High temperature annealing effect on structure, optical property and laser-induced damage threshold of Ta2O5 films

Ta2O5 films were deposited by conventional electron beam evaporation method and then annealed in air at different temperature from 873 to 1273 K. It was found that the film structure changed from amorphous phase to hexagonal phase when annealed at 1073 K, then transformed to orthorhombic phase after annealed at 1273 K. The transmittance was improved after annealed at 873 K, and it decreased as the annealing temperature increased further. The total integrated scattering (TIS) tests and AFM results showed that both scattering and root mean square (RMS) roughness of films increased with the annealing temperature increasing. X-ray photoelectron spectroscopy (XPS) analysis showed that the film obtained better stoichiometry and the O/Ta ratio increased to 2.50 after annealing. It was found that the laser-induced damage threshold (LIDT) increased to the maximum when annealed at 873 K, while it decreased when the annealing temperature increased further. Detailed damaged models dominated by different parameters during annealing were discussed. (C) 2008 Elsevier B. V. All rights reserved.

Laser induced damage threshold at 355 and 1064 nm of Ta2O5 films of different phases

Ta2O5 films are deposited on fused silica substrates by conventional electron beam evaporation method. By annealing at different temperatures, Ta2O5 films of amorphous, hexagonal and orthorhombic phases are obtained and confirmed by x-ray diffractometer ( XRD) results. X-ray photoelectron spectroscopy ( XPS) analysis shows that chemical composition of all the films is stoichiometry. It is found that the amorphous Ta2O5 film achieves the highest laser induced damage threshold ( LIDT) either at 355 or 1064 nm, followed by hexagonal phase and finally orthorhombic phase. The damage morphologies at 355 and 1064 nm are different as the former shows a uniform fused area while the latter is centred on one or more defect points, which is induced by different damage mechanisms. The decrease of the LIDT at 1064nm is attributed to the increasing structural defect, while at 355nm is due to the combination effect of the increasing structural defect and decreasing band gap energy.

Effects of deposition rates on laser damage threshold of TiO2/SiO2 high reflectors

TiO2 single layers and TiO2/SiO2 high reflectors (HR) are prepared by electron beam evaporation at different TiO2 deposition rates. It is found that the changes of properties of TiO2 films with the increase of rate, such as the increase of refractive index and extinction coefficient and the decrease of physical thickness, lead to the spectrum shift and reflectivity bandwidth broadening of HR together with the increase of absorption and decrease of laser-induced damage threshold. The damages are found of different morphologies: a shallow pit to a seriously delaminated and deep crater, and the different amorphous-to-anatase-to-rutile phase transition processes detected by Raman study. The frequency shift of Raman vibration mode correlates with the strain in. film. Energy dispersive X-ray analysis reveals that impurities and non-stoichiometric defects are two absorption initiations resulting to the laser-induced transformation. (C) 2008 Elsevier B. V. All rights reserved.

Structure-function analysis of USP1: insights into the role of Ser313 phosphorylation site and the effect of cancer-associated mutations on autocleavage

