Circadian rhythms in the urinary excretion of heavy metals and organic substances in metal workers in relation to renal excretory mechanism: profile analysis

Summary Circadian rhythms in the urinary excretion of eleven heavy metals and organic substances were examined under free, water-restrictive and water-loading conditions for 6 d (2 d for each of the three conditions) in twenty metal workers exposed to lead, zinc and copper. Circadian rhythms were found for all heavy metals and organic substances as well as for urinary flow (UF) rate, creatinine (Cn) and total urinary solutes (TUS). The Cn rhythm was significantly unparallel to the OF rhythm under the water-loading condition, indicating that the two rhythms were essentially different from each other. Circadian rhythms of the eleven urinary substances were then related to the Cn and OF rhythms, using profile analysis. The results indicated that the rhythms in the manganese, chromium, copper and beta-2-microglobulin excretion depend on the Cn rhythm, i.e. the rhythm of glomerular filtration; the rhythms in the hippuric acid, delta-aminolevulinic acid and TUS excretion are on the OF rhythm, i.e. the rhythm of reabsorption by the distal tubule and collecting duct. On the other hand, the rhythms in the lead, inorganic mercury, cadmium, zinc and coproporphyrin excretion were considered as reflecting complex renal excretory mechanisms.     

Effects of water restriction and water loading on daily urinary excretion of heavy metals and organic substances in metal workers.

The effects of urinary volume on daily urinary excretion of seven heavy metals and four organic substances were examined in relation to the changes in their plasma and erythrocyte concentrations and urinary creatinine excretion in 19 metal workers. The examination was conducted under the conditions of water restriction and loading for six days. The major findings were as follows: (1) urinary excretion of all heavy metals and organic substances except mercury, together with creatinine excretion, significantly decreased under the water restrictive condition whereas under the water loading condition their excretion significantly increased and (2) daily variations in urinary excretion of lead, cadmium, chromium, copper, hippuric acid, delta-aminolaevulinic acid, and coproporphyrin did not differ significantly from the variation in urinary excretion of creatinine (profile analysis, p greater than 0.05). It is suggested that glomerular filtration is the major factor determining renal excretory mechanisms of the four heavy metals and three organic substances examined.     

Effects of water restriction and water loading on daily urinary excretion of heavy metals and organic substances in metal workers

The effects of urinary volume on daily urinary excretion of seven heavy metals and four organic substances were examined in relation to the changes in their plasma and erythrocyte concentrations and urinary creatinine excretion in 19 metal workers. The examination was conducted under the conditions of water restriction and loading for six days. The major findings were as follows: (1) urinary excretion of all heavy metals and organic substances except mercury, together with creatinine excretion, significantly decreased under the water restrictive condition whereas under the water loading condition their excretion significantly increased and (2) daily variations in urinary excretion of lead, cadmium, chromium, copper, hippuric acid, delta-aminolaevulinic acid, and coproporphyrin did not differ significantly from the variation in urinary excretion of creatinine (profile analysis, p greater than 0.05). It is suggested that glomerular filtration is the major factor determining renal excretory mechanisms of the four heavy metals and three organic substances examined.     

Filterable plasma concentration, glomerular filtration, tubular balance, and renal clearance of heavy metals and organic substances in metal workers

To estimate "filterable" plasma concentration (FPx), glomerular filtration, tubular balance, and renal clearance of heavy metals and organic substances, the authors examined the regressions of the 24-hr urinary excretion on glomerular filtration rate [GFR, 24-hr endogenous creatinine (Cn) clearance] in 19 gun-metal foundry workers with blood lead (Pb) concentrations of 25-59 micrograms/dl. It was estimated that the proportion of FPx to total plasma concentration was on average 15, 7, 3, 0.6, 0.06, and 0.008% for Pb, cadmium (Cd), manganese (Mn), zinc (Zn), chromium (Cr), and copper (Cu), respectively. The estimated FPx value was 2.8 X 10(2), 4, 0.08, and 2.8 X 10(4) micrograms/dl for hippuric acid (HA), delta-aminolevulinic acid (ALA), coproporphyrin (CP), and total urinary solutes (TUS), respectively. The estimated glomerular filtration was significantly greater than the zero level for all substances but inorganic mercury (Hg). Similarly, the estimated net tubular secretion was significantly greater than the zero level for Cr, Cu, and TUS; the net tubular reabsorption was significantly greater than the zero level for Pb, ALA, and CP. The renal clearance of "filterable" plasma substance was significantly greater than GFR for Cr, Cu, and TUS and was significantly smaller for Pb, ALA, and CP. Thus the renal excretory mechanisms of substances were classified into four major categories: glomerular filtration for Cd, Mn, Zn, HA, and Cn; glomerular filtration and net tubular secretion for Cr, Cu, and TUS; glomerular filtration and net tubular reabsorption for Pb, ALA, and CP; and no glomerular filtration, i.e., suspected tubular secretion, for Hg.     

