Body Inhalation Exposure Research Articles

Mutagenicity evaluation of methyl tertiary- butyl ether in multiple tissues of transgenic rats following whole body inhalation exposure.

Methyl tertiary-butyl ether (MTBE) is used as a component of motor vehicle fuel to enhance combustion efficiency and to reduce emissions of carbon monoxide and nitrogen oxides. Although MTBE was largely negative in the in vitro and in vivo genotoxicity studies, isolated reports of positive findings along with the observation of tumors in the rat cancer bioassays raised concern for its in vivo mutagenic potential. To investigate this, transgenic male Big Blue Fischer 344 rats were exposed to 0 (negative control), 400, 1000, and 3000 ppm MTBE via whole body inhalation for 28 consecutive days, 6 h/day. Mutant frequencies (MF) at the cII locus of the transgene in the nasal epithelium (portal of entry tissue), liver (site of primary metabolism), bone marrow (rapidly proliferating tissue), and kidney (tumor target) were analyzed (5 rats/exposure group) following a 3-day post-exposure manifestation period. MTBE did not induce a mutagenic response in any of the tissues investigated. The adequacy of the experimental conditions to detect induced mutations was confirmed by utilizing tissue samples from animals treated with the known mutagen ethyl nitrosourea. These data provide support to the conclusion that MTBE is not an in vivo mutagen and male rat kidney tumors are not likely the result of a mutagenic mode of action.

Lung carcinogenicity by whole body inhalation exposure to Anatase-type Nano-titanium Dioxide in rats.

To investigate the carcinogenicity of anatase-type nano-titanium dioxide (aNTiO2), F344/DuCrlCrlj rats were exposed to aNTiO2 aerosol at concentrations of 0, 0.5, 2, and 8 mg/m3. The rats were divided into 2 groups: carcinogenicity study groups were exposed for two years, and satellite study groups were exposed for one year followed by recovery for 1 day, 26 weeks, and 52 weeks after the end of exposure. In the carcinogenicity groups, bronchiolo-alveolar carcinomas were observed in two 8 mg/m3-exposed males, showing an increasing trend by Peto's test. However, this incidence was at the upper limit of JBRC's historical control data. Bronchiolo-alveolar adenomas were observed in 1, 2, 3, and 4 rats of the 0, 0.5, 2, and 8 mg/m3-exposed females and were not statistically significant. However, the incidence in the 8 mg/m3-exposed females exceeded JBRC's historical control data. Therefore, we conclude there is equivocal evidence for the carcinogenicity of aNTiO2 in rats. No lung tumors were observed in the satellite groups. Particle-induced non-neoplastic lesions (alveolar epithelial hyperplasia and focal fibrosis) were observed in exposed males and females in both the carcinogenicity and satellite groups. Increased lung weight and neutrophils of bronchoalveolar lavage fluid were observed in the 8 mg/m3-exposed carcinogenicity groups. The aNTiO2 deposited in the lungs of the satellite group rats was decreased at 26 weeks after the end of exposure compared to 1 day after the end of exposure. At 52 weeks after the end of exposure, the decreased level was the same at 26 weeks after the end of exposure.

Incense smoke (IS) inhalation exposure system: Physicochemical characterization, IS particle deposition and clearance in human airway using MPPD model

