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.

Ballasted & electrically steerable carbon nanotube field emitters

Here we present our on-going efforts toward the development of stable ballasted carbon nanotube-based field emitters employing hydrothermally synthesized zinc oxide nanowires and thin film silicon-on-insulator substrates. The semiconducting channel in each controllably limits the emission current thereby preventing detrimental burn-out of individual emitters that occurs due to unavoidable statistical variability in emitter characteristics, particularly in their length. Fabrication details and emitter characterization are discussed in addition to their field emission performance. The development of a beam steerable triode electron emitter formed from hexagonal carbon nanotube arrays with central focusing nanotube electrodes, is also described. Numerical ab-initio simulations are presented to account for the empirical emission characteristics. Our engineered ballasted emitters have shown some of the lowest reported lifetime variations (< 0.7%) with on-times of < 1 ms, making them ideally-suited for next-generation displays, environmental lighting and portable x-rays sources. © 2012 SPIE.

Near infrared lights emission of Ar-40(16+) impacting on metal surfaces

The highly charged ion Ar-40(16+) with the velocity (kinetic energy E (K)=150 keV, velocity V=8.5x10(5) m/s) smaller than Bohr velocity (V (Bohr)=2.9x10(6) m/s) was found to hove impacts on the surfaces of metals Ni, Mo, Au and Al, and the Ar atomic infrared light lines and X-rays spectra were simultaneously measured. The experimental results show that the highly charged ion that captures electrons is neutralized, and the multiply-excited hollow atom forms. The hollow atom cascade decay radiates lights from infrared to X-ray spectrum. The intensity of infrared lights shows that the metallic work functions play an important role in the neutralization process of highly charged ions during their interaction with metallic surfaces, which verifies the classical over-the-barrier model.

Linear energy transfer dependence of the effects of carbon ion beams on adventitious shoot regeneration from in vitro leaf explants of Saintpaulia ionahta

Purpose: To determine the effects of carbon ion beams with five different linear energy transfer (LET) values on adventitious shoots from in vitro leaf explants of Saintpaulia ionahta Mauve cultivar with regard to tissue increase, shoots differentiation and morphology changes in the shoots. Materials and methods: In vitro leaf explant samples were irradiated with carbon ion beams with LET values in the range of 31 similar to 151 keV/mu m or 8 MeV of X-rays (LET 0.2 keV/mu m) at different doses. Fresh weight increase, surviving fraction and percentage of the explants with regenerated malformed shoots in all the irradiated leaf explants were statistically analysed. Results: The fresh weight increase (FWI) and surviving fraction (SF) decreased dramatically with increasing LET at the same doses. In addition, malformed shoots, including curliness, carnification, nicks and chlorophyll deficiency, occurred in both carbon ion beam and X-ray irradiations. The induction frequency with the former, however, was far more than that with the X-rays. Conclusions: This work demonstrated the LET dependence of the relative biological effectiveness (RBE) of tissue culture of Saintpaulia ionahta according to 50% FWI and 50% SF. After irradiating leaf explants with 5 Gy of a 221 MeV carbon ion beam having a LET value of 96 keV/mu m throughout the sample, a chlorophyll-deficient (CD) mutant, which could transmit the character of chlorophyll deficiency to its progeny through three continuous tissue culture cycles, and plantlets with other malformations were obtained.

Survivin expressions in human hepatoma HepG2 cells exposed to ionizing radiation of different LET

Survivin is a member of the inhibitors of apoptosis (IAP) protein family that interferes with post-mitochondrial events including activation of caspases. To examine the regulation of survivin expression in response to irradiation with different linear energy transfer (LET), human hepatoma HepG2 cells were irradiated in vitro with X-rays and carbon ions. Cellular sensitivities to low- and high-LET radiation were determined by colony formation. Survivin expression at mRNA and protein level were measured with RT-PCR and Western blot analyses, respectively. Radiation-induced cell cycle arrest and apoptosis were investigated with flow cytometry. We found that low-LET X-rays induced dose-dependent increases in survivin expression. After exposure to high-LET carbon ions, survivin expression gradually increased from 0 to 4 Gy, and then declined at 6 Gy. More pronounced survivin expression, stronger G(2)/M phase arrest was observed after exposure to carbon ions in comparison with X-rays at doses from 0 to 4 Gy. These observations indicate that there is a differential survivin expression in response to different LET radiations and the cycle arrest mechanism may be associated with it. In addition, our data on induction of apoptosis are compatible with the assumption that survivin expression induced by low-LET X-rays radiation may play a critical role in inhibiting apoptosis. However, after irradiation with ions, an anti-apoptotic function of survivin is not evident, possibly because of the serious damage produced by densely ionizing radiation.

