Biomarkers of cadmium and arsenic interactions.

Advances in proteomics have led to the identification of sensitive urinary biomarkers of renal dysfunction that are increasingly used in toxicology and epidemiology. Recent animal data show that combined exposure to inorganic arsenic (As) and cadmium (Cd) gives rise to more pronounced renal toxicity than exposure to each of the agents alone. In order to examine if similar interaction occurs in humans, renal dysfunction was studied in population groups (619 persons in total) residing in two metal contaminated areas in China: mainly a Cd contaminated area in Zhejiang province (Z-area) and mainly a As contaminated area in Guizhou province (G-area). Nearby control areas without excessive metal exposure were also included. Measurements of urinary beta(2)-microglobulin (UB2MG), N-acetyl-beta-glucosaminidase (UNAG), retinol binding protein (URBP) and albumin (UALB) were used as markers of renal dysfunction. Urinary Cd (UCd) and total As (UTAs) were analyzed by graphite-furnace atomic absorption spectrometry. Urinary inorganic As and its mono- and di-methylated metabolites (UIAs) were determined by Hydride generation. Results. As expected, the highest UCd values occurred in Z-area (Geometric mean, GM 11.6 microg/g crea) while the highest UTAs values occurred in G-area (GM = 288 microg/g crea). Statistically significant increases compared to the respective control area were present both for UTAs, UCd and for UB2MG, UNAG and UALB in Z-area as well as in G-area. UIAs was determined only in Z area. In G-area, there was a clear dose-response pattern both in relation to UTAs and UCd for each of the biomarkers of renal dysfunction. An interaction effect between As and Cd was demonstrated at higher levels of a combined exposure to As and Cd enhancing the effect on the kidney. In Z-area an increased prevalence of B2MG-uria, NAG-uria and ALB-uria was found in relation to UCd, but no relationship to UTAs was found. A statistically significant relationship between UIAs and UB2MG was found among women in this area and an interaction between As and Cd was indicated for B2MG. Conclusion. The present studies, which employed sensitive biomarkers of renal dysfunction, give support to the idea that human co-exposure to Cd and inorganic arsenic gives rise to more pronounced renal damage than exposure to each of the elements alone, but further studies are needed to establish and clarify this interaction.

Toxicological aspects of metallothionein.

Metallothionein (MT) is expressed to a certain extent in almost all mammalian tissues. The biological significance of MT is related to its various forms MT-1, MT-2, MT-3 and MT4. For MT-1 several isoforms of the protein exist and it is likely that these isoforms are related to various functions involved in developmental processes occurring at various stages of gestation. Toxicokinetics and biochemistry of essential and toxic metals such as cadmium, zinc, mercury and copper in organs e.g. kidney, CNS, are often related to metallothionein. It is debated whether there is a relation or not for other metals e.g. selenium and bismuth. For the toxicokinetics of cadmium, MT plays an important role. By expanding techniques from experimental toxicology and biochemistry to include molecular biology methods, more specific and relevant studies can be performed of the actual role and biological function of MT. The present paper on toxicological aspects of metallothionein, presents an overview and evaluation of present knowledge concerning differences among organs and within organs of the expression of MT and how this affects tissue sensitivity to toxicity.

Differential calcium transport disturbances in renal membrane vesicles after cadmium-metallothionein injection in rats.

Cadmium-metallothionein (CdMT) induced calciuria may result from disturbed calcium (Ca) transport through the renal tubular epithelium. The present study aimed at defining time of onset and the degree of disturbed calcium transport. Kidneys were obtained from rats at 4, 12 and 24 h after a single injection of CdMT (dose 0.4 mg Cd/kg b.w.), and compared to saline injected controls. Rapid-filtration 45Ca-assays were performed on basolateral and luminal membrane vesicles, isolated from kidney cortex using a sequential ultracentrifugation procedure. Luminal 45Ca uptake was increased at 4 h and then declined to about 80% of controls, suggesting an early phase perturbation of Ca absorption. Basolateral 45Ca uptake was reduced to less than 50% of controls, starting already at 4 h while 45Ca binding was reduced at 8 h. This may reflect an inhibited basolateral Ca pump mechanism after the binding step. Since the Ca pump normally expels Ca from the cell, an accumulation of intracellular calcium was indicated. Metal analysis verified a four-fold increase of Ca in kidney cortex at 24 h. This suggests that Cd impact on tubular cells involves disturbances on cellular absorption as well as expulsion of Ca.

