A population-based urinary and plasma metabolomics study of environmental exposure to cadmium.

BACKGROUND: The application of metabolomics-based profiles in environmental epidemiological studies is a promising approach to refine the process of health risk assessment. We aimed to identify potential metabolomics-based profiles in urine and plasma for the detection of relatively low-level cadmium (Cd) exposure in large population-based studies. METHOD: We analyzed 123 urinary metabolites and 94 plasma metabolites detected in fasting urine and plasma samples collected from 1,412 men and 2,022 women involved in the Tsuruoka Metabolomics Cohort Study. Regression analysis was performed for urinary N-acetyl-beta-D-glucosaminidase (NAG), plasma, and urinary metabolites as dependent variables, and urinary Cd (U-Cd, quartile) as an independent variable. The multivariable regression model included age, gender, systolic blood pressure, smoking, rice intake, BMI, glycated hemoglobin, low-density lipoprotein cholesterol, alcohol consumption, physical activity, educational history, dietary energy intake, urinary Na/K ratio, and uric acid. Pathway-network analysis was carried out to visualize the metabolite networks linked to Cd exposure. RESULT: Urinary NAG was positively associated with U-Cd, but not at lower concentrations (Q2). Among urinary metabolites in the total population, 45 metabolites showed associations with U-Cd in the unadjusted and adjusted models after adjusting for the multiplicity of comparison with FDR. There were 12 urinary metabolites which showed consistent associations between Cd exposure from Q2 to Q4. Among plasma metabolites, six cations and one anion were positively associated with U-Cd, whereas alanine, creatinine, and isoleucine were negatively associated with U-Cd. Our results were robust by statistical adjustment of various confounders. Pathway-network analysis revealed metabolites and upstream regulator changes associated with mitochondria (ACACB, UCP2, and metabolites related to the TCA cycle). CONCLUSION: These results suggested that U-Cd was associated with metabolites related to upstream mitochondrial dysfunction in a dose-dependent manner. Our data will help develop environmental Cd exposure profiles for human populations.

Risk of bladder cancer in male Japanese workers exposed to ortho-toluidine and other aromatic amines.

PURPOSE: Nine bladder cancer (BCa) cases were reported among aromatic amine-exposed male workers at a factory manufacturing organic dye/pigment intermediates in Japan. We aimed to evaluate the characteristics of aromatic amine-exposed workers by cross-sectional observation, and the risk of BCa by assessing the standardized incidence ratio (SIR). METHODS: In the cross-sectional study, our subjects were: 9 BCa patients, 36 aromatic amine-exposed non-patients, and 79 non-exposed workers from 3 factories. We evaluated the subjects' medical history, urinalysis, qualitative determination of nuclear matrix protein 22, and urinary cytology. For SIR assessment, 98 aromatic amine-exposed workers from 1 factory were included, and the Japanese general male population was used as a referent population. Since no direct aromatic amine-exposure data were available, we calculated surrogate exposure levels using information on job sites, exposure potency, and duration. RESULTS: Coexistent aromatic amines were ortho-toluidine (OT), aniline, para-toluidine, ortho-anisidine, 2,4-xylidine, and ortho-chloroaniline. The prevalence rates of cystitis and bladder lesion-related symptoms in both BCa patients and aromatic amine-exposed non-patient workers were significantly higher than those of non-exposed workers. Overall, the SIR for BCa in OT-exposed workers was 56.8 (95% CI 27.7-104.3) and apparent dose-response relationships were revealed between the SIR and the surrogate exposure level in the 0-10-year lagged analyses. Overall, SIRs in other aromatic amine-exposed workers were also significantly high but no or unclear dose-response relationships were observed. CONCLUSIONS: We conclude that OT may be responsible for the increased risk of BCa. Regular monitoring of bladder lesion-related symptoms is essential for the early identification of BCa.

Biological monitoring of o-toluidine in urine pretreated by an enzymatic deconjugation method.

