Influence of experimental type 1 diabetes on the pulmonary effects of diesel exhaust particles in mice

Epidemiologically, exposure to particulate air pollution is associated with increases in morbidity and mortality, and diabetics are especially vulnerable to effects of particles. This study was carried out to determine the respiratory effect of diesel exhaust particles (DEP; 0.4 mg/kg) on mice rendered diabetic by the injection of streptozotocin or vehicle (control). Four weeks following induction of diabetes, the animals were intratracheally instilled (i.t.) with DEP (0.4 mg/kg) or saline. 24 h later, the measurement of airway reactivity to methacholine in vivo by a forced oscillation technique showed a significant and dose-dependent increase in airway resistance in non-diabetic mice exposed to DEP versus non-diabetic mice exposed to saline. Similarly, the airway resistance was significantly increased in diabetic mice exposed to DEP versus diabetic mice exposed to saline. Nevertheless, there was no difference in the airway resistance between diabetic and non-diabetic mice after i.t. administration of DEP. Following DEP administration there were neutrophil polymorphs infiltration of pulmonary interalveolar septae and the alveolar spaces with many macrophages containing DEP in both diabetic and non-diabetic mice. Interestingly, apoptotic cells were only found in the examined lung sections from diabetic mice exposed to DEP. Total proteins and albumin concentrations in bronchoalveolar lavage (BAL) fluid, markers for increase of epithelial permeability, were significantly increased in diabetic mice exposed to DEP compared to saline-treated diabetic and DEP-treated non diabetic mice. Superoxide dismutase activity and reduced glutathione concentration in BAL were significantly decreased in diabetic mice exposed to DEP compared to saline-treated diabetic and DEP-treated non diabetic mice. Moreover, tumor necrosis factor α (TNFα) concentrations were significantly increased in diabetic mice exposed to DEP compared to saline-treated diabetic and DEP-treated non diabetic mice. We conclude that, at the dose and time point investigated, DEP equally increased airway resistance and caused infiltration of inflammatory cells in the lung of both diabetic and non-diabetic mice. However, the occurrence of oxidative stress, the presence lung apoptotic cells and the increase of total proteins, albumin and TNFα in BAL fluid were only seen in DEP-exposed diabetic mice suggesting an increased respiratory susceptibility to particulate air pollution.     

Airway resistance, inflammation and oxidative stress following exposure to diesel exhaust particle in angiotensin II-induced hypertension in mice

Exposure to particulate matter is a risk factor for respiratory and cardiovascular diseases. However, the mechanisms underlying these effects are not well understood. Here, we compared the impact of diesel exhaust particles (DEP) on airway resistance, inflammation and oxidative stress in normal mice, or mice made hypertensive by implanting osmotic minipump infusing angiotensin II. On day 13 after the onset of infusion, angiotensin II induced significant increase in heart rate (P<0.05) and systolic blood pressure (P<0.0001). On the same day, mice were intratracheally instilled with either DEP (15 μg/mouse) or saline. Twenty-four hour later, the measurement of airway reactivity to methacholine (0-10mg/ml) in vivo by a forced oscillation technique showed a significant and dose dependent increase in airway resistance in normotensive mice exposed to DEP compared to those exposed to saline. In hypertensive mice, there was no difference in airway resistance in DEP versus saline exposed mice. However, following exposure to DEP, airway resistance significantly increased in normotensive versus hypertensive mice. Bronchoalveolar lavage (BAL) fluid analysis showed a significant increase in macrophage numbers in normotensive mice exposed to DEP compared to those exposed to saline, and to hypertensive mice exposed to DEP. Neutrophil numbers were significantly increased in both normotensive and hypertensive mice exposed to DEP compared with their respective control groups. Superoxide dismutase activity was significantly decreased following DEP exposure in both normotensive and hypertensive mice compared to their respective controls. However, total proteins, a marker for increase of epithelial permeability, and malondialdehyde, a reflection of lipid peroxidation, were only increased in normotensive mice exposed to DEP. Therefore, our data suggest that DEP do not aggravate airway resistance and inflammation in angiotensin II-induced hypertensive mice. On the contrary, at the dose of DEP and time point investigated, airway resistance, inflammation and oxidative stress are increased in normotensive compared to hypertensive mice.     

