Molecular manipulation of keratin 8/18 intermediate filaments: modulators of FAS-mediated death signaling in human ovarian granulosa tumor cells

Background: Granulosa cell tumors (GCT) are a rare ovarian neoplasm but prognosis is poor following recurrence. Keratin intermediate filaments expressed in these tumors are a diagnostic marker, yet paradoxically, may also constitute a target for therapeutic intervention. In the current study, we evaluated keratin 8/18 (K8/18) filament expression as a mechanism of resistance to apoptosis in GCT, specifically focusing on regulation of the cell surface death receptor, Fas (FAS). Methods: The GCT cell line, KGN, was transiently transfected with siRNA to KRT8 and KRT18 to reduce K8/18 filament expression. Expression of K8/18, FAS, and apoptotic proteins (PARP, cleaved PARP) were evaluated by fluorescence microscopy, flow cytometric analysis, and immunoblotting, respectively. The incidence of FAS-mediated apoptosis in KGN cells was measured by caspase 3/7 activity. All experiments were performed independently three to six times, using a fresh aliquot of KGN cells for each experiment. Quantitative data were analyzed by one- or two-way analysis of variance (ANOVA), followed by a Tukey’s post-test for multiple comparisons; differences among means were considered statistically significant at P \u3c 0.05. Results: Control cultures of KGN cells exhibited abundant K8/18 filament expression (~90 % of cells), and minimal expression of FAS (\u3c25 % of cells). These cells were resistant to FAS-activating antibody (FasAb)-induced apoptosis, as determined by detection of cleaved PARP and measurement of caspase 3/7 activity. Conversely, siRNA-mediated knock-down of K8/18 filament expression enhanced FAS expression (\u3e 70 % of cells) and facilitated FasAb-induced apoptosis, evident by increased caspase 3/7 activity (P \u3c 0.05). Additional experiments revealed that inhibition of protein synthesis, but not MEK1/2 or PI3K signaling, also prompted FasAb-induced apoptosis. Conclusions: The results demonstrated that K8/18 filaments provide resistance to apoptosis in GCT by impairing FAS expression. The abundance of keratin filaments in these cells and their role in apoptotic resistance provides a greater mechanistic understanding of ovarian tumorgenicity, specifically GCT, as well as a clinically-relevant target for potential therapeutic intervention

Small quantum networks operating as quantum thermodynamic machines

We show that a 3-qubit system as studied for quantum information purposes can alternatively be used as a thermodynamic machine when driven in finite time and interfaced between two split baths. The spins are arranged in a chain where the working spin in the middle exercises Carnot cycles the area of which defines the exchanged work. The cycle orientation (sign of the exchanged work) flips as the difference of bath temperatures goes through a critical value.Comment: RevTeX, 4 pages, 7 figures. Replaced by version accepted for publication in EP

The Effect of Surface Roughness on the Universal Thermal Conductance

We explain the reduction of the thermal conductance below the predicted universal value observed by Schwab et al. in terms of the scattering of thermal phonons off surface roughness using a scalar model for the elastic waves. Our analysis shows that the thermal conductance depends on two roughness parameters: the roughness amplitude $\delta$ and the correlation length $a$. At sufficiently low temperatures the conductance decrease from the universal value quadratically with temperature at a rate proportional to $\delta ^{2}a$. Values of $\delta$ equal to 0.22 and $a$ equal to about 0.75 of the width of the conduction pathway give a good fit to the data.Comment: 10 pages, 5 figures. Ref. added, typo correcte

Multiresolution kernel matrix algebra

We propose a sparse arithmetic for kernel matrices, enabling efficient scattered data analysis. The compression of kernel matrices by means of samplets yields sparse matrices such that assembly, addition, and multiplication of these matrices can be performed with essentially linear cost. Since the inverse of a kernel matrix is compressible, too, we have also fast access to the inverse kernel matrix by employing exact sparse selected inversion techniques. As a consequence, we can rapidly evaluate series expansions and contour integrals to access, numerically and approximately in a data-sparse format, more complicated matrix functions such as $A^\alpha$ and $\exp(A)$. By exploiting the matrix arithmetic, also efficient Gaussian process learning algorithms for spatial statistics can be realized. Numerical results are presented to quantify and quality our findings

