Mononuclear phagocytes in the pathogenesis of neurodegenerative diseases.

Brain mononuclear phagocytes (MP, bone marrow monocyte-derived macrophages, perivascular macrophages, and microglia) function to protect the nervous system by acting as debris scavengers, killers of microbial pathogens, and regulators of immune responses. MP are activated by a variety of environmental cues and such inflammatory responses elicit cell injury and death in the nervous system. MP immunoregulatory responses include secretion of neurotoxic factors, mobilization of adaptive immunity, and cell chemotaxis. This incites tissue remodelling and blood-brain barrier dysfunction. As disease progresses, MP secretions engage neighboring cells in a vicious cycle of autocrine and paracrine amplification of inflammation leading to tissue injury and ultimately destruction. Such pathogenic processes tilt the balance between the relative production of neurotrophic and neurotoxic factors and to disease progression. The ultimate effects that brain MP play in disease revolves "principally" around their roles in neurodegeneration. Importantly, common functions of brain MP in neuroimmunity link highly divergent diseases (for example, human immunodeficiency virus type-one associated dementia, Alzheimer's disease and Parkinson's disease). Research into this process from our own laboratories and those of others seek to harness MP inflammatory processes with the intent of developing therapeutic interventions that block neurodegenerative processes and improve the quality of life in affected people.

Expression profiling of small cellular samples in cancer: less is more.

Expression profiling of tumours from cancer patients has uncovered several genes that are critically important in the progression of a normal cell to an oncogenic phenotype. Leading the way in these discoveries is the use of microarrays, a technology that is currently in transition from basic science applications to use in the clinic. Microarrays can determine the global gene regulation of an individual cancer, which may be useful in formulating an individualised therapy for the patient. Currently, cells used in breast cancer microarray studies often come from either homogenous cultures or heterogeneous biopsy samples. Both cell sources are at a disadvantage in determining the most accurate gene profile of cancer, which often consists of multiple subspecies of cancerous cells within a background of normal cells. Therefore, acquisition of small, but highly specific biopsies for analysis may be required for an accurate expression analysis of the disease. Amplification methods, such as polymerase chain reaction (PCR) and amplified antisense RNA (aRNA) amplification, have been used to amplify the mRNA signal from very small samples, which can then be used for microarray analysis. In this study, we describe the acquisition, amplification, and analysis of very small samples (<10000 cells) for expression analysis and demonstrate that the ultimate resolution of cancer expression analysis, one cell, is both feasible and practical.

Wild-type and Ras-transformed fibroblasts display differential mitogenic responses to transient sodium arsenite exposure.

Arsenic is a human carcinogen whose mechanism of action remains undefined. Based on the hypothesis that arsenic sensitizes cells to mitogenic stimulation by affecting the receptor tyrosine kinase (RTK) signal transduction pathway, these studies first examined the response of fibroblasts to specific mitogens using a defined media system. In both rodent and human fibroblasts, DNA synthesis was found to be stimulated in cells exposed to a transient, sub-lethal concentration of sodium arsenite followed by stimulation with known RTK pathway activators. This effect is observed for up to 32 h after removal of arsenic, suggesting that the RTK pathway is affected in a sustained manner. In contrast, transient arsenic exposure of ras-transformed cells results in decreased mitogen-stimulated DNA synthesis. Flow cytometry indicates that arsenic increases the percentage of wild-type cells in the S-phase of the cell cycle; conversely, the percentage of ras-transformed cells in S-phase is decreased by arsenic. No evidence of arsenic-induced cytotoxicity was detected using the neutral red assay, ensuring that decreased DNA synthesis in ras-transformed cells is not due to cell death. Taken together, the results of experiments presented herein indicate that arsenic produces sustained alterations in the growth characteristics of rodent and human fibroblasts. It is postulated that the proliferation-enhancing effect of arsenic on wild-type cells contributes to its ability to cause cancer.