Simultaneous quantification of pyrethroid metabolites in urine of non-toilet-trained children in Japan

BACKGROUND@#Pyrethroid (PYR) insecticides are widely used for controlling various pests. There are two types that differ in terms of usage: agricultural-purpose PYR (agriculture-PYR) and hygiene purpose PYR (hygiene-PYRs). Few studies exist on the exposure to these chemicals in small children. In this study, we conducted biomonitoring of urinary pyrethroid metabolites in 1.5-year-old children throughout the year.@*METHODS@#Study subjects were 1075 children participating in an Aichi regional sub-cohort of the Japan Environment and Children's Study as of 18-month health check-up. The concentrations of four specific hygiene-PYR metabolites including 2,3,5,6-tetrafluoro-1,4-benzenedimethanol (HOCH2-FB-Al), and five common metabolites of hygiene- and agriculture-PYRs including 3-phenoxybenzoic acid (3PBA) and cis- and trans-3-(2,2-dichlorovinyl)-2,2-dimethylcyclopropane-1-carboxylic acid (DCCA), were measured in urine samples extracted from soiled diapers using a triple quadrupole gas chromatograph-mass spectrometer.@*RESULTS@#The highest detection frequencies were for 3PBA, followed by DCCA, 1R-trans-chrysanthemum dicarboxylic acid, and HOCH2-FB-Al. Among the six metabolites, urinary concentrations were seasonally varied. However, this variation was not observed in the most studied PYR metabolite, 3PBA. Spearman's correlation analysis demonstrated a significant positive correlation between FB-Al and DCCA (r = 0.56) and HOCH2-FB-Al and 4-methoxymethyl-2,3,5,6-tetrafluorobenzyl alcohol (r = 0.60).@*CONCLUSIONS@#This biomonitoring survey found widespread and seasonally specific exposure to multiple hygiene- and agriculture-PYRs in 1.5-year-old Japanese children.

Optimization and validation of a highly sensitive method for determining glyphosate in human urine by solid-phase extraction and liquid chromatography with tandem mass spectrometry: a methodological study

BACKGROUND@#Glyphosate and its salt formulations are nonselective herbicides that have been extensively used worldwide, both for residential and agricultural purposes. The possible carcinogenicity and teratogenicity of glyphosate remain to be elucidated. We developed a sensitive and high-throughput analytical method for urinary glyphosate using liquid chromatography-tandem mass spectrometry with the aim of contributing to glyphosate exposure assessment in epidemiological studies.@*METHODS@#After urine dilution (creatinine matching dilution to 0.05 g creatinine/L), glyphosate was extracted using two types of solid phase extraction columns (SCX and NH2) with automated sample preparation instruments. The eluate was dried and dissolved in the mobile phase, followed by liquid chromatography-tandem mass spectrometry analysis. The optimized method was applied to urine samples obtained from 54 Japanese adults and children.@*RESULTS@#The results from the validation study demonstrated good recoveries (91.0-99.6%), within- and between-run precisions (< 15%), low detection limits (0.1 μg/L), and lower limit of quantification (0.3 μg/L). The detection frequency and median concentration of the urinary glyphosate in Japanese subjects were 59% and 0.25 μg/L (0.34 μg/g creatinine).@*CONCLUSIONS@#Our reliable determination method was successful in measuring urinary glyphosate concentration. Moreover, this is the first biomonitoring report of urinary glyphosate levels in the Japanese general population.

Intra-individual variations of organophosphate pesticide metabolite concentrations in repeatedly collected urine samples from pregnant women in Japan