A high-reflectivity high-Q micromechanical Bragg-mirror

We report on the fabrication and characterization of a micromechanical oscillator consisting only of a free-standing dielectric Bragg mirror with high optical reflectivity and high mechanical quality. The fabrication technique is a hybrid approach involving laser ablation and dry etching. The mirror has a reflectivity of 99.6%, a mass of 400ng, and a mechanical quality factor Q of approximately 10^4. Using this micromirror in a Fabry Perot cavity, a finesse of 500 has been achieved. This is an important step towards designing tunable high-Q high-finesse cavities on chip.Comment: 3 pages, 2 figure

Inverse Spin Hall Effect and Anomalous Hall Effect in a Two-Dimensional Electron Gas

We study the coupled dynamics of spin and charge currents in a two-dimensional electron gas in the transport diffusive regime. For systems with inversion symmetry there are established relations between the spin Hall effect, the anomalous Hall effect and the inverse spin Hall effect. However, in two-dimensional electron gases of semiconductors like GaAs, inversion symmetry is broken so that the standard arguments do not apply. We demonstrate that in the presence of a Rashba type of spin-orbit coupling (broken structural inversion symmetry) the anomalous Hall effect, the spin Hall and inverse spin Hall effect are substantially different effects. Furthermore we discuss the inverse spin Hall effect for a two-dimensional electron gas with Rashba and Dresselhaus spin-orbit coupling; our results agree with a recent experiment.Comment: 5 page

Self-cooling of a micro-mirror by radiation pressure

We demonstrate passive feedback cooling of a mechanical resonator based on radiation pressure forces and assisted by photothermal forces in a high-finesse optical cavity. The resonator is a free-standing high-reflectance micro-mirror (of mass m=400ng and mechanical quality factor Q=10^4) that is used as back-mirror in a detuned Fabry-Perot cavity of optical finesse F=500. We observe an increased damping in the dynamics of the mechanical oscillator by a factor of 30 and a corresponding cooling of the oscillator modes below 10 K starting from room temperature. This effect is an important ingredient for recently proposed schemes to prepare quantum entanglement of macroscopic mechanical oscillators.Comment: 11 pages, 9 figures, minor correction

Statistical mechanics of transcription-factor binding site discovery using Hidden Markov Models

Hidden Markov Models (HMMs) are a commonly used tool for inference of transcription factor (TF) binding sites from DNA sequence data. We exploit the mathematical equivalence between HMMs for TF binding and the "inverse" statistical mechanics of hard rods in a one-dimensional disordered potential to investigate learning in HMMs. We derive analytic expressions for the Fisher information, a commonly employed measure of confidence in learned parameters, in the biologically relevant limit where the density of binding sites is low. We then use techniques from statistical mechanics to derive a scaling principle relating the specificity (binding energy) of a TF to the minimum amount of training data necessary to learn it.Comment: 25 pages, 2 figures, 1 table V2 - typos fixed and new references adde

Dissipation in nanocrystalline-diamond nanomechanical resonators

We have measured the dissipation and frequency of nanocrystalline-diamond nanomechanical resonators with resonant frequencies between 13.7 MHz and 157.3 MHz, over a temperature range of 1.4–274 K. Using both magnetomotive network analysis and a time-domain ring-down technique, we have found the dissipation in this material to have a temperature dependence roughly following T^(0.2), with Q^(–1) ≈ 10^(–4) at low temperatures. The frequency dependence of a large dissipation feature at ~35–55 K is consistent with thermal activation over a 0.02 eV barrier with an attempt frequency of 10 GHz