OBJECTIVES: To establish an enzymatic deconjugation method to separately quantify urinary o-toluidine (OT), its six metabolites, another six chemicals present in an OT-processing plant, and one metabolite of p-toluidine, and to propose optimal urinary biological monitoring items of OT exposure. METHODS: Thirty-six urine samples of an OT-processing plant's workers were obtained and pretreated by an enzymatic deconjugation method employing ß-glucuronidase/arylsulfatase for 3 hours at 37°C and measured by liquid chromatograph-mass spectrometry (LC-MS). An alkaline hydrolytic pretreatment and 1-chlorobutane extraction procedure was also examined as a widely used urinary OT measurement method. RESULTS: The 14 chemicals were separated by LC-MS condition set by us and 13 chemicals other than 2-chloroaniline showed satisfiable linearity and limits of determination. Standard substances of six OT metabolites decomposed after the alkaline heating. In the 36 urine samples, OT, N-(4-hydroxy-2-methylphenyl) acetamide (NHM), and 4-amino-m-cresol (ACR) accounted for approx. 90% of the total OT and OT metabolites, but inter-individual variation of the three substance excretion seemed to be wide. Time course of urinary excretion revealed that concentration of the three substances was higher 24 hours after the work shift's end rather than just after the work shift. CONCLUSIONS: OT and its six metabolites can each be determined with LC-MS. The alkaline method is not so optimal for exact biological monitoring. Rather, the sum of urinary OT, NHM, and ACR measured by the enzymatic method is a better index, and "end of the workweek" is a good urine-sampling time for the biological monitoring of OT exposure.

Evaluation of personal exposure of workers to indium concentrations in total dust and its respirable fraction at three Japanese indium plants.

This study aimed to evaluate personal exposures of 27 workers to indium compounds as "total" dust and its "respirable" fraction in their breathing zones at 3 Japanese indium plants. Eight-hour time-weighted average (TWA) indium concentrations of personal exposure to dust collected in sampling periods of 6 h or longer were determined by ICP-MS. The arithmetic means of exposure concentrations were 0.095 mg indium (In)/m3, when sampled as total dust, and 0.059 mg In/m3, as respirable fraction. ACGIH's TLV-TWA of 0.1 mg In/m3 for total particulate matter and Acceptable Exposure Concentration Limit (AECL) of 3×10-4 mg In/m3 for the respirable fraction notified by the Japanese Ministry of Health, Labour and Welfare were used to evaluate the exposure concentrations. Twenty-five out of 27 workers were exposed to indium concentrations lower than TLV-TWA, while all of the workers were exposed to the indium concentrations higher than AECL. We noted that there was a large discrepancy between the two occupational exposure limits referred to in this report, and these differences were attributed to the sampling strategies and health effects used as the prevention targets. Carcinogenicity of the respirable fraction of indium-containing particulates was considered in setting AECL, whereas it was not in ACGIH's TLV.

Quantitative assessment of occupational exposure to total indium dust in Japanese indium plants.

This study quantitatively assessed personal exposure of 86 workers to indium compounds as total dust at 11 Japanese indium plants. The personal exposures to indium concentrations in the breathing zone during an 8 h work-shift were determined by ICP-MS. The arithmetic mean indium concentration of all the workers was 0.098 mg Indium (In)/m3, with individual values ranging from 0.0001 to 1.421 mg In/m3. There were 11 workers whose exposure to indium concentrations exceeded the American Conference of Governmental Industrial Hygienists' Threshold Limit Value-Time Weighted Average (TLV-TWA) of 0.1 mg In/m3. Based on the condition TLV-TWA

The feasibility of aromatherapy massage to reduce symptoms of Idiopathic Environmental Intolerance: a pilot study.

OBJECTIVES: Idiopathic Environmental Intolerance (IEI) is an acquired disorder with multiple recurrent symptoms, which is associated with diverse environmental factors that are tolerated by the majority of people. IEI is an illness of uncertain aetiology, making it difficult to treat using conventional medicine. Therefore, there is a need for novel therapies to control the symptoms of IEI. The objective of this study was to evaluate the feasibility and impact of aromatherapy massage for individuals with IEI. DESIGN: Non-blinded crossover trial. SETTING: IEI patients who attended a clinic in Sapporo city were recruited, and sixteen patients were enrolled. Participants were clinically examined by an experienced medical doctor and met the criteria included in the working definition of IEI disorder. INTERVENTIONS: During the active period, participants received four one-hour aromatherapy massage sessions every two weeks. During the control period, the participants did not receive any massages. MAIN OUTCOME MEASUREMENTS: Scores on the IEI-scales trigger checklist, symptoms, life impact, and the State Anxiety Inventory were assessed before and after each period. Short-term mood enhancement was evaluated using the Profiles of Mood Status (POMS) before and after sessions. RESULTS: Due to period effects, evaluation of the results had to be restricted to the first period, and the result showed no effect of intervention. All six sub-scales of the POMS improved after each session (mean score differences: 4.89-1.33, P<0.05). CONCLUSIONS: Aromatherapy was well tolerated by subjects with IEI; however, aromatherapy, as applied in this study, did not suggest any specific effects on IEI condition.

