Yoshinari Fukui

Comparative evaluation of four urinary tubular dysfunction markers, with special references to the effects of aging and correction for creatinine concentration

Comparative evaluation was made on alpha(1)-microglobulin (alpha(1)-MG), beta(2)-microglobulin (beta(2)-MG), retinol binding protein (RBP) and N-acetyl-beta-D-glucosaminidase (NAG), as a marker of renal tubular dysfunction after environmental exposure to cadmium (Cd), with special references to the effects of aging and correction for creatinine concentration. For this purpose, a previously established database of 817 never-smoking Japanese women (at the ages of 20 to 74 years) on hematological [hemoglobin, serum ferritin (FE), etc.] and urinary parameters [alpha(1)-MG, beta(2)-MG, creatinine (cr), and a specific gravity] was revisited. For the present analysis, the database was supplemented by the data on RBP and NAG in urine. The exposure of the women to Cd was such that the geometric mean Cd in urine was 1.3 microg/g cr. Among the four tubular dysfunction markers, NAG showed the closest correlation with Cd, followed by alpha(1)-MG and then beta(2)-MG, and RBP was least so although the correlations were all statistically significant. The observed values of the markers gave the best results, whereas correction for a urine specific gravity gave poorer correlation, and it was the worst when correction for creatinine concentration was applied. Age was the most influential confounding factor. The effect of age appeared to be attributable at least in part to the fact that both creatinine and, to a lesser extent, the specific gravity decreased as a function of age. Iron deficiency anemia of sub-clinical degree as observed among the women did not affect any of the four tubular dysfunction markers. In conclusion, NAG and alpha(1)-MG, rather beta(2)-MG or RBP, are more sensitive to detect Cd-induced tubular dysfunction in mass screening. The use of uncorrected observed values of the markers rather than traditional creatinine-corrected values is recommended when comparison covers people of a wide range of ages.

Bias induced by the use of creatinine-corrected values in evaluation of β2-microgloblin levels

The present study was initiated to examine if the correction for creatinine (CR or cr) is the best approach among the three methods of correction for CR, correction for a specific gravity (SG or sg) and the use of observed values in managing difference in urine density. For this purpose, a database previously developed on 10,753 adult women in 10 non-polluted areas in Japan was re-visited for information on age, urinary levels of Cd, Mg, Ca, Zn, beta(2)-MG, and creatinine, and urine specific gravity as well as smoking habits. Never-smoking women with various urine density counted 8975 cases (the various urine density group). From these cases, 7081 cases with adequate urine density (i.e. 0.5 g/l < or = CR < or = 3.0 g/l and 1.010 < or = SG < or = 1.030) were selected (the adequate urine density group). When a beta(2)-MG level of 400 microg/g CR or 400 microg/l was taken as a cut-off value for beta(2)-MG-uria, both the prevalence of beta(2)-MG(cr)-uria [i.e. cases with beta(2)-MG (as corrected for CR) in excess of 400 microg/g cr] and that of beta(2)-MG(sg)-uria increased as a function of the decrease in Cd(cr) or Cd(sg). The prevalence of beta(2)-MG(ob)-uria also varied as a function of CR and SG, especially of CR, but its range of variation was smaller than the corresponding changes in beta(2)-MG(cr)-uria prevalence. A noteworthy advantage for the use of observed values over that of SG-corrected values was the minimum effect of age. In over-all evaluation, therefore, the recommended approach appeared to be the use of non-corrected observed values (after selection of urine samples for adequate urine density if desired) or correction for SG, rather than correction for CR.