Background: In complex with its cofactor UAF1, the USP1 deubiquitinase plays an important role in cellular processes related to cancer, including the response to DNA damage. The USP1/UAF1 complex is emerging as a novel target in cancer therapy, but several aspects of its function and regulation remain to be further clarified. These include the role of the serine 313 phosphorylation site, the relative contribution of different USP1 sequence motifs to UAF1 binding, and the potential effect of cancer-associated mutations on USP1 regulation by autocleavage. Methods: We have generated a large set of USP1 structural variants, including a catalytically inactive form (C90S), non-phosphorylatable (S313A) and phosphomimetic (S313D) mutants, deletion mutants lacking potential UAF1 binding sites, a mutant (GG/AA) unable to undergo autocleavage at the well-characterized G670/G671 diglycine motif, and four USP1 mutants identified in tumor samples that cluster around this cleavage site (G667A, L669P, K673T and A676T). Using cell-based assays, we have determined the ability of these mutants to bind UAF1, to reverse DNA damage-induced monoubiquitination of PCNA, and to undergo autocleavage. Results: A non-phosphorylatable S313A mutant of USP1 retained the ability to bind UAF1 and to reverse PCNA ubiquitination in cell-based assays. Regardless of the presence of a phosphomimetic S313D mutation, deletion of USP1 fragment 420-520 disrupted UAF1 binding, as determined using a nuclear relocation assay. The UAF1 binding site in a second UAF1-interacting DUB, USP46, was mapped to a region homologous to USP1(420-520). Regarding USP1 autocleavage, co-expression of the C90S and GG/AA mutants did not result in cleavage, while the cancer-associated mutation L669P was found to reduce cleavage efficiency. Conclusions: USP1 phosphorylation at S313 is not critical for PCNA deubiquitination, neither for binding to UAF1 in a cellular environment. In this context, USP1 amino acid motif 420-520 is necessary and sufficient for UAF1 binding. This motif, and a homologous amino acid segment that mediates USP46 binding to UAF1, map to the Fingers sub-domain of these DUBs. On the other hand, our results support the view that USP1 autocleavage may occur in cis, and can be altered by a cancer-associated mutation.

Estudo dos efeitos da radioterapia no tecido cardíaco e sua associação com o metabolismo energético

Durante o tratamento radioterápico para tumores localizados na região torácica, parte do coração frequentemente é incluída no campo de tratamento e pode receber doses de radiação ionizante, significativas em relação à terapêutica. A irradiação do coração é capaz de causar importantes complicações cardíacas ao paciente, caracterizadas por alterações funcionais progressivas cerca de 10 a 20 anos após a exposição do órgão. Devido ao seu alto grau de contração e grande consumo energético, o tecido cardíaco é altamente dependente do metabolismo oxidativo que ocorre nas mitocôndrias. Danos as estas organelas podem levar ao decréscimo da produção de energia, tendo um impacto direto sobre a performance cardíaca. Ainda, ao interagir com as células, a radiação ionizante pode gerar uma série de eventos bioquímicos que conduzem a uma resposta celular complexa, em que muitas proteínas parecem estar envolvidas. Tendo em vista tais conhecimentos, o objetivo do estudo foi avaliar o aspecto ultraestrutural do tecido cardíaco, a bioenergética mitocondrial e a expressão diferencial de proteínas após irradiação. Os ensaios foram realizados em amostras de tecido cardíaco de ratos Wistar irradiados com dose única de 20 Gy direcionada ao coração. As análise tiveram início 4 e 32 semanas após irradiação. A análise ultraestrutural foi realizada através de microscopia eletrônica de transmissão. A respiração mitocondrial foi mensurada em oxígrafo, a partir das taxas de consumo de oxigênio pelas fibras cardíacas. A identificação de proteínas diferencialmente expressas foi investigada através de duas técnicas proteômicas: 2D-DIGE (2-D Fluorescence Difference Gel Electrophoresis) e uma abordagem label-free seguida de espectrometria de massas. Os resultados mostraram que os efeitos tardios da radiação incluem a degeneração das mitocôndrias e das unidades contráteis do tecido cardíaco, disfunções na cadeia respiratória mitocondrial e expressão diferencial de proteínas envolvidas no metabolismo energético de carboidratos, lipídeos e da fosfocreatina. De forma geral, o estudo mostrou que a irradiação cardíaca prejudica o processo de síntese energética, conduzindo a um déficit da taxa respiratória mitocondrial como efeito tardio. Tal evento pode culminar em disfunções mecânicas no coração, caracterizando o desenvolvimento de doenças cardíacas radioinduzidas.