Incense smoke (IS) is source of indoor air pollution and key risk for diverse human diseases. Less information is available regarding controlled IS rodent inhalation exposure system and IS particulate matter (PM) deposition in human airways. Study aimed to demonstrate stable ISPM physicochemical parameters of 10 incense products inside the customized whole body inhalation exposure chamber (without animal) connected to smoke generation unit via aerosol mixing device. IS analyzed for size segregated PM emission, ISPM in vitro aerodynamics (MMAD and GSD determination), fine and ultrafine particle's SEM, SEM-EDX and PAH analysis. Using real life exposure scenario by utilizing MMAD, GSD and PM concentration after Tier 1 exposure assessment as key input parameters, ISPM dosimetry in infant (3 months) and adult (21 years male and female) human airways was calculated using multiple-path particle dosimetry (MPPD 3.04) modeling. Mass median aerodynamic diameter (MMAD) and geometric standard deviation (GSD) ranged between 0.55 and 2.10 μm and 1.22 to 1.77 (polydisperse) respectively. PM1.0 and PM0.1 showed multiple morphology and presence of heavy and trace elements. PAH like acenaphthylene, anthracene, fluorene, naphthalene and phenanthrene were detected (0.84–143.17 μg/g). MPPD results showed higher ISPM deposition in pulmonary region and lowest in trachea bronchial region. ISPM deposition in tissue was higher in lower, peripheral lung as compared to upper and central lung tissue. Whole body inhalation exposure system showed stable IS atmosphere (physicochemical parameters) indicating the device suitability in future inhalation studies. MPPD ISPM deposition fraction and clearance data showed deep lung penetrating and retention behavior with higher risk in infant followed by female and then male. These modeled particle deposition and clearance data may be useful in risk assessment analysis of IS.

Investigation of the potential mutagenicity of ethyl tertiary-butyl ether in the tumor target tissue using transgenic Big Blue Fischer 344 rats following whole body inhalation exposure.

Ethyl tertiary-butyl ether (ETBE) is a fuel oxygenate used for the efficiency of motor vehicle fuels and their octane ratings. ETBE has been reported to induce liver adenomas in male rats in a 2-year bioassay at the highest inhalation concentration tested of 5000 ppm. To investigate the potential mutagenicity of ETBE in the liver, male Big Blue Fischer 344 rats were exposed for 28 consecutive days (6h/day) to 0, 500, 1500, and 5000 ppm ETBE. The treated rats were sacrificed 3 days post-exposure and the frequencies of cII mutants were evaluated in the liver and bone marrow tissues. The mutant frequency (MF) of the liver in the negative control group was 36.3 × 10-6 and this value was not significantly differentin ETBE-exposed animals (39.4, 37.3, and 45.9 × 10-6 in 500, 1500, and 5000 ppm groups, respectively). In the bone marrow, the mean MF in the negative control was 32.9 × 10-6 which was not different from the means of the exposed groups (33.8, 22.6, and 32.0 × 10-6 for groups exposed to 500, 1500 and 5000 ppm, respectively). These data, along with consistent negative response reported in the literature for other apical genotoxicity endpoints informs that mutagenicity is not likely the initial key event in the mode of action for ETBE-induced hepatocarcinogenesis in the rat.

Toxicokinetic evaluation of the common indoor air pollutant, α-pinene, and its potential reactive metabolite, α-pinene oxide, following inhalation exposure in rodents

The toxicokinetic behavior of α-pinene and its potential reactive metabolite, α-pinene oxide, was investigated following whole body inhalation exposure to 50 and 100 ppm α-pinene in rats and mice for 6 h per day for 7d. In both species and sexes, the maximum blood concentration (Cmax) increased more than proportionally while the increase in area under the concentration time curve (AUC) was proportional to the exposure concentration. When normalized to the calculated dose (D), both Cmax/D (male rats, 12.2–54.5; female rats, 17.4–74.1; male mice, 7.41–14.2; female mice, 6.59–13.0 (ng/mL)/(mg/kg)) and AUC/D (male rats, 28.9–31.1; female rats, 55.8–56.8; male mice, 18.1–19.4; female mice, 19.2–22.5 (h*ng/mL)/(mg/kg)) in rats were higher than in mice and in female rats were higher than in male rats; no sex difference was observed in mice. α-Pinene was eliminated from blood with half-lives between 12.2 and 17.4 h in rats and 6.18–19.4 h in mice. At the low dose, the ratio of α-pinene oxide to α-pinene, based on Cmax and AUC, respectively, was 0.200–0.237 and 0.279–0.615 in rats and 0.060–0.086 and 0.036–0.011 in mice demonstrating lower formation of the oxide in mice than in rats. At the high dose, the ratio decreased considerably in both species pointing to saturation of pathways leading to the formation of α-pinene oxide. α-Pinene and the oxide were quantified in the mammary glands of rats and mice with tissue to blood ratios of ≥23 demonstrating retention of these analytes in mammary glands. The findings of epoxide formation and species- and sex-differences in systemic exposure may be important in providing context and relating animal findings to human exposures.

