Relationship between delta-aminolevulinic acid dehydratase genotypes and heme precursors in lead workers

BACKGROUND: The objective of this study was to investigate the relationships between genotypes of delta-aminolevulinic acid (ALA) dehydratase (ALAD) and disturbances in the heme biosynthetic pathway by lead exposure. METHODS: The subjects were 192 male lead workers and 125 control subjects. Blood lead concentrations (Pb-B), plasma ALA concentrations (ALA-P), and ALAD genotypes were determined for all subjects. In lead workers, ALAD activity, ALA in urine (ALA-U), and erythrocyte zinc protoporphyrin (ZP) were also determined. RESULTS: The frequency of ALAD2 (minor type of ALAD allele) was calculated to be 0.087 in all subjects. No significant relationship was found between ALAD2 frequency and Pb-B levels in lead workers. ALAD1 homozygotes showed significantly higher levels of ZP and ALA-P in comparison with those of ALAD2 carriers at Pb-B levels more than 20 microg/dL and 40 microg/dL, respectively. CONCLUSIONS: ALAD1 homozygotes might be more susceptible than ALAD2 carriers to disturbances in heme metabolism caused by lead exposure.     

delta-Aminolevulinic acid dehydratase genotype modifies four hour urinary lead excretion after oral administration of dimercaptosuccinic acid.

OBJECTIVES: Previous research suggests that binding of lead by delta-aminolevulinic acid dehydratase (ALAD) may vary by ALAD genotype. This hypothesis was tested by examining whether ALAD genotype modifies urinary lead excretion (DMSA chelatable lead) after oral administration of dimercaptosuccinic acid (DMSA). METHODS: 57 South Korean lead battery manufacturing workers were given 5 mg/kg oral DMSA and urine was collected for four hours. Male workers were randomly selected from two ALAD genotype strata (ALAD1-1, ALAD1-2) from among all current workers in the two plants (n = 290). Subjects with ALAD1-1 (n = 38) were frequency matched with subjects with ALAD1-2 (n = 19) on duration of employment in the lead industry. Blood lead, zinc protoporphyrin, and plasma aminolevulinic acid concentrations, as well as ALAD genotype, duration of exposure, current tobacco use, and weight were examined as predictors or effect modifiers of levels of DMSA chelatable lead. RESULTS: Blood lead concentrations ranged from 11 to 53 micrograms/dl, with a mean (SD) of 25.4 (10.2) micrograms/dl. After 5 mg/kg DMSA orally, the workers excreted a mean (SD) 85.4 (45.0) micrograms lead during a four hour urine collection (range 16.5-184.1 micrograms). After controlling for blood lead concentrations, duration of exposure, current tobacco use, and body weight, subjects with ALAD1-2 excreted, on average, 24 micrograms less lead during the four hour urine collection than did subjects with ALAD1-1 (P = 0.05). ALAD genotype seemed to modify the relation between plasma delta-aminolevulinic acid (ALA) and DMSA chelatable lead. Workers with ALAD1-2 excreted more lead, after being given DMSA, with increasing plasma ALA than did workers with ALAD1-1 (P value for interaction = 0.01). CONCLUSIONS: DMSA chelatable lead may partly reflect the stores of bioavailable lead, and the current data indicate that subjects with ALAD1-2 have lower stores than those with ALAD1-1. These data provide further evidence that the ALAD genotype modifies the toxicokinetics of lead-for example, by differential binding of current lead stores or by differences in long-term retention and deposition of lead.     

Polymorphism of delta-aminolevulinic acid dehydratase and its effect on blood lead levels in Thai workers

To determine the prevalence of delta-aminolevulinic acid dehydratase (ALAD) polymorphism and its effects on blood lead levels (BLLs) in Thai workers, the authors performed a cross-sectional analysis of 389 Thai workers who were exposed to lead in a battery plant. The authors collected blood for BLLs and genotypic study, and they found that the allele frequencies of ALAD1 and ALAD2 were 0.98 and 0.02, respectively. They made a comparison of BLLs between genotype by dividing the levels into 2 categories of lead exposure (high and medium magnitude of exposure) and using length of employment as a covariance. Their results showed no significant difference of BLLs between the ALAD1-1 and ALAD1-2/ALAD2-2 groups in both levels of exposure. The frequency of ALAD2 in Thai workers was low; the authors found that ALAD polymorphism had a small or only modest effect on BLLs.     

