Positional stability and radial dynamics of sonoluminescent bubbles under bi-harmonic driving: Effect of the high-frequency component and its relative phase.

The use of bi-frequency driving in sonoluminescence has proved to be an effective way to avoid the spatial instability (pseudo-orbits) developed by bubbles in systems with high viscous liquids like sulfuric or phosphoric acids. In this work, we present extensive experimental and numerical evidence in order to assess the effect of the high frequency component (PAc(HF)) of a bi-harmonic acoustic pressure field on the dynamic of sonoluminescent bubbles in an aqueous solution of sulfuric acid. The present study is mainly focused on the role of the harmonic frequency (Nf0) and the relative phase between the two frequency components (φb) of the acoustic field on the spatial, positional and diffusive stability of the bubbles. The results presented in this work were analyzed by means of three different approaches. First, we discussed some qualitative considerations about the changes observed in the radial dynamics, and the stability of similar bubbles under distinct bi-harmonic drivings. Later, we have investigated, through a series of numerical simulations, how the use of high frequency harmonic components of different order N, affects the positional stability of the SL bubbles. Furthermore, the influence of φb in their radius temporal evolution is systematically explored for harmonics ranging from the second to the fifteenth harmonic (N=2-15). Finally, a multivariate analysis based on the covariance method is performed to study the dependences among the parameters characterizing the SL bubble. Both experimental and numerical results indicate that the impact of PAc(HF) on the positional instability and the radial dynamics turns to be progressively negligible as the order of the high frequency harmonic component grows (i.e. N ≫ 1), however its effectiveness on the reduction of the spatial instability remains unaltered or even improved.

Stable tridimensional bubble clusters in multi-bubble sonoluminescence (MBSL).

In the present work, stable clusters made of multiple sonoluminescent bubbles are experimentally and theoretically studied. Argon bubbles were acoustically generated and trapped using bi-frequency driving within a cylindrical chamber filled with a sulfuric acid aqueous solution (SA85w/w). The intensity of the acoustic pressure field was strong enough to sustain, during several minutes, a large number of positionally and spatially fixed (without pseudo-orbits) sonoluminescent bubbles over an ellipsoidally-shaped tridimensional array. The dimensions of the ellipsoids were studied as a function of the amplitude of the applied low-frequency acoustic pressure (PAc(LF)) and the static pressure in the fluid (P0). In order to explain the size and shape of the bubble clusters, we performed a series of numerical simulations of the hydrodynamic forces acting over the bubbles. In both cases the observed experimental behavior was in excellent agreement with the numerical results. The simulations revealed that the positionally stable region, mainly determined by the null primary Bjerknes force (F→Bj), is defined as the outer perimeter of an axisymmetric ellipsoidal cluster centered in the acoustic field antinode. The role of the high-frequency component of the pressure field and the influence of the secondary Bjerknes force are discussed. We also investigate the effect of a change in the concentration of dissolved gas on the positional and spatial instabilities through the cluster dimensions. The experimental and numerical results presented in this paper are potentially useful for further understanding and modeling numerous current research topics regarding multi-bubble phenomena, e.g. forces acting on the bubbles in multi-frequency acoustic fields, transient acoustic cavitation, bubble interactions, structure formation processes, atomic and molecular emissions of equal bubbles and nonlinear or unsteady acoustic pressure fields in bubbly media.

Deposition in St. Mark's Basilica of Venice.

Atmospheric pollutants may cause damage to monuments and historical buildings. Besides air contaminants, soluble salts are also responsible for stone deterioration and decay in outdoor and indoor monuments. The problem of how to conserve works of arts thus requires a deep knowledge of contaminants' concentration and distribution inside buildings. In this work, water-soluble ions inside St. Mark's Basilica in Venice were studied, with the aim of understanding their principal source and distribution inside the building. With the aid of Fourier transform infrared spectroscopy and scanning electron microscopy analysis, the interaction between ions and surface's material was also investigated. Ion chromatographic analysis of depositions highlighted a large amount of "deteriorating agents" such as sulphates and chlorides. A possible source in the innermost area of the basilica has been found for formates and nitrates. On the contrary, a decrease of chloride, from the entrance to the innermost area, has been found, which indicates that the source is outside the building. It is emphasized that different contaminants behave differently on different material, and the effect of pollution inside churches and monuments is not easy to predict. Wood and brick seem to react differently than stone and mortar to the damaging action of salts and pollutants. The present work should be considered a useful tool for the future preservation of St. Mark's Basilica in Venice.

