Novel C-Terminal Hsp90 Inhibitor for Head and Neck Squamous Cell Cancer (HNSCC) with in vivo Efficacy and Improved Toxicity Profiles Compared with Standard Agents

The final publication is available at Springer via http://dx.doi.org/10.1245/s10434-011-1971-1.Background - Current therapies for HNSCC, especially platinum agents, are limited by their toxicities and drug resistance. This study evaluates a novel C-terminal Hsp90 inhibitor (CT-Hsp90-I) for efficacy and toxicity in vitro and in vivo in an orthotopic HNSCC model. Our hypothesis is that C-terminal inhibitors exhibit improved toxicity/efficacy profiles over standard therapies and may represent a novel group of anticancer agents. Methods - MDA-1986 HNSCC cells were treated with doses of 17-AAG or KU363 (a CT-Hsp90-I) and compared for antiproliferation by GLO-Titer and trypan blue exclusion and for apoptosis by PARP cleavage and caspase-3 inactivation by Western analysis. In vivo studies in Nu/Nu mice examined an orthotopic model of MDA-1986 cells followed by drug dosing intraperitoneally for a 21-day period (mg/kg/dose: cisplatin = 3.5, low-dose KU363 = 5, high-dose KU363 = 25, 17-AAG = 175). Tumor size, weight, and toxicity (body score) were measured 3×/week. Results - The IC50 levels for KU363 = 1.2–2 μM in MDA-1986. KU363 induces apoptosis at 1 μM with cleavage of PARP and inactivation of caspase-3 levels after 24 h. Client proteins Akt and Raf-1 were also downregulated at 1–3 μM of drug. In vivo, 100% of controls had progressive disease, while 100% of cisplatin animals showed some response, all with significant systemic toxicity. High-dose KU363 showed 88% of animals responding and low-dose KU363 showed 75% responding. KU363 animals showed significantly less toxicity (P < 0.01) than cisplatin or 17-AAG. Conclusion - This novel CT-Hsp90-I KU363 manifests potent anticancer activity against HNSCC, showing excellent in vivo efficacy and reduced toxicity compared with standard agents justifying future translational evaluation

Alternative Stacking Sequences in Hexagonal Boron Nitride

The relative orientation of successive sheets, i.e. the stacking sequence, in layered two-dimensional materials is central to the electronic, thermal, and mechanical properties of the material. Often different stacking sequences have comparable cohesive energy, leading to alternative stable crystal structures. Here we theoretically and experimentally explore different stacking sequences in the van der Waals bonded material hexagonal boron nitride (h-BN). We examine the total energy, electronic bandgap, and dielectric response tensor for five distinct high symmetry stacking sequences for both bulk and bilayer forms of h-BN. Two sequences, the generally assumed AA' sequence and the relatively unknown (for h-BN) AB (Bernal) sequence, are predicted to have comparably low energy. We present a scalable modified chemical vapor deposition method that produces large flakes of virtually pure AB stacked h-BN; this new material complements the generally available AA' stacked h-BN

Infrared point source variability between the Spitzer and MSX surveys of the Galactic mid-plane

We present a list of 552 sources with suspected variability, based on a comparison of mid-infrared photometry from the GLIMPSE I and MSX surveys, which were carried out nearly a decade apart. We were careful to address issues such as the difference in resolution and sensitivity between the two surveys, as well as the differences in the spectral responses of the instruments. We selected only sources where the IRAC 8.0 and MSX 8.28 micron fluxes differ by more than a factor of two, in order to minimize contamination from sources where the difference in fluxes at 8 micron is due to a strong 10 micron silicate feature. We present a subset of 40 sources for which additional evidence suggests variability, using 2MASS and MIPSGAL data. Based on a comparison with the variability flags in the IRAS and MSX Point-Source Catalogs we estimate that at least a quarter of the 552 sources, and at least half of the 40 sources are truly variable. In addition, we tentatively confirm the variability of one source using multi-epoch IRAS LRS spectra. We suggest that most of the sources in our list are likely to be Asymptotic Giant Branch stars.Comment: 47 pages, 12 Figures, 3 Tables, accepted for publication in A

Absolute diffuse calibration of IRAC through mid-infrared and radio study of HII regions

We investigate the diffuse absolute calibration of the InfraRed Array Camera on the Spitzer Space Telescope at 8.0microns using a sample of 43 HII regions with a wide range of morphologies near GLON=312deg. For each region we carefully measure sky-subtracted,point-source- subtracted, areally-integrated IRAC 8.0-micron fluxes and compare these with Midcourse Space eXperiment (MSX) 8.3-micron images at two different spatial resolutions, and with radio continuum maps. We determine an accurate median ratio of IRAC 8.0-micron/MSX\8.3-micron fluxes, of 1.55+/-0.15. From robust spectral energy distributions of these regions we conclude that the present 8.0-micron diffuse calibration of the SST is 36% too high compared with the MSX validated calibration, perhaps due to scattered light inside the camera. This is an independent confirmation of the result derived for the diffuse calibration of IRAC by the Spitzer Science Center (SSC). From regression analyses we find that 843-MHz radio fluxes of HII regions and mid-infrared (MIR) fluxes are linearly related for MSX at 8.3-microns and Spitzer at 8.0 microns, confirming the earlier MSX result by Cohen & Green. The median ratio of MIR/843-MHz diffuse continuum fluxes is 600 times smaller in nonthermal than thermal regions, making it a sharp discriminant. The ratios are largely independent of morphology up to a size of ~24 arcsec. We provide homogeneous radio and MIR morphologies for all sources. MIR morphology is not uniquely related to radio structure. Compact regions may have MIR filaments and/or diffuse haloes, perhaps infrared counter- parts to weakly ionized radio haloes found around compact HII regions. We offer two IRAC colour-colour plots as quantitative diagnostics of diffuse HII regions.Comment: 29 pages, LaTeX (aastex), incl. 31 PostScript (ps,eps) figures and 5 tables. Accepted by MNRAS (main journal). Replaced an unused file and added this URL for people wishing to download a version with high-resolution images: http://www.astro.wisc.edu/sirtf/martin.hii.accepted.pd