The relationship between exposure to microbial volatile organic compound and allergy prevalence in single-family homes.

Microbial volatile organic compounds (MVOCs) are a type of VOCs produced by microorganisms. Exposure to 1-octen-3-ol, one of the known MVOCs, has been reported to reduce nasal patency and increase nasal lavage myeloperoxidase, eosinophil cationic proteins, and lysozymes in both experimental and field studies. We reported in a previous paper that 1-octen-3-ol exposure at home is associated with mucosal symptoms. In this study, our aim was to investigate the relationship between asthma and allergies and MVOC exposure in single-family homes. The subjects were 624 inhabitants of 182 detached houses in six regions of Japan. Air samples were collected using diffusive samplers, and the concentrations of eight selected MVOCs were analyzed using gas chromatography/mass spectrometry in selected-ion-monitoring mode. Each inhabitant of each of the dwellings was given a self-administered questionnaire. Among the 609 subjects who answered all of the questions about allergies, history of the medical treatment for asthma, atopic dermatitis, allergic rhinitis, and allergic conjunctivitis within the preceding two years was 4.8%, 9.9%, 18.2%, and 7.1%, respectively. A significant association between 1-octen-3-ol (per log(10) unit) and allergic rhinitis odds ratio (OR): 4.10, 95% confidence interval (CI): 1.71 to 9.80 and conjunctivitis (OR: 3.54, CI: 1.17 to 10.7) was found after adjusting for age, sex, tobacco, wall-to-wall carpeting, signs of dampness, history of treatment for hay fever, and other potentially relevant environmental factors. No relationships were found between any MVOCs and asthma or atopic dermatitis after the adjustment. The levels of MVOCs and airborne fungi were only weakly correlated. These results are consistent with previous studies that have associated higher levels of 1-octen-3-ol exposure with increased irritation of nasal and ocular mucosae. Although the indoor-air concentrations of 1-octen-3-ol found in this study were relatively low, we conclude that exposure to MVOC may be related to rhinitis and conjunctivitis.

Assessment of workplace air concentrations of indium dust in an indium-recycling plant.

OBJECTIVES: Suspended indium dust in an indium-recycling plant was quantified, in order to improve the work environment and to reduce workers' exposure to the dust. METHODS: Assessment of indium dust in the workplace air by multipoint area sampling and personal breathing zone sampling was conducted twice in 2004 and 2008. RESULTS: In 2004, all recycling processes except for purity analysis were classified into control class III according to the 2004 Notification. Two out of 5 workers were exposed to total dust with indium concentrations exceeding the ACGIH's TLV-TWA of 0.1 mg In/m(3). In 2008, the indium-contaminated workplace air was improved by local exhaust ventilation systems installed in some processes, resulting in control class I. According to the 2010 Technical Guideline, however, all the processes were classified into stage II or III, indicating that the first assessment value or Measurement B-based concentrations exceeded the acceptable exposure concentration limit of 0.0003 mg In/m(3) of respirabe dust. Exposure of almost all the workers to indium dust was below the TLV-TWA. CONCLUSIONS: The first field survey showed that almost all workplaces were classified into control class III, and that some workers were exposed to dust with indium concentrations exceeding the TLV-TWA. It was found in the second survey that workplace air contamination was improved by the local exhaust ventilation system, but was not reduced sufficiently to a level that meets the new Guideline.

Exposure assessment of ETBE in gas station workers and gasoline tanker truck drivers.

