Saliva DHEA and cortisol responses following short-term corticosteroid intake.

BACKGROUND: Given the high correlation between the serum and saliva hormone values demonstrated at rest, saliva provides a convenient non-invasive way to determine dehydroepiandrosterone (DHEA) and cortisol concentrations. However, to our knowledge, pituitary adrenal recovery following short-term suppression with corticosteroids has never been investigated in saliva. The aim of this study was therefore to examine how steroid hormone concentrations in saliva are influenced by short-term corticosteroid administration. MATERIALS AND METHODS: We studied saliva DHEA and cortisol concentrations before, during (day 1-day 7) and following (day 8-day 16) the administration of oral therapeutic doses of prednisone (50 mg daily for 1 week) in 11 healthy recreationally trained women. RESULTS: Mean saliva DHEA and cortisol concentrations decreased immediately after the start of prednisone treatment (P < 0.05). Three days after concluding prednisone administration, both saliva DHEA and cortisol had returned to pretreatment levels. CONCLUSIONS: These data are consistent with previous studies on blood samples and suggest that non-invasive saliva samples may offer a practical approach to assessing pituitary-adrenal function continuously during and after short-term corticosteroid therapy.

Cortisol, DHEA, and testosterone concentrations in saliva in response to an international powerlifting competition.

The purpose of this study was to examine salivary cortisol, dehydroepiandrosterone (DHEA), and testosterone responses to the bench press in an international powerlifting competition and to determine whether these salivary hormone concentrations could be used to predict performance. Twenty-six elite athletes (13 females and 13 males) provided saliva samples during the official weighing-in and after the last attempt at the bench press, as well as at baseline on a non-competition day. Performance index was determined with the Wilks formula, which adjusts powerlifting scores according to body mass. Salivary cortisol concentrations were significantly increased in all subjects after the bench press (p < 0.01), whereas DHEA concentrations were significantly increased in women (p < 0.01) but not in men after the bench press. No significant change in testosterone concentrations was observed during the experiment in either men or women, which resulted in a marked decrease in the testosterone/cortisol ratio. The performance index showed no significant correlation with any of the hormone responses to competition. In conclusion, despite the increase in stress adrenocortical hormone responses to an international powerlifting competition, these hormone concentrations alone are not predictors of bench press performance in elite powerlifting athletes.

Isotope ratio mass spectrometry analysis of the oxidation products of the main and minor metabolites of hydrocortisone and cortisone for antidoping controls.

Metabolites of hydrocortisone (HC) and cortisone (C), namely tetrahydrocortisol (THF), tetrahydrocortisone (THE), allo-THF, allo-THE for the main metabolites and 11-hydroxyandrosterone, 11-hydoxyetiocholanolone, 11-ketoandrosterone, and 11-ketoetiocholanolone for the minor metabolites, as well as the two main metabolites of testosterone, androsterone and etiocholanolone, were separated from each other using HPLC fractionation of urine extracts. An isotopic ratio mass spectrometry (IRMS) analysis determined the absolute delta(13)C values of 5alpha-androstanetrione (5alpha-AT) and 5beta-androstanetrione (5beta-AT) as the oxidation products (ox-products) of the HC and C metabolites and as target compounds (TCs). We also performed IRMS analysis of 5alpha-androstanedione (5alpha-AD) and 5beta-androstanedione (5beta-AD) as the ox-products of etiocholanolone and androsterone and as endogenous reference compounds (ERCs). Urine samples came from two male volunteers treated with a single 10-mg oral dose and a single 100-mg intramuscular dose of HC hemisuccinate, a male volunteer treated with a single 25-mg oral dose of C acetate, and a control group of 30 drug-free athletes. The mean -3SD of delta(13)C depletion values from the controls were -1.46, -1.98, -1.78 and -2.42 for 5beta-AT-5beta-AD, 5alpha-AT-5beta-AD, 5beta-AT-5alpha-AD and 5alpha-AT-5alpha-AD, respectively, indicating -3 per thousand as a safe cut-off value for differentiating the pharmaceutical from the natural form. In the main metabolite fraction, delta(13)C depletion values peaked around -5 per thousand and -9 per thousand after oral and intramuscular administration of HC, respectively, and around -6 per thousand after oral administration of C. In comparison, less impressive results were obtained when IRMS analysis focused on the ox-products of the minor metabolites.

Salbutamol intake and substrate oxidation during submaximal exercise.