Effects of single exposure to cadmium on the primary humoral antibody response

Effects of a single intraperitoneal (i.p.) injection of cadmium (Cd) on the primary humoral antibody responses against sheep red blood cells (SRBC) in mice were studied by assaying splenic plaque forming cells (PFC). PFC responses in mice were suppressed when exposed to Cd 2 days after immunization, and inconsistently stimulated when exposed before immunization. Dose-response relationships were observed in the suppressive effect of Cd exposure 2 days after immunization, but not consistently in the stimulative effect of Cd exposure before immunization. Thymus weights and cell numbers decreased markedly 4 days after Cd exposure with or without the antigenic stimulus. Splenic weights increased 2 days after Cd exposure, while the number of spleen cells was dramatically decreased 1 days after Cd exposure and still remained below normal 2 days after exposure.     

Effect of diesel exhaust particulate on bacillus Calmette-Guerin lung infection in mice and attendant changes in lung interstitial lymphoid subpopulations and IFNgamma response.

The effect of exposure to diesel exhaust particulate (DEP) on bacillus Calmette-Guerin (BCG) lung infection in mice was studied. C57Bl/6J female mice were infected with BCG (2.5 x 104 bacteria/mouse) by intrapulmonary instillation, with or without coadministration of DEP (100 microg/mouse). Five weeks later, mice exposed to DEP + BCG had about a four-fold higher BCG load in the lungs than mice exposed only to BCG (p < 0.05). DEP treatment alone had no effect on the total number of lung lymphocytes or numbers of T, B, or NK cells recovered from lungs. In contrast, BCG infection significantly increased (p< 0.05) recovery levels of all types of lymphocytes from lungs. Coexposure to DEP + BCG further increased the recovery of lymphocytes from lungs of BCG-infected mice. The pulmonary lymphocyte subpopulation expressing the greatest levels of mRNA for IFNgamma after BCG infection was CD4+ T cells. Expression levels were similar in mice exposed to BCG or BCG + DEP and were elevated as compared to noninfected mice and mice treated with DEP alone. Recovery of IFNgamma-secreting lymphocytes and IFNgamma-secreting T cells was significantly higher (p < 0.05) from lungs of BCG-infected mice as compared to control or DEP-exposed mice. BCG and BCG + DEP groups of mice did not differ significantly in the numbers of IFNgamma-secreting lymphocytes in lungs. Taken together, these results indicated that coexposure to DEP + BCG did not significantly affect the level of IFNgamma response of mice to BCG infection. However, DEP treatment was found to inhibit IFNgamma-induced nitric oxide (NO) production by mouse alveolar macrophages in vitro. Our results indicate that DEP exposure did not alter the IFNgamma response to BCG infection, but reduced responsiveness of alveolar macrophages to IFNgamma. Reduced sensitivity of DEP-exposed alveolar macrophages to IFNgamma may contribute to a greater load of BCG in the lungs of BCG-infected mice given DEP.     

Sensitivity to ozone, diesel exhaust particles, and standardized ambient particulate matter in rats with a listeria monocytogenes-induced respiratory infection

Ambient particulate matter may increase respiratory allergic skewing of the T-cell-mediated immune response toward a T-helper-2 (Th2) response, with the consequence that the Th1 response develops less well. Successful clearing of a respiratory bacterial infection depends on an adequate Th1 immune response; therefore, the subject would not control the infection as well if exposed to particulate matter. To substantiate this hypothesis, we examined the effect of exposure to diesel exhaust particles (DEP) and urban particulate matter (EHC-93, Ottawa dust) on rats with a Listeria monocytogenes respiratory infection. Since this hypothesis has been confirmed for ozone, we used it as a positive control. Wistar rats were exposed to ozone (2 mg/m3 for 24 h/day for 7 days) and to DEP or to EHC-93 (50 microg/rat intranasally daily for 7 consecutive days). Twenty-four hours after the last exposure, the rats were infected intratracheally with 1 x 10(6) L. monocytogenes bacteria. The number of L. monocytogenes was determined after 3, 4 and 5 days. Statistically significant increases of the number of L. monocytogenes in rats exposed to ozone were observed in the lungs and spleen at all three times. However, we found no significant differences in the numbers of bacteria that were found in rats exposed to DEP or EHC-93 compared to the saline-treated group at any of the three times. In conclusion, the results of this study do not support the hypothesis that exposure to DEP or EHC-93 reduces subsequent resistance to a respiratory infection in rats.     