MicroRNAs in breast cancer progression and DNA damage response

Os tumores de mama são caracterizados pela sua alta heterogeneidade. O câncer de mama é uma doença complexa, que possui o seu desenvolvimento fortemente influenciado por fatores ambientais, combinada a uma progressiva acumulação de mutações genéticas e desregulação epigenética de vias críticas. Alterações nos padrões de expressão gênica podem ser resultado de uma desregulação no controle de eventos epigenéticos, assim como, na regulação pós-transcricional pelo mecanismo de RNA de interferência endógeno via microRNA (miRNA). Estes eventos são capazes de levar à iniciação, à promoção e à manutenção da carcinogênese, como também ter implicações no desenvolvimento da resistência à terapia Os miRNAs formam uma classe de RNAs não codificantes, que durante os últimos anos surgiram como um dos principais reguladores da expressão gênica, através da sua capacidade de regular negativamente a atividade de RNAs mensageiros (RNAms) portadores de uma seqüencia parcialmente complementar. A importância da regulação mediada por miRNAs foi observada pela capacidade destas moléculas em regular uma vasta gama de processos biológicos incluindo a proliferação celular, diferenciação e a apoptose. Para avaliar a expressão de miRNAs durante a progressão tumoral, utilizamos como modelo experimental a série 21T que compreende 5 linhagens celulares originárias da mesma paciente diagnosticada com um tumor primário de mama do tipo ErbB2 e uma posterior metástase pulmonar. Essa série é composta pela linhagem obtida a partir do tecido normal 16N, pelas linhagens correspondentes ao carcinoma primário 21PT e 21NT e pelas linhagens obtidas um ano após o diagnóstico inicial, a partir da efusão pleural no sítio metastatico 21MT1 e 21MT2. O miRNAoma da série 21T revelou uma redução significativa nos níveis de miR-205 e nos níveis da proteina e-caderina e um enriquecimento do fator pró-metastático ZEB-1 nas células 21MT. Considerando a importância dos miRNAs na regulação da apoptose, e que a irradiação em diferentes espectros é comumente usada em procedimentos de diagnóstico como mamografia e na radioterapia, avaliamos a expressão de miRNAs após irradiação de alta e baixa energia e do tratamento doxorrubicina. Para os ensaios foram utilizados as linhagens não tumorais MCF-10A e HB-2 e as linhagens de carcinoma da mama MCF-7 e T-47D. Observou-se que raios-X de baixa energia são capazes de promover quebras na molécula do DNA e apoptose assim como, alterar sensivelmente miRNAs envolvidos nessas vias como o let-7a, miR-34a e miR-29b. No que diz respeito à resposta a danos genotóxicos, uma regulação positiva sobre a expressão de miR-29b, o qual em condições normais é regulado negativamente foi observada uma regulação positiva sobre miR-29b expressão após todos os tratamentos em células tumorais. Nossos resultados indicam que miR-29b é um possível biomarcador de estresse genotóxico e que miR-205 pode participar no potencial metastático das células 21T.

Effect of analytical conditions in wavelength dispersive electron microprobe analysis on the measurement of strontium-to-calcium (Sr/Ca) ratios in otoliths of anadromous salmonids

The use of strontium-to-calcium (Sr/Ca) ratios in otoliths is becoming a standard method to describe life history type and the chronology of migrations between freshwater and seawater habitats in teleosts (e.g. Kalish, 1990; Radtke et al., 1990; Secor, 1992; Rieman et al., 1994; Radtke, 1995; Limburg, 1995; Tzeng et al. 1997; Volk et al., 2000; Zimmerman, 2000; Zimmerman and Reeves, 2000, 2002). This method provides critical information concerning the relationship and ecology of species exhibiting phenotypic variation in migratory behavior (Kalish, 1990; Secor, 1999). Methods and procedures, however, vary among laboratories because a standard method or protocol for measurement of Sr in otoliths does not exist. In this note, we examine the variations in analytical conditions in an effort to increase precision of Sr/Ca measurements. From these findings we argue that precision can be maximized with higher beam current (although there is specimen damage) than previously recommended by Gunn et al. (1992).

Feasibility of damage detection using vibration data in a population of cylinders

The feasibility of vibration data to identify damage in a population of cylindrical shells is assessed. Vibration data from a population of cylinders were measured and modal analysis was employed to obtain natural frequencies and mode shapes. The mode shapes were transformed into the Coordinate Modal Assurance Criterion (COMAC). The natural frequencies and the COMAC before and after damage for a population of structures show that modal analysis is a viable route to damage identification in a population of nominally identical cylinders. Modal energies, which are defined as the integrals of the real and imaginary components of the frequency response functions over various frequency ranges, were extracted and transformed into the Coordinate Modal Energy Assurance Criterion (COMEAC). The COMEAC before and after damage show that using modal energies is a viable approach to damage identification in a population of cylinders.