低水平铅暴露儿童的GH/IGF-1轴变化及其与ALAD基因多态性的关系

目的:探讨低水平铅暴露儿童的生长激素/胰岛素样生长因子-1(GH/IGF-1)轴的变化及其与δ-氨基-γ-酮戊酸脱氢酶(ALAD)基因多态性的关系。方法:用钨舟原子吸收光谱法测定来自深圳市区242例学龄前儿童静脉血铅水平(BLL)。按血铅水平将患儿分为两组,A组BLL<50μg/L,B组BLL≥50μg/L。对两组对象的身高(H)、血红蛋白水平(Hb)、ALAD基因、生长激素(GH)、胰岛素样生长因子-1(IGF-1)和胰岛素样生长因子结合蛋白-3(IGFBP-3)水平进行比较。身高测量和血红蛋白检测用常规方法进行;ALAD基因多态性检测采用聚合酶链反应-限制性片段长度多态性(PCR-RFLP)法;生长激素测定采用化学发光法;IGF-1和IGFBP-3测定采用ELISA法。结果:242例儿童的血铅水平范围为8～146μg/L,几何均数47μg/L。BLL≥50μg/L者占43%,≥100μg/L者占0.8%。两组儿童的IGFBP-3水平差异有统计学意义,B组明显低于A组。身高和IGF-1略有差异,但无统计学意义。基因多态性分析显示,223例为ALAD1-1型,19例为ALAD1-2型,未发现ALAD2-2型。ALAD2基因出现的频率为7.85%。两组儿童突变基因的频率分布差异无统计学意义。结论:本研究发现即使低水平铅暴露也与儿童血IGFBP-3水平明显降低有关,表明铅中毒损害儿童GH/IGF-1轴功能。在低水平铅暴露状态下,ALAD基因的多态性对儿童血铅水平并无明显影响。可能只有在高水平铅暴露时,ALAD基因变异才会对儿童铅中毒易感性发挥作用。     

Influence of the delta-aminolevulinic acid dehydratase (ALAD) polymorphism on biomarkers of lead exposure in Turkish storage battery manufacturing workers

BACKGROUND: The relationship between delta-aminolevulinic acid dehydratase polymorphism (ALAD) and biomarkers of exposure was investigated in Turkish lead workers in this study. METHODS: Seventy two male lead battery manufacturing workers were selected for the study. Blood lead (BPb) and urinary lead (UPb) concentrations were determined by atomic absorption spectrometry. Erythrocyte ALAD activity and urinary 5-aminolevulinic acid (UALA) were measured spectrophotometrically. The polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to determine the genotype of the ALAD gene. RESULTS: In total, 51 workers (70.8%) had the ALAD 1-1 genotype, whereas 21 workers (29.2%) had the ALAD 1-2 genotype. No significant relationships were found between the two genotypes and BPb, UPb, and ALAD activity. ALAD1 homozygotes showed significantly higher levels of UALA in comparison with those ALAD2 carriers. CONCLUSIONS: ALAD 1-1 individuals might be an increased risk compared to ALAD2 carriers to disturbance in heme biosynthetic pathway in high lead exposure.     

Critical dose of lead affecting delta-aminolevulinic acid levels

To estimate the critical dose of the association between the blood lead concentration (BPb) and delta-aminolevulinic acid (ALA) levels, ALA levels in plasma (ALA-P), blood (ALA-B), and urine (ALA-U), and the activity of delta-aminolevulinic acid dehydratase (ALAD) were determined in 186 Japanese lead workers, aged 18-62 yr, with BPb levels of 2.1-62.9 g/dl. For this purpose, the benchmark dose (BMD) method, recently used in the environmental health field in place of the no-observed-adverse-effect level, was introduced into this study. The BMD was defined as the BPb level that resulted in an increased probability of abnormal change in ALA-related parameters by an excess risk (BMR) of 5% in exposed workers i.e., from P0 (abnormal probability of 5% in unexposed workers) to P0+BMR for exposed workers at the BMD. ALA-related parameters were significantly correlated with BPb. The BMDs computed from the 186 workers, after controlling for age, were 15.3-20.9 microg/dl for ALA levels, and 2.9 microg/dl for ALAD; likewise, the BMDs from the 154 workers with BPb levels of less than 40 microg/dl were 3.3-8.8 microg/dl for ALA levels, and 2.7 microg/dl for ALAD. Since the cutoff value of ALA-P, computed from the latter workers, seems to be closer to the upper normal limit in unexposed adults than does that from the former workers, it is suggested that the critical dose of BPb causing the increased levels of ALA is below 10 microg/dl. Such critical doses are necessary to promote preventive activities of adverse effects of lead.     

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.     