Biological monitoring of cadmium exposure and renal effects in a population group residing in a polluted area in China.

In an area of China, not previously studied in detail concerning cadmium pollution and possible adverse effects on the kidney of exposed populations, concentrations of cadmium in urine as an indicator of renal accumulation of cadmium was studied and related to indicators of renal dysfunction in order to examine if a relationship could be documented. Cadmium concentrations in urine were analysed by graphite furnace atomic absorption spectrometry and urinary beta-2 microglobulin (UBM) and albumin (UALB) were measured as indicators of renal dysfunction, Rice samples and urine samples were obtained from three areas in Zhejiang province, China, representing a highly exposed area, a medium exposed area and a control area, respectively. Cadmium concentrations in rice were 3.70, 0.51 and 0.072 mg/kg for the heavily, medium polluted areas and the control area, respectively. Cadmium concentrations in urine (geometric means) were 10.7, 1.62 and 0.40 micrograms/l in the high, medium and control areas respectively. There was a clear increase in UBM and UALB in the heavily exposed group in comparison to the control group and a slight increase in the medium exposed group. There was a statistically significant dose-response relationship between cadmium in urine and beta 2-microglobulin excretion in urine, which is similar to what has previously been reported in other countries. The findings constitute the first report concerning a dose-response relationship in this population group in Zhejiang province in China.

Cytotoxicity, zinc protection, and stress protein induction in rat proximal tubule cells exposed to cadmium chloride in primary cell culture.

Primary cell culture was utilized to study the relationships between stress protein induction by zinc in vivo and cadmium toxicity in vitro. Effects of cadmium on cell viability were evaluated by the alamar blue assay, in conjunction with the ultrastructural morphology of cells by transmission electron microscopy. The expression of stress protein gene products was evaluated by 35S two-dimensional gel electrophoresis. The results showed cytotoxicity of CdCl2 at and above 129 microM (14.55 micrograms cadmium/mL medium) following 4 h of exposure. Prior zinc administration (20 mg zinc/kg, s.c., two daily doses) in vivo significantly protected the cells in vitro as demonstrated by improved cell viability. The 35S labeling of proteins induced by CdCl2 exposure clearly demonstrated for the first time that gene product of the 70-kDa family was induced in cultured rat proximal tubule cells which are the target cells for cadmium toxicity in vivo. Zinc in vivo pretreatment of animals induced proteins in the 90-, 70-, and 38-kDa families, which may act together with metallothionein to protect cells against cadmium toxicity. The results also indicate that the protective effect of zinc remains after the cells have been put in culture and thus provides a system in which we can study the changes that occur as a result of zinc exposure that decreases cadmium toxicity.

Increased blood lead and decreased calcium levels during pregnancy: a prospective study of Swedish women living near a smelter.

OBJECTIVES: The purpose of this study was to monitor blood lead in a northern Swedish cohort of mothers and children during pregnancy and at birth. METHODS: Blood lead was analyzed during pregnancy and in the umbilical cords of 290 women living near a smelter and in 194 control subjects. RESULTS: During pregnancy, there were statistically significant overall increases in blood lead concentrations by 20% and 15% in the smelter and reference areas, respectively. Mean maternal blood lead concentrations at delivery were 0.15 mumol/L (3.11 micrograms/dL) in the smelter area and 0.13 mumol/L (2.69 micrograms/dL) in the control area. Umbilical cord blood lead levels were 80% to 87% of the maternal levels. Blood lead levels were influenced by place of residence, employment at the smelter, smoking, and wine consumption. Maternal serum calcium levels decreased during pregnancy and were significantly lower than those of the newborns. CONCLUSIONS: An increase in blood lead concentrations was found during pregnancy, despite increased blood volume and unchanged or decreasing environmental lead levels. The mobilization of lead from bone during pregnancy may explain the increase.

Is placenta a good indicator of cadmium and lead exposure?