The effect of beam interruptions on the integrity of ADSR fuel pin cladding: A thermo-mechanical analysis

During its lifetime in the core, the cladding of an Accelerator Driven Subcritical Reactor (ADSR) fuel pin is expected to experience variable stresses due to frequent interruptions in the accelerator proton beam. This paper investigates the thermal fatigue damage in the cladding due to repetitive and unplanned beam interruptions under certain operational conditions. Beam trip data was obtained for four operating high power proton accelerators, among which the Spallation Neutron Source (SNS) superconducting accelerator was selected for further analysis. 9Cr-1Mo-Nb-V (T91) steel was selected as the cladding material because of its proven compatibility with proposed ADSR design concepts. The neutronic, thermal and stress analyses were performed using the PTS-ADS, a code that has been specifically developed for studying the dynamic response to beam-induced transients in accelerator driven subcritical systems. The lifetime of the fuel cladding in the core was estimated for three levels of allowed pin power and specific operating conditions. © 2012 Elsevier Ltd. All rights reserved.

Modelling and Fragility Analysis of Non-Ductile Reinforced Concrete Buildings in Low-to-Moderate Seismic Zones

Reinforced concrete buildings in low-to-moderate seismic zones are often designed only for gravity loads in accordance with the non-seismic detailing provisions. Deficient detailing of columns and beam-column joints can lead to unpredictable brittle failures even under moderate earthquakes. Therefore, a reliable estimate of structural response is required for the seismic evaluation of these structures. For this purpose, analytical models for both interior and exterior slab-beam-column subassemblages and for a 1/3 scale model frame were implemented into the nonlinear finite element platform OpenSees. Comparison between the analytical results and experimental data available in the literature is carried out using nonlinear pushover analyses and nonlinear time history analysis for the subassemblages and the model frame, respectively. Furthermore, the seismic fragility assessment of reinforced concrete buildings is performed on a set of non-ductile frames using nonlinear time history analyses. The fragility curves, which are developed for various damage states for the maximum interstory drift ratio are characterized in terms of peak ground acceleration and spectral acceleration using a suite of ground motions representative of the seismic hazard in the region.

Reliability analysis of deep excavation based on a semi-empirical approach

The design and construction of deep excavations in urban environment is often governed by serviceability limit state related to the risk of damage to adjacent buildings. In current practice, the assessment of excavation-induced building damage has focused on a deterministic approach. This paper presents a component/system reliability analysis framework to assess the probability that specified threshold design criteria for multiple serviceability limit states are exceeded. A recently developed Bayesian probabilistic framework is used to update the predictions of ground movements in the later stages of excavation based on the recorded deformation measurements. An example is presented to show how the serviceability performance for excavation problems can be assessed based on the component/system reliability analysis. © 2011 ASCE.

Building damage assessment for deep excavations in Singapore and the influence of building stiffness

One of the biggest issues for underground construction in a densely built-up urban environment is the potentially adverse impact on buildings adjacent to deep excavations. In Singapore, a building damage assessment is usually carried out using a three-staged approach to assess the risk of damage caused by major underground construction projects. However, the tensile strains used for assessing the risk of building damage are often derived using deflection ratios and horizontal strains under 'greenfield' conditions. This ignores the effects of building stiffness and in many cases may be conservative. This paper presents some findings from a study on the response of buildings to deep excavations. Firstly, the paper discusses the settlement response of an actual building - the Singapore Art Museum - adjacent to a deep excavation. By comparing the monitored building settlement with the adjacent ground settlement markers, the influence of building stiffness in modifying the response to excavation-induced settlements is observed. Using the finite element method, a numerical study on the building response to movements induced by deep excavations found a consistent relationship between the building modification factor and a newly defined relative bending stiffness of the building. This relationship can be used as a design guidance to estimate the deflection ratio in a building from the greenfield condition. By comparing the case study results with the design guidance developed from finite element analysis, this paper presents some important characteristics of the influence of building stiffness on building damages for deep excavations.