In order to test the hypothesis that salbutamol would change substrate oxidation during submaximal exercise, eight recreationally trained men twice performed 1 h at 60% VO(2) peak after ingestion of placebo or 4 mg of salbutamol. Gas exchange was monitored and blood samples were collected during exercise for GH, ACTH, insulin, and blood glucose and lactate determination. With salbutamol versus placebo, there was no significant difference in total energy expenditure and substrate oxidation, but the substrate oxidation balance was significantly modified after 40 min of exercise. ACTH was significantly decreased with salbutamol during the last 10 min of exercise, whereas no difference was found between the two treatments in the other hormonal and metabolic parameters. The theory that the ergogenic effect of salbutamol results from a change in substrate oxidation has little support during relatively short term endurance exercise, but it is conceivable that longer exercise duration can generate positive findings.

Effects of short-term corticoid ingestion on food intake and adipokines in healthy recreationally trained men.

In order to test the hypothesis that short-term corticoid intake alters food intake, body composition and adipokines secretion in healthy volunteers with regular sport practice, nutrient intake was assessed in eight male athletes with and without prednisolone (PRED, 60 mg/day for 1 week) ingestion in a random, double blind, crossover design. Body weight, body composition, adipokines (i.e., leptin, adiponectin and TNF-alpha), insulin and blood glucose were determined before and at the end of each treatment. PRED did not induce any significant change in body weight, body composition or food intake. Insulin and TNF-alpha were not significantly altered with PRED compared to placebo but blood glucose, leptin and adiponectin concentrations at rest appear significantly increased after PRED treatment (P < 0.05). Our data show that 1 week glucocorticoid treatment does not promote obesity in recreationally trained men but further studies are necessary to understand its effects on the metabolically active hormones, leptin and adiponectin.

Short-term glucocorticoid intake combined with intense training on performance and hormonal responses.

OBJECTIVE: To investigate the effects of short-term prednisolone ingestion combined with intense training on exercise performance, hormonal (adrenocorticotrophic hormone (ACTH), prolactin, luteinising hormone (LH), growth hormone (GH), thyroid-stimulating hormone (TSH), dehydroepiandrosterone (DHEA), testosterone, insulin) and metabolic parameters (blood glucose, lactate, bicarbonate, pH). METHODS: Eight male recreational athletes completed four cycling trials at 70-75% peak O(2) consumption until exhaustion just before (1) and after (2) either oral placebo or prednisolone (60 mg/day for 1 week) treatment coupled with standardised physical training (2 hours/day), according to a double-blind and randomised protocol. Blood samples were collected at rest, during exercise and passive recovery for the hormonal and metabolic determinations. RESULTS: Time of cycling was not significantly changed after placebo but significantly increased (p<0.05) after prednisolone administration (50.4 (6.2) min for placebo 1, 64.0 (9.1) min for placebo 2, 56.1 (9.1) min for prednisolone 1 and 107.0 (20.7) min for prednisolone 2). There was no significant difference in any measured parameters after the week of training with placebo but a decrease in ACTH, DHEA, PRL, GH, TSH and testosterone was seen with prednisolone treatment during the experiment (p<0.05). No significant change in basal, exercise or recovery LH, insulin, lactate, pH or bicarbonate was found between the two treatment, but blood glucose was significantly higher under prednisolone (p<0.05) at all time points. CONCLUSION: Short-term glucocorticoid administration induced a marked improvement in endurance performance. Further studies are needed to determine whether these results obtained in recreational male athletes maintaining a rigorous training schedule are gender-dependent and applicable to elite athletes.

Effects of acute prednisolone administration on exercise endurance and metabolism.

OBJECTIVE: To examine whether acute glucocorticoid (GC) intake alters performance and selected hormonal and metabolic variables during submaximal exercise. METHODS: In total, 14 recreational male athletes completed two cycling trials at 70-75% maximum O(2) uptake starting 3 h after an ingestion of either a lactose placebo or oral GC (20 mg of prednisolone) and continuing until exhaustion, according to a double-blind randomised protocol. Blood samples were collected at rest, after 10, 20, 30 minutes, and at exhaustion and recovery for measurement of growth hormone (GH), adrenocorticotropic hormone (ACTH), dehydroepiandrosterone (DHEA), prolactin, insulin, blood glucose, lactate and interleukin (IL)-6 determination. RESULTS: Cycling duration was not significantly changed after GC or placebo administration (55.9 (5.2) v 48.8 (2.9) minutes, respectively). A decrease in ACTH and DHEA (p<0.01) was observed with GC during all of the experiments and in IL-6 after exhaustion (p<0.05). No change in basal, exercise or recovery GH, prolactin, insulin or lactate was found between the two treatments but blood glucose was significantly higher with GC (p<0.05) at any time point. CONCLUSION: From these data, acute systemic GC administration does seem to alter some metabolic markers but did not influence performance during submaximal exercise.