A large-scale (19)F MRI-based cell migration assay to optimize cell therapy.

Adoptive transfer of cells for therapeutic purposes requires efficient and precise delivery to the target organ whilst preserving cell function. Therefore, therapeutically applied cells need to migrate and integrate within their target tissues after delivery, e.g. dendritic cells (DCs) need to migrate to lymph nodes to elicit an antigen-specific immune response. Previous studies have shown that inappropriate cell delivery can hinder DC migration and result in insufficient immune induction. As migration can be extremely difficult to study quantitatively in vivo, we propose an in vitro assay that reproduces key in vivo conditions to optimize cell delivery and migration in vivo. Using DC migration along a chemokine gradient, we describe here a novel (19)F MR-based, large-scale, quantitative assay to measure cell migration in a three-dimensional collagen scaffold. Unlike conventional migration assays, this set-up is amenable to both large and small cell numbers, as well as opaque tissue samples and the inclusion of chemokines or other factors. We labeled primary human DCs with a (19)F label suitable for clinical use; (0.5-15) × 10(6) cells in the scaffolds were imaged sequentially, and migration was assessed using two independent methods. We found no migration with larger numbers of cells, but up to 3% with less than one million cells. Hence, we show that the cell density in cell bolus injections has a decisive impact on migration, and this may explain the limited migration observed using large cell numbers in the clinic.

Impedance analysis of cultured cells: a mean-field electrical response model for electric cell-substrate impedance sensing technique.

In this paper we present a model to describe the electrical properties of a confluent cell monolayer cultured on gold microelectrodes to be used with electric cell-substrate impedance sensing technique. This model was developed from microscopic considerations (distributed effects), and by assuming that the monolayer is an element with mean electrical characteristics (specific lumped parameters). No assumptions were made about cell morphology. The model has only three adjustable parameters. This model and other models currently used for data analysis are compared with data we obtained from electrical measurements of confluent monolayers of Madin-Darby Canine Kidney cells. One important parameter is the cell-substrate height and we found that estimates of this magnitude strongly differ depending on the model used for the analysis. We analyze the origin of the discrepancies, concluding that the estimates from the different models can be considered as limits for the true value of the cell-substrate height.

Proton field-cycling nuclear magnetic resonance relaxometry in the smectic A mesophase of thermotropic cyanobiphenyls: Effects of sonication.

Proton field-cycling nuclear magnetic resonance relaxometry is used to study the spin-lattice relaxation dispersion of selected standard smectic A liquid crystals at different temperatures. Relaxation features at both, in the presence and absence of a monochromatic ultrasonic field are considered. We show that the laboratory-frame spin-lattice relaxation time is mainly governed by translational diffusion. Order director fluctuations (ODF) are less important while rotational diffusion seems to be only relevant near the clearing point. Our study suggests that sonication enhances the ODF contribution in the SmA mesophase. Within the framework of the approach we have outlined, different features associated with the ODF mechanism can be investigated.

Spin-lattice dispersion in nematic and smectic-A mesophases in the presence of ultrasonic waves: a theoretical approach.

We present a theoretical study of the Larmor frequency dependence of the nuclear spin-lattice relaxation caused by order director fluctuations for both nematic and smectic-A mesophases. The analysis is focused on the case where the molecular system is subjected to sonication during the relaxation process. The departure from the nonsonicated case is discussed for various values of the involved parameters. Two different approaches are discussed for the smectic case.

Apparent low-field spin-lattice dispersion in the smectic-A mesophase of thermotropic cyanobiphenyls.

Proton field-cycling spin-lattice relaxometry T1 of the smectic-A mesophase in cyanobiphenyls revealed the presence of steep dispersions in the low-frequency regime. We clearly show that the strong dispersion characteristic of smectic organizations cannot be attributed to the collective molecular dynamics (order director fluctuations), as it is usually interpreted. We present two independent experimental evidences: the dependence of the dispersion with the slew rate of the magnetic field cycle and the dependence of the dispersion with the presence and power of an ultrasonic field.