OBJECTIVES: In order to measure occupational exposure concentrations of ethyl tertiary-butyl ether (ETBE), we developed a diffusive sampling method for monitoring ETBE and performed an ETBE exposure assessment. METHODS: The applicability of diffusive samplers was examined by exposing the samplers to ETBE vapor in test chambers. The personal exposure levels of workers and airborne concentrations were measured at 4 gas stations. RESULTS: The ETBE sampling rate for the diffusive samplers (VOC-SD, Sigma-Aldrich Japan) was 25.04 ml/min (25°C). Compared with the active sampling method, the diffusive samplers could be used for short-term measurements and in environments containing a mixture of organic solvents. The geometric mean (GM) of TWA-8h ETBE was 0.08 ppm (0.02-0.28 ppm) in 28 gas station workers and 0.04 ppm (0.01-0.21 ppm) in 2 gasoline tanker truck drivers. With regard to ETBE airborne concentrations, the GM was 4.12 ppm (0.93-8.71 ppm) at the handles of hanging pumps but dropped to less than 0.01 ppm (less than 0.01-0.01 ppm) at the side of a public road. CONCLUSION: The diffusive sampling method can be used for the measurement of occupational ETBE exposure. The threshold limit of TLV-TWA 5 ppm recommended by the ACGIH was not exceeded in any of the workers in this study.

Inhalation carcinogenicity and chronic toxicity of indium-tin oxide in rats and mice.

OBJECTIVES: Carcinogenicity and chronic toxicity of indium-tin oxide (ITO) were examined by inhalation exposure of rats and mice to ITO aerosol. METHODS: Fifty mice of both sexes were exposed to ITO at 0 (control), 0.01, 0.03 or 0.1 mg/m(3) for 6 h/day, 5 day/wk for 104 wk, and 50 rats of both sexes were exposed to 0, 0.01 or 0.03 mg/m(3) ITO for the same time period. The repeated exposure of 50 rats of both sexes to 0.1 mg/m(3) ITO was discontinued at the 26th wk, followed by clean air exposure for the remaining 78 wk. RESULTS: In rats, incidences of bronchiolo-alveolar adenomas and carcinomas, bronchiolo-alveolar hyperplasia, alveolar wall fibrosis and thickened pleural wall, alveolar proteinosis and infiltrations of alveolar macrophages and inflammatory cells were significantly increased. Combined incidences of malignant lung tumors in male rats and total lung tumors in male and female rats were significantly increased at exposure to 0.01 mg/m(3) ITO. In mice, no carcinogenic response occurred, but thickened pleural wall, alveolar proteinosis and alveolar macrophage infiltration were induced. Mice were less susceptible to ITO than rats. The lung content of indium was the greatest, followed by the spleen, kidney and liver. Blood indium levels increased dose-dependently. CONCLUSIONS: There was clear evidence of carcinogenicity of inhaled ITO in male and female rats but not clear evidence in mice, together with occurrence of the chronic pulmonary lesions in both rats and mice.

Pulmonary toxicity in mice by 2- and 13-week inhalation exposures to indium-tin oxide and indium oxide aerosols.

OBJECTIVES: Inhalation toxicities of indium-tin oxide (ITO) and indium oxide (IO) in mice were characterized in comparison with those previously reported in rats. METHODS: B6C3F(1) mice of both sexes were exposed by inhalation to ITO or IO aerosol for 6 h/day, 5 day/wk for 2 wk at 0, 0.1, 1, 10 or 100 mg/m(3) or 13 wk at 0, 0.1or 1 mg/m(3). RESULTS: ITO and IO particles were deposited in the lung, mediastinal lymph node (MLN) and nasal-associated lymphoid tissue. Alveolar proteinosis, infiltrations of alveolar macrophages and inflammatory cells and increased lung weight were induced by 2- and 13-week exposures to ITO and IO, while alveolar epithelial hyperplasia occurred only in the 2-week exposures. Thickened pleural wall, hyperplastic MLN, extramedullary hematopoiesis in the spleen and increased levels of erythrocyte parameters were induced by 13-week exposure to ITO. The ITO- and IO-induced pulmonary lesions were milder in mice than those previously reported in rats, and the fibrotic lesions were different between these two species. Indium levels in the lung and pooled blood were analyzed in the mice exposed to ITO and IO for 13 wk. In the 13-week inhalation exposure of mice to ITO, alveolar proteinosis and significantly increased lung weight were induced at the same exposure concentration as the current threshold limit value for indium and its compounds.

