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OBJECTIVES: To assess environmental exposure to polycyclic aromatic hydrocarbons (PAHs) in the vicinity of a Söderberg aluminium reduction plant in Shawinigan, Canada with urinary 1-hydroxypyrene (1-OHP) as a biomarker. METHODS: Urine samples were collected from 20 non-occupationally exposed subjects living less than 500 m from the plant and from 20 controls living in Trois-Rivières, another industrial town 40 km from Shawinigan. Concentrations of 1-OHP were measured by high performance liquid chromatography (HPLC). RESULTS: Among controls, geometric mean (range) 1-OHP concentrations were 0.046 (0.012-0.116) mumol/mol creatinine in non-smokers and 0.125 (0.051-0.282) mumol/mol creatinine in smokers. Among exposed subjects, values were 0.103 (0.056-0.196) mumol/mol creatinine in non-smokers and 0.250 (0.112-0.448) mumol/mol creatinine in smokers. Excretion of 1-OHP was significantly higher in exposed subjects than in controls among non-smokers and smokers (P < 0.05). CONCLUSION: Based on urinary 1-OHP as a biomarker, it seems that living near an industrial point source of PAHs is associated with higher exposure. The health significance of this finding will require further investigation.

beta 2-microglobulin (beta 2-m) was measured in the urine of rats by a specific immunoassay based on latex particles agglutination. The excretion of this protein was compared to the excretion of the enzyme beta-N-acetyl-D-glucosaminidase (NAG), albumin and amino acids in rats treated with either a single dose of sodium chromate (5 and 10 mg kg-1), repeated doses of gentamicin (5 and 20 mg kg-1), or cadmium (1 mg kg-1), and in aging rats (from 2 to 20 months). All treatments resulted in an early increased excretion of beta 2-m indicative of functional alterations of the proximal tubular cells. An increased NAG excretion was observed only at the highest dose of chromate and in the cadmium model but the relative increases of beta 2-m were much larger (up to 200 times the control values against four times the control values for NAG). From 2 to 20 months of age, urinary beta 2-m increases by a factor of four. Aminoacids excretion showed little sensitivity in the various models. Albumin showed little variations in purely tubular or in the tubular phase of renal injury but the chronic progressive nephrosis of aging rats caused a 40-fold increase in its excretion between 2 and 20 months of age. Therefore urinary beta 2-m, albumin and albumin/beta 2-m ratio provide useful tools in the assessment of nephrotoxicity and of its mechanisms in various experimental models.

The urinary excretion of beta 2-microglobulin, retinol-binding protein and albumin was measured in 65 workers exposed to styrene at levels averaging 50 percent of the current threshold limit value (215 mg/m2) for 1-13 years (mean: 6 years). By comparison with a control group matched for age and socioeconomic status, no significant difference was observed in the urinary excretion of proteins. In rats, styrene was weakly nephrotoxic. No functional or morphological renal change could be disclosed in rats exposed to 565 mg of styrene/m3, 5 days/week for 13 weeks. The repeated i.p. injection of 1 g styrene/kg (1/5 of oral LD50) for 10 days induced only a slight tubular dysfunction as evidenced by a 5-fold increase in beta 2-microglobulinuria. Altogether, these epidemiological and experimental data suggest that the current threshold limit value for styrene (215 mg/m3) proposed by the American Conference of Governmental and Industrial Hygienists does not entail any risk of renal toxicity.

Inhalation exposure of male Sprague-Dawley rats to an industrial white-spirit solvent composed of a C10–C12 mixture of branched-chain unsubstituted satured aliphatic hydrocarbons at 6500 mg/m3 for over 9 months resulted in decreased urinary concentrating ability, decreased net acid excretion following a mild ammonium chloride load and increased urinary lactate dehydrogenase (LDH) activity, whereas urinary s-N- acetyl- d -glycosaminidase (NAG) activity remained normal. These observations suggest the existence of distal tubular alteration in the rat kidney.