Static analysis of Euler-Bernoulli beams with multiple unilateral cracks under combined axial and transverse loads

The paper deals with the static analysis of pre-damaged Euler-Bernoulli beams with any number of unilateral cracks and subjected to tensile or compression forces combined with arbitrary transverse loads. The mathematical representation of cracks with a bilateral behaviour (i.e. always open) via Dirac delta functions is extended by introducing a convenient switching variable, which allows each crack to be open or closed depending on the sign of the axial strain at the crack centre. The proposed model leads to analytical solutions, which depend on four integration constants (to be computed by enforcing the boundary conditions) along with the Boolean switching variables associated with the cracks (whose role is to turn on and off the additional flexibility due to the presence of the cracks). An efficient computational procedure is also presented and numerically validated. For this purpose, the proposed approach is applied to two pre-damaged beams, with different damage and loading conditions, and the results so obtained are compared against those given by a standard finite element code (in which the correct opening of the cracks is pre-assigned), always showing a perfect agreement. © 2013 Elsevier Ltd. All rights reserved.

First Identification of the Hepatotoxic Microcystins in the Serum of a Chronically Exposed Human Population Together with Indication of Hepatocellular Damage

Hepatotoxic microcystins (MCs) are the most commonly reported cyanotoxins in eutrophic freshwaters. In 1996, human intoxications by MCs caused deaths of 76 patients at Caruaru dialysis centers in Brazil. So far, there have been no direct evidences of MC occurrence in human tissue in consequence of exposure to MC. In this study, we improved cleanup procedures for detecting MCs in serum sample using liquid chromatographymass spectrometry, and confirmed for the first time the presence of MCs in serum samples (average 0.39 ng/ml, which amounts to ca. 1/87 of the concentrations found in tissue samples of the Caruaru victims) of fishermen at Lake Chaohu. Daily intake by the fishermen was estimated to be in the range of 2.2-3.9 mu g MC-LReq, whereas the provisional World Health Organization tolerable daily intake (TDI) for daily lifetime exposure is 0.04 mu g/kg or 2-3 mu g per person. Moreover, statistical analysis showed closer positive relationships between MC serum concentrations and concentrations of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and lactate dehydrogenase than between the MC concentrations and other biochemical indicators. Thus, the data raise the question whether extended exposure in the range of the TDI or up to a factor of 10 above it may already lead to indication of liver damage. The results also demonstrate a risk of health effects from chronic exposure to MCs at least for populations with high levels of exposure, like these fishermen.

Field and laboratory studies on pathological and biochemical characterization of microcystin-induced liver and kidney damage in the phytoplanktivorous bighead carp

Field and experimental studies were conducted to investigate pathological characterizations and biochemical responses in the liver and kidney of the phytoplanktivorous bighead carp after intraperitoneal (i.p.) administration of microcystins (MCs) and exposure to natural cyanobacterial blooms in Meiliang Bay, Lake Taihu. Bighead carp in field and laboratory studies showed a progressive recovery of structure and function in terms of histological, cellular, and biochemical features. In laboratory study, when fish were i.p. injected with extracted MCs at the doses of 200 and 500 mu g MC- LReq/kg body weight, respectively, liver pathology in bighead carp was observed in a time dose-dependent manner within 24 h postinjection and characterized by disruption of liver structure, condensed cytoplasm, and the appearance of massive hepatocytes with karyopyknosis, karyorrhexis, and karyolysis. In comparison with previous studies on other fish, bighead carp in field study endured higher MC doses and longer-term exposure, but displayed less damage in the liver and kidney. Ultrastructural examination in the liver revealed the presence of lysosome proliferation, suggesting that bighead carp might eliminate or lessen cell damage caused by MCs through lysosome activation. Biochemically, sensitive responses in the antioxidant enzymes and higher basal glutathione concentrations might be responsible for their powerful resistance to MCs, suggesting that bighead carp can be used as biomanipulation fish to counteract cyanotoxin contamination.