Two- and 13-week inhalation toxicities of indium-tin oxide and indium oxide in rats.

OBJECTIVES: Two- and 13-week inhalation toxicities of indium-tin oxide (ITO) and indium oxide (IO) were characterized for risk assessments of workers exposed to ITO. METHODS: F344 rats of both sexes were exposed by inhalation to ITO or IO aerosol for 6 h/day, 5 day/wk for 2 wk at 0, 0.1, 1, 10 or 100 mg/m(3) or 13 wk at 0, 0.1 or 1 mg/m(3). An aerosol generator and inhalation exposure system was constructed. RESULTS: Blood and lung contents of indium were elevated in a dose-related manner in the ITO- and IO-exposed rats. ITO and IO particles were deposited in the lung, mediastinal lymph node and nasal-associated lymphoid tissue. Exposures to ITO and IO induced alveolar proteinosis, infiltrations of alveolar macrophages and inflammatory cells and alveolar epithelial hyperplasia in addition to increased lung weight. ITO affected the lung more severely than IO did. Fibrosis of alveolar wall developed and some of these lesions worsened at the end of the 26-week post-exposure period. CONCLUSIONS: Persistent pulmonary lesions including alveolar proteinosis and macrophage infiltration occurred after 2- and 13-week inhalation exposures of rats to ITO and IO. Fibrosis of alveolar wall developed later. These lesions occurred after ITO exposure at the same concentration as the current occupational exposure limit in the USA and at blood indium levels below the biological exposure index in Japan for indium.

Validation of urine density correction in cases of hippuric acid and un-metabolized toluene in urine of workers exposed to toluene.

To investigate if it is appropriate to apply urine density correction when a urine sample is dense or dilute. Data on hippuric acid (HA-U), toluene (Tol-U), creatinine (CR) and specific gravity (SG) in end-of-shift urine samples and exposure to air-borne toluene were cited from previous publications. In practice, 837 cases were available, and they were classified into dense, intermediate and dilute groups taking 0.3 and 3.0 g/l of CR and 1.010 and 1.030 of SG as cut-off points. Lines of regression of HA-U and Tol-U (as observed, CR-corrected or SG-corrected) with air-borne toluene were calculated for each density groups, and correlation coefficients (CCs) were compared. The dense groups gave CCs similar to those of the intermediate groups. Dilute versus intermediate group comparison also gave promising results. These conclusions were however based primarily on the findings with observed values, because the numbers of cases in the dilute or dense group were limited when CR- or SG-correction was applied. Literature survey showed that urine density correction does not always improve the correlation between solvents in air and exposure makers in urine. It was concluded that no correction for urine density may be necessary in evaluating HA-U and Tol-U in dense (and probably also dilute) urine samples as markers of occupational toluene exposure. Just in case when correction for urine density is desired for any reason, SG-correction may be recommended.

Relationship between selected indoor volatile organic compounds, so-called microbial VOC, and the prevalence of mucous membrane symptoms in single family homes.

Microorganisms are known to produce a range of volatile organic compounds, so-called microbial VOC (MVOC). Chamber studies where humans were exposed to MVOC addressed the acute effects of objective and/or subjective signs of mucosal irritation. However, the effect of MVOC on inhabitants due to household exposure is still unclear. The purpose of this epidemiological study was to measure indoor MVOC levels in single family homes and to evaluate the relationship between exposure to them and sick building syndrome (SBS). All inhabitants of the dwellings were given a self-administered questionnaire with standardized questions to assess their symptoms. Air samples were collected and the concentrations of eight selected compounds in indoor air were analyzed by gas chromatography/mass spectrometry - selective ion monitoring mode (GC/MS-SIM). The most frequently detected MVOC was 1-pentanol at a detection rate of 78.6% and geometric mean of 0.60 microg/m(3). Among 620 participants, 120 (19.4%) reported one or more mucous symptoms; irritation of the eyes, nose, airway, or coughing every week (weekly symptoms), and 30 (4.8%) reported that the symptoms were home-related (home-related symptoms). Weekly symptoms were not associated with any of MVOC, whereas significant associations between home-related mucous symptoms and 1-octen-3-ol (per log(10)-unit: odds ratio (OR) 5.6, 95% confidence interval (CI): 2.1 to 14.8) and 2-pentanol (per log(10)-unit: OR 2.3, 95% CI: 1.0 to 4.9) were obtained after adjustment for gender, age, and smoking. Associations between home-related symptoms and 1-octen-3-ol remained after mutual adjustment. However, concentrations of the selected compounds in indoors were lower than the estimated safety level in animal studies. Thus, the statistically significant association between 1-octen-3-ol may be due to a direct effect of the compounds or the associations may be being associated with other offending compounds. Additional studies are needed to evaluate these possibilities.

