Sulfur mustard bis(2-chloroethyl)sulfide; SM is a chemical warfare agent that produces edema and blister formation with a severe inflammatory reaction. The mouse ear vesicant model for SM injury has ...been used to evaluate pharmacological agents for countering SM dermal injury. The vanilloid olvanil reduces SM-induced edema and mRNA expression of cytokines and chemokines, suggesting that blocking the inflammatory effects of neuropeptides, such as substance P (SP), may provide protection against SM-induced dermal injury. This study examined SP expression in mice exposed to SM (0.16
mg) on the inner surface of the right ear, with or without olvanil pretreatment at 1, 10, 30, 60, and 360
min following exposure. In naïve skin, SP mRNA localization was associated with blood vessels and sebaceous glands. In SM-exposed skin, SP mRNA was also detected in perivascular dermal cells. Immunohistochemical localization of SP protein was observed in the ear skin of naïve, SM-, olvanil/SM-, and vehicle-treated mice. Quantification of SP
+ perivascular dermal cells revealed that SM exposure led to a significant increase (
P ≤ 0.05) in SP
+ cells over the observed time period. Olvanil pretreatment significantly reduced (
P ≤ 0.05) the mean number of SP
+ cells at 60 and 360
min. This study demonstrates that SP expression could provide an additional endpoint for evaluating the effectiveness of vanilloid drugs on SM-induced skin inflammation.
Infection with pathogenic influenza viruses is associated with intense inflammatory disease. Here, we investigated the innate immune response in mice infected with H5N1 A/Vietnam/1203/04 and with ...reassortant human H1N1 A/Texas/36/91 viruses containing the virulence genes hemagglutinin (HA), neuraminidase (NA) and NS1 of the 1918 pandemic virus. Inclusion of the 1918 HA and NA glycoproteins rendered a seasonal H1N1 virus capable of inducing an exacerbated host innate immune response similar to that observed for highly pathogenic A/Vietnam/1203/04 virus. Infection with 1918 HA/NA:Tx/91 and A/Vietnam/1203/04 were associated with severe lung pathology, increased cytokine and chemokine production, and significant immune cell changes, including the presence of CD11b super(+)Gr-1 super(+) cells in the blood, lung and bone marrow. Significant differential gene expression in the lung included pathways for cell death, apoptosis, production and response to reactive oxygen radicals, as well as arginine and proline metabolism and chemokines associated with monocyte and neutrophil/granulocyte accumulation and/or activation. Arginase was produced in the lung of animals infected with A/Vietnam/1204. These results demonstrate that the innate immune cell response results in the accumulation of CD11b super(+)Gr-1 super(+) cells and products that have previously been shown to contribute to T cell suppression.
Infection with pathogenic influenza viruses is associated with intense inflammatory disease. Here, we investigated the innate immune response in mice infected with H5N1 A/Vietnam/1203/04 and with ...reassortant human H1N1 A/Texas/36/91 viruses containing the virulence genes hemagglutinin (HA), neuraminidase (NA) and NS1 of the 1918 pandemic virus. Inclusion of the 1918 HA and NA glycoproteins rendered a seasonal H1N1 virus capable of inducing an exacerbated host innate immune response similar to that observed for highly pathogenic A/Vietnam/1203/04 virus. Infection with 1918 HA/NA:Tx/91 and A/Vietnam/1203/04 were associated with severe lung pathology, increased cytokine and chemokine production, and significant immune cell changes, including the presence of CD11b^sup +^Gr-1^sup +^ cells in the blood, lung and bone marrow. Significant differential gene expression in the lung included pathways for cell death, apoptosis, production and response to reactive oxygen radicals, as well as arginine and proline metabolism and chemokines associated with monocyte and neutrophil/granulocyte accumulation and/or activation. Arginase was produced in the lung of animals infected with A/Vietnam/1204. These results demonstrate that the innate immune cell response results in the accumulation of CD11b^sup +^Gr-1^sup +^ cells and products that have previously been shown to contribute to T cell suppression.PUBLICATION ABSTRACT
