Donna Pack

Time course of gene expression of inflammatory mediators in rat lung after diesel exhaust particle exposure.

Diesel exhaust particles (DEPs) at three concentrations (5, 35, and 50 mg/kg body weight) were instilled into rats intratracheally. We studied gene expression at 1, 7, and 30 days postexposure in cells obtained by bronchoalveolar lavage (BAL) and in lung tissue. Using real-time reverse transcriptase-polymerase chain reaction (RT-PCR), we measured the mRNA levels of eight genes [interleukin (IL)-1β, IL-6, IL-10, iNOS (inducible nitric oxide synthase), MCP-1 (monocyte chemoattractant protein-1), MIP-2 (macrophage inflammatory protein-2), TGF-β1 (transforming growth factor-β1), and TNF-α (tumor necrosis factor-α)] in BAL cells and four genes [IL-6, ICAM-1 (intercellular adhesion molecule-1), GM-CSF (granulocyte/macrophage-colony stimulating factor), and RANTES (regulated upon activation normal T cell expressed and secreted)] in lung tissue. In BAL cells on day 1, high-dose exposure induced a significant up-regulation of IL-1β, iNOS, MCP-1, and MIP-2 but no change in IL-6, IL-10, TGF-β1, and TNF-α mRNA levels. There was no change in the mRNA levels of IL-6, RANTES, ICAM-1, and GM-CSF in lung tissue. Nitric oxide production and levels of MCP-1 and MIP-2 were increased in the 24-hr culture media of alveolar macrophages (AMs) obtained on day 1. IL-6, MCP-1, and MIP-2 levels were also elevated in the BAL fluid. BAL fluid also showed increases in albumin and lactate dehydrogenase. The cellular content in BAL fluid increased at all doses and at all time periods, mainly due to an increase in polymorphonuclear leukocytes. In vitro studies in AMs and cultured lung fibroblasts showed that lung fibroblasts are a significant source of IL-6 and MCP-1 in the lung.

Enhanced Pulmonary Response to the Inhalation of Freshly Fractured Silica as Compared with Aged Dust Exposure

Abstract We have reported previously that grinding crystalline silica generates radicals on its cleavage planes and that this fresh dust is more cytotoxic in vitro than aged silica. The objective of the present study was to determine if freshly fractured silica was also more toxic and inflammatory in vivo than aged silica of the same composition and particle size. Fresh α-quartz was generated using a jet mill, while aged dust was milled and then stored for 2 months before use. Analysis of surface radicals by electron spin resonance spectroscopy verified the enhanced surface activity of this fresh silica compared with aged dust. Male Fischer 344 rats were exposed to fresh or aged α-quartz by inhalation (20 mg/m3, 5 hours per day, 5 days per week for 2 weeks) and pulmonary responses were determined 1 to 3 days after exposure. Exposure to aged silica resulted in an increase in total cells, red blood cells, lymphocytes, and granulocytes harvested by bronchoalveolar lavage, and in elevated acellular lavage pro...

Comparative in vitro toxicity of grape- and citrus-farm dusts.

Agricultural workers are exposed to a variety of airborne dusts, including crystalline silica and other inorganic minerals. This study was designed to characterize the organic and inorganic components of agricultural dusts in California grape- and citrus-farm fields and to compare their cytotoxicity using in vitro toxicity bioassays as predictors of pathogenicity. Aerosolized dusts collected from farm fields were characterized by scanning-electron-microscopic energy-dispersive x-ray analysis, x-ray diffraction, trace metal analysis by plasma emission spectroscopy, and surface area measurements. As indicators of cytotoxicity, cell viability, release of alveolar enzymes activities (lactate dehydrogenase, N-acetyl glucosaminidase), production of reactive oxygen species (ROS), such as H2O2 and hydroxyl radical (OH), and lipid peroxidation were monitored after exposure of cells to grape- and citrus-farm dusts or inorganic components of these dusts. In addition, activation of nuclear factor κ B and activator protein-1 were evaluated at the peak time for response of 36 h postexposure. All toxicity studies were done in comparison with crystalline silica of similar particle size and diameter using the same mass concentrations as farm dusts. The results showed that inorganic minerals in the aerosolized farm dust fractions were mostly composed of aluminum silicates, crystalline silica, and free iron. Crystalline silica used in these studies was more cytotoxic than grape- and citrus-farm dusts. However, in general, citrus farm dust exhibited the greatest ability to generate ROS and induce lipid peroxidation. These results support human epidemiologic studies, reporting an increased incidence of pulmonary fibrosis in farm workers, by documenting the potential of farm dusts to induce oxidative stress and initiate disease development.