Short term salbutamol ingestion and supramaximal exercise in healthy women.

OBJECTIVE: To test the hypothesis that chronic salbutamol intake improves performance during supramaximal exercise and to estimate the effects of this treatment on body composition, bone mass, and metabolic indices in healthy women. METHODS: Fourteen female volunteers (seven sedentary and seven recreationally trained) performed a 30 second Wingate test with and without salbutamol ingestion (12 mg/day for four weeks) in a random, double blind, crossover design. Blood samples were collected at rest, at the end of the test, and during passive recovery for lactate measurement. Body composition and bone mass were determined by dual energy x ray absorptiometry. RESULTS: Peak power appeared significantly earlier and was significantly (p<0.05) increased after salbutamol intake in all subjects. There was no difference in total work performed and fatigue indices with salbutamol compared with placebo. No significant alterations in lean or fat body mass and bone variables were observed with salbutamol treatment in either trained or untrained subjects during the trial. In contrast, blood lactate was significantly (p<0.05) increased during the recovery period after salbutamol ingestion compared with placebo. CONCLUSION: As in men, chronic administration of therapeutic concentrations of salbutamol did not induce an anabolic effect in women but increased maximal anaerobic power. Further studies are necessary to clarify the mechanisms involved.

Effects of short-term oral salbutamol administration on exercise endurance and metabolism.

The present study examined whether oral short-term administration of salbutamol (Sal) modifies performance and selected hormonal and metabolic variables during submaximal exercise. Eight recreational male athletes completed two cycling trials at 80-85% peak O(2) consumption until exhaustion after either gelatin placebo (Pla) or oral Sal (12 mg/day for 3 wk) treatment, according to a double-blind and randomized protocol. Blood samples were collected at rest, after 5, 10, and 15 min, and at exhaustion to determine growth hormone (GH), cortisol, testosterone, triiodothyronine (T(3)), C peptide, free fatty acid (FFA), blood glucose, lactate, and blood urea values. Time of cycling was significantly increased after chronic Sal intake (Sal: 30.5 +/- 3.1 vs. Pla: 23.7 +/- 1.6 min, P < 0.05). No change in any variable was found before cycling except a decrease in blood urea concentration and an increase in T(3) after Sal that remained significant throughout the exercise test (P < 0.05). Compared with rest, exercise resulted in a significant increase in GH, cortisol, testosterone, T(3), FFAs, and lactate and a decrease in C peptide after both treatments with higher exercise FFA levels and exhaustion GH concentrations after Sal (P < 0.05). Sal but not Pla significantly decreased exercise blood glucose levels. From these data, short-term Sal intake did appear to improve performance during intense submaximal exercise with concomitant increase in substrate availability and utilization, but the exact mechanisms involved need further investigation.

Effects of prenatal methylmercury exposure on urinary proximal tubular enzyme excretion in neonatal rats.

The basal developmental pattern of excretion of 3 proximal tubular enzymes was determined in 8-h urinary specimens from neonatal rats. Gammaglutamyltransferase (GGT), alkaline phosphatase (ALP), and N-acetyl-beta-glucosaminidase (NAG) activities were measured at 3, 6, 9 and 12 days after birth. Subsequently, methylmercury chloride (CH3HgCl), known to induce foetotoxic changes in the proximal tubule was administered on days 8, 10 and 12 of gestation at 3 or 6 mg/kg and its effects on the enzyme activities were examined. Dose-related increases in the 3 enzyme activities occurred at dose levels that produced no maternal or postnatal toxicity, nor overt morphological malformation of the kidney. The peak of enzyme activities averaged about 200% and 130% of the control values for GGT, ALP, and NAG respectively, and occurred on days 3 and 6 in the treated groups. Urinary enzyme activities returned to the control levels from days 6 to 12. Our data point to the possibility of detecting CH3HgCl-induced prenatal effect on the kidney by measuring the 8-h urinary excretion of enzymes by rats in the early postnatal period.