Stability in urine of authentic phenylglyoxylic and mandelic acids as urinary markers of occupational exposure to styrene.

Phenylglyoxylic acid (PhGA) and mandelic acid (MA) are two popular urinary markers of occupational exposure to styrene, but PhGA has been considered to be relatively unstable when urine samples are stored. This study was initiated to examine the stability of PhGA in urine under two storage conditions, i.e., at room temperature (at 25 degrees C) and in refrigeration (at 4 degrees C) for up to 14 days. The experiments showed that no substantial decrease was observed in either PhGA or MA even at room temperature within one day, but, depending on urine samples, a gradual decrease in PhGA took place both at 4 degrees C and more markedly at 25 degrees C when kept for a week. Further reduction was observed in two weeks even at 4 degrees C. No reduction was observed in MA up to two weeks both at 4 degrees C and at 25 degrees C. The observation on stability of MA and limited stability of PhGA (i.e., no significant decrease for 4 days when stored as refrigerated) was confirmed by a repeated experiment. Further analyses disclosed that PhGA when stored at 4 degrees C tended to be more unstable when urine samples were alkaline (e.g., at pH 8) rather than acidic (e.g., at pH 6 or below), but the trend varied subject to individual samples. Thus, the practical recommendation is that urine samples should be analyzed on the day of collection if all possible, or kept at 4 degrees C, or more preferably at -20 degrees C. Refrigeration will allow storage of up to 4 days without substantial decrease in either PhGA or MA.

Urinary excretion of an oxidative stress marker, 8-hydroxyguanine (8-OH-Gua), among nickel-cadmium battery workers.

The relationship between oxidative stress and carcinogenic metals including nickel and cadmium is a matter of interest. To assess the oxidative stress status of workers exposed to nickel and cadmium simultaneously, we determined urinary excretion of 8-hydroxyguanine (8-OH-Gua), a urinary oxidative stress marker. Our subjects were 66 (64 males and 2 females) nickel-cadmium battery workers. Spot urine and blood samples were collected. The levels of cadmium in blood (Cd-B) and nickel in urine (Ni-U) were determined by graphite furnace atomic absorption spectrophotometry. 8-OH-Gua in urine was analyzed using a high performance liquid chromatography-electrochemical detector (HPLC-ECD) system. Data on age, sex, duration of present work and smoking status were also obtained from each subject. Creatinine-adjusted 8-OH-Gua was significantly correlated with age, Ni-U and Cd-B in univariate analysis, while multivariate analysis revealed that Ni-U and Cd-B were significant independent variables and that these two biological exposure indices were positively correlated with 8-OH-Gua. The data were also analyzed in the context of mixture toxicity. The subjects were divided into groups based on median level of Ni-U and Cd-B (2.86 mug/g creatinine and 0.23 mug/dl, respectively). Workers with high Ni-U/high Cd-B (Group IV) had the highest levels of 8-OH-Gua levels (GM (GSD), 21.7(2.0)), followed by those with high Ni-U/low Cd-B (11.5(1.6) Group III), those with low Ni-U/high Cd-B (8.9(1.9) Group II), and those with low Ni-U/low Cd-B (8.5(1.5) Group I). The p values of Students' t-tests between Group I and Group II, III and IV were 0.847, 0.050 and <0.001, respectively. The combined effect of Cd and Ni on the urinary excretion of 8-OH-Gua departed from additivity.