N-Acetyl-β-D-glucosaminidase (NAG) as the most sensitive marker of tubular dysfunction for monitoring residents in non-polluted areas

Alpha(1)-microglobulin (alpha(1)-MG), beta(2)-microglobulin (beta(2)-MG) and N-acetyl-beta-D-glucosaminidase (NAG) are tubular dysfunction markers often used in cadmium (Cd) epidemiology. The purpose of the present study was to identify the best maker among the above-referred three that correlates most closely with Cd in urine of residents with no known Cd pollution. Survey was conducted in 2007-2008 in three prefectures in Japan. Adult women, 2163 in total, participated in the survey; they provided informed consents, offered spot urine samples and filled questionnaires on possible confounders of Cd burden. Urine samples were analyzed for Cd, alpha(1)-MG, beta(2)-MG and NAG together with calcium (Ca), magnesium (Mg), creatinine (CR) and specific gravity (SG). The analyte levels, as observed (e.g., Cd(ob)) or after correction for CR (e.g., Cd(cr)) or SG (e.g., Cd(sg)) were subjected to simple and multiple regression analysis. Correlation matrix analysis with observed values for total cases showed that the coefficients of correlation with Cd were highest for NAG, followed by alpha(1)-MG, and lowest for beta(2)-MG. Multiple regression analysis by three prefectures either separately or in combination (thus four analyses) disclosed that Cd was the independent variable most influential to NAG (as the dependent variable) throughout the four analysis conditions with high R(2) values (>0.3), whereas the most influential variables were not the same depending on the analysis conditions in cases with alpha(1)-MG and beta(2)-MG. When coefficients of correlation for the three dysfunction markers with Cd were compared among the observed, CR- and SG-corrected values, the coefficients for the observed values were higher than the counterpart values for CR- or SG-corrected values. In conclusion, NAG rather than alpha(1)-MG or beta(2)-MG should be recommended for monitoring Cd exposure-related tubular effects among general populations. Observed (i.e., un-corrected) values rather than CR- or SG-corrected values should be used.

Closer correlation of cadmium in urine than that of cadmium in blood with tubular dysfunction markers in urine among general women populations in Japan

ObjectivesThe objectives of the present study are to investigate whether cadmium in blood (Cd-B) and cadmium in urine (Cd-U) correlate with each other irrespective of age among general populations and which one of Cd-B or Cd-U correlates more closely with three renal tubular dysfunction markers in urine of α1-microglobulin (α1-MG-U), β2-microglobulin (β2-MG-U) and N-acetyl-β-d-glucosaminidase (NAG-U).MethodsData on two exposure markers (Cd-B and Cd-U) and three effect markers (α1-MG-U, β2-MG-U and NAG-U) were collected for 1,403 adult women in non-polluted areas all over Japan. Possible significance of correlation between the parameters and dependency on age was examined by simple and multiple regression analysis.ResultsBoth Cd-B and Cd-U increased as a function of age. The two exposure markers correlated significantly with each other, and the Cd-U over Cd-B ratio also increased as a function of age. Although both Cd-B and Cd-U correlated significantly with the three effect markers, the correlation was closer for Cd-U than for Cd-B.ConclusionsCd-U rather than Cd-B should be recommended as an exposure marker of choice in Cd biological monitoring of general populations. Effects of aging should be taken into account when evaluating study results.

α1-Microglobulin levels and correlation with cadmium and other metals in urine of non-smoking women among general populations in Japan

The present study was initiated to examine urinary α1-microglobulin (α1-MG-U) levels among non-smoking women in the general population in Japan. A previously established database on spot urine samples from adult woman volunteers in 10 non-polluted areas all over Japan was re-examined. The data examined were on α1-MG-U, cadmium, calcium, magnesium and zinc levels in urine (Cd-U, Ca-U, Mg-U and Zn-U, respectively), urinary creatinine (CR or cr), urine specific gravity (SG or sg), smoking habits and age. Thus, 8975 never-smoking women were selected for statistical analyses. The grand geometric mean (GM) for α1-MG-U among the population was 2.1 mg/L or 2.5 mg/g cr, depending on the correction for urine density. It was 1.1 µg/L or 1.3 µg/g cr for GM Cd-U. The inter-area difference in α1-MG-U was <1.5 mg/g cr or <0.7 mg/L; the area with the highest or lowest GM Cd-U was not always highest or lowest in GM α1-MG-U. The correlation coefficient (0.53) between log Cd-U and log α1-MG-U (both without urine density correction) became substantially smaller when the analyte levels were corrected for CR (0.25) or SG (0.26). In multiple regression analysis, the power of influence of the five independent variables (log Cd-U, Ca-U, Mg-U, Zn-U and age) in combination was small (R 2 ≦ 0.13). In contrast, logistic regression analysis suggested that α1-MG-U might be elevated as a function of an increase in Cd-U, depending on the cut-off values. Discussion was made on dose (Cd-Ucr) and response (α1-MG-Ucr) relationship based on information available in literatures to show that the increment in α1-MG-Ucr per Cd-Ucr was much greater when Cd-Ucr was large, e.g., in excess of 10 µg/g cr.