BACKGROUND@#Low-dose exposure to organophosphate (OP) insecticides during pregnancy may adversely affect neurodevelopment in children. To evaluate the OP exposure levels, single urine sampling is commonly adopted to measure the levels of dialkylphosphates (DAPs), common OP metabolites. However, the inter-day variations of urinary DAP concentrations within subjects are supposed to be large due to the short biological half-lives of the metabolites, and it is thus considered difficult to accurately assess OP exposure during pregnancy with single sampling. This study aimed to assess intra-individual variations of DAP concentrations and the reproducibility of the exposure dose categorization of OPs according to DAP concentration ranges in pregnant women in Japan.@*METHODS@#Urine samples were collected from 62 non-smoking pregnant women (12-22 weeks of gestation) living in Aichi Prefecture, Japan. First morning void (FMV) and spot urine samples taken between lunch and dinner on the same day were collected on five different days during 2 weeks. The concentrations of DAP and creatinine in urine samples were measured using an ultra performance liquid chromatography with tandem mass spectrometry. Creatinine-adjusted and unadjusted concentrations were used for the intraclass correlation coefficient (ICC) calculations and surrogate category analyses.@*RESULTS@#For all DAP metabolites, the creatinine-adjusted single ICCs exceeded 0.4, indicating moderate reliability. Overall, ICCs of spot urine samples taken in the afternoon were better than those taken as FMV. Surrogate category analyses showed that participants were categorized accurately into four exposure dose groups according to the quartile points.@*CONCLUSION@#This study indicated that a single urine sample taken in the afternoon may be useful in assessing OP exposure as long as the exposure is categorized into quartiles when conducting epidemiological studies in early to mid-pregnant women in Japan.

Association between exposure to household smoking and dental caries in preschool children: a cross-sectional study

BACKGROUND@#We aimed to examine the association of exposure to environmental tobacco smoke with dental caries among preschool children. Exposure to environmental tobacco smoke was assessed in terms of urinary cotinine concentrations and pack-years of exposure to smoking by parents and other family members at home.@*METHODS@#This cross-sectional study included 405 preschool children aged 3-6 years from two preschools in Japan in 2006. Information on the smoking habits of family members living with the child was obtained from parent-administered questionnaires. Dental examination was conducted to assess dental caries, that is, decayed and/or filled teeth. Urinary cotinine levels were measured using first-void morning urine samples.@*RESULTS@#Overall, 31.1% of the children had dental caries, and 29.5% had decayed teeth. Exposure to current maternal and paternal smoking was positively associated with the presence of dental caries after controlling for covariates. More than three pack-years of exposure to maternal smoking and more than five pack-years of exposure to smoking by all family members were significantly associated with the presence of dental caries as compared with no exposure (odds ratio [OR] = 5.55, 95% confidence interval [CI] = 2.17-14.22, P for trend < 0.001 and OR = 2.00, 95% CI = 1.12-3.58, P for trend = 0.004, respectively). These exposure variables were similarly associated with the presence of decayed teeth (OR = 2.92, 95% CI = 1.23-6.96, P for trend = 0.01 and OR = 1.75, 95% CI = 0.96-3.20, P for trend = 0.03, respectively). As compared with lowest tertile of the urinary cotinine level, the highest tertile of the urinary cotinine level was significantly associated with the presence of dental caries as well as decayed teeth; the ORs for the highest vs. lowest tertile of urinary cotinine levels were 3.10 (95% CI = 1.71-5.63, P for trend = 0.012) and 2.02 (95% CI = 1.10-3.70, P for trend = 0.10), respectively.@*CONCLUSIONS@#These data suggest that exposure to tobacco smoke may have a dose-dependent influence on the development of caries.

Oxidized human serum albumin as a possible correlation factor for atherosclerosis in a rural Japanese population: the results of the Yakumo Study

BACKGROUND@#The effect of the redox state of human serum albumin (HSA) on the antioxidant properties of the entire body has been a focus of recent research. The usefulness of HSA redox state as a biomarker for reducing oxidative stress has been investigated in clinical settings; however, evidence for its significance as a health index in non-clinical settings is yet to be established. This study aimed to examine the associations between HSA redox state and the atherosclerotic indices of carotid intima-media thickness (IMT) and plaque formation in a rural Japanese population.@*METHODS@#We conducted a cross-sectional study as part of a health check-up program in the rural area of Hokkaido, Japan, at the end of August 2013. A total of 281 residents (124 men and 157 women) were included in the final analysis. Lifestyle-related data were obtained through a self-reported questionnaire, and ultrasound examinations were performed to measure IMT and determine plaque formation. The high-performance liquid chromatography postcolumn bromocresol green method was used to separate HSA into human nonmercaptalbumin and human mercaptalbumin (HMA).@*RESULTS@#We found a significant negative relationship between the fraction of HMA [f(HMA)] and IMT (standardized β = - 0.132, p = 0.03). Moreover, f(HMA) was significantly associated with plaque formation (p < 0.01) with an odds ratio of 0.89 (95% confidence interval, 0.81-0.97) for every 10% increment in f(HMA).@*CONCLUSIONS@#We found that the HSA redox state, as determined by f(HMA), was associated with atherosclerotic indices in Japanese subjects. These results suggest that the HSA redox state indicates the risk of developing atherosclerosis.

