SET protein accumulates in HNSCC and contributes to cell survival: Antioxidant defense, Akt phosphorylation and AVOs acidification

Objectives: Determination of the SET protein levels in head and neck squamous cell carcinoma (HNSCC) tissue samples and the SET role in cell survival and response to oxidative stress in HNSCC cell lineages. Materials and Methods: SET protein was analyzed in 372 HNSCC tissue samples by immunohistochemistry using tissue microarray and HNSCC cell lineages. Oxidative stress was induced with the pro-oxidant tert-butylhydroperoxide (50 and 250 mu M) in the HNSCC HN13 cell lineage either with (siSET) or without (siNC) SET knockdown. Cell viability was evaluated by trypan blue exclusion and annexin V/propidium iodide assays. It was assessed caspase-3 and -9, PARP-1, DNA fragmentation, NM23-H1, SET, Akt and phosphorylated Akt (p-Akt) status. Acidic vesicular organelles (AVOs) were assessed by the acridine orange assay. Glutathione levels and transcripts of antioxidant genes were assayed by fluorometry and real time PCR, respectively. Results: SET levels were up-regulated in 97% tumor tissue samples and in HNSCC cell lineages. SiSET in HN13 cells (i) promoted cell death but did not induced caspases, PARP-1 cleavage or DNA fragmentation, and (ii) decreased resistance to death induced by oxidative stress, indicating SET involvement through caspase-independent mechanism. The red fluorescence induced by siSET in HN13 cells in the acridine orange assay suggests SET-dependent prevention of AVOs acidification. NM23-H1 protein was restricted to the cytoplasm of siSET/siNC HN13 cells under oxidative stress, in association with decrease of cleaved SET levels. In the presence of oxidative stress, siNC HN13 cells showed lower GSH antioxidant defense (GSH/GSSG ratio) but higher expression of the antioxidant genes PRDX6, SOD2 and TXN compared to siSET HN13 cells. Still under oxidative stress, p-Akt levels were increased in siNC HN13 cells but not in siSET HN13, indicating its involvement in HN13 cell survival. Similar results for the main SET effects were observed in HN12 and CAL 27 cell lineages, except that HN13 cells were more resistant to death. Conclusion: SET is potential (i) marker for HNSCC associated with cancer cell resistance and (ii) new target in cancer therapy. (C) 2012 Elsevier Ltd. All rights reserved.

A new evolutionary clustering search for a no-wait flow shop problem with set-up times

This paper addresses the m-machine no-wait flow shop problem where the set-up time of a job is separated from its processing time. The performance measure considered is the total flowtime. A new hybrid metaheuristic Genetic Algorithm-Cluster Search is proposed to solve the scheduling problem. The performance of the proposed method is evaluated and the results are compared with the best method reported in the literature. Experimental tests show superiority of the new method for the test problems set, regarding the solution quality. (c) 2012 Elsevier Ltd. All rights reserved.

SET overexpression decreases cell detoxification efficiency: ALDH2 and GSTP1 are downregulated, DDR is impaired and DNA damage accumulates

Alcohol and tobacco consumption are risk factors for head and neck squamous cell carcinoma (HNSCC). Aldehyde dehydrogenase 2 (ALDH2) and glutathione Stransferase pi 1 (GSTP1) are important enzymes for cellular detoxification and low efficiencies are implicated in cancer. We assessed the potential role of SET protein overexpression, a histone acetylation modulator accumulated in HNSCC, in gene regulation and protein activity of ALDH2 and GSTP1. SET was knocked down in HN13, HN12 and Cal27, and overexpressed in HEK293 cells; ethanol and cisplatin were the chemical agents. Cells with SET overexpression (HEK293/SET, HN13 and HN12) showed lower ALDH2 and GSTP1 mRNA levels and trichostatin A increased them (real-time PCR). Ethanol upregulated GSTP1 and ALDH2 mRNAs, whereas cisplatin upregulated GSTP1 in HEK293 cells. SET-chromatin binding revealed SET interaction with ALDH2 and GSTP1 promoters, specifically via SET NAP domain; ethanol and cisplatin abolished SET binding. ALDH2 and GSTP1 efficiency was assessed by enzymatic and comet assay. A lower ALDH2 activity was associated with greater DNA damage (tail intensity) in HEK293/SET compared with HEK293 cells, whereas HN13/siSET showed ALDH2 activity higher than HN13 cells. HN13/siSET cells showed increased tail intensity. Cisplatin-induced DNA damage response showed negative relationship between SET overexpression and BRCA2 recruitment. SET downregulated repair genes ATM, BRCA1 and CHEK2 and upregulated TP53. Cisplatin-induced cell-cycle arrest occurred in G0/G1 and S in HEK293 cells, whereas HEK293/SET showed G2/M stalling. Overall, cisplatin was more cytotoxic for HN13 than HN13/siSET cells. Our data suggest a role for SET in cellular detoxification, DNA damage response and genome integrity.