Distribution of lead and cadmium was studied in 25 placentas. Samples were taken from 6 different lobuli, and lead and cadmium concentrations were not determined by graphite furnace atomic absorption spectrometry. Lead and cadmium were not distributed uniformly, and the concentrations differed by a factor > or = 2 among different lobuli within the same placenta in 36% and 52% of the placentas, respectively. Placental lead and cadmium concentrations were also determined in homogenized samples from smelter (n = 49) and control (n = 53) areas in northern Sweden. Mean lead and cadmium concentrations were low, even in the smelter area (geometric means = 10 ng/g and 3 ng/g wet weight, respectively). The significant differences observed (i.e., higher blood lead concentrations in the smelter area during pregnancy and in umbilical cord blood) were not reflected in the placenta. We concluded, therefore, that the placenta is not a suitable organ to use for the monitoring of environmental exposure to lead. It could be used to monitor cadmium exposure, but if pregnancy outcome is to be studied, consideration should be given to the sampling procedure.

Nephrotoxic impact of multiple short-interval cadmium-metallothionein injections in the rat.

The cadmium-metallothionein (CdMT) injection model was used to examine whether multiple short-interval injections of CdMT, instead of a single dose, could better reproduce the features of chronic exposure to inorganic cadmium. Male Wistar rats were given an initial CdMT dose and four subsequent doses subcutaneously at 2-h intervals. A control group, given saline, was compared with a low dose group (0.2 + 4 x 0.1 mg Cd/kg b.w.) and high dose group (0.4 + 4 x 0.1 mg Cd/kg b.w.). Nephrotoxic effects were seen at the high dose. A marked proteinuria began 6-12 h after the first injection and extended to day 9. A progressive, unreversed calciuria appeared at 6 h and reached its maximum at day 13. This was a marked increase in duration compared with the transient peaks of proteinuria and calciuria observed in previous single dose studies. The unreversed calciuria and the marked proteinuria are suggestive of residual tubular damage, which may be irreversible. In conclusion, the model with multiple short-interval CdMT injections more closely reproduces the situation in long-term exposure to inorganic cadmium, compared to the single dose models previously employed.

Aluminium accumulation in some tissues of rats with compromised kidney function induced by cadmium-metallothionein.

Two experiments (I and II) were performed to study aluminium accumulation in brain as well as in several other tissues in male Wistar rats. A single intraperitoneal injection of cadmium-metallothionein (CdMT, 0.1-0.4 mg Cd/kg b.wt.) was used to compromise kidney function 12 hr before the final aluminium injection in both experiments. In experiment I, rats were maintained on diets deficient (0.01%, w/w) in calcium (-Ca) or providing adequate (+Ca) dietary calcium (0.9%) for 6 weeks. Among animals given a daily intraperitoneal dose of aluminium chloride (10.8 mg Al/kg per day) on 6 consecutive days there was a tendency towards higher aluminium level in brains of rats with compromised kidney function from CdMT (in -Ca rats: the geometric mean [G] = 288 versus 205 ng/g wet weight [w., wt.], P = 0.07, and in +Ca rats: G = 242 versus 164, P < 0.05) as compared to animals given no CdMT. The results from experiment II (all rats were given aluminium 5.6 mg Al/kg 2 and 12 hr after CdMT injection) demonstrated a higher level of aluminium (G: 41 ng/g w. wt., P < 0.05) in brains of rats with only slightly damaged kidney function (0.1 mg Cd/kg) than in those given no CdMT (G: 29 ng/g w. wt.). It was also observed that 1) calcium deficiency had a statistically significant effect (P < 0.05) in increasing kidney retention of intraperitoneal aluminium (G: 327 micrograms/g w. wt.) as compared to rats with a normal calcium supply in the diet (G: 54 micrograms/g w. wt.); 2) when aluminium concentration in kidney was at and above 54 micrograms/g wet tissue, kidney damage was observed. The above results indicate that compromised kidney function including tubular damage induced by a low-dose of CdMT may play a crucial role in the accumulation of aluminium in brain and other tissues. Since tubular function decreases with age in human populations, these findings in rats may be of considerable importance if a similar phenomenon would occur in humans. Therefore, the possibility of increased aluminium retention in persons with low calcium and high aluminium intakes may need to be further investigated.

Nephrotoxicities of aluminium and/or cadmium-metallothionein in rats: creatinine excretion and metabolism of selected essential metals.