Relationship between dietary habits and urinary concentrations of 3-phenoxybonzoic acid in a middle-aged and elderly general population in Japan

<p><b>OBJECTIVES</b>The ingestion of pesticides in the daily diet is assumed to be the main modality of pesticide exposure for most people. A widely used class of pesticides in agricultural or residential settings is pyrethroid. We have examined the relationship between the intake frequency of selected items of vegetables and fruits and urinary metabolites of pyrethroid pesticides in a healthy general population.</p><p><b>METHODS</b>A total of 535 residents (184 men and 351 women) who attended a healthcare checkup program conducted in a rural area of Hokkaido, Japan, in August 2005 provided informed consent for their spot urine samples to be used for the determination of 3-phenoxybenzoic acid (3-PBA) levels. They also completed a self-administered questionnaire regarding the intake frequency of 12 food items. The concentrations of creatinine-corrected 3-PBA were predicted by the intake frequency of each item, using analysis-of-covariance models to adjust for age, sex, body mass index, and drinking and smoking status.</p><p><b>RESULTS</b>Both a significant association between the 3-PBA concentration and the frequency of tomato consumption and a significant positive linear trend was found in female subjects. In contrast, no such association was found in the male subjects.</p><p><b>CONCLUSIONS</b>The frequency of tomato consumption was confirmed to strongly predict the urinary pyrethroid metabolite levels in the general population-presumably because tomatoes are most often consumed raw and unpeeled (more so than all other vegetables and fruits analyzed in the current study). However, it should be noted that the 3-PBA levels, even among those subjects with the highest consumption of tomatoes, were far below the levels of toxicological significance, although the health consequences from long-term low-level exposure to pyrethroid requires further exploration.</p>

Association of serum NO( x ) level with clustering of metabolic syndrome components in middle-aged and elderly general populations in Japan

<p><b>OBJECTIVES</b>The aim of this study was to determine whether the serum nitrite plus nitrate (NO( x )) level correlates with biomarkers that are known components of the metabolic syndrome (MetS).</p><p><b>METHODS</b>Serum NO( x ) levels were measured using a commercial kit in 608 Japanese men and women between the ages of 39 and 85 years. Multivariate adjustments for age, smoking status, alcohol consumption and exercise were made in the analysis of covariance (ANCOVA). The components of the metabolic syndrome were defined based on the following criteria: body mass index (BMI) >/=25.0 kg/m(2), glycated hemoglobin (HbA1c) >/=5.6%, systolic blood pressure >/=130 mmHg or diastolic blood pressure >/=85 mmHg, high-density lipoprotein-cholesterol (HDL-C) </=1.03 mmol/l for men and </=1.29 mmol/l for women and triglyceride >/=1.69 mmol/l.</p><p><b>RESULTS</b>The logarithmically transformed age-adjusted serum NO( x ) (lnNO( x )) value was significantly higher in the low HDL-C group (1.76 +/- 0.05 mumol/l; p < 0.05) than MetS component groups (1.65 +/- 0.01 mumol/l) in men, but no difference was found in women. The means of serum lnNO( x ) after multivariate adjustment were 1.64, 1.65, 1.64, 1.66, and 1.81 mumol/l for 0, 1, 2, 3, and 4-5 MetS components for all subjects, respectively. The results of ANCOVA confirmed that the serum lnNO( x ) level was significantly correlated with the clustering of MetS components in both men and women (p < 0.0001 for trend).</p><p><b>CONCLUSION</b>Our results suggest that an increase in the clustering of MetS components was associated with the increase in serum NO levels in our general population.</p>