Properties of stationary nonequilibrium states in the thermostatted periodic Lorentz gas: the multiparticle system.

We study the stationary nonequilibrium states of N-point particles moving under the influence of an electric field E among fixed obstacles (disk) in a two-dimensional torus. The total kinetic energy of the system is kept constant through a Gaussian thermostat that produces a velocity dependent mean field interaction between the particles. The current and the particle distribution functions are obtained numerically and compared for small /E/ with analytic solutions of a Boltzmann-type equation obtained by treating the collisions with the obstacles as random independent scatterings. The agreement is surprisingly good for both small and large N. The latter system in turn agrees with a self-consistent one-particle evolution expected to hold in the N-->infinity limit.

Thermodynamic entropy production fluctuation in a two-dimensional shear flow model.

We investigate fluctuations in the momentum flux across a surface perpendicular to the velocity gradient in a stationary shear flow maintained by either thermostated deterministic or by stochastic boundary conditions. In the deterministic system the fluctuation relation for the probability of large deviations, which holds for the phase space volume contraction giving the Gibbs ensemble entropy production, never seems to hold for the flux which gives the hydrodynamic entropy production. In the stochastic case the fluctuation relation is found to hold for the total flux, as predicted by various exact results, but not for the flux across part of the surface. The latter appear to satisfy a modified fluctuation relation. Similar results are obtained for the heat flux in a steady state produced by stochastic boundaries at different temperatures.

Retinoblastoma family proteins induce differentiation and regulate B-myb expression in neuroblastoma cells.

BACKGROUND: The expression of several genes is modulated during neuroblastoma differentiation. The retinoblastoma family proteins, pRb, p107 and pRb2/p130, act in the repression of proliferation genes, interacting mainly with the E2F transcription factors. PROCEDURE AND RESULTS: In this study, we found that, in neuroblastoma cell lines, pRb and p107 proteins decreased, undergoing progressive dephosphorylation, whereas pRb2/p130 increased at late stages of differentiation. B-myb expression was down-regulated in association with the up-regulation of pRb2/p130, the major partner of E2F on the E2F site of the B-myb promoter in differentiated cells. Transfection of each of the retinoblastoma family genes in neuroblastoma cells was able to induce neural differentiation, to inhibit 3H-thymidine incorporation, and to down-regulate B-myb promoter activity. CONCLUSIONS: In conclusion, our data suggest a major contribution of retinoblastoma proteins, and especially of pRb2/p130, in B-myb promoter regulation and demonstrate the induction of neural differentiation by p107 and pRb2/p130, suggesting a role of these proteins in triggering differentiation-specific genes.

The retinoblastoma-related Rb2/p130 gene is an effector downstream of AP-2 during neural differentiation.

Rb2/p130, a member of the Retinoblastoma family of growth and tumour suppressor genes, is extensively implicated in the control of cell cycle and differentiation. The minimal promoter region of Rb2/p130 in T98G human glioblastoma cells was identified and its analysis revealed the presence of a KER1 palindromic sequence able to bind the transcription factor AP-2, a regulatory protein that plays a crucial role in ectodermal differentiation. This KER1 site interacted in vitro with AP-2, and AP-2 overexpression increased Rb2/p130 transcription and translation. We also found that rat PC12 pheochromocytoma cells, when induced to differentiate by NGF, displayed an increase of AP-2 protein levels and of Rb2/p130 transcription and protein levels. AP-2-transfected PC12 cells displayed enhanced transcription and translation of Rb2/p130 and of the cdk inhibitor p21(WAF1/CIP1), a gene known to be under the control of AP-2, but unable by itself to elicit PC12 differentiation. Overexpression of either AP-2 or Rb2/p130 elicited per se cell differentiation in the absence of NGF, while coexpression of AP-2B, a negative regulator of AP-2 transcriptional activity, inhibited only AP-2-induced differentiation. Altogether, these results indicate that Rb2/p130 is a critical effector of AP-2 in sustaining ectodermal differentiation.

Interaction between the pRb2/p130 C-terminal domain and the N-terminal portion of cyclin D3.