Pregnancy-associated changes in renal toxicity of cadmium-metallothionein: possible role of intracellular metallothionein.

Nulliparous female, 10-day and 20-day pregnant rats were injected intraperitoneally with saline or labelled cadmium-metallothionein (109Cd-MTh) at a single dose of 25 or 250 micrograms Cd as cadmium-metallothionein (Cd-MTh)/kg and sacrificed at 24 h. The renal toxicity was manifested by increased 24-h urinary excretion of beta 2-microglobulin (beta 2-m) and the increased number of damaged convoluted proximal tubules at 24 h. The renal excretion of 109Cd and 109Cd content in the maternal liver and kidney and in the foeto-placental unit were determined. The binding of 109Cd to kidney proteins and the level of intracellular metallothionein (MTh) in livers and kidneys were also determined. It was found that the nephrotoxicity of injected Cd-MTh did not differ in nulliparous and 10-day pregnant rats. This result was consistent with the absence of difference in the renal uptake of 109Cd, its binding to kidney proteins and in the content of endogenous MTh in the kidneys between nulliparous and 10-day pregnant rats. In contrast, 20-day pregnant rats exhibited much more nephrotoxicity than nulliparous rats. The most prominent finding in relation to the extreme sensitivity of 20-day pregnant rats was a lower basal level of intracellular MTh in the kidneys and the accumulation of 109Cd in the high molecular weight proteins in the soluble fraction. It is suggested that the decrease of intracellular MTh in the kidneys of 20-day pregnant rats is the reason for the low protection against the renal toxicity of injected Cd-MTh.

Guinea pig pulmonary response to sensitization by five preformed monoisocyanate-ovalbumin conjugates.

The ability of 5 dissimilar monoisocyanates conjugated to ovalbumin (OA) as a carrier protein to induce pulmonary hypersensitivity towards the hapten specific component was assessed by using a previously described method based on the determination of a respiratory index (RI) in the guinea pig. The test chemicals included the commercially available p-tolyl and hexyl monoisocyanates (TMI and HMI), with 4-isocyanoto-4'-diphenylmethane (IDM), 4-isocyanoato-4'-methyldiphenylmethane (IMDM) and 1-isocyanato-methyl-1,3,3-trimethylcyclohexane (IMTC) as synthetized monoisocyanates. Guinea pigs were exposed daily to an aerosol of the OA conjugate of each monoisocyanate up to day 15. Increases in respiratory rate and/or respiratory collapse occurred in the guinea pigs exposed to TMI-OA and HMI-OA conjugates by days 9 and 15, with RI values of 155 and 177, respectively, being recorded. The greatest mean RI values in guinea pigs exposed to IDM-OA, IMDM-OA and IMTC-OA conjugates up to day 15 were 20, 25 and 22, respectively, and were not indicative of any pulmonary reaction. Guinea pigs exposed in parallel to each test conjugate did not exhibit any pulmonary reaction when they were exposed to OA on the challenge days. All these findings evidence pulmonary hypersensitivity as the result of exposure to TMI-OA and HMI-OA conjugates and suggest a high degree of conjugation and strong linkage of all the monoisocyanates with OA.(ABSTRACT TRUNCATED AT 250 WORDS)

An attempt to explain interindividual variability in 24-h urinary excretion of inorganic arsenic metabolites by C57 BL/6J mice.

Forty C57 BL/6J mice, injected subcutaneously with 0.5 mg/kg arsenic as sodium arsenite, were examined for 24-h urinary excretion of total arsenic metabolites, creatinine and S-adenosylmethionine (SAM) and for 24-h faecal excretion of arsenic and levels of arsenic in the blood, liver, kidneys, lung, skin, spleen and bone at 24-h post-dose. Total urinary arsenic metabolites were calculated by summing up the inorganic (Asi), monomethylated (MMA) and dimethylated (DMA) derivatives directly measured by selective arsine generation-atomic absorption spectrometry (AG-AAS) or were measured by AG-AAS following complete mineralization. Both sets of results showed interindividual differences varying by as much as 7-fold and correlated with the 24-h urinary excretion of both SAM (r = 0.84 and r = 0.86, respectively) and creatinine (r = 0.82 and r = 0.87, respectively). There was interindividual variability of about a 30-fold range in 24-h faecal excretion of arsenic which correlated inversely with 24-h urinary excretion of arsenic metabolites (r = -0.69) and 24-h urinary excretion of both creatinine (r = -0.70) and SAM (r = -0.67). Body tissue levels of arsenic were low and not related to 24-h urinary excretion of arsenic metabolites, SAM and creatinine. Taken together, the results indicate that differences in the profile of urinary arsenic excretion and in the retention of arsenic in a particular organ do not contribute to interindividual variability in 24-h urinary excretion of arsenic metabolites by C57 BL/6J mice, but that variability in faecal excretion does, at least in part. It is speculated that there is most likely a predominant contribution from a diffuse tissue retention of arsenic or from a third route of arsenic elimination, i.e. respiratory, to this phenomenon in view of the small faecal contribution.