Tidal perturbations of the rotation of the moon: the creep tide approach

In this communication we report results from the application to the study of the rotation of the Moon of the creeping tide theory just proposed (Ferraz-Mello, Cel. Mech. Dyn. Astron., submitted. ArXiv astro-ph 1204.3957). The choice of the Moon for the first application of this new theory is motivated by the fact that the Moon is one of the best observed celestial bodies and the comparison of the theoretical predictions of the theory with observations i may validate the theory or point out the need of further improvements. Particularly, the tidal perturbations of the rotation of the Moon - the physical libration of the Moon - have been detected in the Lunar Laser Ranging measurements (Williams et al. JGR 106, 27933, 2001). The major difficulty in this application comes from the fact that tidal torques in a planet-satellite system are very sensitive to the distance between the two-bodies, which is strongly affected by Solar perturbations. In the case of the Moon, the main solar perturbations - the Evection and the Variation - are more important than most of the Keplerian oscillations, being smaller only than the first Keplerian harmonic (equation of the centre). Besides, two of the three components of the Moon's libration in longitude whose tidal contributions were determined by LLR are related to these perturbations. The results may allow us to determine the main parameter of a possible Moon's creeping tide. The preliminary results point to a relaxation factor (gamma) 2 to 4 times smaller than the one predicted from the often cited values of thr Moon's quality factor Q (between 30 and 40), and points to larger Q values.

Regularizing a set of unstructured 3D points from a sequence of stereo images

[EN] In this paper we present a method for the regularization of a set of unstructured 3D points obtained from a sequence of stereo images. This method takes into account the information supplied by the disparity maps computed between pairs of images to constraint the regularization of the set of 3D points. We propose a model based on an energy which is composed of two terms: an attachment term that minimizes the distance from 3D points to the projective lines of camera points, and a second term that allows for the regularization of the set of 3D points by preserving discontinuities presented on the disparity maps. We embed this energy in a 2D finite element method. After minimizing, this method results in a large system of equations that can be optimized for fast computations. We derive an efficient implicit numerical scheme which reduces the number of calculations and memory allocations.

Multiproperty modeling for a set of binary systems. Evaluation of a model to correlate simultaneously several mixing properties of methyl ethanoate+alkanes and new experimental data

[EN] This work makes a theoretical–experimental contribution to the study of ester and alkane solutions. Experimental data of isobaric vapor–liquid equilibria (VLE) are presented at 101.3 kPa for binary systems of methyl ethanoate with six alkanes (from C5 to C10), and of volumes and mixing enthalpies, vE and hE.

Viola-Jones based detectors: How much affects the training set?

[EN]This paper presents a study on the facial feature detection performance achieved using the Viola-Jones framework. A set of classi- ers using two di erent focuses to gather the training samples is created and tested on four di erent datasets covering a wide range of possibili- ties. The results achieved should serve researchers to choose the classi er that better ts their demands.

'We deliberately set high demands for the gold standard'… Inclusive and exclusive pronouns in two social science cultures

[EN] This study analyses the use of inclusive and exclusive pronominal signals in English and Spanish research articles and investigates whether there are differences between the two languages in terms of pronominal signals frequency and usage.

A comprehensive study of the 26th December 2004 Sumatra earthquake: possible implications on Earth rotation and investigations on the coseismic and postseismic stress diffusion associated with the seismic rupture

In this work a multidisciplinary study of the December 26th, 2004 Sumatra earthquake has been carried out. We have investigated both the effect of the earthquake on the Earth rotation and the stress field variations associated with the seismic event. In the first part of the work we have quantified the effects of a water mass redistribution associated with the propagation of a tsunami wave on the Earth’s pole path and on the length-of-day (LOD) and applied our modeling results to the tsunami following the 2004 giant Sumatra earthquake. We compared the result of our simulations on the instantaneous rotational axis variations with some preliminary instrumental evidences on the pole path perturbation (which has not been confirmed yet) registered just after the occurrence of the earthquake, which showed a step-like discontinuity that cannot be attributed to the effect of a seismic dislocation. Our results show that the perturbation induced by the tsunami on the instantaneous rotational pole is characterized by a step-like discontinuity, which is compatible with the observations but its magnitude turns out to be almost one hundred times smaller than the detected one. The LOD variation induced by the water mass redistribution turns out to be not significant because the total effect is smaller than current measurements uncertainties. In the second part of this work of thesis we modeled the coseismic and postseismic stress evolution following the Sumatra earthquake. By means of a semi-analytical, viscoelastic, spherical model of global postseismic deformation and a numerical finite-element approach, we performed an analysis of the stress diffusion following the earthquake in the near and far field of the mainshock source. We evaluated the stress changes due to the Sumatra earthquake by projecting the Coulomb stress over the sequence of aftershocks taken from various catalogues in a time window spanning about two years and finally analyzed the spatio-temporal pattern. The analysis performed with the semi-analytical and the finite-element modeling gives a complex picture of the stress diffusion, in the area under study, after the Sumatra earthquake. We believe that the results obtained with the analytical method suffer heavily for the restrictions imposed, on the hypocentral depths of the aftershocks, in order to obtain the convergence of the harmonic series of the stress components. On the contrary we imposed no constraints on the numerical method so we expect that the results obtained give a more realistic description of the stress variations pattern.