An association between cyclin D3 and the C-terminal domain of pRb2/p130 was demonstrated using the yeast two-hybrid system. Further analysis restricted the epitope responsible for the binding within the 74 N-terminal amino acids of cyclin D3, independent of the LXCXE amino acid motif present in the D-type cyclin N-terminal region. In a coprecipitation assay in T98G cells, a human glioblastoma cell line, the C-terminal domain of pRb2/p130 was able to interact solely with cyclin D3, while the corresponding portion of pRb interacted with either cyclin D3 or cyclin D1. In T98G cells, endogenous cyclin D3-associated kinase activity showed a clear predisposition to phosphorylate preferentially the C-terminal domain of pRb2/p130, rather than that of pRb. This propensity was also confirmed in LAN-5 human neuroblastoma cells, where phosphorylation of the pRb2/p130 C-terminal domain and expression of cyclin D3 also decreased remarkably in the late neural differentiation stages.

BCR-ABL antisense oligodeoxynucleotide in vitro purging and autologous bone marrow transplantation for patients with chronic myelogenous leukemia in advanced phase.

BCR-ABL antisense oligodeoxynucleotides (ODN) have provided evidence of antileukemia effect when tested in vitro against Philadelphia-positive (Ph-pos) cells and in vivo when injected into leukemic mice. On the basis of the results obtained in vitro at diagnosis, eight patients with chronic myelogenous leukemia (CML) were selected and submitted to autologous bone marrow transplantation (ABMT) with bone marrow (BM) cells purged in vitro with junction-specific (J-sp) BCR-ABL antisense ODN at the time of transformation in accelerated phase or during second chronic phase. Mononuclear BM cells were treated in vitro for 24 or 72 hours with 150 micro/mL of antisense ODN yielding a median recovery of 47.6% mononuclear cells, 48.8% CD34(+) cells, and 20.3% clonogenic cells. After a conditioning regimen including busulphan and etoposide, the reinfused treated cells allowed engraftment and hematologic reconstitution in all patients. Evaluation of the antileukemic effect by standard cytogenetic analysis and fluorescence in situ hybridization showed a complete karyotypic response in two cases and a minimal or no response in the other six. The patient autografted in second chronic phase died in blast crisis 7 months after ABMT; of the seven patients autografted in transformation, three developed blast crisis 21 to 39 months after reinfusion, one died from unrelated BMT complications 30 months after ABMT, and three are in persistent second chronic phase 14 to 26 months after autograft. The low toxicity of the protocol and the hemopoietic reconstitution observed in all patients make this approach feasible; the marked karyotypic response observed in some patients and the duration of the second chronic phase show that ODN-mediated BM purging and autograft is a promising treatment for this high-risk group of CML.

The RB-related gene Rb2/p130 in neuroblastoma differentiation and in B-myb promoter down-regulation.

The retinoblastoma family of nuclear factors is composed of RB, the prototype of the tumour suppressor genes and of the strictly related genes p107 and Rb2/p130. The three genes code for proteins, namely pRb, p107 and pRb2/p130, that share similar structures and functions. These proteins are expressed, often simultaneously, in many cell types and are involved in the regulation of proliferation and differentiation. We determined the expression and the phosphorylation of the RB family gene products during the DMSO-induced differentiation of the N1E-115 murine neuroblastoma cells. In this system, pRb2/p130 was strongly up-regulated during mid-late differentiation stages, while, on the contrary, pRb and p107 resulted markedly decreased at late stages. Differentiating N1E-115 cells also showed a progressive decrease in B-myb levels, a proliferation-related protein whose constitutive expression inhibits neuronal differentiation. Transfection of each of the RB family genes in these cells was able, at different degrees, to induce neuronal differentiation, to inhibit [3H]thymidine incorporation and to down-regulate the activity of the B-myb promoter.

(Global and local) fluctuations of phase space contraction in deterministic stationary nonequilibrium.

