ABSTRACT Drug screening in the immediate term will be best accomplished by early use of primary cells in which the target of the screen is a network of proteins measured in populations of single cells.
"failure of a specific pathway to activate, hyper/hyposensitivity of the pathway to physiological modulators, altered kinetics of response, and rewiring of canonical pathways). The technology is based on the assumption that a significant amount of information about a signaling network can be gained by tracking signaling activity as it occurs under different conditions . Generally, this is achieved experimentally by exposing live cells to external modulators, quantifying the state of each signaling node and then comparing it to the basal state. "
[Show abstract][Hide abstract] ABSTRACT: Autoimmune diseases are complex and heterogeneous in nature and show varying responses to therapeutic treatment. A more accurate biological characterization of individual patients would assist in disease classification, prognosis, and treatment decisions. This characterization ideally would incorporate cellular, biochemical, and molecular information that contribute to the inflammatory state. The advent of single-cell network profiling (SCNP) using phospho-flow multiparametric flow cytometry allows insight into the complexity of signaling networks in various cell types. Here, we describe the potential of SCNP to inform on the biological characterization of autoimmune disease, the application in clinical medicine, and the utility in drug development.
"This is particularly the case for microscopy studies, including large-scale siRNA screens with imaging read out. In contrast, there is substantial controversy of how well cell lines, which are often established from late stage cancer, preserve aspects of the disease and whether or not they should be used in cancer drug development (3–5). Thus animal experiments or studies in primary cell lines are often preferred despite their added complexity. "
[Show abstract][Hide abstract] ABSTRACT: Biological experiments are most often performed with immortalized cell lines because they are readily available and can be expanded without limitation. However, cell lines may differ from the in vivo situation in important aspects. Here we introduce a straightforward methodology to compare cell lines to their cognate primary cells and to derive a comparative functional phenotype. We used SILAC (stable isotope labeling by amino acids in cell culture) for quantitative, mass spectrometry-based comparison of the hepatoma cell line Hepa1-6 with primary hepatocytes. The resulting quantitative proteome of 4,063 proteins had an asymmetric distribution, with many proteins down-regulated in the cell line. Bioinformatic analysis of the quantitative proteomics phenotypes revealed that Hepa1-6 cells were deficient in mitochondria, reflecting re-arrangement of metabolic pathways, drastically up-regulate cell cycle-associated functions and largely shut down drug metabolizing enzymes characteristic for the liver. This quantitative knowledge of changes provides an important basis to adapt cell lines to more closely resemble physiological conditions.
[Show abstract][Hide abstract] ABSTRACT: Helicon waves are excited in an argon plasma discharge by a twisted double helix antenna operating at 13.56 MHz. Two optical probes are used. One can be inserted into a I cm diameter glass tube in the center of the plasma chamber (about 11 cm diameter). The other observes the plasma from the outside. The optical probes are measuring emission from the short-lifetime At II states. Software is used to correlate the optical emission to the phase of the RF source. The Ar II emission is measured along with B-dot and Langmuir probe measurements at different axial positions. 105 GHz and 10 GHz; interferometry are used to verify Langmuir probe measurements. The result is to compare Ar II emission to B<sub>z</sub> phase velocity and plasma density to determine the traveling wave particle and background Maxwellian interactions. The background level and modulation index of the Ar II emission is measured for this reason.
Plasma Science, 2002. ICOPS 2002. IEEE Conference Record - Abstracts. The 29th IEEE International Conference on; 02/2002
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