A Synthesis of Rates and Controls on Elemental Mercury Evasion in the Great Lakes Basin

Rates of surface-air elemental mercury (Hgo) fluxes in the literature were synthesized for the Great Lakes Basin (GLB). For the majority of surfaces, fluxes were net positive (evasion). Digital land-cover data were combined with representative evasion rates and used to estimate annual Hgo evasion for the GLB (7.7 Mg/yr). This value is less than our estimate of total Hg deposition to the area (15.9 Mg/yr), suggesting the GLB is a net sink for atmospheric Hg. The greatest contributors to annual evasion for the basin are agricultural (~55%) and forest (~25%) land cover types, and the open water of the Great Lakes (~15%). Areal evasion rates were similar across most land cover types (range: 7.0 to 21.0 μg/m2-yr), with higher rates associated with urban (12.6 μg/m2-yr) and agricultural (21.0 μg/m2-yr) lands. Uncertainty in these estimates could be partially remedied through a unified methodological approach to estimating Hgo fluxes

A novel C-terminal HSP90 inhibitor KU135 induces apoptosis and cell cycle arrest in melanoma cells

Heat shock protein 90 (Hsp90) is differentially expressed in tumor cells including melanoma and involved in proper folding, stabilization and regulation of cellular proteins. We investigated a novobiocin-derived Hsp90 C-terminal inhibitor, KU135, for anti-proliferative effects in melanoma cells. The results indicate that KU135 reduced cell viability and cell proliferation in melanoma cells and IC50 values for A735(DRO), M14(NPA), B16F10 and SKMEL28 cells were 0.82, 0.92, 1.33 and 1.30 M respectively. KU135 induced a more potent anti-proliferative effect in most melanoma cells versus N-terminal Hsp90 inhibitor 17AAG. KU135 induced apoptosis in melanoma cells, as indicated by annexin V/PI staining, reduction in the mitochondrial membrane potential, mitochondrial cytochrome C release and caspase 3 activation. KU135 reduced levels of Hsp90 client proteins Akt, BRAF, RAF-1, cyclin B and cdc25 proteins. Additionally, it reduced Hsp70, Hsp90 paralog, GRP94 and HSF1 levels. KU135 induced strong G2/M cell cycle arrest, associated with decreased expression of cdc25c, cyclin B and increased phosphorylation of cdc25c. These finding show that KU135 reduced cell survival, proliferation, and induces apoptosis in melanoma cells. We suggest that KU135 may be a potential candidate for cancer therapy against melanoma

A Toolbox for Spatiotemporal Analysis of Voltage-Sensitive Dye Imaging Data in Brain Slices

Voltage-sensitive dye imaging (VSDI) can simultaneously monitor the spatiotemporal electrical dynamics of thousands of neurons and is often used to identify functional differences in models of neurological disease. While the chief advantage of VSDI is the ability to record spatiotemporal activity, there are no tools available to visualize and statistically compare activity across the full spatiotemporal range of the VSDI dataset. Investigators commonly analyze only a subset of the data, and a majority of the dataset is routinely excluded from analysis. We have developed a software toolbox that simplifies visual inspection of VSDI data, and permits unaided statistical comparison across spatial and temporal dimensions. First, the three-dimensional VSDI dataset (x,y,time) is geometrically transformed into a two-dimensional spatiotemporal map of activity. Second, statistical comparison between groups is performed using a non-parametric permutation test. The result is a 2D map of all significant differences in both space and time. Here, we used the toolbox to identify functional differences in activity in VSDI data from acute hippocampal slices obtained from epileptic Arx conditional knock-out and control mice. Maps of spatiotemporal activity were produced and analyzed to identify differences in the activity evoked by stimulation of each of two axonal inputs to the hippocampus: the perforant pathway and the temporoammonic pathway. In mutant hippocampal slices, the toolbox identified a widespread decrease in spatiotemporal activity evoked by the temporoammonic pathway. No significant differences were observed in the activity evoked by the perforant pathway. The VSDI toolbox permitted us to visualize and statistically compare activity across the spatiotemporal scope of the VSDI dataset. Sampling error was minimized because the representation of the data is standardized by the toolbox. Statistical comparisons were conducted quickly, across the spatiotemporal scope of the data, without a priori knowledge of the character of the responses or the likely differences between them