Biomedical subjects

T A Freeman

Publications and source records attributed to T A Freeman.

4 recordsLinked to original sources

PubMed2006

Involvement of hydrogen peroxide in the differentiation and apoptosis of preosteoclastic cells exposed to arsenite.

Long-term exposure to sodium arsenite (AsO(2)) promotes the development of various cancers. Paradoxically, arsenic also induces pro-myelomonocytic leukemia cell differentiation, and at higher concentrations, apoptosis. The present study investigated the effects of AsO(2) on preosteoclasts. When treated with 2.5-5microM AsO(2), RAW264.7 cells underwent osteoclast differentiation as evidenced by an increase in the number of multinucleate cells expressing tartrate resistant acid phosphatase (TRAP). The appearance of these phenotypic markers was preceded by a low level increase in extracellular production of H(2)O(2) and was prevented by the addition of catalase (4.5microg/ml), an enzyme that removes H(2)O(2). Only at high concentrations (10-25microM) of AsO(2) was a significant loss of cell viability and a high level increase in H(2)O(2) production (1.5microM) observed. Apoptosis was blocked by pretreatment with diphenylene iodonium chloride (2microM), a NAD(P)H-flavoprotein inhibitor, suggesting the involvement of NADPH-oxidase. The data show that AsO(2), dose-dependently, stimulates increasing amounts of H(2)O(2) production. Moreover, at concentrations found in tissues of individuals exposed to geochemical AsO(2), osteoclasts underwent an H(2)O(2)-dependent differentiation. Therefore, chronic exposure to low-level amounts of AsO(2) could result in increased bone resorption and contribute to bone related pathologies.

Acid Phosphatase↗

PubMed2006

Active caspase-3 is required for osteoclast differentiation.

Based on our earlier observation that caspase-3 is present in osteoclasts that are not undergoing apoptosis, we investigated the role of this protein in the differentiation of primary osteoclasts and RAW264.7 cells (Szymczyk KH, et al., 2005, Caspase-3 activity is necessary for RANKL-induced osteoclast differentiation. The Proceedings of the 8th ICCBMT). We noted that osteoclast numbers are decreased in long bones of procaspase-3 knockout mice and that receptor activator of NF-kappaB ligand (RANKL) does not promote differentiation of isolated preosteoclasts. In addition, after treatment with inhibitors of caspase-3 activity, neither the wild-type primary nor the RAW264.7 cells express TRAP or became multinucleated. We found that immediately following RANKL treatment, procaspase-3 is cleaved and the activated protein is localized to lipid regions of the plasma membrane and the cytosol. We developed RAW264.7 procaspase-3 knockdown clonal cell lines using RNAi technology. Again, treatment with RANKL fails to induce TRAP activity or multinucleation. Finally, we evaluated NF-kappaB in procaspase-3 silenced cells. We found that RANKL treatment prevented activation and nuclear translocation of NF-kappaB. Together these findings provide direct support for the hypothesis that caspase-3 activity is required for osteoclast differentiation.

Animals↗

PubMed1994

Membrane-associated tubulin in the rat anterior pituitary and GH3 cells.

Plasma membranes from anterior pituitary lobes of rats and GH3 cells were exposed to monoclonal anti-beta tubulin antibodies or normal mouse IgG, and treated with FITC-labeled anti-mouse IgG. Specimens were visualized with a microscope equipped with epifluorescence optics. Tubulin-specific fluorescence was very prominent in membranes incubated with tubulin antibody and was negligible in membranes incubated with normal mouse IgG or FITC-labeled anti-mouse IgG alone. Two dimensional electrophoresis revealed that 27% of the rat pituitary tubulin is beta-tubulin and 73% is alpha-tubulin; 17% of the GH3 cell tubulin is beta-tubulin and 83% is alpha-tubulin. These results suggest that tubulin is a component of plasma membranes of the rat pituitary and GH3 cells.

Animals↗

PubMed1992

Lesioning a connectionist model of visual search: selective effects on distractor grouping.

A connectionist model of visual search is presented, in which search is determined by patterns of grouping between distractors and between the target and the distractors. Grouping is based on conjunctive relations between simple form elements (corners, line-end terminators), and uses principles of similarity and spatial proximity. In its normal, 'unlesioned' state, the model simulates the search data generated by human subjects when they search for simple form conjunctions amongst either homogeneous or heterogeneous distractors (Humphreys & Muller, in press). In this paper, the performance of the model is examined after the model is subject to various types of 'lesion'. 'Lesioning' is produced either by increasing the internal noise on the activation functions governing the interactions between processing units, or by eliminating processing units from different loci in the model. Increasing the internal noise within the model, or lesioning units within 'high-level' processing stages, can generate the selective effects on search found in a detailed single case study of an agnosic patient (Humphreys, Riddoch, Quinlan, Price & Donnelly, this volume): Namely, there is selective disruption of search for a form conjunction amongst homogeneous distractors relative to search for the same target amongst heterogeneous distractors. This selective effect can be attributed to the 'lesion' disrupting grouping between distractors, which normally facilitates search with homogeneous distractors. The simulations demonstrate that (1) search processes can be disrupted by adding internal noise to search functions, (2) that there can be selective effects on grouping processes, and (3) that these selective effects can be associated with different types of lesion to different stages in the model. The implications of the simulations for understanding visual processing impairments in agnosia are discussed.

Agnosia↗