http://hdl.handle.net/2144/2429 Sun, 19 May 2013 22:08:42 GMT 2013-05-19T22:08:42Z http://hdl.handle.net/2144/3370 Quantitative Observations of Fever and Its Treatment before the Advent of Short Clinical Thermometers. Estes, J W Mon, 01 Apr 1991 00:00:00 GMT http://hdl.handle.net/2144/3370 1991-04-01T00:00:00Z http://hdl.handle.net/2144/3371 Signaling Pathways Required for Macrophage Scavenger Receptor-Mediated Phagocytosis: Analysis by Scanning Cytometry Sulahian, Timothy H; Imrich, Amy; DeLoid, Glen; Winkler, Aaron R; Kobzik, Lester BACKGROUND. Scavenger receptors are important components of the innate immune system in the lung, allowing alveolar macrophages to bind and phagocytose numerous unopsonized targets. Mice with genetic deletions of scavenger receptors, such as SR-A and MARCO, are susceptible to infection or inflammation from inhaled pathogens or dusts. However, the signaling pathways required for scavenger receptor-mediated phagocytosis of unopsonized particles have not been characterized. METHODS. We developed a scanning cytometry-based high-throughput assay of macrophage phagocytosis that quantitates bound and internalized unopsonized latex beads. This assay allowed the testing of a panel of signaling inhibitors which have previously been shown to target opsonin-dependent phagocytosis for their effect on unopsonized bead uptake by human in vitro-derived alveolar macrophage-like cells. The non-selective scavenger receptor inhibitor poly(I) and the actin destabilizer cytochalasin D were used to validate the assay and caused near complete abrogation of bead binding and internalization, respectively. RESULTS. Microtubule destabilization using nocodazole dramatically inhibited bead internalization. Internalization was also significantly reduced by inhibitors of tyrosine kinases (genistein and herbimycin A), protein kinase C (staurosporine, chelerythrine chloride and Gö 6976), phosphoinositide-3 kinase (LY294002 and wortmannin), and the JNK and ERK pathways. In contrast, inhibition of phospholipase C by U-73122 had no effect. CONCLUSION. These data indicate the utility of scanning cytometry for the analysis of phagocytosis and that phagocytosis of unopsonized particles has both shared and distinct features when compared to opsonin-mediated phagocytosis. Thu, 07 Aug 2008 00:00:00 GMT http://hdl.handle.net/2144/3371 2008-08-07T00:00:00Z http://hdl.handle.net/2144/3369 Cell Density Plays a Critical Role in Ex Vivo Expansion of T Cells for Adoptive Immunotherapy Ma, Qiangzhong; Wang, Yawen; Lo, Agnes Shuk-Yee; Gomes, Erica M.; Junghans, Richard P. The successful ex vivo expansion of a large numbers of T cells is a prerequisite for adoptive immunotherapy. In this study, we found that cell density had important effects on the process of expansion of T cells in vitro. Resting T cells were activated to expand at high cell density but failed to be activated at low cell density. Activated T cells (ATCs) expanded rapidly at high cell density but underwent apoptosis at low cell density. Our studies indicated that low-cell-density related ATC death is mediated by oxidative stress. Antioxidants N-acetylcysteine, catalase, and albumin suppressed elevated reactive oxygen species (ROS) levels in low-density cultures and protected ATCs from apoptosis. The viability of ATCs at low density was preserved by conditioned medium from high-density cultures of ATCs in which the autocrine survival factor was identified as catalase. We also found that costimulatory signal CD28 increases T cell activation at lower cell density, paralleled by an increase in catalase secretion. Our findings highlight the importance of cell density in T cell activation, proliferation, survival and apoptosis and support the importance of maintaining T cells at high density for their successful expansion in vitro. Wed, 30 Jun 2010 00:00:00 GMT http://hdl.handle.net/2144/3369 2010-06-30T00:00:00Z http://hdl.handle.net/2144/3368 Pharmacological Properties of DOV 315,090, an Ocinaplon Metabolite Berezhnoy, Dmytro; Gravielle, Maria C; Downing, Scott; Kostakis, Emmanuel; Basile, Anthony S; Skolnick, Phil; Gibbs, Terrell T; Farb, David H BACKGROUND. Compounds targeting the benzodiazepine binding site of the GABAA-R are widely prescribed for the treatment of anxiety disorders, epilepsy, and insomnia as well as for pre-anesthetic sedation and muscle relaxation. It has been hypothesized that these various pharmacological effects are mediated by different GABAA-R subtypes. If this hypothesis is correct, then it may be possible to develop compounds targeting particular GABAA-R subtypes as, for example, selective anxiolytics with a diminished side effect profile. The pyrazolo[1,5-a]-pyrimidine ocinaplon is anxioselective in both preclinical studies and in patients with generalized anxiety disorder, but does not exhibit the selectivity between α1/α2-containing receptors for an anxioselective that is predicted by studies using transgenic mice. RESULTS. We hypothesized that the pharmacological properties of ocinaplon in vivo might be influenced by an active biotransformation product with greater selectivity for the α2 subunit relative to α1. One hour after administration of ocinaplon, the plasma concentration of its primary biotransformation product, DOV 315,090, is 38% of the parent compound. The pharmacological properties of DOV 315,090 were assessed using radioligand binding studies and two-electrode voltage clamp electrophysiology. We report that DOV 315,090 possesses modulatory activity at GABAA-Rs, but that its selectivity profile is similar to that of ocinaplon. CONCLUSION. These findings imply that DOV 315,090 could contribute to the action of ocinaplon in vivo, but that the anxioselective properties of ocinaplon cannot be readily explained by a subtype selective effect/action of DOV 315,090. Further inquiry is required to identify the extent to which different subtypes are involved in the anxiolytic and other pharmacological effects of GABAA-R modulators. Fri, 13 Jun 2008 00:00:00 GMT http://hdl.handle.net/2144/3368 2008-06-13T00:00:00Z