To study the effect of gentamicin on the renal uptake of proteins, Sprague-Dawley female rats were intravenously injected with solutions containing unlabeled human beta 2-microglobulin (beta 2-m), retinol-binding protein, and increasing amounts of gentamicin (from 0.063 up to 31.5 mg/kg). The concentrations of human proteins and that of endogenous beta 2-m, albumin, and IgG in the urine collected during the 2 hr following the injection were determined by immunoassays. Gentamicin transiently increased the urinary excretion of rat and human beta 2-m in a dose-dependent manner. The mean relative increase of rat beta 2-m excretion ranged from 2 at a gentamicin dose of 0.06 mg/kg up to 500 at a gentamicin dose of 31.5 mg/kg. By contrast, the urinary excretion of other proteins was only increased by a factor of 2 to 5 at the highest dose of gentamicin. The relative increase of the urinary excretion of proteins was positively correlated with the fractional reabsorption of the proteins by the rat kidney. The inhibitory effect of gentamicin on the renal uptake of protein was very similar to that observed in rats injected with polycationic proteins like lysozyme and cytochrome C. These observations, combined with the fact that gentamicin, like proteins, enters the tubular cell by adsorptive endocytosis, strongly suggest that this drug competes with proteins for common binding sites on the apical tubular membrane and for subsequent endocytosis. Furthermore, the iv injection of large amounts of gentamicin and polycationic proteins induces a lysosomal enzymuria which very likely is a manifestation of an increased exocytosis.

The renal uptake of proteins: A nonselective process in conscious rats. The selectivity of the renal reabsorption of proteins has been investigated by competition experiments in conscious rats. The animals were intravenously injected with increasing doses of proteins over a wide range of net charge and size, including lysozyme, cytochrome C, metallothionein, β2-microglobulin, retinol-binding protein, albumin and IgG. The urinary excretion of exogenous proteins injected concomitantly (humanβ2-microglobulin, retinol-binding protein, albumin and/or egg white lysozyme depending on the experiment) and of rat β2-microglobulin, albumin and IgG was determined with specific immunoassays. The results show that low molecular weight cationic proteins and low or high molecular weight anionic proteins can increase each other's urinary excretion. Several observations strongly suggest that these effects result from a competitive inhibition of renal uptake. The phenomenon is dose-related in most cases and, as evidenced by cytochrome C injection, transient, reproducible and saturable. In addition, the injected proteins induce a tubular type proteinuria irrespective of their net charge and size. In the case of cationic proteins, this finding excludes the possibility of an enhanced glomerular permeability due to a partial neutralization of the glomerular polyanion which, as demonstrated with protamine sulfate, entails a glomerular type proteinuria. These quantitative data on the mutual inhibition of renal uptake of a wide spectrum of specific proteins lead us to challenge the concept of charge- and size-selective tubular reabsorption of proteins, and to postulate that proteins filtered through the glomeruli are taken up by common tubular endocytotic sites irrespectively of their physicochemical features. As demonstrated by the ability of β2-microglobulin and IgG to inhibit the uptake of lysozyme, the affinity of a protein for reabsorption sites is not simply related to its size and net positive charge. Evidence is also presented that proteins, when administered intravenously at high doses, induce a lysosomal enzymuria most likely reflecting a stimulated exocytosis.

The effect of a previous chronic exposure to cadmium, lead or inorganic mercury on the nephrotoxic potential of gentamicin was investigated in female Sprague-Dawley rats. A daily dose of 10 mg gentamicin/kg body weight/day was administered for 21 days to rats having a renal load of 168 micrograms Cd, 35 micrograms Pb or 129 micrograms Hg/g whole kidney. Urine analysis suggests an attenuation of the nephrotoxic potential of gentamicin while a microscopical examination of kidneys indicates a superimposition of the effects of the metals and the antibiotics. The only clear interaction observed consists in a reduction of gentamicin accumulation in the cortex of cadmium-treated animals. It is concluded that none of the metal pretreatments potentiates the nephrotoxic effects of gentamicin.

Polychlorinated dibenzo-p-dioxins (PCDDs) and other organic compounds are solvent extracted from 10- to 20-g samples of fly ash from municipal incinerators with 200 mL of benzene using ultrasonic agitation for 1 h. A convenient filtering device is used to separate fly ash and solvent which is then concentrated to 100 ..mu..L using a rotary evaporator. Extracts are analyzed directly by gas chromatography/mass spectrometry, including selected ion monitoring for PCDDs. Results from five replicate analyses of a fly ash sample yielded averages and standard deviations (ng/g) for the tetra-to octachlorinated dibenzo-p-dioxins of 8.6 +- 2.2, 15.0 +- 4.0, 13.0 +- 3.4, 3.2 +- 1.0, and 0.4 +- 0.1, respectively. 2 tables, 3 figures.