Diesel exhaust particles in blood trigger systemic and pulmonary morphological alterations

Short-term exposure to elevated levels of particulate matter is associated with increased respiratory and cardiovascular mortality and morbidity. However, the mechanisms underlying these effects are still unclear. Recent studies have suggested that inhaled ultrafine particles are able to translocate into the bloodstream. To gain more insight into this potential mechanism, we studied the effect of diesel exhaust particles (DEP, 0.02 and 0.1mg/kg), 48h following their intravenous administration, on systemic inflammation and both pulmonary and cardiac morphological alterations in rats. The intravenous administration of DEP (0.1mg/kg) triggered systemic inflammation characterized by an increase of monocyte and granulocyte numbers. Both doses of DEP caused a reduction of the number of red blood cells (RBC) and haemoglobin concentration. Transmission electron microscopy analysis of RBC after in vitro incubation (5microg/ml) or in vivo administration of DEP, revealed the presence of ultrafine-sized aggregates of DEP within the RBC. Larger aggregates were also taken up by the RBC. Moreover, while the myocardial morphology and capillary bed were not affected by DEP exposure, the lungs of rats exposed to DEP (0.02 and 0.1mg/kg) showed clear evidence of inflammation, characterized by neutrophils infiltration. Stereological analysis revealed an increase in interalveolar wall thickness and a decrease in numbers of alveolar sacs per unit area of lung parenchyma of rat exposed to DEP. We conclude that 48h after their systemic administration, DEP cause systemic and pulmonary morphological alterations.     

Inflammatory and immunological responses to subchronic exposure to endotoxin-contaminated metalworking fluid aerosols in F344 rats

Rats were exposed for 6 h per day in inhalation chambers to a 10 mg/m(3) concentration of metalworking fluid (MWF) contaminated with endotoxin at concentrations of 1813 (low dose) and 20,250 eu/m(3) (high dose) 5 days per week for 8 weeks. It was found that 94.7% of the MWF aerosol particles had diameters in the range of 0.42-4.6 microm, with geometric mean diameter of 1.56 microm. The body weight and pulmonary function parameters were measured every week during the 8 weeks of exposure, whereas bronchoalveolar lavage (BAL) fluid was prepared to measure the inflammatory markers and cytokines after the 8 weeks of exposure. There were no changes in body weight and respiratory function (tidal volume and respiratory frequency) during the 8 weeks of exposure to the MWF containing endotoxins, yet lung weight increased significantly (P < 0.05) in the rats exposed to the MWF both with and without endotoxins. The number of polymorphonuclear (PMN) cells in the BAL fluid increased significantly (P < 0.05) in the rats exposed to MWF with endotoxins, and the levels of cytokines such as IL-4, INF-gamma, IL-1beta, and TNF-alpha also were significantly increased (P < 0.05) compared to the control. The NOx production activity of the BAL cells increased significantly (P < 0.05) in the rats exposed to the MWF with and without endotoxins. Increases in lung weight, number of PMN cells, and levels of extracellular cytokines and NOx were all more significant in the rats exposed to the MWF with endotoxins rather than in those exposed to MWF without endotoxins. In spleen cell cultures, T-cell proliferation activity was decreased, yet cytokine levels (INF-gamma, IL-1beta, IL-4, and TNF-alpha) remained unchanged after repeated exposure to MWF with and without endotoxins. Although the levels of total IgG(1), IgG(2a), and IgE antibodies in the serum were not changed, the levels of endotoxin-specific antibodies, including IgG(2a) and IgE, were increased significantly (P < 0.05) in the rats exposed to endotoxins, but there was not a significant increase in endotoxin-specific IgG(1). When taken together, the results indicate that lung inflammatory responses can be induced without changing pulmonary function after repeated exposure to MWFs contaminated with endotoxins. In addition, endotoxin-specific IgG(2a) and IgE may be effective biomarkers for workers exposed to MWFs contaminated with endotoxins in the workplace.     

