Maja Blanuša

Chelators as antidotes of metal toxicity: therapeutic and experimental aspects.

The effects of chelating drugs used clinically as antidotes to metal toxicity are reviewed. Human exposure to a number of metals such as lead, cadmium, mercury, manganese, aluminum, iron, copper, thallium, arsenic, chromium, nickel and platinum may lead to toxic effects, which are different for each metal. Similarly the pharmacokinetic data, clinical use and adverse effects of most of the chelating drugs used in human metal poisoning are also different for each chelating drug. The chelating drugs with worldwide application are dimercaprol (BAL), succimer (meso-DMSA), unithiol (DMPS), D-penicillamine (DPA), N-acetyl-D-penicillamine (NAPA), calcium disodium ethylenediaminetetraacetate (CaNa(2)EDTA), calcium trisodium or zinc trisodium diethylenetriaminepentaacetate (CaNa(3)DTPA, ZnNa(3)DTPA), deferoxamine (DFO), deferiprone (L1), triethylenetetraamine (trientine), N-acetylcysteine (NAC), and Prussian blue (PB). Several new synthetic homologues and experimental chelating agents have been designed and tested in vivo for their metal binding effects. These include three groups of synthetic chelators, namely the polyaminopolycarboxylic acids (EDTA and DTPA), the derivatives of BAL (DMPS, DMSA and mono- and dialkylesters of DMSA) and the carbodithioates. Many factors have been shown to affect the efficacy of the chelation treatment in metal poisoning. Within this context it has been shown in experiments using young and adult animals that metal toxicity and chelation effects could be influenced by age. These findings may have a bearing in the design of new therapeutic chelation protocols for metal toxicity.

Placental cadmium and progesterone concentrations in cigarette smokers

Cadmium and progesterone concentrations were evaluated in term placentas collected from 56 healthy parturients in the city of Zagreb. Concentrations of lead, iron, zinc, and copper in placentas were analyzed. Data collected by questionnaire identified 29 nonsmoking and 27 smoking women. From each placenta, three samples from different locations were taken. Metals were measured by atomic absorption spectrometry. Progesterone was determined by specific radioimmunoassay in homogenized and lyophilized tissue samples after steroid extraction with ethanol. No effect of sample location was found. In placentas of smoking women an increase in cadmium, reduced progesterone and a decrease in iron concentrations were found. Placental copper and zinc concentrations were not altered. In conclusion, the results present new evidence that maternal smoking reduces placental progesterone content and support the established association of smoking with placental cadmium.

Mercury, arsenic, lead and cadmium in fish and shellfish from the Adriatic Sea

The aim was to measure concentrations of total mercury, total arsenic, lead and cadmium in common edible fresh fish and shellfish from various areas of the Adriatic Sea. Estimates of intake of these elements were made through seafood consumption by the general population. Samples were either wet digested for mercury and arsenic, or dry ashed for lead and cadmium analysis. Mercury was measured by cold vapour atomic absorption spectrometry (CV AAS) and arsenic, lead and cadmium by electrothermal atomic absorption spectrometry (ET AAS). Quality control procedures of analytical methods, which included analyses of dogfish muscle-certified reference material DORM-2, confirmed the acceptability of methods. The highest mercury and arsenic concentrations were found in hake (Merluccius merluccius) and the lowest in mackerel (Scomber scombrus). The respective values in hake were 0.373 ± 0.075 and 23.3 ± 3.6, and in mackerel 0.153 ± 0.028 and 1.06 ± 0.29 mg kg-1 fresh weight (mean ± SD). Lead and cadmium concentrations were about 10 times higher in shellfish than in analysed fish. The highest lead and cadmium concentrations were found in mussel (Mytilus galloprovincialis) and the lowest in hake. Respective lead and cadmium values in mussel were 0.150 ± 0.009 and 0.142 ± 0.017, and in hake were 0.007 ± 0.004 and 0.002 ± 0.001 mg kg-1 fresh weight. The concentrations of analysed elements were below acceptable levels for human consumption set by the Croatian Ministry of Health, except for total arsenic. The estimated intake of those trace elements included in this study through seafood consumption by the general population did not exceed the provisional tolerable weekly intake recommended by the Joint FAO/WHO Expert Committee on Food Additives.

