Transforming growth factor-beta1 mediates epithelial to mesenchymal transdifferentiation through a RhoA-dependent mechanism.

Transforming growth factor-beta1 (TGF-beta) can be tumor suppressive, but it can also enhance tumor progression by stimulating the complex process of epithelial-to-mesenchymal transdifferentiaion (EMT). The signaling pathway(s) that regulate EMT in response to TGF-beta are not well understood. We demonstrate the acquisition of a fibroblastoid morphology, increased N-cadherin expression, loss of junctional E-cadherin localization, and increased cellular motility as markers for TGF-beta-induced EMT. The expression of a dominant-negative Smad3 or the expression of Smad7 to levels that block growth inhibition and transcriptional responses to TGF-beta do not inhibit mesenchymal differentiation of mammary epithelial cells. In contrast, we show that TGF-beta rapidly activates RhoA in epithelial cells, and that blocking RhoA or its downstream target p160(ROCK), by the expression of dominant-negative mutants, inhibited TGF-beta-mediated EMT. The data suggest that TGF-beta rapidly activates RhoA-dependent signaling pathways to induce stress fiber formation and mesenchymal characteristics.

A strong negative transcriptional regulatory region between the human cytomegalovirus UL127 gene and the major immediate-early enhancer.

The region of the human cytomegalovirus (CMV) genome between the UL127 open reading frame and the major immediate-early (MIE) enhancer is referred to as the unique region. DNase I protection analysis with human cell nuclear extracts demonstrated multiple protein binding sites in this region of the viral genome (P. Ghazal, H. Lubon, C. Reynolds-Kohler, L. Hennighausen, and J. A. Nelson, Virology 174:18-25, 1990). However, the function of this region in the context of the viral genome is not known. In wild-type human CMV-infected human fibroblasts, cells permissive for viral replication, there is little to no transcription from UL127. We determined that the unique region prevented transcription from the UL127 promoter but had no effect on the divergent MIE promoter. In transient-transfection assays, the basal level of expression from the UL127 promoter increased significantly when the wild-type unique sequences were mutated. In recombinant viruses with similar mutations in the unique region, expression from the UL127 promoter occurred only after de novo viral protein synthesis, typical of an early viral promoter. A 111-bp deletion-substitution of the unique sequence caused approximately a 20-fold increase in the steady-state level of RNA from the UL127 promoter and a 245-fold increase in the expression of a downstream indicator gene. This viral negative regulatory region was also mutated at approximately 50-bp regions proximal and distal to the UL127 promoter. Although some repressive effects were detected in the distal region, mutations of the region proximal to the UL127 promoter had the most significant effects on transcription. Within the proximal and distal regions, there are potential cis sites for known eucaryotic transcriptional repressor proteins. This region may also bind unknown viral proteins. We propose that the unique region upstream of the UL127 promoter and the MIE enhancer negatively regulates the expression from the UL127 promoter in permissive human fibroblast cells. This region may be a regulatory boundary preventing the effects of the very strong MIE enhancer on this promoter.

Computer-assisted measurement of lumbar spine radiographs.

RATIONALE AND OBJECTIVES: The authors evaluated a method for obtaining reproducible, reliable measurements from standard lumbar spine radiographs for determining the degree of spondylolisthesis, vertebral body height, intervertebral disk space height, disk space angle, and degree of vertebral body wedging. MATERIALS AND METHODS: Four to six easily defined points were identified on each vertebral body on anteroposterior and lateral plain radiographs of the lumbosacral spine of patients. From these points, the degree of spondylolisthesis, the vertebral body height, the intervertebral disk space height, the disk space angle, and the degree of vertebral body wedging were easily calculated by using well-known geometric relationships. This method requires the use of a personal computer and a standard spreadsheet program but does not require the use of any other specialized radiographic equipment, computer hardware, or custom software. RESULTS: Calculations of intra- and interobserver variability for the measurement of spondylolisthesis, disk space height, disk space angle, and vertebral body height measurement showed that the technique is extremely reproducible. CONCLUSION: This technique may prove useful in the prospective evaluation of potential candidates for lumbar spinal stenosis surgery.

Dialysis-related Arthropathy in Patients on Long-term Hemodialysis: Radiographic Features.

The purpose of this study was to evaluate the character and evolution of bone lesions attributable to amyloid deposition in patients on long-term dialysis. Thirty-five patients who were treated with hemodialysis for 5 to 22 years were studied by a review of medical records and hand radiographs. The frequency, distribution, character, and evolution of skeletal cyst-like lesions believed to be secondary to amyloid deposition were evaluated in relation to dialysis duration. The number and size of these lesions increased with dialysis duration, present in 28% of the patients after 5 through 9 years of hemodialysis and in 91% after 15 through 22 years. In contrast, the changes of hyperparathyroidism decreased. Of patients with skeletal wrist lesions, radiographs of symptomatic large joints were available in 15; five had bone abnormalities. Skeletal amyloid deposition was verified pathologically in nine sites (five patients). It is concluded that skeletal lesions believed to be due to amyloid deposition increase with dialysis duration and most commonly affect the wrists. They have a distinctive character, distribution, and evolution and are often associated with carpal tunnel syndrome.

Outcome after laminectomy for lumbar spinal stenosis. Part II: Radiographic changes and clinical correlations.

The pre- and postoperative lumbar spine radiographs of 119 patients who underwent decompressive lumbar laminectomy were studied to evaluate radiographic changes and to correlate them with clinical outcome. An accurate and reproducible method was used for measuring pre- and postoperative radiographs that were separated by an average interval of 4.6 years. Levels of the spine that underwent laminectomy showed greater change in spondylolisthesis, disc space angle, and disc space height than unoperated levels. Outcome correlated with radiographic changes at operated and unoperated levels. This study demonstrates that radiographic changes are greater at operated than at unoperated levels and that some postoperative symptoms do correlate with these changes. Lumbar fusion should be considered in some patients who undergo decompressive laminectomy. The efficacy of and unequivocal indications for lumbar fusion can only be determined from randomized, prospective, controlled trials, however, and these studies have not yet been undertaken.

Consort 1 flight results: a synopsis.

Consort 1 was the first low gravity materials processing payload to be launched by a commercially licensed rocket in the U.S.A. It carried six experiments which operated as planned during approx. 7 min of suborbital, low gravity flight (10(-5) g) and were returned in excellent condition to the investigators within 4 h of launch. Nearly 150 physical samples supported by measurements and photographs made during the flight were obtained for analysis. In addition to the experimental data returned, the success of Consort 1 demonstrated the ability of industry, working with university centers and government agencies, to rapidly prepare and launch payloads. A brief description of the rocket flight and payload configuration is given. Experiment objectives and methods are described and preliminary results and conclusions are presented.