The effects of exposure to aluminium (Al) and cadmium (Cd) on urinary creatinine and protein excretion, and the concentrations of calcium, magnesium and copper in kidney and urine were studied in 32 male adult Wistar rats. The animals were divided into 8 groups, groups 1-4 given a calcium-deficient diet (0.01%, i.e. 0.01 g calcium/100 g diet weight) and groups 5-8 a calcium-adequate diet (0.9%) for 6 weeks. Single daily intraperitoneal injections of AlCl3 (10.8 mg Al/kg body weight, per day) were done on 6 consecutive days to groups 3, 4, 7 and 8 during the last week of the experiment. One single intraperitoneal injection of cadmium-metallothionein (Cd-MT, 0.4 mg Cd/kg) was administered 12 hr before the final Al dose to groups 2, 4, 6, and 8 and the rats were sacrificed 47 hr after the Cd-MT injection. The rate of creatinine clearance was significantly lower in rats injected intraperitoneally with either Cd-MT or Al, and the concentrations of magnesium and calcium in urine were lower in rats administered both Al and Cd-MT as compared to those in control groups. Histological examination showed that Al was toxic to the kidney tubule cells of rats, however, an adequate supply of calcium in food protected to some extent the renal tubules from Al toxicity as indicated by a higher creatinine clearance, and there was also less tubule damage as shown by histological examination. The copper concentrations in kidney tissue were lower in groups treated with either Al or Cd-MT.(ABSTRACT TRUNCATED AT 250 WORDS)

Subcellular targets of cadmium nephrotoxicity: cadmium binding to renal membrane proteins in animals with or without protective metallothionein synthesis.

Nephrotoxic effects of cadmium exposure are well established in humans and experimental animals. An early manifestation of such toxicity is calciuria a few hours after injection of CdMT in rats. Protection against calciuria and other adverse effects such as proteinuria (occurring later) is offered by pretreatment with Cd, which effectively induces metallothionein synthesis. In the present experiment, one group of animals was given pretreatment with CdCl2 to induce metallothionein synthesis. The comparison group was left without pretreatment. The distribution of Cd from a normally nephrotoxic dose of 109CdMT was studied by gel chromatography in subcellular fractions of kidney cortex in both groups. In the pretreated animals, 109Cd in the plasma membrane and microsome fractions of renal cortical cells was mainly bound to metallothionein and other low molecular weight proteins at 4 hr. In nonpretreated animals the major part of 109Cd was bound to high molecular weight proteins. These findings indicate that membrane proteins may be important targets for Cd when inducing nephrotoxicity and that sequestering of Cd by metallothionein (and other low molecular weight proteins) may be a mechanism of protection.

Influence of zinc and copper administration on metal disposition in rats with cadmium-metallothionein-induced nephrotoxicity.

Only a few studies have investigated the effects of the interaction between Cu, Zn, and Cd on the toxicity of Cd in vivo and the metabolism of these metals. The present study is concerned with these effects. Observations of the distribution of these metals in liver and renal cortex and of their excretion in urine were made. Sixty male Wistar rats were divided into 10 groups of six (a full factorial design complemented by a center point and a control group) and were administered Zn (0 or 25 mg/kg body mass), Cu (0 or 12.5 mg/kg), and CdMT (cadmium--metallothionein; 0.1 or 0.4 mg Cd/kg) by sc injection. The rats were given Zn and/or Cu 24 hr prior to the CdMT injection. The data were analyzed by linear multiple regression. After CdMT injection, the Cd retention level was markedly reduced in renal cortex and increased in liver by Cu pretreatment, while the urinary excretion of Cd was significantly lower in these rats. The levels of endogenous Zn in renal cortex and liver increased significantly in rats pretreated with Cu. The production of MT in liver and renal cortex was induced more efficiently by Cu than by Zn. The results obtained may be of importance in understanding the mechanism of CdMT toxicity and the potential influence of Zn and Cu on the chronic nephrotoxicity of cadmium.

The susceptibility of spontaneously diabetic mice to cadmium-metallothionein nephrotoxicity.

Cadmium metallothionein (CdMT) was injected subcutaneously into obese hyperglycaemic Umeå ob/ob mice or their lean litter mates (normal mice) at doses of 0, 0.1 and 0.4 mg Cd/kg. Proteinuria and calciuria were induced in both types of mice, but in the ob/ob mice this condition developed at a lower dose of CdMT (0.1 mg Cd/kg) than in the normal mice (0.4 mg Cd/kg). These results show, therefore, that Umeå ob/ob mice are particularly susceptible to CdMT-induced nephrotoxicity. The mechanism underlying this phenomenon needs to be further investigated. After the administration of CdMT, a dose-related increase in glycosuria was observed in both types of mice, in spite of decreased levels of serum insulin and glucose. It is suggested that such glycosuria induced by CdMT could be one of the signs of cadmium nephrotoxicity. The results of the present study thus indicate that metabolic changes like those in diabetes may increase susceptibility to cadmium-induced renal tubular damage.