Sensitivity to Ozone,Diesel Exhaust Particles,and Standardized Ambient Particulate Matter in Rats with a Listeria Monocytogenes-Induced Respiratory Infection

Ambient particulate matter may increase respiratory allergic skewing of the T-cell-mediated immune response toward a T-helper-2 (Th2) response, with the consequence that the Th1 response develops less well. Successful clearing of a respiratory bacterial infection depends on an adequate Th1 immune response; therefore, the subject would not control the infection as well if exposed to particulate matter. To substantiate this hypothesis, we examined the effect of exposure to diesel exhaust particles (DEP) and urban particulate matter (EHC-93, Ottawa dust) on rats with a Listeria monocytogenes respiratory infection. Since this hypothesis has been confirmed for ozone, we used it as a positive control. Wistar rats were exposed to ozone (2 mg/m³ for 24 h/day for 7 days) and to DEP or to EHC-93 (50 μg/rat intranasally daily for 7 consecutive days). Twenty-four hours after the last exposure, the rats were infected intratracheally with 1 × 10⁶ L. monocytogenes bacteria. The number of L. monocytogenes was determined after 3, 4 and 5 days. Statistically significant increases of the number of L. monocytogenes in rats exposed to ozone were observed in the lungs and spleen at all three times. However, we found no significant differences in the numbers of bacteria that were found in rats exposed to DEP or EHC-93 compared to the saline-treated group at any of the three times. In conclusion, the results of this study do not support the hypothesis that exposure to DEP or EHC-93 reduces subsequent resistance to a respiratory infection in rats.     

Systemic immunotoxicity in AJ mice following 6-month whole body inhalation exposure to diesel exhaust

The purpose of these studies was to determine the effects of subchronic diesel exposure on indicators of systemic immunity in mice. AJ mice were exposed daily for 6 months (6 h/day) to atmospheres containing one of four concentrations (30, 100, 300, and 1000 microg/m(3)) of diluted diesel exhaust (DE) in whole body exposure chambers. The effects of DE were compared to chamber exposure controls receiving fresh air. DE was assessed for effects on systemic immunity by measuring the proliferative response of spleen cells following stimulation with T cell (phytohemagglutinin, or PHA) or B cell (lipopolysaccharide, or LPS) mitogens. The results showed that DE at all exposure levels suppressed the proliferative response of T cells. B cell proliferation was increased at 30 microg/m(3) and was unaffected at the 100, 300, and 1000 microg/m(3) exposures. Polycyclic aromatic hydrocarbons (PAHs) are known to suppress spleen cell mitogenic responses, and it has been hypothesized by several groups that PAHs and perhaps benzo(a)pyrene (BaP)-quinones (BPQs) may be responsible for the effects of DE or diesel exhaust particles (DEP). Therefore, a second purpose of these studies was to determine the effects of in vitro BPQs on AJ mouse spleen cell mitogenic responses and compare to DE in preliminary studies. Unlike DE, BPQs were found to increase T cell proliferation. In addition, analysis of chamber atmospheres showed that there was little if any PAH and BPQs in DE. Therefore, these results demonstrate that because of the absence of BPQs in DE, they are likely not responsible for the immunosuppressive effect of DE on murine spleen cell responses.     