Bisphenol A triggers the malignancy of nasopharyngeal carcinoma cells via activation of Wnt/β-catenin pathway

It is critical to understand the risk factors responsible for the tumorigenesis and progression of nasopharyngeal carcinoma (NPC). Bisphenol A (BPA) can regulate the estrogenic signals to modulate cancer progression, while its roles in NC were not investigated. Our present study revealed that the BPA can increase proliferation and migration of NPC cells while decrease the chemosensitivity to doxorubicin (Dox). The inhibitor of GSK-3β/β-catenin (LiCl) can restore BPA-induced cell proliferation of NPC cells, which is due to that BPA can decrease phosphorylation while increase expression and nucleus localization of β-catenin. Mechanistically, BPA can increase the mRNA stability of β-catenin (encoded by CTNNB1) via suppressing the expression of miR-214-3p, which can direct target the 3’UTR of β-catenin mRNA. Further, BPA can decrease phosphorylation of β-catenin via repressing the expression of CK1α. Collectively, our data showed that BPA can trigger the proliferation and malignancy of NPC cells via activation of Wnt/β-catenin pathway. It indicated that body accumulation and inhalation exposure of BPA might be a risk factor for NPC development.

Carcinogenicity of butyl 2,3-epoxypropyl ether in rats and mice by whole body inhalation for two years.

Butyl 2,3-epoxypropyl ether (CAS No. 2426-08-6, synonym: n-butylglycidyl ether, BGE) was exposed by whole body inhalation to F344 rats and BDF1 mice of both sexes (50 animals per group) 6 hours per day, 5 days per week for 104 weeks at targeted concentrations of 0, 10, 30 or 90 ppm (v/v) for rats and 0, 5, 15 or 45 ppm for mice. In rats, 90 ppm of BGE increased the incidences of nasal squamous cell carcinomas in both sexes. Nasal adenomas and splenic mononuclear cell leukemia were increased in male rats exposed to 30 ppm. Splenic mononuclear cell leukemia was increased in female rats by trend test. Non-neoplastic nasal lesions, such as squamous cell hyperplasia with atypia, squamous cell metaplasia and the inflammation of the respiratory region and atrophy of the olfactory epithelium were increased in both sexes in a dose-dependent manner. In mice, the incidences of histiocytic sarcomas of the uterus in female mice were increased in a dose-dependent manner and the incidences of nasal hemangiomas in both sexes were increased in a dose-dependent manner. Nasal squamous cell carcinoma, a rare tumor, was observed, although not statistically significant, in both sexes. Non-neoplastic lesions such as nodular hyperplasia of the transitional epithelium and cuboidal changes of the respiratory epithelium in the nasal cavity, were increased both in males and females in a dose-dependent manner. The present study demonstrated clear evidence of carcinogenicity of BGE in both rats and mice by the 2-year whole body inhalation exposure.

Toxicity of humidifier disinfectant polyhexamethylene guanidine hydrochloride by two-week whole body-inhalation exposure in rats.