Cutaneous exposure to sulfur mustard bis(2-chloroethyl) sulfide (SM) produces a delayed inflammatory skin response that is followed by severe dermal injury. Assessment of anti-inflammatory therapies ...against SM-induced skin injury has mainly relied on qualitative histopathological evaluation. The goal of this study was to identify proinflammatory biomarkers in the hairless mouse vesicant model that could be used as additional indicators of SM-induced skin injury for evaluating anti-inflammatory treatment. SM-induced inflammation was determined at 2, 6, and 24 hr postexposure by changes in edema. Ribonuclease protection assay (RPA) was used to determine changes in gene expression of inflammatory mediators. At 2, 6, and 24 hr postexposure, a time-dependent increase in edema was observed in SM-exposed skin, which was significant at 6 and 24 hr when compared to unexposed controls. Ribonuclease protection assay analysis revealed a two-fold or greater increase in monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-2 (MIP-2), MIP-1α, tumor necrosis factor-α, and interleukin (IL)-1β following exposure to SM when compared to unexposed controls. A significant time-dependent increase was observed in MCP-1, MIP-1α, and IL-1β over the 24 hr time period. At 24 hr postexposure, skin treated with the anti-inflammatory drug olvanil showed a significant decrease in SM-induced edema. Additionally, mRNA levels of MCP-1, MIP-2, and IL-1β were decreased when compared to skin exposed to SM alone. In this study, we identified molecular biomarkers at the mRNA level, up-regulated in skin exposed to SM, which can be partially suppressed by olvanil. Further characterization of the mRNA and protein expression patterns of proinflammatory biomarkers may enable the use of other classes of anti-inflammatory drugs or therapeutic treatments against SM dermal injury.
Abstract only
Time‐related changes in the histopathology of mouse skin occur after topical exposure to the vesicant agent, bis(2‐chloroethyl) sulfide (SM) using the mouse ear vesicant model (MEVM). ...Some of these changes include increasing edema, detachment of the epidermis from the dermis, and upregulation of laminin‐332. Endoplasmic reticulum stress response (ESR) was observed by confocal microscopy. Microarray analysis and RT‐PCR experiments indicated there was upregulation of several heat shock proteins and folding chaperones such glucose‐regulated protein 78 and GRP94 in the ER. It confirmed the confocal observations and suggested that ER stress occurs very quickly in the MEVM. The accumulation of laminin γ2 protein, one of the chains of laminin‐332, in the ER was visualized by confocal microscopy and co localized with the ER chaperones. This accumulation appeared specifically in the migrating, but not proliferating cells. These observations are consistent with recent reports that laminin‐332 is found in migrating, transformed epithelial cells that have left the cell cycle, but not in proliferating cells. These results suggest that laminin γ2 is misfolded after SM treatment, resulting in decreased secretion and reduced overall amounts of laminin‐332 in the extracellular matrix. This would explain the observation that there is a delayed wound healing response evident in this wound model.
Grant Funding Source
ES005022
,
ES004738
, EY09056, and the NIH CounterACT Program through NIAMS U54AR055073
Sulfur mustard bis(2-chloroethyl)sulfide, SM is a chemical warfare agent that penetrates the skin rapidly and causes extensive blistering. Using the mouse ear vesicant model (MEVM), we evaluated the ...effect of topically applied anti-inflammatory agents (octyl homovanillamide and heptyl isovanillamide) on ear edema formation and gene expression following SM exposure. Relative ear weight and real-time reverse transcriptase polymerase chain reaction of GM-CSF, IL-1β, and IL-6 were used to evaluate the effects of octyl homovanillamide and heptyl isovanillamide. Both vanilloids significantly reduced SM-induced edema. At the single dose and number of animals/group tested, octyl homovanillamide produced a trend of reduced mRNA levels; however, the reduction was not significant for GM-CSF, IL-1β, or IL-6. Heptyl isovanillamide significantly reduced (p ≤ 0.05) GM-CSF, IL-1β, and IL-6 mRNA levels. These results show that octyl homovanillamide and heptyl isovanillamide reduce skin edema and heptyl isovanillamide significantly reduced cytokine mRNA expression following SM exposure. In addition to measuring edema formation, monitoring expression of biomarkers such as GM-CSF, IL-1β, and IL-6 may also serve to evaluate therapeutic treatments against SM-induced dermal injury.