Probenecid-induced protection against acute hexachloro-1,3-butadiene and methyl mercury toxicity to the mouse kidney.

Male Swiss OF1 mice received a single oral dose of either 80 mg/kg hexachloro-1,3-butadiene (HCBD) or 40 mg/kg methyl mercury (MeHg). Examination of cryostat kidney sections stained for alkaline phosphatase (APP) revealed damage to about 50% of the proximal tubules after 8 h. Treatment with the organic anion transport inhibitor probenecid (i.p., 3 x 0.75 mmol/kg) did not have any renal effect in normal mice but reduced the number of damaged tubules by 80 and 90% in mice treated with HCBD and MeHg respectively. The results support the conclusion that the toxicity of HCBD and MeHg to the mouse kidney is related to a probenecid-sensitive transport process. It cannot be stated from the present investigation whether the inhibition nephrotoxicity data are related to classic organic anion secretion by the kidney.

Role of gamma-glutamyltranspeptidase and beta-lyase in the nephrotoxicity of hexachloro-1,3-butadiene and methyl mercury in mice.

Male Swiss OF1 mice received a single oral dose of either 80 mg/kg hexachloro-1,3-butadiene (HCBD) or 80 mg/kg methyl mercury (MeHg). Examination of cryostat kidney sections stained for alkaline phosphatase (APP) revealed damage to about 50% of the proximal tubules after 8 h. Pretreatment with the gamma-glutamyltranspeptidase (gamma-GT) inactivator AT-125 (Acivin, 50 mg/kg i.p., plus 50 mg/kg p.o., reduced the number of damaged tubules by 59 and 58% in mice treated with HCBD and MeHg, respectively. Pretreatment with the two beta-lyase inhibitors, amino-oxyacetic acid (AOAA, 3 x 100 mg/kg p.o.) and DL-propargylglycine (PPG, 300 mg/kg i.p. plus 300 mg/kg p.o.), reduced HCBD nephrotoxicity by 46 and 59%, respectively, but did not protect against MeHg nephrotoxicity. The results support a role for gamma-GT and beta-lyase in the mouse renal toxicity of HCBD and implicate gamma-GT but not beta-lyase in MeHg-induced nephrotoxicity in mice.

Effects of probenecid and acivicin on potassium dichromate-induced acute nephrotoxicity in mice.

Male OF1 mice were injected subcutaneously with 80 mg/kg potassium dichromate (K2Cr2O7). Examination of cryostat kidney sections stained for alkaline phosphatase (APP) revealed damage to about 40-70% of the proximal tubules after 8 h. Pretreatment with the organic anionic transport inhibitor probenecid (i.p., 3 x 0.75 mmol/kg) reduced the number of damaged tubules by 60% in mice treated with potassium dichromate. Pretreatment with the gamma-glutamyltranspeptidase (gamma-GT) inactivator acivicin (AT-125, 50 mg/kg p.o., plus 50 mg/kg i.p.) failed to prevent chromate-induced renal toxicity. These results support the conclusion that a probenecid-sensitive transport process, but not a gamma-GT-catalyzed degradation, is involved in the mouse renal toxicity of potassium dichromate.

Assessment of urinary rat beta 2-microglobulin by enzyme immunoassay.

An enzyme immunoassay (EIA) was developed using unlabelled and peroxidase-labelled rabbit antibodies to assess urinary rat beta 2-microglobulin (beta 2-m) excretion. It consisted in the adsorption of rabbit anti-rat beta 2-m immunoglobulin to a polystyrene surface, the addition of beta 2-m samples or standard and the addition of peroxidase-labelled rabbit anti-rat beta 2-m immunoglobulin. After addition of hydrogen peroxide and o-phenylenediamine, the enzyme activity of the solid phase was measured photometrically at 490 nm. Analytical recovery of pure beta 2-m in urine was 102%. From determinations made on normal female and male rats, the ranges of 24-h urine beta 2-m individually excreted were found to be 0.84-4.77 and 3.7-17.3 micrograms, respectively. The means +/- SEM were 2.49 +/- 0.17 and 10.2 +/- 0.55 micrograms. EIA was of value in evidencing the tubule-damaging properties of sodium chromate and hexachloro-1,3-butadiene in rats.