The effects of water restriction and water loading on urinary excretion of lead, delta-aminolevulinic acid and coproporphyrin.

Alterations in daily urinary excretion of lead, delta-aminolevulinic acid (ALA), coproporphyrin, creatinine and total solutes following water restriction and water loading were examined in nine lead workers. Excretion of lead, ALA and total solutes was significantly decreased when urinary volume was reduced, showing that these values are dependent on urinary volume: conversely, excretion of coproporphyrin and creatinine was independent of urinary volume. Excretion of lead and total solutes was also dependent on creatinine excretion. The renal excretory mechanism of lead, ALA and coproporphyrin is discussed in the light of these findings.     

Circadian rhythms of seven heavy metals in plasma, erythrocytes and urine in men: observation in metal workers

To elucidate circadian rhythms (variation within a day) of 7 toxic or essential metals in plasma and erythrocytes in relation to the rhythms in urine in men, 19 male metal foundry workers were examined; they were exposed to lead (Pb), zinc (Zn) and copper (Cu) occupationally but separated from the exposure during the study. Circadian rhythms were found for plasma concentration of Pb, cadmium (Cd), Zn, Cu and chromium (Cr) in the workers. Circadian rhythms were also found for Pb, inorganic mercury (Hg), Zn and Cr in erythrocytes and for all metals except Zn in urine. Both the plasma and urinary levels of Pb, Cd, Cu and Cr together with urinary excretion of Mn and creatinine tended to decrease during night hours; both the erythrocyte and urinary levels of Hg together with both the plasma and erythrocyte levels of Zn tended to increase during morning hours. The results of profile analysis suggested that the circadian rhythm of Pb in urine was affected more strongly by its plasma and erythrocyte rhythms than by the rhythm of creatinine in urine, i.e. the rhythm of glomerular filtration; the urinary rhythms of Cd, Cr, and Mn were affected more by the creatinine rhythm; and the urinary rhythm of Cu was affected by both its plasma and creatinine rhythms. On the other hand, the urinary rhythm of Hg was assumed to be independent of the creatinine rhythm and be affected by its erythrocyte rhythm. The present study suggested that different biological limit values might be needed for different hours of the day especially for shift workers who are exposed to various heavy metals. Instead, further studies should be conducted to find the adjustment methods by which no circadian rhythms are discerned.     

Filterable Plasma Concentration,Glomerular Filtration,Tubular Balance,and Renal Clearance of Heavy Metals and Organic Substances in Metal Workers

To estimate “filterable” plasma concentration (FPx), glomerular filtration, tubular balance, and renal clearance of heavy metals and organic substances, the authors examined the regressions of the 24-hr urinary excretion on glomerular filtration rate [GFR, 24-hr endogenous creatinine (Cn) clearance] in 19 gun-metal foundry workers with blood lead (Pb) concentrations of 25–59 μg/dl. It was estimated that the proportion of FPx to total plasma concentration was on average 15,7,3,0.6,0.06, and 0.008% for Pb, cadmium (Cd), manganese (Mn), zinc (Zn), chromium (Cr), and copper (Cu), respectively. The estimated FPx value was 2.8 × 10², 4,0.08, and 2.8 × 10⁴ μg/dl. for hippuric acid (HA), delta-aminolevulinic acid (ALA), coproporphyrin (CP), and total urinary solutes (TUS), respectively. The estimated glomerular filtration was significantly greater than the zero level for all substances but inorganic mercury (Hg). Similarly, the estimated net tubular secretion was significantly greater than the zero level for Cr, Cu, and TUS; the net tubular reabsorption was significantly greater than the zero level for Pb, ALA, and CP. The renal clearance of “filterable” plasma substance was significantly greater than GFR for Cr, Cu, and TUS and was significantly smaller for Pb, ALA, and CP. Thus the renal excretory mechanisms of substances were classified into four major categories: (1) glomerular filtration for Cd, Mn, Zn, HA, and Cn; (2) glomerular filtration and net tubular secretion for Cr, Cu, and TUS; (3) glomerular filtration and net tubular reabsorption for Pb, ALA, and CP; and (4) no glomerular filtration, i.e., suspected tubular secretion, for Hg.     

