstrated a statistically significant difference between the materials used to join the dentures for repair, where the dentures were joined with sticky wax presented a larger variation in the distances between the points.Conclusion:The variation in distances between the points is influenced by the agent of repair.

Influence of surface treatments on the flexural strength of denture base repair

Objective: The purpose of this study was to evaluate the flexural strength of repairs made with autopolymerising acrylic resin after different treatments of joint surfaces.Material and Methods: Fifty rectangular specimens were made with heat-polymerised acrylic resin and 40 were repaired with autopolymerising acrylic resin following joint surface treatments: group 1 (intact specimens), group 2 (chemical treatment: wetting with methyl-methacrylate for 180 s), group 3 (abraded with silicon carbide paper), group 4 (abraded and wetting with methyl-methacrylate for 180 s) and group 5 (without surface treatment). The flexural strength was measured by a three-point bending test using a universal testing machine with a 100 Kgf load cell in the centre of repair at 5 mm/min cross-head speed. All data were analysed using one-way ANOVA and Tukey HSD test for multiple comparisons (p < 0.05).Results: Among repaired specimens, groups 2 and 4 had 66.53 +/- 3.4 and 69.38 +/- 1.8 MPa mean values and were similar. These groups had superior flexural strength than groups 3 and 5 that were similar and had 54.11 +/- 3.4 and 51.24 +/- 2.8 MPa mean values, respectively. Group 1 had a mean value of 108.30 +/- 2.8 MPa being the highest result.Conclusion: It can be concluded that the treatment of the joint surfaces with methyl-methacrylate increases the flexural strength of denture base repairs, although the strength is still lower than that observed for the intact denture base resin. Abrasion with sandpaper was not able to influence the flexural strength of repaired denture bases.

Repair Welding Effects on the Bending Fatigue Strength of AISI 4130 Aeronautical Steel used in a Critical to the Flight-Safety Structure

The aim of this study was to analyze the effect of successive TIG (tungsten inert gas) welding repairs on the reverse bending fatigue strength of AISI 4130 steel, which is widely used in components critical to the flight-safety. In order to simulate the abrupt maneuvers, wind bursts, motor vibration and helixes efforts, which generate cyclic bending loadings at the welded joints of a specific aircraft component called motor cradle, experimental reverse bending fatigue tests were carried out on specimens made from hot-rolled steel plate, 1.10 mm (0.043 in) thick, by mean of a SCHENK PWS equipment, with load ratio R = -1, under constant amplitude, at 30 Hz frequency and room temperature. It was observed that the bending fatigue strength decreases after the TIG (Tungsten Inert Gas) welding process application on AISI 4130 steel, with subsequent decrease due to re-welding sequence as well. Microstructural analyses and microhardness measurements on the base material, heat-affected zone (HAZ) and weld metal, as well as the effects of the weld bead geometry on the obtained results, have complemented this study.

Considerations about the welding repair effects on the structural integrity of an airframe critical to the flight-safety

Structures critical to the flight-safety are commonly submitted to several maintenance repairs at the welded joints in order to prolong the in-service life of aircrafts. The aim of this study is to analyze the effects of Tungsten Inert Gas (TIG) welding repair on the structural integrity of the AISI 4130 aeronautical steel by means of experimental fatigue crack growth tests in base-material, heat-affected zone (HAZ) and weld metal. The tests were performed on hot-rolled steel plate specimens, 0.89 mm thick, with load ratio R = 0.1, constant amplitude, at 10 Hz frequency and room temperature. Increase of the fracture resistance was observed in the weld metal but decreasing in the HAZ after repair. The results were associated to microhardness and microstructural changes with the welding sequence. (C) 2010 Published by Elsevier Ltd.

Proliferation of the vascular endothelium of the iris following total debridement of the corneal epithelium and limbal excision of rabbits

Background Damage to the corneal epithelium causes not only a reaction for its repair but also affects other parts of the cornea as well as different components of the anterior segment of the eye. The purpose of this investigation was to analyze the consequences, following epithelial and limbal damage, to the iris of rabbits (Oryctolagus cuniculus).Methods The corneal epithelium was thoroughly scraped followed by surgical excision of the limbus. Next, (3)H-thymidine ((3)H-TdR) was injected intravitreally both into the right (experimental) and left (control) eyes which had their anterior segments processed for autoradiography at intervals of 2, 7 and 21 days after surgery (three rabbits per interval). The irises were also examined with scanning-electron and confocal microscopy after Evans blue injection.Results There was a high frequency of labeling in the cells of the iris blood vessels in the experimental eye, particularly the endothelial ones. The ratio of labeled cells between experimental and control irises was 40:1, with a population of nuclei increasing by 25% and remaining labeled up to 21 days. There was also an increase in the volume of the iris vasculature as shown by confocal microscopy. The high labeling frequencies of the vascular cells were observed throughout the iris from the ciliary to the pupillary regions.Conclusions The lesions on the corneal epithelium elicit proliferation of the iris vascular cells, mainly its endothelium, as well as an early breakdown of the blood-aqueous barrier. The daughter cells resulting from the damage to the eye surface were detected up to 21 days after a single injection of (3)H-TdR, most likely due to their slow turnover. As a consequence of this proliferation, the vasculature of the iris increased in volume.

Pomada a base de óleo de semente de maracujá no tratamento de lesões cutâneas em coelhos

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influência do curativo de demora à base de hidróxido de cálcio na reparação dos tecidos apicais e periapicais de dentes sem vitalidade pulpar com ou sem lesão periapical visível radiograficamente: estudo histopatológico em dentes de cães

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)