Structural and Electrical Characterization of 2" Ammonothermal Free-Standing GaN Wafers. Progress toward Pilot Production.

Free-standing gallium nitride (GaN) substrates are in high demand for power devices, laser diodes, and high-power light emitting diodes (LEDs). SixPoint Materials Inc. has begun producing 2" GaN substrates through our proprietary Near Equilibrium AmmonoThermal (NEAT) growth technology. In a single 90 day growth, eleven c-plane GaN boules were grown from free-standing hydride vapor phase epitaxy (HVPE) GaN substrates. The boules had an average X-ray rocking curve full width at half maximum (FWHM) of 33 ± 4 in the 002 reflection and 44 ± 6 in the 201 reflection using 0.3 mm divergence slits. The boules had an average radius of curvature of 10.16 ± 3.63 m. The quality of the boules was highly correlated to the quality of the seeds. A PIN diode grown at Georgia Tech on a NEAT GaN substrate had an ideality factor of 2.08, a high breakdown voltage of 1430 V, and Baliga's Figure of Merit of >9.2 GW/cm2. These initial results demonstrate the suitability of using NEAT GaN substrates for high-quality MOCVD growth and fabrication of high-power vertical GaN switching devices.

Quantitative proteomics identifies a beta-catenin network as an element of the signaling response to Frizzled-8 protein-related antiproliferative factor.

Antiproliferative factor (APF), a Frizzled-8 protein-related sialoglycopeptide involved in the pathogenesis of interstitial cystitis, potently inhibits proliferation of normal urothelial cells as well as certain cancer cells. To elucidate the molecular mechanisms of the growth-inhibitory effect of APF, we performed stable isotope labeling by amino acids in cell culture analysis of T24 bladder cancer cells treated with and without APF. Among over 2000 proteins identified, 54 were significantly up-regulated and 48 were down-regulated by APF treatment. Bioinformatic analysis revealed that a protein network involved in cell adhesion was substantially altered by APF and that ß-catenin was a prominent node in this network. Functional assays demonstrated that APF down-regulated ß-catenin, at least in part, via proteasomal and lysosomal degradation. Moreover, silencing of ß-catenin mimicked the antiproliferative effect of APF whereas ectopic expression of nondegradable ß-catenin rescued growth inhibition in response to APF, confirming that ß-catenin is a key mediator of APF signaling. Notably, the key role of ß-catenin in APF signaling is not restricted to T24 cells, but was also observed in an hTERT-immortalized human bladder epithelial cell line, TRT-HU1. In addition, the network model suggested that ß-catenin is linked to cyclooxygenase-2 (COX-2), implying a potential connection between APF and inflammation. Functional assays verified that APF increased the production of prostaglandin E(2) and that down-modulation of ß-catenin elevated COX-2 expression, whereas forced expression of nondegradable ß-catenin inhibited APF-induced up-regulation of COX-2. Furthermore, we confirmed that ß-catenin was down-regulated whereas COX-2 was up-regulated in epithelial cells explanted from IC bladder biopsies compared with control tissues. In summary, our quantitative proteomics study describes the first provisional APF-regulated protein network, within which ß-catenin is a key node, and provides new insight that targeting the ß-catenin signaling pathway may be a rational approach toward treating interstitial cystitis.

An hTERT-immortalized human urothelial cell line that responds to anti-proliferative factor.

Studies of the urothelium, the specialized epithelial lining of the urinary bladder, are critical for understanding diseases affecting the lower urinary tract, including interstitial cystitis, urinary tract infections and cancer. However, our understanding of urothelial pathophysiology has been hampered by a lack of appropriate model systems. Here, we describe the isolation and characterization of a non-transformed urothelial cell line (TRT-HU1), originally explanted from normal tissue and immortalized with hTERT, the catalytic subunit of telomerase. We demonstrate responsiveness of the cells to anti-proliferative factor (APF), a glycopeptide implicated in the pathogenesis of interstitial cystitis. TRT-HU1 carries a deletion on the short arm of chromosome 9, an early genetic lesion in development of bladder cancer. TRT-HU1 urothelial cells displayed growth and migration characteristics similar to the low-grade papilloma cell line RT4. In contrast, we observed marked differences in both phenotype and gene expression profiles between TRT-HU1 and the highly malignant T24 cell line. Together, these findings provide the first demonstration of a non-transformed, continuous urothelial cell line that responds to APF. This cell line will be valuable for studies of both benign and malignant urothelial cell biology.

A signaling network in phenylephrine-induced benign prostatic hyperplasia.

Benign prostatic hyperplasia (BPH) is an age-related disease of unknown etiology characterized by prostatic enlargement and coinciding with distinctive alterations in tissue histomorphology. To identify the molecular mechanisms underlying the development of BPH, we conducted a DNA microarray study using a previously described animal model in which chronic alpha(1)-adrenergic stimulation by repeated administration of phenylephrine evokes histomorphological changes in the rat prostate that resemble human BPH. Bioinformatic tools were applied to microarray data obtained from prostate tissue to construct a network model of potentially relevant signal transduction pathways. Significant involvement of inflammatory pathways was demonstrable, including evidence for activation of a TGF-beta signaling cascade. The heterodimeric protein clusterin (apolipoprotein J) was also identified as a prominent node in the network. Responsiveness of TGF-beta signaling and clusterin gene and protein expression were confirmed independently of the microarray data, verifying some components of the model. This is the first attempt to develop a comprehensive molecular network for histological BPH induced by adrenergic activation. The study also implicated clusterin as a novel biochemical target for therapy.