In vivo measurements of lead in bone in long-term exposed lead smelter workers.

In-vivo measurements of lead concentrations in calcaneus (mainly trabecular bone) and tibia (mainly cortical bone) were performed by x-ray fluorescence (XRF) in 70 active and 30 retired lead smelter workers who had long-term exposure to lead. Comparison was made with 31 active and 10 retired truck assembly workers who had no known occupational exposure to lead. After physical examination, all participants provided blood and urine samples and answered a computerized questionnaire. Since 1950, blood lead has been determined repeatedly in lead workers at the smelter, which made it possible to calculate a time-integrated blood lead index for each worker. Lead concentrations in blood, urine, calcaneus, and tibia in active and retired lead workers were significantly higher than in the corresponding control groups (p < .001). The highest bone lead concentrations were found among retired lead workers (p < .001), which was the result of considerably higher lead exposure during 1940 to 1960. Lead concentrations in calcaneus in active lead workers were significantly higher than in tibia when expressed in ug of lead per gram of bone mineral, which suggests a quicker absorption over time in this mainly trabecular bone. The estimated biological half-times were 16 y in calcaneus (95% confidence interval [95% CI] = 11-29 y) and 27 y in tibia (95% CI = 16-98 y). A strong positive correlation was found between lead concentrations in calcaneus and tibia for all lead workers (r = 0.54; p < .001). A strong positive correlation was also found between the bone lead concentrations and the cumulative blood lead index. Blood lead, at the time of study, correlated well with bone lead concentrations in retired--but not in active--workers, reflecting the importance of the endogenous (skeletal) lead exposure. The findings in this study indicate that bone lead measurements by XRF can give a good index of long-term lead exposure. Tibia measurements offer a higher precision than calcaneus measurements. The method is of particular interest in epidemiologic studies of adverse health effects caused by long-term lead exposure.

Cadmium carcinogenesis and its relationship to other health effects in humans.

Several different adverse health effects can be caused by cadmium exposure to humans and animals. In environmental and occupational health it is important to identify effects that occur at relatively low exposures (ie., the critical effects that are crucial for preventive action). In long-term human exposures to cadmium, effects of cadmium on the kidney have been considered to be critical effects and quantitative risk assessments of these effects have been performed on the basis of both risk modeling and direct epidemiologic observation. However, experimental and epidemiologic studies are providing increasing evidence that cadmium is carcinogenic, and this effect, which is considered to be stochastic in character, can be considered to be the critical effect. A quantitative evaluation of the cancer risk is currently difficult to make, but the preventive action against such effects is usually to limit the exposure as much as possible.

International project for producing reference values for concentrations of trace elements in human blood and urine--TRACY.

In assessing the concentrations of toxic metals, such as cadmium, chromium, and mercury, in human blood and urine samples to determine whether they are abnormal or not, reliable reference values are needed from populations of nonoccupationally exposed subjects. Numerous publications present concentrations claimed to be typical for the study populations, but they can differ by up to an order of magnitude for a particular element. This is the consequence of general problems that are related to the definition of the reference groups, and the sampling and analytical procedures used, and that make it difficult to define typical and unbiased values. An international group of experts now establishes criteria and procedures to evaluate publications containing information on the concentrations of metals in tissues and body fluids for reference populations. These evaluations have been compiled in a data base (TRACY).

Biological monitoring of arsenic, lead and cadmium in occupationally and environmentally exposed pregnant women.

Lead and cadmium in blood (B-Pb and B-Cd, respectively) and arsenic in urine (U-As) were analyzed three times during pregnancy for women living around a metal smelter and women living in a reference town. The B-Pb levels were significantly higher in the smelter town. In the women of both towns, the B-Pb levels increased during pregnancy. Women who were employed at the smelter had higher B-Pb levels than women in the surrounding area. There were no significant differences in the B-Cd levels between the smelter and reference towns, except for non- and ex-smokers at the onset of pregnancy. No difference between the two areas was seen among the smokers, whose cadmium levels were twice those of non- and ex-smokers. There were no significant differences in the U-As levels, which were comparable with previously reported values in Sweden.

Lung cancer in smelter workers--interactions of metals as indicated by tissue levels.