A study on set variable representations in constraint programming

Il lavoro presentato in questa tesi si colloca nel contesto della programmazione con vincoli, un paradigma per modellare e risolvere problemi di ricerca combinatoria che richiedono di trovare soluzioni in presenza di vincoli. Una vasta parte di questi problemi trova naturale formulazione attraverso il linguaggio delle variabili insiemistiche. Dal momento che il dominio di tali variabili può essere esponenziale nel numero di elementi, una rappresentazione esplicita è spesso non praticabile. Recenti studi si sono quindi focalizzati nel trovare modi efficienti per rappresentare tali variabili. Pertanto si è soliti rappresentare questi domini mediante l'uso di approssimazioni definite tramite intervalli (d'ora in poi rappresentazioni), specificati da un limite inferiore e un limite superiore secondo un'appropriata relazione d'ordine. La recente evoluzione della ricerca sulla programmazione con vincoli sugli insiemi ha chiaramente indicato che la combinazione di diverse rappresentazioni permette di raggiungere prestazioni di ordini di grandezza superiori rispetto alle tradizionali tecniche di codifica. Numerose proposte sono state fatte volgendosi in questa direzione. Questi lavori si differenziano su come è mantenuta la coerenza tra le diverse rappresentazioni e su come i vincoli vengono propagati al fine di ridurre lo spazio di ricerca. Sfortunatamente non esiste alcun strumento formale per paragonare queste combinazioni. Il principale obiettivo di questo lavoro è quello di fornire tale strumento, nel quale definiamo precisamente la nozione di combinazione di rappresentazioni facendo emergere gli aspetti comuni che hanno caratterizzato i lavori precedenti. In particolare identifichiamo due tipi possibili di combinazioni, una forte ed una debole, definendo le nozioni di coerenza agli estremi sui vincoli e sincronizzazione tra rappresentazioni. Il nostro studio propone alcune interessanti intuizioni sulle combinazioni esistenti, evidenziandone i limiti e svelando alcune sorprese. Inoltre forniamo un'analisi di complessità della sincronizzazione tra minlex, una rappresentazione in grado di propagare in maniera ottimale vincoli lessicografici, e le principali rappresentazioni esistenti.

Elf magnetic field influence on ION Channels studied by Patch Clamp Technique: exposure set up and "Whole Cell" measurements on Potassium currents

The aim of this thesis was to study the effects of extremely low frequency (ELF) electromagnetic magnetic fields on potassium currents in neural cell lines ( Neuroblastoma SK-N-BE ), using the whole-cell Patch Clamp technique. Such technique is a sophisticated tool capable to investigate the electrophysiological activity at a single cell, and even at single channel level. The total potassium ion currents through the cell membrane was measured while exposing the cells to a combination of static (DC) and alternate (AC) magnetic fields according to the prediction of the so-called ‘ Ion Resonance Hypothesis ’. For this purpose we have designed and fabricated a magnetic field exposure system reaching a good compromise between magnetic field homogeneity and accessibility to the biological sample under the microscope. The magnetic field exposure system consists of three large orthogonal pairs of square coils surrounding the patch clamp set up and connected to the signal generation unit, able to generate different combinations of static and/or alternate magnetic fields. Such system was characterized in term of field distribution and uniformity through computation and direct field measurements. No statistically significant changes in the potassium ion currents through cell membrane were reveled when the cells were exposed to AC/DC magnetic field combination according to the afore mentioned ‘Ion Resonance Hypothesis’.