Fly-ash samples from municipal incinerators in Canada and Norway were analyzed for polychlorinated dibenzo-p-dioxins using gas chromatography/mass spectrometry preceded by fractionation on column chromatography. Qualitative and quantitative differences in the organic compounds identified were found between the two samples. Many of the same dioxin congeners were present in both samples; however, tetrachlorodibenzo-p-dioxins were only detected in the Canadian sample.

Prolonged inhalation exposure to 6.5 mg/l of an isoparaffinic solvent consisting of saturated aliphatic hydrocarbons (SAHC) resulted in both functional and morphological renal changes in male rats to the exclusion of female or castrated rats. Functionally, the increased excretion of lactate dehydrogenase in the absence of an increased beta-N-acetyl-D-glucosaminidase excretion together with a decreased urinary concentrating ability upon water deprivation and slower antinatriuretic response when the sodium intake is abruptly reduced, suggest a distal tubular alteration. beta 2-Microglobulin excretion is unchanged indicating good proximal tubular cell function. The increased excretion of albumin and slightly lowered glomerular filtration rate suggest a moderate glomerular impairment. Light microscopy shows prominent hyaline droplet accumulation in proximal tubular cells and a few scattered foci of regenerative epithelia in both proximal and distal cells of the deep cortex. The urinary clearance of the major male rat urinary protein, alpha 2u-globulin, is similar in control and exposed rats but the latter have a 10-fold greater renal accumulation of this protein while the hepatic levels are identical in both groups. It is concluded that SAHC exposure causes moderate and reversible tubular and also glomerular changes in the male rat kidney.

In workers chronically exposed to cadmium and without signs of renal insufficiency, plasma proteins with molecular weight ranging from 11,800 to 450,000 are excreted in greater amount in urine. Increased urinary excretion of low and high molecular weight proteins can occur independently. Because of its greater stability in urine and provided a sensitive immunological technique is used, the determination of retinol-binding protein is a more practical and reliable test of proximal tubular function than beta 2-microglobulin. The evaluation of renal function of workers removed from cadmium exposure indicates that cadmium-induced renal lesions, albeit of slow progression, are not reversible when exposures ceases. In workers chronically exposed to cadmium or removed from cadmium exposure, metallothionein in urine is directly correlated with cadmium in urine but not with cadmium in blood or years of cadmium exposure. The association between cadmium in urine and metallothionein in urine is independent of the status of renal function and the intensity of current exposure to cadmium. Whereas the repeated IP injection of high doses of cadmium to rat gives rise to a mixed or tubular type proteinuria, the prolonged oral administration of cadmium results mainly in the development of a glomerular type proteinuria. The former is usually reversible after cessation of treatment whereas the latter is not. Circulating antiglomerular basement membrane antibodies have been found in man and in rat chronically exposed to cadmium. The pathogenic significance of this finding deserves further investigation. Images FIGURE 5.

j Summary: A sensitive immunoassay based on latex particle agglutination for the measurement of circulating j IgE-containing complexes is described. In this method, the anti-IgE-coated particles are incubated with !' diluted serum and the resulting agglutination is quantified by turbidimetry or particle counting. In the latter \ Version, the assay is fully automated in a continuous flow System. IgE-containing complexes were detected i in all tested sera. Increased concentrations were observed in about 80% of the subjects with elevated serum j IgE. However, high levels of IgE-complexes may also be found in subjects with a normal or even a very low j serum concentration of IgE. The same latex immunoassay can be used for the determination of total IgE, after pepsin digestion of the -globulin fraction of the serum. The results obtained correlate well with those found with a sandwich radioimmunoassay (r = 0.91, n = 83). The present method, however, yields a greater number of significantly positive results than the radioimmunoassay, probably because of its ability to detect IgE entrapped in circulating complexes.