We studied numerically the validity of the fluctuation relation introduced in Evans et al. [Phys. Rev. Lett. 71, 2401-2404 (1993)] and proved under suitable conditions by Gallavotti and Cohen [J. Stat. Phys. 80, 931-970 (1995)] for a two-dimensional system of particles maintained in a steady shear flow by Maxwell demon boundary conditions [Chernov and Lebowitz, J. Stat. Phys. 86, 953-990 (1997)]. The theorem was found to hold if one considers the total phase space contraction sigma occurring at collisions with both walls: sigma=sigma( upward arrow )+sigma( downward arrow ). An attempt to extend it to more local quantities sigma( upward arrow ) and sigma( downward arrow ), corresponding to the collisions with the top or bottom wall only, gave negative results. The time decay of the correlations in sigma( upward arrow, downward arrow ) was very slow compared to that of sigma. (c) 1998 American Institute of Physics.

Retinoblastoma-related protein pRb2/p130 and its binding to the B-myb promoter increase during human neuroblastoma differentiation.

Neuroblastoma cells can undergo neural differentiation upon treatment with a variety of chemical inducers and growth factors. During this process, many cell cycle-related genes are downregulated while differentiation-specific genes are triggered. The retinoblastoma family proteins, pRb, p107, and pRb2/p130, are involved in transcriptional repression of proliferation genes, mainly through their interaction with the E2F transcription factors. We report that pRb2/p130 expression levels increased during differentiation of neuroblastoma cell line LAN-5. On the other hand, both pRb and p107 decreased and underwent progressive dephosphorylation at late differentiation times. The expression of B-myb and c-myb, two targets of the retinoblastoma family proteins, were downregulated in association with the increase of pRb2/p130, which was detected as the major component of the complex with E2F on the E2F site of the B-myb promoter in differentiated cells. Interestingly, E2F4, a preferential partner of p107 and pRb2/p130, was upregulated and underwent changes in cellular localization during differentiation. In conclusion, our data suggest a major role of pRb2/p130 in the regulation of B-myb promoter during neural differentiation despite the importance of cofactors in modulating the function of the retinoblastoma family proteins.

Retinoblastoma protein family in cell cycle and cancer: a review.

Two genes, p107 and Rb2/p130, are strictly related to RB, the most investigated tumor suppressor gene, responsible for susceptibility to retinoblastoma. The products of these three genes, namely pRb, p107, and pRb2/p130 are characterized by a peculiar steric conformation, called "pocket," responsible for most of the functional interactions characterizing the activity of these proteins in the homeostasis of the cell cycle. The interest in these genes and proteins springs from their ability to regulate cell cycle processes negatively, being able, for example, to dramatically slow down neoplastic growth. So far, among these genes, only RB is firmly established to act as a tumor suppressor, because its lack-of-function is clearly involved in tumor onset and progression. It has been found deleted or mutated in most retinoblastomas and sarcomas, but its inactivation is likely to play a crucial role in other types of human cancers. The two other members of the family have been discovered more recently and are currently under extensive investigation. We review analogies and differences among the pocket protein family members, in an attempt to understand their functions in normal and cancer cells.

Cloning of a novel human RNA polymerase II subunit downregulated by doxorubicin: new potential mechanisms of drug related toxicity.

Using the differential display PCR method, we have isolated an mRNA downregulated in doxorubicin resistant human cell lines. The full length cDNA clone was identified as the human homologue of yeast RPB11 subunit of RNA polymerase II. Northern blot analysis of normal tissues detected a particularly high expression of RPB11 mRNA in heart and skeletal muscle. Reduction of this mRNA expression was observed in all the cell lines tested after drug treatment and was paralleled by a similar decrease of the protein levels. These findings suggest that doxorubicin may exert in vivo specific inhibitory effects on a major component of the transcription machinery.

The retinoblastoma gene product in acute myeloid leukemia: a possible involvement in promyelocytic leukemia.

The retinoblastoma susceptibility gene in leukemia and lymphoma has been investigated using different approaches involving either gene or protein analysis. In this study, a novel method, which evaluates the functional status of the retinoblastoma gene product by a binding assay to an in vitro-translated viral oncoprotein, has been applied to leukemic cells from acute myeloid leukemia patients. One hundred twenty-two cases were considered, and 42 of them were also analyzed by Western blot. Results obtained with the two methods were comparable, with the exception of few cases, where the retinoblastoma protein appeared detectable but unable to bind to the viral oncoprotein. The retinoblastoma protein has been found defective mostly in the M3 promyelocytic subtype.