There is evidence for immunotoxicity of aflatoxin B1 (AFB1) in chronic animal feeding studies; however, little information is available as to the effects of inhalation exposure. This study evaluated ...the acute affects of aerosolized AFB1 on systemic immune function of female C57BL/6N mice following a single aerosol exposure. Mice were exposed in nose-only inhalation tubes to 0, 2.86, 6.59 and 10 μg AFB1 aerosol/L air for 90 minutes. A negative control group of untreated mice and a positive control group of cyclophosphamide-treated mice were included to account for day to day variation. Three days following exposure, mice were sacrificed and body, liver, lung, thymus and spleen weights, and complete blood counts and white blood cell differentials were measured. Splenocytes were isolated for flow cytometric analysis of CD4+ and CD8+ lymphocytes, CD19+ B-cells and natural killer cells (NK 1.1+). The effect of AFB1 on humoral immunity was assessed by measuring serum anti-keyhole limpet hemocyanin (KLH) IgM levels. Of the tissues examined, only the thymus weight of AFB1 exposed mice decreased significantly compared to naïve mice; however, the decrease was not dose related and was also observed in the 0 AFB1 aerosol control group. A decrease in the mean white blood cell count of treated vs. naïve mice was observed at all dose levels but was clearly not dose related and was statistically significant only in the 0 and 2.86 μg/L groups. Red blood cell and platelet counts and white blood cell differentials were not significantly affected by AFB1. The number of CD4+ (helper T-cells), CD8+ (cytotoxic T-cells) and CD19+ (B-cells) decreased in spleens of AFB1 aerosol exposed mice compared to naïve mice; however, the decrease was not dose-related and was also observed in the 0 AFB1 exposure group. Dose-related changes in the CD4+/CD8+ T-lymphocyte ratios were not observed. The IgM response to KLH was not significantly different in AFB1 compared to naïve mice, suggesting that AFB1 did not effect antigen-specific antibody production. Based on the results of this study, a single AFB1 inhalation exposure up to 10 μg/L for 90 minutes (CxT = 900 μg ·min/L) did not significantly alter the immune parameters measured in this study. The aerosol vehicle (ethanol) and/or stress could have masked subtle AFB1-dependent changes in thymus and spleen weights, and in splenic lymphocyte subpopulations. However, for other immunological parameters, such as the IgM response to KLH, there was clearly no significant effect of AFB1 aerosol exposure.
Despite the large number of pharmaceutically active compounds found in natural environments little is known about their transport behavior in groundwater, which is complicated by their wide range of ...physical and chemical properties. The transport behavior of five widely used and often detected pharmaceutical compounds and one lifestyle drug has therefore been investigated, using a set of three column experiments. The investigated compounds were the anticonvulsant carbamazepine, the lifestyle drug caffeine, the antibiotic sulfamethoxazole, the lipid regulator gemfibrozil, and the nonsteroidal anti-inflammatories ibuprofen and naproxen. The columns were filled with three different types of sand. The substrates consisted of artificially prepared iron-coated sand, artificially prepared organic carbon sand (with 5% leaf compost), and natural aquifer sand from Long Point, Ontario (Canada). The experiments were conducted simultaneously under the same hydraulic conditions and with the same input solution of about 1μg·L−1 of each compound. The transport behavior of the organic compounds differed significantly between both the different columns and the different compounds. A strong correlation was observed between the retardation factors for carbamazepine, gemfibrozil, and ibuprofen and the organic carbon content of the substrate. While the retardation increased with increasing organic carbon content, no direct relationship was observed between the organic carbon content and the removal of these compounds. In contrast, the retardation factors for sulfamethoxazole and naproxen showed no correlation with the organic carbon content but these compounds were significantly removed in the presence of organic matter. The influence of the Fe3+ surfaces in the iron-coated sand was less significant than expected, with all compounds except for sulfamethoxazole having retardation factors <1.8. Caffeine was so strongly removed during transport through those substrates containing organic carbon that no reliable retardation factor could be determined.
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•Investigation of a mixture of six widely used and reported micropollutants•Identical saturated column flow through experiment with but different substrates•Investigation of two model substrates in comparison to a natural aquifer sediment•Retardation and degradation depend of same micropollutants different for different substrates.•The most important factor controlling transport is the organic carbon content.
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