The influence of fractionation on cell survival and premature differentiation after carbon ion irradiation

Carbon ion radiotherapy/Fractionated irradiation/R-BE/Premature terminal differentiation. To investigate the influence of fractionation on cell survival and radiation induced premature differentiation as markers for early and late effects after X-rays and carbon irradiation. Normal human fibroblasts NHDF, AG1522B and WI-38 were irradiated With 250 kV X-rays, or 266 MeV/u, 195 MeV/u and I I MeV/u carbon ions. Cytotoxicity was measured by a clonogenic survival assay or by determination of the differentiation pattern. Experiments with high-energy carbon ions show that fractionation induced repair effects are similar to photon irradiation. The RBE10 values for clonogenic survival are 1.3 and 1.6 for irradiation in one or two fractions for NHDF cells and around 1.2 for AG1522B cells regardless of the fractionation scheme. The RBE for a doubling of post mitotic fibroblasts (PMF) in the population is I for both single and two fractionated irradiation of NHDF cells. Using I I MeV/u carbon ions, no repair effect can be seen in WI-38 cells. The RBE10 for clonogenic survival is 3.2 for single irradiation and 4.9 for two fractionated irradiations. The RBE for a doubling of PMF is 3.1 and 5.0 for single and two fractionated irradiations, respectively. For both cell lines the effects of high-energy carbon ions representing the irradiation of the skin and the normal tissue in the entrance channel are similar to the effects of X-rays. The fractionation effects are maintained. For the lower energy, which is representative for the irradiation of the tumor region. RBE is enhanced for clonogenic survival as well as for premature terminal differentiation. Fractionation effects are not detectable. Consequently, the therapeutic ratio is significantly enhanced by fractionated irradiation with carbon ions.

The experimental measurement of the relative biological effectiveness of carbon ions with different qualities

The relative biological effectiveness (RBE) of carbon ions with linear energy transfer (LET) of 172 keV/mu m and 13.7 keV/mu m were determined in this study. The clonogenic survival and premature terminal differentiation were measured on normal human. broblasts AG01522C and NHDF after exposure of the cells to 250 kV X-rays and carbon ions with different qualities. RBE was determined for these two biological end points. The results showed that the measured RBE10 with a survival fraction of 10% was 3.2 for LET 172 keV/mu m, and 1.33 for LET 13.7 keV/mu m carbon ions. RBE for a doubling of post-mitotic. broblasts (PMF) in the population was 2.8 for LET 172 keV/mu m, and 1 for LET 13.7 keV/mu m carbon ions. For the carbon ion therapy, a high RBE value on the Bragg peak results in a high biological dose on the tumour. The tumour cells can be killed effectively. At the same time, the dose on healthy tissue would be reduced accordingly. This will lighten the late effect such as fibrosis on normal tissue.

Early effects of carbon-ion irradiation on murine lymphocytes and thymocytes

To estimate the biological risks from space radiation encountered by cosmonauts in outer space, the effects from whole-body exposure to carbon ions or X-rays irradiations at 0, 0.39, 0.55 and 1 Gy at a dose rate of 0.2 Gy/min were investigated in BALB/c mice. The relative thymus and spleen weights were measured at 24 h after exposure, and the cell cycle distribution and percentage of apoptosis of thymocytes and spleen and peripheral blood lymphocytes were determined by flow cytometry. The data showed that exposure to carbon ions delayed cell progression of peripheral blood lymphocytes in S-phase, and delayed thymocytes and spleen lymphocytes in G(0)/G(1)-phase. Apoptosis of thymocytes and peripheral blood lymphocytes induced by carbon ions increased more rapidly with dose than was the case for X-rays. There were some differences between the relative weight loss of the thymus and the spleen with increasing dose of either carbon ions or X-rays. The results obtained provide evidence of dose- and organ-specific differences induced by carbon ions compared to X-rays, with increased apoptosis in peripheral blood lymphocytes and thymocytes, but not spleen lymphocytes. Our data may suggest that further work would be of interest to estimate risk of changes in immune function during particle radiation exposures in space travel. (c) 2007 COSPAR

Chromatid-type aberrations following irradiation in G0 lymphocytes with heavy ions

We describe a low level of chromatid-type aberrations which included the relatively rare isochromatid/chromatid triradial in peripheral blood lymphocytes that were irradiated, ostensibly in GO, with accelerated heavy C-12 ions. These were produced only at the energies of 69 MeV/n (34.6 keV/ mu m), almost absent at the energy of either 58.6 McV/n (46.07 keV/mu m) or 19.3 MeV/n (97 keV/mu m), nor were they found after low-LET X-rays. Mechanisms potentially responsible for their formation are discussed.

Protective effects of melatonin against low- and high-LET irradiation

To investigate the protective effects of melatonin against high-LET ionizing radiation, V79 Chinese hamster cells were irradiated with 100 keV/mu m carbon beam. Parallel experiments were performed with 200 kV X-rays. To avoid the impact from extra solvents, melatonin was dissolved directly in culture medium. Cells were cultured in melatonin medium for 1 hr before irradiation. Cell inactivation was measured with conventional colony forming assay, medium containing 6-thioguanine was used for the selection of mutants at hprt locus, and the cell cycle was monitored by flow cytometry. Both carbon beam and X-rays induced cell inactivation, hprt gene mutation and cell cycle G2 block dose-dependently. But carbon beam showed stronger effects as indicated by all three endpoints and the relative biological effectiveness (RBE) was 3.5 for cell killing (at 10% survival level) and 2.9 for mutation induction (at 5 x 10(-5) mutants/ cell level). Melatonin showed protective effects against ionizing radiation in a dose-dependent manner. In terms of cell killing, melatonin only increased the survival level of those samples exposed to 8Gy or larger of X-rays or 6 Gy or larger of carbon beam. In the induction of hprt mutation and G2 block, melatonin reduced such effects induced by carbon beam but not by X-rays. The results suggest that melatonin reduces the direct interaction of particles with cells rather than an indirect interaction. Further studies are required to disclose the underlying mechanisms.