Diesel exhaust affects the abnormal delivery in pregnant mice and the growth of their young

To clarify the toxic effects of diesel exhaust (DE) on delivery in mice and on growth of young, C57Bl-strain females were exposed to 0.3, 1.0, or 3.0 mg diesel exhaust particles (DEP)/m3 or filtered clean air (control) for 4 mo (12 h/day, 7 days/wk). After exposure, some females from each group were examined by necropsy, and the remainders were mated with unexposed males. Estrous females for necropsy who had been exposed to 1.0 mg DEP/m3 had significantly lower uterine weights than the control estrous females. In the mated females, 9.1, 10.0, or 25.0% (0.3, 1.0, or 3.0 mg DEP/m3 of the pregnancies resulted in abnormal deliveries (abortion and unable delivery), but this was not significant. The rate of good nest construction by delivered females exposed to 3.0 mg DEP/m3 was significantly lower. Young were weighed at 11, 14, and 21 days, and weekly from wk 4 to 9 after birth. Body weights of male young of dams exposed to 1.0 or 3.0 mg DEP/m3 were significantly lower at 6 and 8 wk of age. Body weights of female young of dams exposed to 1.0 or 3.0 mg DEP/m3 were also significantly lower at 6, 8, and 9 wk. Vaginal orifices of young female mice whose dams were exposed to 0.3 and 1.0 mg DEP/m3 opened significantly earlier. The young were killed at 30 or 70 days during deep anesthesia, and their body weights, organ weights, and body lengths were measured. Anogenital distance (AGD) of 30-day-old males whose dams were exposed to 0.3 mg DEP/m3 was significantly shorter than that of the controls. Weights of thymus and ovary in 30-day-old females whose dams were exposed to 3.0 mg DEP/m3 were significantly lower. In 70-day-old males of dams exposed to 3.0 mg DEP/m3, body weights were significantly lower and AGD was significantly shorter. Weights of adrenals, testes, and seminal vesicles in 70-day-old males with dams exposed to 1.0 mg DEP/m3 were significantly lower. In 70-day-old females with dams exposed to DE, body weights in the 3.0-mg DEP/m3 group were significantly lower, and weights of adrenals, liver, and thymus in the 1.0-mg DEP/m3 group were significantly lower. Thymus weights in 70-day-old females with dams exposed to 0.3 mg DEP/m3 were significantly lower. Crown-rump length (CR) in 70-day-old females with dams exposed to 1.0 or 3.0 mg DEP/m3 was significantly shorter. These results show that toxic substances in DE might cause abnormal delivery in mice, and that exposed females affected the growth and sexual maturation of their young.     

DIESEL EXHAUST AFFECTS THE ABNORMAL DELIVERY IN PREGNANT MICE AND THE GROWTH OF THEIR YOUNG

To clarify the toxic effects of diesel exhaust (DE) on delivery in mice and on growth of young, C57Bl-strain females were exposed to 0.3, 1.0, or 3.0 mg diesel exhaust particles (DEP)/m 3 or filtered clean air (control) for 4 mo (12 h/day, 7 days/wk). After exposure, some females from each group were examined by necropsy, and the remainders were mated with unexposed males. Estrous females for necropsy who had been exposed to 1.0 mg DEP/m 3 had significantly lower uterine weights than the control estrous females. In the mated females, 9.1, 10.0, or 25.0% (0.3, 1.0, or 3.0 mg DEP/m 3) of the pregnancies resulted in abnormal deliveries (abortion and unable delivery), but this was not significant. The rate of good nest construction by delivered females exposed to 3.0 mg DEP/m 3 was significantly lower. Young were weighed at 11, 14, and 21 days, and weekly from wk 4 to 9 after birth. Body weights of male young of dams exposed to 1.0 or 3.0 mg DEP/m 3 were significantly lower at 6 and 8 wk of age. Body weights of female young of dams exposed to 1.0 or 3.0 mg DEP/m 3 were also significantly lower at 6, 8, and 9 wk. Vaginal orifices of young female mice whose dams were exposed to 0.3 and 1.0 mg DEP/m 3 opened significantly earlier. The young were killed at 30 or 70 days during deep anesthesia, and their body weights, organ weights, and body lengths were measured. Anogenital distance (AGD) of 30-day-old males whose dams were exposed to 0.3 mg DEP/m 3 was significantly shorter than that of the controls. Weights of thymus and ovary in 30-day-old females whose dams were exposed to 3.0 mg DEP/m 3 were significantly lower. In 70-day-old males of dams exposed to 3.0 mg DEP/m 3, body weights were significantly lower and AGD was significantly shorter. Weights of adrenals, testes, and seminal vesicles in 70-day-old males with dams exposed to 1.0 mg DEP/m 3 were significantly lower. In 70-day-old females with dams exposed to DE, body weights in the 3.0-mg DEP/m 3 group were significantly lower, and weights of adrenals, liver, and thymus in the 1.0-mg DEP/m 3 group were significantly lower. Thymus weights in 70-day-old females with dams exposed to 0.3 mg DEP/m 3 were significantly lower. Crown-rump length (CR) in 70-day-old females with dams exposed to 1.0 or 3.0 mg DEP/m 3 was significantly shorter. These results show that toxic substances in DE might cause abnormal delivery in mice, and that exposed females affected the growth and sexual maturation of their young.     