The concentrations of the elements antimony, arsenic, cadmium, chromium, cobalt, lanthanum, lead, selenium, and zinc were determined in lung tissue of 85 decreased smelter workers by neutron activation analysis and atomic absorption spectrophotometry. The concentrations of all these elements, except zinc, were significantly higher among the workers as compared with rural referents. Workers who died from lung cancer (N = 7) had the lowest lung selenium content relative to other metals, both compared with workers with other diseases and with rural (N = 15) and urban (N = 10) referents. The low lung tissue levels may have influenced the development of lung cancer. The highest lung cadmium concentrations were observed in the lung cancer group, in which, however, smokers and ex-smokers were over-represented. The observations make it likely that the excess lung cancer risk in this smelter environment is multifactorial in character, involving interactions between both carcinogenic and anticarcinogenic factors.

Modulation of calciuria by cadmium pretreatment in rats with cadmium-metallothionein-induced nephrotoxicity.

One group of male Wistar rats (Group B) was pretreated by a daily subcutaneous injection with CdCl2 during 5 days with increasing doses (0.5, 1, 1, 2 and 2 mg Cd/kg). Another group of rats (Group A) was daily given normal saline subcutaneously for 5 days. On the second day after the last injection, a single s.c. injection of 109Cd-metallothionein (CdMT, 0.4 mg Cd/kg) was given to each animal in both groups. Urinary calcium, protein, metallothionein (MT), N-acetyl-beta-D-glucosaminidase (NAG) and gamma glutamyltransferase (gamma-GT) were measured. In Group A, calciuria, proteinuria, metallothioneinuria and enzymuria was induced by CdMT. Calciuria reached a peak during 0-6 h after the administration of CdMT, thus appearing earlier than other effects. Enzymuria was displayed at 6-12 h for gamma-GT and 12-24 h for NAG. A prominent increase of proteinuria appeared at 24-48 h after the challenge of CdMT. In Group B, no significant increase of urinary calcium, protein, or NAG was observed after the CdMT injection and urinary gamma-GT was only slightly elevated, thus demonstrating the protective action of pretreatment. This study demonstrates for the first time that calciuria, one of the signs of cadmium nephrotoxicity, can be prevented by cadmium pretreatment. Urinary MT increased slightly during the 4-5 days of CdCl2 pretreatment. This is in accordance with previous observations that cadmium pretreatment induces new synthesis of MT which is likely to constitute the background for the resistance to the CdMT challenge to the kidney.

A multivariate study of protective effects of Zn and Cu against nephrotoxicity induced by cadmium metallothionein in rats.

Factorial experimental design was used to study the protective effects of Zn and Cu on cadmium-metallothionein(CdMT)-induced nephrotoxicity in male Wistar rats. In the factorial design two levels of Zn (0 and 25 mg/kg body weight), two levels of Cu (0 and 12.5 mg/kg), and two levels of CdMT (0.1 and 0.4 mg of Cd/kg) were used as varied factors. The factorial design was complemented with a center point with all three variables at an intermediate setting, i.e., Zn at 12.5 mg/kg, Cu at 6.25 mg/kg, and CdMT at 0.25 mg Cd/kg. Each of the nine combinations of settings was administered to one of nine groups with six rats in each. Zn and Cu were injected sc 24 hr prior to the injection of CdMT. The concentrations of protein and Ca in urine and Ca in renal cortex were used as effects. The relationship between the experimental design settings and the effects were modeled with multiple regression. The multiple regression analysis revealed that for the high dose of CdMT (i) the enhanced values of protein in urine caused by CdMT injection could be more efficiently reduced by Zn than by Cu, and (ii) excessive Ca in urine and renal cortex could be more efficiently reduced by Cu than by Zn. No significant synergism or antagonism between Cu and Zn was found. These models can be used to estimate the dose levels of Zn and Cu which will reduce the toxic effects of CdMT. The treatment of 20.4 mg/kg Zn, for example, will reduce the effects of 0.4 mg Cd/kg as CdMT on protein in urine, and 2.8 mg/kg Cu will reduce the Ca in urine to the levels of those caused by 0.25 mg Cd/kg (no Zn and Cu). Similarly, the effect of 0.4 mg Cd/kg on Ca level in renal cortex can be reduced to that of 0.28 mg Cd/kg as CdMT by 7.98 mg Cu/kg, which is three times as efficient as Zn. The obtained results might be of importance in understanding the mechanism of cadmium toxicity and the potential risk to the health of the population exposed to cadmium occupationally or environmentally.