Toxic and essential metal concentrations in four tissues of red deer (Cervus elaphus) from Baranja, Croatia

Kidney (n  = 297), liver (n  = 52), jawbone (n  = 80) and muscle (n  = 48) samples collected from red deer (Cervus elaphus) from north-eastern Croatia in the 2002–05 hunting season were analysed for cadmium (Cd), copper (Cu), iron (Fe), mercury (Hg), lead (Pb), selenium (Se) and zinc (Zn) using atomic absorption spectrometry. Statistical evaluation of results showed age-related accumulations of renal cortex Cd and Zn, bone Pb, and muscle Zn. Renal cortex Cd and Zn were significantly associated. In addition, concentrations of Cd and Pb in muscle tissue were significantly correlated with Fe content. Found levels of toxic metals were not likely to affect the health status of animals. A total of 49% of the muscle, 60% of the kidney and 6% of the liver samples were unsuitable for human consumption according to Croatian regulations for Cd in food. However, the calculated intake of Cd through deer meat consumption is small and represents no health risk when consumption is moderate.

Dietary calcium and blood lead levels in women

Nutritional factors are known to influence metabolism and toxicity of several metals in animal experiments, but relevant human data are scarce and inconclusive. In this work, we tested the hypothesis that dietary calcium influences lead metabolism in humans. Blood lead concentrations were used as indicators of lead exposure and metabolism. Two groups of peasant women living in similar conditions in two different regions in Yugoslavia (100 in each) were chosen as subjects for this purpose. In region A, the dietary calcium intake was about 940 mg, and in region B about two times lower, i.e., 450 mg/day. The average blood lead concentration was significantly lower in women from region A (69 μg/L) than from region B (83 μg/L). Our results support the assumption that adequate calcium intake might be one of the preventive measures for decreasing lead absorption. This new evidence, sought for some time by nutritionists and toxicologists, needs further international confirmation.

Age and intestinal retention of mercury and cadmium in rats

The site of cadmium and mercury retention in the intestine was determined in 6-day-old sucklings and 6-week-old weaned rats 6 days after oral administration of 115mCd and 203Hg. The ileum was found to be the main site of intestinal retention of both cations in sucklings but not in weaned rats. Other age- and element-specific differences in the site of metal retention in the intestine were also found. These differences indicate that even in neonates, metal absorption might be a more specific process than previously assumed.

Simultaneous reduction of radioactive strontium, caesium and iodine retention by single treatment in rats

The effect of a mixture of calcium alginate, iron (III) ferrocyanide and potassium iodide added to rat diet on 85Sr, 137Cs and 131I metabolism and health was investigated in female rats after four weeks of treatment. The retention of these radioisotopes was determined in the whole body and critical organs six days after 85 Sr and 137CS and one day after 131I oral administration. The health effect of the mixture was evaluated by measuring body weights, haematological parameters, concentrations of iron, zinc and manganese in the kidneys, liver and femur, bone parameters (femur composition and morphometry) and by a histopathological examination. The mixture reduced 85Sr retention in the femur 11 times, 137Cs retention in the thigh muscle 102 times and 131I retention in the thyroid 134 times. Treated animals were in good health and the only differences found between the control and experimental rats were slightly lower haemoglobin values in the blood and a slightly lower iron concentration in the liver. It is concluded that the mixture was very efficient for decreasing body retention of three important fission products and that it can be used over long periods without causing adverse health effects.