The renal handling of human beta 2-microglobulin (beta 2-m) was investigated in normal rat and in rat with cadmium-induced renal damage. Cadmium was administered either in drinking water at a concentration of 100 ppm for up to 16 months or by i.p. injection of 1 mg Cd/kg, five times a week for up to 4 months. When renal dysfunction has developed, namely after 2 and 10 months of the i.p. and oral treatment respectively, unlabelled human beta 2-m was injected intravenously and its disappearance in serum and its urinary excretion were studied by means of a sensitive immunoassay. In serum, the level of beta 2-m drops by about 90% during the 10 first min, then declines more slowly with a half life around 20 min. Serum disappearance curves of beta 2-m in normal and cadmium-treated rats did not differ markedly. The amount of beta 2-m recovered in urine during the 4 h following the injection averaged 0.03% of the injected dose in normal rats. It increased on the average to 10% in rats treated i.p. with 1 mg Cd/kg for 3 months. However, in rats given 100 ppm Cd per os for 10 months, this amount averaged only 0.14%. A similar value was observed 5 months later, although at that stage, the critical level of cadmium in kidney cortex had been reached for 6-7 months. These data which were in accordance with the disturbances of the other renal parameters measured in cadmium-treated rats indicate that: 1) human beta 2-m is reabsorbed by rat kidney at a similar rate as by human kidney; 2) if the occurrence of cadmium tubulopathy is concomitant with the saturation of cadmium-binding sites in kidney, its severity depends greatly on the rate at which cadmium reaches the saturated kidneys.

Rat beta 2-microglobulin has been isolated from the urine of rats pretreated with sodium chromate. The purified protein was used to raise antisera in two rabbits. Treatment of the antisera with ammonium sulphate followed by gel chromatography led to an immunoglobulin fraction which was used to coat latex particles by physical adsorption. The latex suspension was used in an automated system including an optical particle counter to analyse the protein in serum, urine and cerebrospinal fluid. Serum contains 5.8 mg of beta 2-microglobulin/l, of which 33% is found as a 55000 dalton complex while 66% is present as the 'free' protein. The daily urinary excretion of beta 2-microglobulin in females is about 2 micrograms of which 7% is found to be a 65000-dalton complex while 92% is the free protein. From this, it can be calculated that the fractional urinary excretion of beta 2-microglobulin is about 0.03%. The cerebrospinal fluid contains about 1 mg of beta 2-microglobulin/l. Preliminary tests also suggest that the method can be adapted for non-automated turbidimetric detection. In the automated assay, the within- and between-assay coefficients of variation are less than 10% for the three biological fluids tested. The analytical recovery in the urine is 93%. In urine, beta 2-microglobulin undergoes proteolytic degradation at pH below 6. This does not represent a serious drawback to its use as a sensitive index of tubular function since in most experimental circumstances, rats excrete urine with a pH above this value.

Female Sprague-Dawley rats received phenacetin or aspirin at average daily doses of 135 and 27 mg/kg respectively in the diet and either demineralized water (DMW) or a 100 ppm cadmium (Cd) solution as their drinking water for 12 months. This dose of Cd produced borderline tubular toxicity, as measured by the excretion of IV-injected human beta 2-microglobulin. The kidney accumulation of Cd just reached the critical level of 200 ppm in all groups at the end of the study. The various treatments did not significantly affect growth, creatinine clearance, urine osmolality and the urinary excretion of beta-N-acetyl-D-glucosaminidase and aminoacids. No interaction resulted from the concomitant administration of analgesics and Cd. Both aspirin subgroups (receiving DMW or Cd) showed an attenuation of the age-related decline of the renal function as revealed by a lower urinary excretion of albumin and total protein. The accentuation of the mesangial matrix seen upon aging was also partly inhibited in the aspirin rats.

The relationships between the urinary excretion of ferritin and that of albumin, β2-microglobulin and retinol-binding protein were studied in 60 healthy subjects and in 34 subjects exposed to a nephrotoxic agent (cadmium). The results suggest that the ferritin present in urine originates mainly from the kidney and its concentration depends on both the renal store of ferritin and the importance of tubular cell damage.

Describes the design, use, and advantages of a gas chromatography / mass spectrometer / computerized data system.