Excretion of porphyrin and porphyrin precursor during ethylene oxide inhalation

The effect of chronic inhalation of ethylene oxide on urinary coproporphyrin and delta-aminolevulinic acid were studied. When Wistar male rats were exposed to 500 ppm ethylene oxide three times a week, daily urine volume was increased by 200-300% from the first week to the fifth week of the experimental period. After exposure, daily coproporphyrin excretion and urinary coproporphyrin per mg of creatinine increased by 250% and 141%, respectively. On the other hand, daily excretion of delta-aminolevulinic acid in urine tended to increase but did not increase significantly by creatinine-correction. We think this is the first report of ethylene oxide induced experimental porphyria.     

Exposure to toluene increases the urinary excretion of D-glucaric acid

Workers at a printing plant exposed to low concentrations of toluene (43-401 mg/m3, median 155 mg/m3) had increased urinary D-glucaric acid (3.55-5.12 mmol/mol creatinine) excretion at the end of the shift compared with controls (2.45-3.35 mmol/mol creatinine). No increase was found after the summer holiday (1.92-2.89 mmol/mol creatinine) but excretion had increased two weeks later (4.05-5.55 mmol/mol creatinine). These changes in the excretion of D-glucaric acid were not correlated to levels of exposure, to changes of urinary hippuric acid and o-cresol half lives (three to eight hours), nor to o-cresol/hippuric acid concentration ratios when measured at the end of daily exposure. Since a significant intra and interindividual variability of urinary D-glucaric acid was found in all groups, urinary D-glucaric acid excretion is suitable to monitor group but not individual exposure.     

Exposure to toluene increases the urinary excretion of D-glucaric acid.

Workers at a printing plant exposed to low concentrations of toluene (43-401 mg/m3, median 155 mg/m3) had increased urinary D-glucaric acid (3.55-5.12 mmol/mol creatinine) excretion at the end of the shift compared with controls (2.45-3.35 mmol/mol creatinine). No increase was found after the summer holiday (1.92-2.89 mmol/mol creatinine) but excretion had increased two weeks later (4.05-5.55 mmol/mol creatinine). These changes in the excretion of D-glucaric acid were not correlated to levels of exposure, to changes of urinary hippuric acid and o-cresol half lives (three to eight hours), nor to o-cresol/hippuric acid concentration ratios when measured at the end of daily exposure. Since a significant intra and interindividual variability of urinary D-glucaric acid was found in all groups, urinary D-glucaric acid excretion is suitable to monitor group but not individual exposure.     

Influence of dietary zinc on lead toxicity in the rat.

An investigation of the influence of dietary zinc (8, 35, 200 ppm) on the toxicity of dietary lead (0, 50, 200 ppm) in the young male rat in a seven week period indicated that as dietary zinc increased, the severity of lead toxicity decreased. Evidence included decreased lead concentration in blood, liver, kidneys, and tibias; decreased excretion of urinary delta-aminolevulinic acid; decreased accumulation of free erythrocyte porphyrins; decreased inhibition of kidney delta-aminolevulinic acid dehydrase activity; and a decrease in apparent lead absorption. Infected zinc did not afford protection against lead toxicity. The data indicate that the protective effect of zinc on lead toxicity is largely mediated by an inhibition of lead absorption at the intestinal level.     

Postabsorptive effect of increased dietary zinc on toxicity and removal of tissue lead in rats

Weanling male albino rats were fed a diet containing 12 ppm zinc and 200 ppm lead for 3 weeks. At the end of this time a representative number of samples were collected to determine tissue zinc and lead, inhibition of the lead-sensitive liver enzyme delta-aminolevulinic acid dehydratase (ALAD), and urinary delta-aminolevulinic acid (ALA). Dietary lead exposure was terminated, and the remaining rats were fed diets containing either 12 or 200 ppm zinc. Analyses were repeated at 5-day intervals over a 15-day period after lead exposure. As expected, inhibition of liver ALAD, excretion of urinary ALA and soft tissue lead content rapidly decreased after lead was removed from the diet approaching control levels by day 15. Although high dietary zinc increased the zinc content of plasma, liver and tibia, there was little or no therapeutic effect on recovery of liver ALAD, urinary ALA excretion or on the removal of lead from liver, kidney or tibia. Removal of red blood cell lead, however, was greater for rats fed the high zinc diet. Results of this study indicate that the postabsorptive interaction between zinc and lead is considerably less important than the previously reported intestinal interaction.     

