Evaluation of parameters critical to observing proteins inside living Escherichia coli by in-cell NMR spectroscopy

Our recently developed in-cell NMR procedure now enables one to observe protein conformations inside living cells. Optimization of the technique demonstrates that distinguishing the signals produced by a single protein species depends critically on protein overexpression levels and the correlation time in the cytoplasm. Less relevant is the selective incorporation of (15)N. Poorly expressed proteins, insoluble proteins, and proteins that cannot tumble freely due to associations within the cell cannot yet be observed. We show in-cell NMR spectra of bacterial NmerA and human calmodulin and discuss limitations of the technique as well as prospects for future applications.     

A Realistic Computer-Simulated Brain Phantom for Evaluation of PET Charactenstics

To evaluate accurately the imaging characteristics of positron emission tomography (PET), a realistic computer-simulated brain phantom was developed. A cross-sectional slice from a human cadaver brain was chosen for its combination of gray matter, white matter, and cerebrospinal fluid (CSF) regions. The slice was photographed and digitized into a gray-level image with a video digitizer, boundary edges were located around cerebral structures in the digitized image, and each structural region was assigned a uniform pixel value dependent on both the cerebral parameter (e.g., blood flow, oxygen uptake, metabolic rate) under investigation and the type of structure (gray matter, white matter, CSF). Line integrals through the regions were generated at various angular and transverse positions according to specific physical characteristics (such as detector line-spread function) of a tomographic scanner configuration to create a set of simulated but realistic projection measurements. The set of projection measurements can be processed with any standard reconstruction program to create a tomographic image to reveal the effects of various PET characteristics. Investigations with this computer-simulated brain phantom have demonstrated its usefulness for examining the interrelations among neuroanatomical structure volume, tomographic spatial resolution, partial volume effect, and nonlinear parameter estimation. Transportability of the simulated phantom and the procedure to other medical imaging environments is described, and limitations of this simulation procedure are discussed.