The use of polyhexamethylene guanidine hydrochloride (PHMG·HCl) as a humidifier disinfectant caused an outbreak of pulmonary disease, leading to the deaths of pregnant women and children in South Korea. However, limited information is available on the inhalation toxicity of PHMG·HCl. Therefore, this study aimed to characterize the subacute inhalation toxicity of PHMG·HCl by whole-body exposure in rats. F344 rats were exposed to 0 mg/m3, 1 mg/m3, 5 mg/m3, or 25 mg/m3 of PHMG·HCl for 6 h/day, 5 days/week for two weeks via whole-body inhalation. Emaciation and rale were observed in rats in the 25 mg/m3 PHMG·HCl group. Significant changes in body weight, hematology, serum chemistry and organ weight were observed in all PHMG·HCl-exposed groups. Gross lesions showed ballooning or red focus in the lungs of rats in the PHMG·HCl-exposed groups. In histopathological examination, most of histological lesions (including degeneration, atrophy, ulcer, inflammatory cell infiltration, inflammation, and fibrosis in nasal cavity, larynx, trachea, and lungs) indicated tissue damage by PHMG·HCl in all PHMG·HCl-exposed groups. Additionally, atrophy of the spleen, thymus, and reproductive organs; immaturity of the testes; and cell debris in the epididymides were affected by the reduction in body weight in PHMG·HCl-exposed groups. In conclusion, two-week repeated whole-body inhalation exposure of rats to PHMG·HCl reveled toxic effects on the respiratory system and secondary effects on other organs. The results of this study indicate that the no observable adverse effect level (NOAEL) for PHMG·HCl is below 1 mg/m3.

雌雄ラットを用いたアナターゼ型ナノ酸化チタン（ANN-TiO2）の2週間吸入毒性試験

雌雄ラットを用いたアナターゼ型ナノ酸化チタン（ANN-TiO2）の2週間吸入毒性試験

多層カーボンナノチューブ（MWNT-7）のラット全身吸入ばく露による肺発がん

多層カーボンナノチューブ（MWNT-7）のラット全身吸入ばく露による肺発がん

MWCNTのマウス全身暴露吸入における原末と高分散処理検体（Taquann法）の肺沈着量の比較

MWCNTのマウス全身暴露吸入における原末と高分散処理検体（Taquann法）の肺沈着量の比較

多層カーボンナノチューブ（MWNT-7）の全身吸入暴露によるラットの発がん性試験

多層カーボンナノチューブ（MWNT-7）の全身吸入暴露によるラットの発がん性試験

Evaluation of health risk caused by chloroform in drinking water

In this paper, the results of cancer risk evaluation for chloroform in tap water at Cracow city (Poland) are presented. Cracow is a large city which is mostly supplied with surface water. The raw water seasonally has a high content of organic matter. Since chlorine is the basic disinfectant, organic compounds react with chlorine which results in the disinfection byproducts formation, such as trihalomethanes, in delivered water. Calculations were made, separately in male and female, for average chloroform concentration in the whole city as well as with respect to the season and supply zone. The study is based on the multipathway method which takes into account three ways in which chloroform contained in the tap water enters the human body (oral ingestion, dermal absorption, and inhalation exposure). The total cancer risk is at the acceptable level (between 10−6 and 10−5) because the chloroform concentration in water supply system only incidentally exceeds the maximum allowable concentration regulated by Polish Ministry of Health, i.e. 30 µg/L.

The acute exposure effects of inhaled nickel nanoparticles on murine endothelial progenitor cells

Introduction: The discovery of endothelial progenitor cells (EPCs) may help to explain observed cardiovascular effects associated with inhaled nickel nanoparticle exposures, such as increases in vascular inflammation, generation of reactive oxygen species, altered vasomotor tone and potentiated atherosclerosis in murine species.Methods: Following an acute whole body inhalation exposure to 500 µg/m3 of nickel nanoparticles for 5 h, bone marrow EPCs from C57BL/6 mice were isolated. EPCs were harvested for their RNA or used in a variety of assays including chemotaxis, tube formation and proliferation. Gene expression was assessed for important receptors involved in EPC mobilization and homing using RT-PCR methods. EPCs, circulating endothelial progenitor cells (CEPCs), circulating endothelial cells (CECs) and endothelial microparticles (EMPs) were quantified on a BD FACSCalibur to examine endothelial damage and repair associated with the exposure.Results and conclusions: Acute exposure to inhaled nickel nanoparticles significantly increased both bone marrow EPCs as well as their levels in circulation (CEPCs). CECs were significantly elevated indicating that endothelial damage occurred due to the exposure. There was no significant difference in EMPs between the two groups. Tube formation and chemotaxis, but not proliferation, of bone marrow EPCs was impaired in the nickel nanoparticle exposed group. These results coincided with a decrease in the mRNA of receptors involved in EPC mobilization and homing. These data provide new insight into how an acute nickel nanoparticle exposure to half of the current Occupational Safety & Health Administration (OSHA) permissible exposure limit may adversely affect EPCs and exacerbate cardiovascular disease states.