Combined inhaled diesel exhaust particles and allergen exposure alter methylation of T helper genes and IgE production in vivo.

Changes in methylation of CpG sites at the interleukin (IL)-4 and interferon (IFN)-gamma promoters are associated with T helper (Th) 2 polarization in vitro. No previous studies have examined whether air pollution or allergen exposure alters methylation of these two genes in vivo. We hypothesized that diesel exhaust particles (DEP) would induce hypermethylation of the IFN-gamma promoter and hypomethylation of IL-4 in CD4+ T cells among mice sensitized to the fungus allergen Aspergillus fumigatus. We also hypothesized that DEP-induced methylation changes would affect immunoglobulin (Ig) E regulation. BALB/c mice were exposed to a 3-week course of inhaled DEP exposure while undergoing intranasal sensitization to A. fumigatus. Purified DNA from splenic CD4+ cells underwent bisulfite treatment, PCR amplification, and pyrosequencing. Sera IgE levels were compared with methylation levels at several CpG sites in the IL-4 and IFN-gamma promoter. Total IgE production was increased following intranasal sensitization A. fumigatus. IgE production was augmented further following combined exposure to A. fumigatus and DEP exposure. Inhaled DEP exposure and intranasal A. fumigatus induced hypermethylation at CpG(-45), CpG(-53), CpG(-205) sites of the IFN-gamma promoter and hypomethylation at CpG(-408) of the IL-4 promoter. Altered methylation of promoters of both genes was correlated significantly with changes in IgE levels. This study is the first to demonstrate that inhaled environmental exposures influence methylation of Th genes in vivo, supporting a new paradigm in asthma pathogenesis.     

Altered immune response during cadmium administration in mice

C57BL/6 mice were administered 50 or 200 ppm of Cd as CdCl2 in the drinking water for either 3 to 4 (short term) or 9 to 11 (long term) weeks. In other experimental designs, mice were exposed orally to 300 ppm of Cd or injected with 2.5 mg/kg of Cd ip. The proliferative response to the T cell mitogens Con A and PHA was increased in cultures of spleen cells from orally treated mice in most of the experiments performed. After primary immunization with sheep red blood cells, the number of IgM antibody forming cells per 10(7) spleen cells was also moderately higher in mice exposed to 50 or 200 ppm of Cd for short or long term. In contrast, long-term exposure to 300 ppm of Cd depressed the antibody response to SRBC. Administration of ZnCl2 prevented the enhancement of the PFC response in mice orally administered 50 ppm of Cd. The capacity to suppress the antibody response of spleen cells preincubated with sodium periodate was decreased after short-term oral or ip. Cd administration but was completely or partially recovered after long-term exposure to either 50 or 200 ppm of Cd.     

Pulmonary exposure to diesel exhaust particles induces airway inflammation and cytokine expression in NC/Nga mice