Image reconstruction by an alternating minimisation

This paper focuses on the problem of incomplete data in the applications of the circular cone-beam computed tomography. This problem is frequently encountered in medical imaging sciences and some other industrial imaging systems. For example, it is crucial when the high density region of objects can only be penetrated by X-rays in a limited angular range. As the projection data are only available in an angular range, the above mentioned incomplete data problem can be attributed to the limited angle problem, which is an ill-posed inverse problem. This paper reports a modified total variation minimisation method to reduce the data insufficiency in tomographic imaging. This proposed method is robust and efficient in the task of reconstruction by showing the convergence of the alternating minimisation method. The results demonstrate that this new reconstruction method brings reasonable performance. (C) 2010 Elsevier B.V. All rights reserved.

High strength Ni based composite reinforced by solid solution W(Al) obtained by powder metallurgy

The solid-solution-particle reinforced W(Al)-Ni composites were successfully fabricated by using mechanical alloying (MA) and hot-pressing (HP) technique when the content of Ni is between 45 wt% and 55 wt%. Besides, samples of various original component ratio of Al50W50 to Ni have been fabricated, and the corresponding microcomponents and mechanical properties such as microhardness, ultimate tensile strength and elongation were characterized and discussed. The optimum ultimate tensile strength under the experiment conditions is 1868 MPa with elongation of 10.21 % and hardness of 6.62 GPa. X-ray diffraction (XRD), FE-SEM and energy dispersive analysis of X-rays (EDS) were given to analysis the components and morphology of the composite bulk specimens.

Restauração directa de classes II em pré-molares com compósitos

Tese de Doutoramento apresentada à Universidade Fernando Pessoa como parte dos requisitos para obtenção do grau de Doutor em em Biotecnologia e Saúde, Epidemiologia e Saúde Pública.

Midlatitude daytime D region ionosphere variations measured from radio atmospherics

We measured the midlatitude daytime ionospheric D region electron density profile height variations in July and August 2005 near Duke University by using radio atmospherics (or sferics for short), which are the high-power, broadband very low frequency (VLF) signals launched by lightning discharges. As expected, the measured daytime D region electron density profile heights showed temporal variations quantitatively correlated with solar zenith angle changes. In the midlatitude geographical regions near Duke University, the observed quiet time heights decreased from ∼80 km near sunrise to ∼71 km near noon when the solar zenith angle was minimum. The measured height quantitative dependence on the solar zenith angle was slightly different from the low-latitude measurement given in a previous work. We also observed unexpected spatial variations not linked to the solar zenith angle on some days, with 15% of days exhibiting regional differences larger than 0.5 km. In these 2 months, 14 days had sudden height drops caused by solar flare X-rays, with a minimum height of 63.4 km observed. The induced height change during a solar flare event was approximately proportional to the logarithm of the X-ray flux. In the long waveband (wavelength, 1-8 Å), an increase in flux by a factor of 10 resulted in 6.3 km decrease of the height at the flux peak time, nearly a perfect agreement with the previous measurement. During the rising and decaying phases of the solar flare, the height changes correlated more consistently with the short, rather than the long, wavelength X-ray flux changes. © 2010 by the American Geophysical Union.

Fractures in myelomeningocele.

BACKGROUND: In patients with myelomeningocele (MMC), a high number of fractures occur in the paralyzed extremities, affecting mobility and independence. The aims of this retrospective cross-sectional study are to determine the frequency of fractures in our patient cohort and to identify trends and risk factors relevant for such fractures. MATERIALS AND METHODS: Between March 1988 and June 2005, 862 patients with MMC were treated at our hospital. The medical records, surgery reports, and X-rays from these patients were evaluated. RESULTS: During the study period, 11% of the patients (n = 92) suffered one or more fractures. Risk analysis showed that patients with MMC and thoracic-level paralysis had a sixfold higher risk of fracture compared with those with sacral-level paralysis. Femoral-neck z-scores measured by dual-energy X-ray absorptiometry (DEXA) differed significantly according to the level of neurological impairment, with lower z-scores in children with a higher level of lesion. Furthermore, the rate of epiphyseal separation increased noticeably after cast immobilization. Mainly patients who could walk relatively well were affected. CONCLUSIONS: Patients with thoracic-level paralysis represent a group with high fracture risk. According to these results, fracture and epiphyseal injury in patients with MMC should be treated by plaster immobilization. The duration of immobilization should be kept to a minimum (<4 weeks) because of increased risk of secondary fractures. Alternatively, patients with refractures can be treated by surgery, when nonoperative treatment has failed.