Subacute inhalation toxicity assessment of fly ash from industrial waste incinerators

Fly ash from industrial waste incinerators has been a significant concern because of their constituent toxic heavy metals and organic compounds. The objective of this study was to identify the subacute inhalation toxicity of fly ash from industrial waste incinerators, using whole body inhalation exposure chambers. Male and female groups of Sprague-Dawley rats were exposed to fly ash by inhalation of concentrations of 0, 50, 100, 200 mg/m3, for 6 h/day, 5 days/week for 4 weeks. There was no significant difference in body weight, and relative organ weight to body weight, between the exposure groups and the control group. Hematological examinations revealed a significant increase of monocyte counts in fly ash exposed rats and brown pigment laden macrophage was found in the lungs of rats exposed to high concentration of fly ash. A decrease of blood glucose levels and an increase in glutamate oxaloacetate transaminase activity were observed in fly ash treated rats. There was also a significant increase of lactate dehydrogenase levels in rat blood exposed fly ash. A significant dose-dependent increase of DNA damage was found in lymphocytes, spleen, bronchoalveolar lavage, liver, lung, and thymus of rats exposed to fly ash. In addition, the level of lipid peroxidation was increased in the plasma of rats exposed to a high concentration of fly ash. These results suggest that inhalation of fly ash from industrial waste incinerators can induce histopathologic, hematological, and serum biochemical changes and oxidative damage.

Whole body inhalation exposure of multi-walled carbon nanotube by using an acoustical dust generator and measurements of its body burden in lung

Whole body inhalation exposure of multi-walled carbon nanotube by using an acoustical dust generator and measurements of its body burden in lung

Endosulfan-induced hepatotoxicity is route of exposure independent in rats

Endosulfan is an important hepatotoxic agent that generates free oxygen radicals in liver. With the widespread use of endosulfan in agriculture, human beings are most likely to be exposed to it by eating food contaminated with endosulfan, exposure to its low levels by skin contact with contaminated soil, smoking cigarettes made from tobacco that has endosulfan residues on it, or by nose and whole body inhalation exposure in the farms during its application. Since endosulfan is a frequently used pesticide, and the incidence of toxic injury to the liver tissue in relation to its widespread use reported in the literature, we considered it necessary to investigate whether endosulfan-induced liver injury could be route of exposure dependent. Eighteen mature male albino Wistar rats, weighing between 180 and 220 g, were used in this study. The hepatotoxic effects of oral administration of endosulfan (5 mg/kg body weight) daily for 30 days, and 30 days whole body inhalation exposure to ungraded concentration of endosulfan were investigated in rats using serum liver enzymes and histopathological assay. At the end of the experimental period, serum alanine aminotransferase, aspartate amino transferase, alkaline phosphatase, and creatine kinase activities obtained for the group of rats exposed orally to endosulfan were not significantly different (p ≥ 0.05) from the activities obtained for rats exposed by whole body inhalation. However, the activity of these enzymes obtained for the rats exposed to endosulfan by both oral and inhalation routes were significantly increased (p ≤ 0.05) compared, respectively, to the control. Also, on microscopic examination, the liver tissues of experimental groups exhibited severe damage histopathologically. The results of the enzyme and histological analyses showed that both oral and whole body inhalation exposure to endosulfan may cause liver tissue damage in rats. The exposure to endosulfan in rats caused liver tissue damage independent of the route of exposure.