The effect of iron additive to milk on cadmium, mercury, and managanese absorption in rats

Abstract Six-day-old sucklings and 6-week-old female rats were used in the experiment to estimate whether low iron content of milk is the reason for increased gastrointestinal absorption of metals. The absorption of 115mCd, 203Hg, and 54Mn was determined by measuring the body retention (whole body, carcass, and gut) 6 days after oral administration. To sucklings radioisotopes were administered by artificial feeding with cows milk with or without 100 ppm Fe additive. Iron addition had no influence on metal absorption at this age. The same radioisotopes were administered by stomach tube to older animals fed rats diet or milk with or without 100 ppm Fe additive. Increased iron content of milk decreased the absorption of all metals. For mercury and cadmium most of this effect was due to decreased gut retention and only partly to decreased carcass retention. It is concluded that the high absorption of metals in sucklings is not likely to be due to the low iron content of milk and also that iron does not interact with cadmium, mercury, and manganese in the absorption process at this age.

Combined chelation therapy in reducing tissue lead concentrations in suckling rats

The very young are more prone to lead poisoning than adults, and the treatment with chelating agents, either as monotherapy or combined treatment, is still a matter of dispute. The purpose of this work was to evaluate the efficiency of three chelating agents administered either as monotherapies or as combined treatments in sucklings. Lead acetate (5 mg Pb kg−1 i.p.) was administered to the 7‐day‐old rat pups in eight litters on experimental day 1 and chelating agents on experimental days 2 and 3. Pups were divided into six groups: (1) untreated control; (2) EDTA (calcium disodium ethylendiaminetetraacetate, 0.3 mmol kg−1 i.p. at 4 p.m.); (3) meso‐DMSA (meso‐2,3‐dimeracaptosuccinic acid, 0.5 mmol kg−1 p.o. at 10 a.m.); (4) rac‐DMSA (racemic‐2,3‐dimeracaptosuccinic acid, 0.5 mmol kg−1 p.o. at 10 a.m.); (5) EDTA+meso‐DMSA; and (6) EDTA+rac‐DMSA. Rats were killed on experimental day 5. Tissue element concentrations were analyzed by atomic absorption spectrometry. Treatment with EDTA did not affect tissue Pb, but it reduced Zn in the carcass and liver. Meso‐DMSA reduced Pb in the kidneys and brain, and it did not affect organ essential elements. Rac‐DMSA most efficiently reduced Pb concentrations in the carcass, kidneys and brain, but it also reduced Zn and Cu in the liver and Zn in the kidneys. Combined treatments with EDTA never improved the efficiency of either DMSA isoform in decreasing tissue Pb but they did reduce tissue Zn concentrations. All treatments caused the same decrease in the carcass Ca concentrations. The results do not support combined treatment in this age group, which is especially sensitive to trace element deficiencies, and suggest that meso‐DMSA might be the treatment of choice in acute lead poisoning in infants. Copyright

Dental amalgam mercury exposure in rats.

The aim of this study was to measure the distribution of mercury, in tissues of rats exposed to amalgam over a two months period. Possible interaction of mercury with copper and zinc in organs was also evaluated. Rats were either exposed to mercury from 4 dental amalgams, or fed the diet containing powdered amalgam during two months. Mercury was measured in the kidney, liver and brain, copper in kidney and brain and zinc in kidney. The results showed significantly higher concentrations of mercury in the kidneys and the brains of rats in both exposed groups compared to control. Even after two months of exposure to mercury brain mercury concentration in rats with amalgam fillings was 8 times higher than in the control and 2 times higher than in rats exposed to amalgam supplemented diet. The highest mercury concentration in the latter group was found in the kidneys and it was 5 times higher than in the control group. We found no significant differences between mercury levels in exposed and control rats liver. Exposure to mercury from dental amalgams did not alter the concentrations of copper and zinc in the tissues. Histopathological analyses of rats tissues did not show any pathological changes. These results support previously proposed nose-brain transport of mercury released from dental amalgam fillings.