Association of glycemic profiles with whole blood polyamine among middle-aged Japanese men: colorimetric assay using oat and barley seedling polyamine oxidase

<p><b>OBJECTIVES</b>Polyamines have long been known to have an insulin-like action, but their antiglycating effect has only recently attracted the attention of researchers. The aim of our investigation was to determine the whole blood polyamine concentration in a healthy population in order to examine its relationship with glycemic profiles.</p><p><b>METHODS</b>The study cohort comprised 622 men aged 40-59 who participated in a health checkup program conducted in 1997, when they underwent measurements of fasting plasma glucose (FPG), insulin (FPI), and fructosamine as glycemic indices. Colorimetric assay methods using oat and barley seedling polyamine oxidase were used to determine total polyamine (spermidine + spermine) and spermine concentrations in the whole blood, respectively. Polyamine concentrations adjusted for hemoglobin were quartiled for the analysis of covariance to assess the association with glycemic indices.</p><p><b>RESULTS</b>A significant association was demonstrated between the FPG and total polyamine concentrations. In the trend test, FPG and fructosamine levels increased in accordance with the shift of quartiles of total polyamine concentrations from low to high. In contrast, the association between the spermine and glycemic indices was not statistically significant based on the test for difference of multivariate-adjusted means or trend for linearity.</p><p><b>CONCLUSIONS</b>This is the first epidemiological study to reveal that the concentrations of blood polyamines are related with either FPG or fructosamine level in a healthy population. There may be some feedback mechanism for the elevation of circulating polyamines to quench the glycation reaction under hyperglycemic conditions. In addition, total polyamines, rather than spermine alone, seem to be a sensitive biomarker representing the antiglycation effect of polyamines.</p>

Association of Abdominal Circumference with Serum Nitric Oxide Concentration in Healthy Population

Objectives: We determined the relationship between abdominal circumference and the concentration of nitric oxide (NO), an endothelial cell product known to play an important role in the regulation of vascular tone and thrombocyte activations. Methods: Subjects were 177 men and 339 women aged 40 or over who were free from a history of diabetes or malignancy. Analysis of covariance was applied to examine the gender-specific and smoking-status-specific associations of abdominal fat volume measured as waist circumference, waist-hip ratio, and waist-to-stature ratio, with serum NO level represented by the concentration of NO metabolites (NOx; nitrate plus nitrite). Results: Although men showed no statistical association between abdominal fat accumulation and NOx concentration, abdominal adiposity seemed to inversely affect the serum NOx concentration of never- and current-smoking women. Conclusion: Our results suggest that a reduction in NO bioactivity occurs with abdominal fat accumulation in women. The underlying biological mechanism might involve adipocytokines secreted from visceral fat, but is yet to be elucidated.

Association of abdominal circumference with serum nitric oxide concentration in healthy population

<p><b>OBJECTIVES</b>We determined the relationship between abdominal circumference and the concentration of nitric oxide (NO), an endothelial cell product known to play an important role in the regulation of vascular tone and thrombocyte activations.</p><p><b>METHODS</b>Subjects were 177 men and 339 women aged 40 or over who were free from a history of diabetes or malignancy. Analysis of covariance was applied to examine the gender-specific and smoking-status-specific associations of abdominal fat volume measured as waist circumference, waist-hip ratio, and waist-to-stature ratio, with serum NO level represented by the concentration of NO metabolites (NOx; nitrate plus nitrite).</p><p><b>RESULTS</b>Although men showed no statistical association between abdominal fat accumulation and NOx concentration, abdominal adiposity seemed to inversely affect the serum NOx concentration of never- and current-smoking women.</p><p><b>CONCLUSION</b>Our results suggest that a reduction in NO bioactivity occurs with abdominal fat accumulation in women. The underlying biological mechanism might involve adipocytokines secreted from visceral fat, but is yet to be elucidated.</p>