An Inhalation Exposure System for the Oil Dispersant COREXIT® EC9500A

An automated whole body inhalation exposure system capable of exposing 12 rats was designed to examine the potential health effects of the oil dispersant (COREXIT® EC9500A [CE]) used during the Deepwater Horizon oil spill. A computer‐controlled syringe pump injected the CE into an atomizer where droplets and vapor were formed and mixed with diluent air. The aerosolized CE was passed into a customized exposure chamber where a calibrated light scattering instrument estimated the real‐time particle mass concentration of the aerosol in the chamber. A custom software GUI was developed to record and control parameters during the exposures. Software feedback loops controlled the chamber aerosol concentration and pressure throughout each exposure. Particle size of the dispersant aerosol was measured and shown to have a number peak at 271 nm and a mass peak at 961 nm. The method used to determine the chamber concentration was a modification of the acidified methylene blue spectrophotometric assay for anionic surfactants. Filter samples were extracted in 5 mL water/filter using sonication for 30 min and diluted 1/10 for analyses. CE filter content was extrapolated from a standard plot using the original CE (4–130 nL/mL water) material and adjusted for density. Five automated 5 hour animal exposures were performed that produced controlled and consistent CE concentrations (27.1 ± 2.9 mg/m3).

Whole-body inhalation exposure to 1-bromopropane suppresses the IgM response to sheep red blood cells in female B6C3F1 mice and Fisher 344/N rats

1-Bromopropane (1-BP) is categorized as a high-production-volume chemical and is currently used in the manufacture of pharmaceuticals, pesticides, and other chemicals. Its usage is estimated to be around 5 million pounds per year, resulting in the potential for widespread exposure in the workplace. Case reports and animal studies have suggested exposure to this compound may cause adverse reproductive and neurological effects. Using a battery of immunological assays, the immunotoxicity of 1-BP after whole body inhalation exposure in both mice and rats was evaluated. Significant decreases in the spleen immunoglobulin (Ig) M response to sheep red blood cells (SRBC) were observed in both mice (125–500 ppm) and rats (1000 ppm) after exposure to 1-BP for 10 wk. In addition, total spleen cells and T cells were significantly decreased after approximately 4 wk of 1-BP exposure in both mice (125–500 ppm) and rats (1000 ppm). No change in natural killer (NK) cell activity was observed. The observed alterations in spleen cellularity, phenotypic subsets, and impairment of humoral immune function across species raise further concern about human exposure to 1-BP and demonstrate the need for additional investigations into potential adverse health effects.

Inhalation of tobacco smoke induces increased proliferation of urinary bladder epithelium and endothelium in female C57BL/6 mice

Cigarette smoking is the major environmental risk factor for bladder cancer in humans. Aromatic amines, potent DNA-reactive bladder carcinogens present in cigarette smoke, contribute significantly. However, increased cell proliferation, caused by direct mitogenesis or in response to cytotoxicity, may also play a role since urothelial hyperplasia has been observed in human cigarette smokers. We examined the urothelial effects of cigarette smoke (whole body inhalation exposure (Teague) system) in female C57BL/6 mice at various times in two studies, including reversibility evaluations. In both studies, no urothelial hyperplasia was observed by light microscopy in any group. However, in study 1, the Ki-67 labeling index (LI) of the urothelium was significantly increased in the smoke exposed group compared to controls through 3 months, but was not present at 6, 9 or 12 months even with continued exposures. In the groups that discontinued smoke exposure, it returned to the same levels as controls or lower. In study 2, the bromodeoxyuridine LI was similar to controls on day 1 but significantly increased at 5 days in the smoke exposed group. In the group that discontinued smoke exposure for 2 days, the LI was increased compared to controls but not significantly. Superficial urothelial cell cytotoxicity and necrosis were detectable by scanning electron microscopy at 5 days. Changes in LI of submucosal endothelial cells generally followed those of the urothelium and effects were reversible upon cessation of exposure. The increased urothelial proliferation appeared to be due to superficial cell cytotoxicity with consequent regeneration.