Effects of inhalation exposure to carbon disulfide and its combination with hydrogen sulfide on embryonal and fetal development in rats.

Pregnant rats were exposed to 0, 100, 200, 400 or 800 ppm of carbon disulfide (CS2), 100 ppm of hydrogen sulfide (H2S) alone or in combination with 400 and 800 ppm CS2, 6 h/d during days 6-20 of gestation. Maternal reproduction and fetal parameters were evaluated on gestational day 21. Treatment with 100 or 200 ppm CS2 or with 100 ppm H2S caused no maternal toxicity or adverse effects on the developing embryo or fetus. Exposure to 400 or 800 ppm CS2 resulted in a low incidence of club foot and in a significant reduction of maternal weight gain. Significant increases in unossified sternebrae occurred at 800 ppm CS2 and reduction of fetal body weight at 400 and 800 ppm CS2. The latter effect was enhanced by combination with 100 ppm H2S. These results support the conclusion that, at levels of exposure associated with maternal toxicity, CS2 leads to an increase in incidence of club foot and to fetal toxicity which is enhanced by simultaneous exposure to H2S.

Nasal and pulmonary toxicity of allyl glycidyl ether in mice.

A concentration-dependent expiratory bradypnea, indicative of irritation of the nasal mucosa, occurred during a 15-min oronasal exposure of mice to allyl glycidyl ether (AGE) in the concentration range of 1.9-8.6 ppm. The level of exposure responsible for a 50% decrease in the respiratory rate (RD50) was 5.7 ppm. Non-anaesthetized, tracheally cannulated mice exposed for 120 min to AGE at levels ranging from 105 to 185 ppm showed a concentration-dependent decrease in respiratory rate due to pulmonary toxicity. The level of exposure to AGE which produced a 50% decrease in the respiratory rate of tracheally cannulated mice (RD50TC) was 134 ppm. Mice were subjected to whole body exposure for 4, 9 or 14 days, 6 h/day to 7.1 or 2.5 ppm AGE. The 4-day exposure to 7.1 ppm AGE produced in the nasal cavities of mice lesions consisting of necrosis of the respiratory epithelium and complete erosion of the olfactory epithelium without pulmonary injury. Restorative responses were observed in the nasal cavities of mice exposed for 9 and 14 days to 7.1 ppm AGE. Exposure to 2.5 ppm AGE caused neither nasal nor pulmonary injury. The results indicate that AGE primarily affects the upper airways. They also make it questionable that the occupational standard of 5 ppm assures an adequate margin of protection against AGE-induced nasal effects.

A sensitive cadmium-affinity assay for the determination of thionein in urine.

A sensitive cadmium-affinity assay was developed for measurement of urinary thionein (Th) level. Acidification with HCl (2.5 M) was used to remove metal ions from purified urinary metallothionein (MTh), adsorbed on activated polystyrene tubes. It was found that the amount of 109Cd bound to standard Th (rabbit Th) was proportional to that of Th in the concentration range of 20-6000 ng/ml. The method exhibited a sensitivity below 20 ng/ml and a precision of about 5%. The results of the Cd-affinity assay were unaffected by dilution of, or addition of standard Th to urine samples. Under the latter circumstances, the Cd-affinity assay was performed with a mean analytical recovery of 100.3 +/- 4%. Addition of Cd, Zn, Hg and Cu (50 micrograms each) or glutathione and cysteine (0.1 mmol) to urine specimens (1 ml) did not interfere with the determination of Th. The mean values of urinary Th in healthy subjects were 200 +/- 53 micrograms/g creatinine (n = 9) and 256 +/- 97 micrograms/g creatinine (n = 8) for men and women respectively. The mean daily excretions of Th by non-fasting female and male healthy adult rats were 9.95 +/- 2.7 micrograms (n = 10) and 18.2 +/- 3.9 micrograms, respectively. The Cd-affinity assay succeeded in indicating Cd exposure and/or development of Cd toxicity in rats.