Sistemi liquido cristallini complessi: simulazioni al calcolatore e studi ESR

The aim of this PhD thesis was to study at a microscopic level different liquid crystal (LC) systems, in order to determine their physical properties, resorting to two distinct methodologies, one involving computer simulations, and the other spectroscopic techniques, in particular electron spin resonance (ESR) spectroscopy. By means of the computer simulation approach we tried to demonstrate this tool effectiveness for calculating anisotropic static properties of a LC material, as well as for predicting its behaviour and features. This required the development and adoption of suitable molecular models based on a convenient intermolecular potentials reflecting the essential molecular features of the investigated system. In particular, concerning the simulation approach, we have set up models for discotic liquid crystal dimers and we have studied, by means of Monte Carlo simulations, their phase behaviour and self­-assembling properties, with respect to the simple monomer case. Each discotic dimer is described by two oblate Gay­Berne ellipsoids connected by a flexible spacer, modelled by a harmonic "spring" of three different lengths. In particular we investigated the effects of dimerization on the transition temperatures, as well as on the characteristics of molecular aggregation displayed and the relative orientational order. Moving to the experimental results, among the many experimental techniques that are typically employed to evaluate LC system distinctive features, ESR has proved to be a powerful tool in microscopic scale investigation of the properties, structure, order and dynamics of these materials. We have taken advantage of the high sensitivity of the ESR spin probe technique to investigate increasingly complex LC systems ranging from devices constituted by a polymer matrix in which LC molecules are confined in shape of nano- droplets, as well as biaxial liquid crystalline elastomers, and dimers whose monomeric units or lateral groups are constituted by rod-like mesogens (11BCB). Reflection-mode holographic-polymer dispersed liquid crystals (H-PDLCs) are devices in which LCs are confined into nanosized (50­-300 nm) droplets, arranged in layers which alternate with polymer layers, forming a diffraction grating. We have determined the configuration of the LC local director and we have derived a model of the nanodroplet organization inside the layers. Resorting also to additional information on the nanodroplet size and shape distribution provided by SEM images of the H-PDLC cross-section, the observed director configuration has been modeled as a bidimensional distribution of elongated nanodroplets whose long axis is, on the average, parallel to the layers and whose internal director configuration is a uniaxial quasi- monodomain aligned along the nanodroplet long axis. The results suggest that the molecular organization is dictated mainly by the confinement, explaining, at least in part, the need for switching voltages significantly higher and the observed faster turn-off times in H-PDLCs compared to standard PDLC devices. Liquid crystal elastomers consist in cross-linked polymers, in which mesogens represent the monomers constituting the main chain or the laterally attached side groups. They bring together three important aspects: orientational order in amorphous soft materials, responsive molecular shape and quenched topological constraints. In biaxial nematic liquid crystalline elastomers (BLCEs), two orthogonal directions, rather than the one of normal uniaxial nematic, can be controlled, greatly enhancing their potential value for applications as novel actuators. Two versions of a side-chain BLCEs were characterized: side­-on and end­-on. Many tests have been carried out on both types of LCE, the main features detected being the lack of a significant dynamical behaviour, together with a strong permanent alignment along the principal director, and the confirmation of the transition temperatures already determined by DSC measurements. The end­-on sample demonstrates a less hindered rotation of the side group mesogenic units and a greater freedom of alignment to the magnetic field, as already shown by previous NMR studies. Biaxial nematic ESR static spectra were also obtained on the basis of Molecular Dynamics generated biaxial configurations, to be compared to the experimentally determined ones, as a mean to establish a possible relation between biaxiality and the spectral features. This provides a concrete example of the advantages of combining the computer simulation and spectroscopic approaches. Finally, the dimer α,ω-bis(4'-cyanobiphenyl-4-yl)undecane (11BCB), synthesized in the "quest" for the biaxial nematic phase has been analysed. Its importance lies in the dimer significance as building blocks in the development of new materials to be employed in innovative technological applications, such as faster switching displays, resorting to the easier aligning ability of the secondary director in biaxial phases. A preliminary series of tests were performed revealing the population of mesogenic molecules as divided into two groups: one of elongated straightened conformers sharing a common director, and one of bent molecules, which display no order, being equally distributed in the three dimensions. Employing this model, the calculated values show a consistent trend, confirming at the same time the transition temperatures indicated by the DSC measurements, together with rotational diffusion tensor values that follow closely those of the constituting monomer 5CB.

Messung der paritätsverletzenden Neutronenspin-Rotation in 139 La unter Einsatz von 3 He-Neutronenspin-Filtern

Die Machbarkeit der Detektion einer P,T-Verletzung in der Neutron-Kern-p-Wellenresonanz von La-139 wird untersucht. Als experimentelleVerbesserung wurde der He-3- Neutronenspin- Filter (He-3-NSF) getestet. Mit einem He-3-NSF als Polarisationsanalysator wurde die rein P-verletzende Neutronenspin-Rotation in 139La erstmals mit guter Genauigkeit unmittelbar in der Resonanz bestimmt. Das schwache Matrixelement betrug v=1.51(5)stat(7)syst meV. Es wurde ein adiabatischer He-3-Spin-Rotator entwickelt und erfolgreich getestet, welcher es erlaubt, die Richtung der He-3-Polarisation einzustellen. Dies kann die bei Polarisationsmessungen üblichen Spinpräzessionsspulen ersetzen. Das Problem der Unterdrückung apparativer Falscheffekte während einer P,T-Detektion wird diskutiert und konnte jedoch noch nicht gelöst werden.