Mobilisation of heavy metals into the urine by CaEDTA: relation to erythrocyte and plasma concentrations and exposure indicators.

To investigate the effects of calcium disodium ethylenediamine tetra-acetate (CaEDTA) on the urinary excretion, erythrocyte, and plasma concentrations and exposure indicators of seven heavy metals, CaEDTA was administered by intravenous infusion to 20 workers exposed to lead, zinc, and copper. The workers' blood lead concentrations ranged from 22 to 59 micrograms/dl (mean 38 micrograms/dl (1.8 mumol/l]. The 24 hour urinary excretion of metals after CaEDTA administration (mobilisation yield) was on average 13 times the background excretion for lead, 11 times for zinc, 3.8 times for manganese, 3.4 times for cadmium, 1.3 times for copper, and 1.1 times for chromium; no significant increase was found for mercury. The mobilisation yield of lead (MPb) was significantly correlated with whole blood and erythrocyte concentrations and the urinary excretion of lead but not with its plasma concentration; similarly, the mobilisation yield of cadmium was significantly correlated with its erythrocyte concentration. In addition, MPb was significantly correlated with intra-erythrocytic enzyme delta-aminolaevulinic acid dehydratase activity and urinary coproporphyrin excretion. The relation between the mobilisation yield of heavy metals and their body burden (and toxic signs) is discussed in the light of these findings.     

Mobilisation of heavy metals into the urine by CaEDTA: relation to erythrocyte and plasma concentrations and exposure indicators

To investigate the effects of calcium disodium ethylenediamine tetra-acetate (CaEDTA) on the urinary excretion, erythrocyte, and plasma concentrations and exposure indicators of seven heavy metals, CaEDTA was administered by intravenous infusion to 20 workers exposed to lead, zinc, and copper. The workers' blood lead concentrations ranged from 22 to 59 micrograms/dl (mean 38 micrograms/dl (1.8 mumol/l]. The 24 hour urinary excretion of metals after CaEDTA administration (mobilisation yield) was on average 13 times the background excretion for lead, 11 times for zinc, 3.8 times for manganese, 3.4 times for cadmium, 1.3 times for copper, and 1.1 times for chromium; no significant increase was found for mercury. The mobilisation yield of lead (MPb) was significantly correlated with whole blood and erythrocyte concentrations and the urinary excretion of lead but not with its plasma concentration; similarly, the mobilisation yield of cadmium was significantly correlated with its erythrocyte concentration. In addition, MPb was significantly correlated with intra-erythrocytic enzyme delta-aminolaevulinic acid dehydratase activity and urinary coproporphyrin excretion. The relation between the mobilisation yield of heavy metals and their body burden (and toxic signs) is discussed in the light of these findings.     

Effects of urinary volume on urinary concentrations of lead, delta-aminolaevulinic acid, coproporphyrin, creatinine, and total solutes.

Urinary volume was related to urinary concentrations of lead, delta-aminolaevulinic acid (ALA), coproporphyrin (COPRO), creatinine, and total solutes in nine lead workers. On a log scale, linear relationships were found between urinary volume and the urinary concentrations. There was a certain difference in the extent of the effects of urinary volume on the urinary concentrations. For example, the concentration of creatinine was more affected by urinary volume than those of lead, ALA, and total solutes among the substances examined: similarly, the concentration of COPRO more than that of ALA. An equation was introduced to eliminate the effects of urinary volume on urinary concentrations. The effects of urinary volume on the concentrations adjusted to urinary specific gravity, osmolality, and creatinine are discussed in the light of these findings.     

Immunization Against Poliomyelitis

The δ-aminolevulinic acid (ALA) dehydratase activity in erythrocytes, the cholinesterase (ChE) activity in serum, the urinary output of ALA, coproporphyrin, protein, and mercury, and the level of reduced glutathione (GSH) in erythrocytes were determined in mercury workers. A significant correlation (P < 0.01) was found between the urinary level of mercury and the values for the decreased activities of ALA dehydratase and ChE, and for the urinary level of coproporphyrin, though their values were not so prominent as in lead poisoning. It was concluded that the response of the biological system in vivo to the metals differs greatly from that in vitro, and that the maximum permissible urinary concentration of mercury would be 200μ g/gm creatinine.     