Although several studies have reported that diesel exhaust particles (DEP) affect cardiorespiratory health in animals and humans, the effect of DEP on animal models with spontaneous allergic disorders has been far less intensively studied. The Nc/Nga mouse is known to be a typical animal model for human atopic dermatitis (AD). In the present study, we investigated the effects of repeated pulmonary exposure to DEP on airway inflammation and cytokine expression in NC/Nga mice. The animals were randomized into two experimental groups that received vehicle or DEP by intratracheal instillation weekly for six weeks. Cellular profiles of bronchoalveolar lavage (BAL) fluid and expressions of cytokines and chemokines in both the BAL fluid and lung tissues were evaluated 24 h after the last instillation. The DEP challenge produced an increase in the numbers of total cells, neutrophils, and mononuclear cells in BAL fluid as compared to the vehicle challenge (P<0.01). DEP exposure significantly induced the lung expressions of interleukin (IL)-4, keratinocyte chemoattractant (KC), and macrophage inflammatory protein (MIP)-1 when compared to the vehicle challenge. These results indicate that intratracheal exposure to DEP induces the recruitment of inflammatory cells, at least partially, through the local expression of IL-4 and chemokines in NC/Nga mice.     

Cardiovascular and thermoregulatory responses of unrestrained rats exposed to filtered or unfiltered diesel exhaust

Diesel exhaust has been associated with adverse cardiovascular and pulmonary health effects. The relative contributions of the gas phase and particulate components of diesel exhaust are less well understood. We exposed telemetered Wistar-Kyoto rats to air or diesel exhaust that was either filtered (F) or unfiltered [gas-phase plus diesel exhaust particles (DEP)], containing ~1.9 mg/m3 of particulate matter for 5 h/day; 5 days/week for 4 consecutive weeks. Blood pressure (BP), core temperature (T(c)), heart rate (HR), and cardiac contractility (CC) estimated by the QA interval were monitored by radiotelemetry during exposure as well as during a 2-week period of recovery. Pulmonary injury and inflammation markers were analysed after 2-day, and 4 weeks of exposure, and 2-week recovery. Exposure to F or DEP was associated with a trend for a reduction in BP during weeks 1, 2 and 4. A reduction in HR in the DEP group was apparent during week 4. Exposure to DEP but not F was associated with significant reduction in CC over weeks 1-4. There was also a slight elevation in T(c) during DEP exposure. All telemetry parameters were normal during recovery at night and a 2-week recovery period. Neutrophilic inflammation in bronchoalveolar lavage fluid was evident after 2 days and 4 weeks of exposure to F and DEP. There were no signs of inflammation after 2-week recovery. We found a significant decrease in CC and slight reduction in BP. Exposure to DEP and F is associated with pulmonary inflammation, and mild effects on HR, BP, and T(c) but there is a marked effect of DEP on CC.     

Immunoglobulin and Lymphocyte Responses Following Silica Exposure in New Zealand Mixed Mice

Epidemiological studies have shown strong associations between silica exposure and several autoimmune diseases, including scleroderma and systemic lupus erythematosus.We previously reported that the New Zealand mixed (NZM) mouse develops silicosis and exacerbated autoimmunity following crystalline silica exposure, including increased levels of autoantibodies, proteinuria, circulating immune complexes, pulmonary fibrosis, and glomerulonephritis. In this study, the NZM mouse was used to examine changes in immune activation following silica exposure by measuring levels of immunoglobulin, cytokines and lymphocyte populations. Levels of immunoglobulin (Ig) G1 were significantly decreased from 1124 ± 244 µg/ml in saline exposed mice to 614 ± 204 µg/ml in silica-exposed mice, suggesting a decrease in the Th2 response. The levels of tumor necrosis factor (TNF)-α were significantly increased (1.5-fold) in the bronchoalveolar lavage fluid of the silica-exposed mice as compared to the saline-exposed mice. The number of B1a B cells were significantly increased sixfold within the superficial cervical lymph nodes of silica-exposed mice as compared with saline-exposed mice. Following silica exposure, CD4+T cells significantly increased threefold within the superficial cervical lymph nodes. During this increase in the number of CD4+T cells, the number of CD4+CD25+ regulatory T cells was not significantly changed, altering the ratio of regulatory T cells to T helper cells from 1:5 to 1:8 following silica exposure. Therefore, the silica-induced alterations in immunoglobulin levels, increased TNF-α, increased B1a B cells and CD4+ T cells, with decreased regulatory T cells, may provide an environment that allows for increased autoreactivity. These studies begin to provide possible mechanisms for environmentally induced autoimmune diseases that have been reported in many epidemiological studies.     