No meaningful increase in urinary tubular dysfunction markers in a population with 3μg cadmium/g creatinine in urine

The critical Cd exposure level to induce tubular dysfunctions is a focus of public concern among general populations in Japan. To answer this question, one group each (about 1000 adult women/area) in nonpolluted areas with high (Area H) and low Cd exposure (Area L) was obtained, and 742 strictly age-matched pairs of never-smoking adult women were selected for comparison. Cd, α1-MG (microglobulin) and β2-MG in urine were taken as markers of exposure and tubular dysfunction, respectively. Geometric mean Cd levels as corrected for creatinine (Cdcr) was greater than three times higher in Area H (2.8 μg/g cr) than in Area L (0.8 μg/g cr). Nevertheless, β2-MGcr did not differ between the two areas (125 μg/g cr for Area H vs 118 μg/g cr for Area L). α1-MGcr was only marginally higher in Area H (2.8 mg/g cr) than in Area L (2.1 mg/g cr), with no biomedical significance. Results were essentially the same when analyses were conducted with noncorrected observed values or values corrected for a specific gravity. Thus, the effects of Cd exposure in Area H on renal tubular function should be essentially nil.

Changes in correlation coefficients of exposure markers as a function of intensity of occupational exposure to toluene

This study was initiated to identify a marker of choice to monitor occupational exposure to toluene through quantitative evaluation of changes in correlation coefficients (CCs), taking advantage of a large database. Six known or proposed exposure markers in end-of-shift blood (B) and urine (U) were studied, i.e., toluene in blood (Tol-B) and benzyl alcohol, benzylmercapturic acid, o-cresol, hippuric acid and toluene in urine (BeOH-U, BMA-U, o-CR-U, HA-U, and Tol-U, respectively). To construct a database, data on 8-h time-weighted average intensity of occupational exposure to toluene and resulting levels of the six exposure markers in blood or urine were cited for 901 cases from previous four publications of this study group and combined with 146 new cases. In practice, 874 cases (all men) were available when extremely dilute or dense urine samples were excluded. The 874 cases were classified taking the upper limit (from 120 ppm to 1 ppm) of the toluene exposure concentration, and the CCs for the six markers with TWA toluene exposure intensity were calculated. For further evaluation, the 874 cases were divided into 10 per thousand in terms of TWA toluene exposures, and several 10 per thousand were combined so that sufficient numbers of cases were available for calculation of the CCs at various levels of toluene exposure. Perusal was made to know whether or not and which one of the six makers gave significant CC even at low level of toluene exposure. The CCs for BMA-U, o-CR-U and HA-U with TWA toluene exposure were well >0.7 when toluene exposure was intense (e.g., up to 60-100 ppm as the upper limit of the exposure range), but were reduced when the upper limit of toluene exposure was less than 50 ppm, and the CCs were as small as <==0.2 when the upper limit was about 10 ppm or less. In contrast, Tol-U and Tol-B were correlated with exposure to toluene down to the <or=3 ppm range. The CC for BeOH-U was <0.1 almost throughout the exposure ranges. Further analyses showed that the CCs for all markers (except the CC for BeOH-U) were >0.4 when the cases in the 60th-100th per thousand were examined. The CCs for Tol-U and Tol-B were >0.3 also for cases in the 0th-60th or 30th-70th per thousand, whereas the CCs for other four markers were <0.3. In over-all evaluation, it was concluded that HA and o-CR are among the markers of choice to monitor occupational toluene exposure at high levels, and that only un-metabolized toluene in urine or in blood is recommended when toluene exposure level is low (e.g., 10 ppm or less). Toluene in urine may be preferred rather than that in blood due to practical reasons, such as non-invasiveness.