Adjustment of Creatinine-Adjusted Value to Urine Flow Rate in Lead Workers

Two male lead workers, aged 57 and 51 y, were studied to compare the urinary flow/creatinine-adjusted values published earlier by Araki et al. and by Greenberg and Levine. We collected 24-h urine samples once a month for 31 mo and 16 mo for workers 1 and 2, respectively. The workers' urinary excretions of lead, delta-aminolevulinic acid, and coproporphyrin were measured. No significant correlations between urine flow rate and urinary flow/creatinine-adjusted values published by Araki et al. for the three substances were found for these two workers. However, urinary flow/creatinine-adjusted values presented by Greenberg and Levine for lead and delta-aminolevulinic acid were correlated positively with urine flow rate in the two workers, and their adjusted value for coproporphyrin was correlated positively with urine flow rate in one of the workers. We concluded that use of the urinary flow/creatinine-adjusted value by Greenberg and Levine for biological monitoring poses a problem because of the theoretical fallacy.     

Serum laminin, hydrocarbon exposure, and glomerular damage.

It has been postulated that occupational exposure to hydrocarbons may damage the kidney and lead to glomerulonephritis and chronic renal failure. As laminin is a ubiquitous basement membrane component that seems to play a central part in the structure and function of basement membranes and as the normal renal filtration process is highly dependent on an intact glomerular basement membrane, the serum laminin concentration was examined in a population of workers exposed to hydrocarbons. The hydrocarbon exposure was assessed by exposure surrogates (exposure duration and exposure score). An interaction between occupational exposure to hydrocarbons and hypertension increased the laminin concentration whereas the laminin concentration decreased in workers exposed for a long time probably because of a selection effect. In a subgroup of printers exposed to toluene whose hippuric acid excretion had been recorded for several years this interaction was confirmed when the hippuric acid excretion was substituted for the other exposure indices. In the exposed group, the age-related decline in creatinine clearance was accelerated. These results seem to confirm that occupational exposure to hydrocarbons is a non-specific factor that may promote a deterioration of renal function.     

Influence of dietary protein deficiency on lead-copper interaction in rats

The influence of dietary protein deficiency on the effects of exposure to lead or its combination with copper was investigated in rats. The administration of lead (100 ppm in drinking water) inhibited the activity of blood delta-aminolevulinic acid dehydratase; decreased hemoglobin, brain dopamine, and 5-hydroxytryptamine; and increased urinary excretion of delta-aminolevulinic acid, blood zinc protoporphyrin, and tissue accumulation of lead more markedly in animals fed a protein-deficient diet (10% casein) than in those fed a normal diet (21% casein). The simultaneous supplementation of copper (100 ppm in diet) reduced some of the lead-induced alterations and body uptake of lead more efficiently in animals fed a normal diet than in those fed a protein-deficient diet, which shows that the beneficial effects of copper in lead toxicity are adversely affected by low dietary protein.     

Erythrocyte deformability in workers exposed to lead.

Erythrocyte deformability and other hematological indicators were determined in 17 male workers exposed to lead at a secondary lead refinery and 13 controls. Blood lead, urine lead, urine coproporphyrin, delta-aminolevulinic acid and erythrocyte zinc protoporphyrin were determined to evaluate the degree of lead exposure in the lead workers above. For the measurement of erythrocyte deformability, the microfilter method was used. The results were summarized as follows: 1. The mean values of blood lead, urine lead, urine coproporphyrin, urine delta-aminolevulinic acid and erythrocyte zinc protoporphyrin levels in lead workers were 53.5 micrograms/100g, 141.4 micrograms/l, 115.9 micrograms/l, 12.0 mg/l and 68.9 micrograms/dl respectively, suggesting a moderate influence of lead exposure. 2. The mean values of erythrocyte count, hematocrit and hemoglobin were significantly lower in lead workers than those in controls. No significant differences were found in the mean values of mean corpuscular hemoglobin, mean corpuscular hemoglobin concentration and corpuscular natrium and potassium between lead workers and controls. 3. Erythrocyte deformability was significantly reduced in lead workers compared with controls.