Effects of inhalation exposure to a high-boiling (288 to 454 degrees C) coal liquid

Coal liquids have been evaluated in a variety of short-term toxicological assays; however, few studies have been conducted to determine the systemic effects after inhalation exposure to these materials. To extend the data base on potential health effects from coal liquefaction materials, we performed a study with solvent refined coal (SRC)-II heavy distillate (HD). Fischer-344 rats were exposed for 6 hr/day, 5 days/week for 5 or 13 weeks to an aerosol of HD (boiling range, 288 to 454 degrees C) at concentrations of 0.69, 0.14, 0.03, or 0.0 mg/liter of air for the high, middle, low, and control groups, respectively. Survival through 13 weeks of exposure was greater than 90% for all groups; body weights for exposed animals were decreased in a dose-dependent manner. Significant increases in liver weights and decreases in thymus and ovary weights were observed for treated animals compared with controls. There were also significant treatment-related decreases in erythrocytes, hemoglobin, volume of packed red blood cells, lymphocytes, eosinophils, and total white blood cells. After 5 weeks of exposure serum cholesterol concentrations increased in a dose-dependent manner for both sexes and serum triglyceride amounts decreased for males but not for females. After 13 weeks of exposure, high-dose animals had significant increases in cholesterol (males only), triglycerides, blood urea nitrogen, and serum glutamic pyruvic transaminase (SGPT; males) and significant decreases in albumin, SGPT (females), and lactate dehydrogenase (LDH). Examination of bone-marrow preparations from exposed animals demonstrated consistent decreases in the degree of cellularity, suggesting that this organ is a target for HD. Microscopic evaluation of organ sections indicated exposure-related changes for nasal mucosa, pulmonary macrophages, thymus, liver, kidney, bone marrow, ovaries, and cecum. Results from this study indicated dose-dependent increases in the severity of the lesions observed, with few effects in the low-exposure group that were attributable to the exposure.     

A subchronic inhalation study with unleaded petrol in rats

The effect of intermittent 2 months' exposure to 2 g/m3 unleaded petrol on the hypothalamo-pituitary-thyroid-adrenal system was evaluated by measuring hypothalamic noradrenaline (NA), serum corticosterone (CS), thyroxine (T4) and adrenal catecholamine (CA) levels in male rats. Serum CS and adrenal CA were increased and hypothalamic NA was decreased by exposure. No changes were observed in serum T4. Exposure induced an increase in spleen, kidney, liver and lung weights; weights of adrenals and hypothalamus were not changed. All the petrol-induced effects depended on the length of exposure. Rats exposed to petrol gained less weight than controls. The results suggest a non-specific stress response in the rat.     

Effects of subchronic inhalation exposure to ethyl tertiary butyl ether on splenocytes in mice

Ethyl tertiary-butyl ether (ETBE) is a motor fuel oxygenate used in reformulated gasoline. The current use of ETBE in gasoline or petrol is modest but increasing. To investigate the effects of ETBE on splenocytes, mice were exposed to 0 (control), 500 ppm, 1750 ppm, or 5000 ppm of ETBE by inhalation for 6 h/day for 5 days/wk over a 6- or 13-week period. Splenocytes were harvested from the control and exposed mice, and the following cell phenotypes were quantified by flow cytometry: (1) B cells (PerCP-Cy5.5-CD45R/B220), (2) T cells (PerCP-Cy5-CD3e), (3) T cell subsets (FITC-CD4 and PE-CD8a), (4) natural killer (NK) cells (PE-NK1.1), and (5) macrophages (FITC-CD11b). Body weight and the weight of the spleen were also examined. ETBE-exposure did not affect the weight of the spleen or body weight, while it transiently increased the number of RBC and the Hb concentration. The numbers of splenic CD3+, CD4+, and CD8+ T cells, the percentage of CD4+ T cells and the CD4+/CD8+ T cell ratio in the ETBE-exposed groups were significantly decreased in a dose-dependent manner. However, ETBE exposure did not affect the numbers of splenic NK cells, B cells, or macrophages or the total number of splenocytes. The above findings indicate that ETBE selectively affects the number of splenic T cells in mice.