Lead and delta-aminolevulinic acid dehydratase polymorphism: where does it lead? A meta-analysis

BACKGROUND: Lead poisoning affects many organs in the body. Lead inhibits delta-aminolevulinic acid dehydratase (ALAD), an enzyme with two co-dominantly expressed alleles, ALAD1 and ALAD2. OBJECTIVE: Our meta-analysis studied the effects of the ALAD polymorphism on a) blood and bone lead levels and b) indicators of target organ toxicity. DATA SOURCE: We included studies reporting one or more of the following by individuals with genotypes ALAD1-1 and ALAD1-2/2-2: blood lead level (BLL), tibia or trabecular lead level, zinc protoporphyrin (ZPP), hemoglobin, serum creatinine, blood urea nitrogen (BUN), dimercaptosuccinic acid-chelatable lead, or blood pressure. DATA EXTRACTION: Sample sizes, means, and standard deviations were extracted for the genotype groups. DATA SYNTHESIS: There was a statistically significant association between ALAD2 carriers and higher BLL in lead-exposed workers (weighted mean differences of 1.93 microg/dL). There was no association with ALAD carrier status among environmentally exposed adults with BLLs < 10 microg/dL. ALAD2 carriers were potentially protected against adverse hemapoietic effects (ZPP and hemoglobin levels), perhaps because of decreased lead bioavailability to heme pathway enzymes. CONCLUSION: Carriers of the ALAD2 allele had higher BLLs than those who were ALAD1 homozygous and higher hemoglobin and lower ZPP, and the latter seems to be inversely related to BLL. Effects on other organs were not well delineated, partly because of the small number of subjects studied and potential modifications caused by other proteins in target tissues or by other polymorphic genes.     

Relationship of blood lead levels to blood pressure in exhaust battery storage workers

Several researches has focused the hypothesis that low blood lead levels could be associated with an increased risk of hypertension. To assess the relation between occupational lead exposure and elevated blood pressure a group of 27 workers, age range from 27 to 62 years, mean (SD) 36.52 (+/- 8.16) yr; length of employment mean (DS) 2.97 (+/- 1.67) yr, were recruited as study subjects. The following variables were measured: blood lead concentration (BPb), delta-Aminolevulinic Acid Dehydratase (ALAD) activity, Zinc Protoporphirin (ZPP), creatinine, hematocrit, Body Mass Index (BMI) and Systolic Blood Pressure (SBP) and Diastolic Blood (DBP) Pressure. The results showed that long term occupational exposure was related to a slight increase of systolic and diastolic blood pressure among workers who had been exposed to higher level of lead with respect to workers exposed to lower level of lead. Furthermore, blood lead concentration (BPb) and ZPP resulted higher among workers exposed to higher level of ambient lead, while in the same group of workers ALAD activity resulted more inhibited. The authors concluded long term cumulative lead exposure can significantly increase blood pressure in low level Pb exposed workers.     

Evaluation of genotoxic effects of lead in pottery-glaze workers using micronucleus assay, alkaline comet assay and DNA diffusion assay

Purpose

We investigated genotoxic effects of occupational exposure to lead acetate in pottery-glaze ceramic workers.

Methods

The study was carried out in 30 exposed workers and 30 matched controls, to whom several biochemical parameters—the blood lead (B-Pb; range: exposed, 41.68–404.77; controls, 12–52) and cadmium (B-Cd) level, the activity of delta-aminolevulinic acid dehydratase (ALAD), erythrocyte protoporphyrin (EP), the level of vitamin B₁₂ and folate in serum—were measured. The genotoxic effects were evaluated by the alkaline comet assay, the DNA diffusion assay and micronucleus test in peripheral blood lymphocytes.

Results

Subjects exposed to lead had significantly higher B-Pb level and, consequently, increased values of tail intensity (TI), frequency of apoptotic and necrotic cells, and frequency of micronuclei (MN). In contrast, their activity of ALAD, the level of vitamin B₁₂ and folate in serum were significantly lower compared to controls. Poisson regression analysis showed a significant correlation of profession, duration of exposure, smoking, level of cadmium in blood, ALAD and EP with primary DNA damage. A majority of primary damage repairs in a short period after exposure to a genotoxic agent. In addition, the influence of gender and level of vitamin B₁₂ and folate in serum MN frequency in exposed group was observed.

Conclusions

In this study, DNA diffusion and micronucleus test showed higher influence of tested parameters to DNA damage. The results indicate a need for concomitant use of at least two different biomarkers of exposure when estimating a genetic risk of lead exposure.     

Erythrocyte delta-aminolevulinic acid dehydratase genotype and other mechanisms affecting workers' susceptibility to lead.

In this study, the role of delta-aminolevulinic acid dehydratase (ALAD) variants in lead susceptibility was examined. The study subjects comprised 223 male workers, and the relationship between their blood lead level and erythrocyte ALAD activity or plasma/urine delta-aminolevulinic acid level was studied. Leukocyte specimens from 11 workers, whose erythrocyte ALAD activities were as low as one-fifth that of the other normal workers, were subjected to analyses of their ALAD and ALAD alleles. Further, the entire exon fragment of the ALAD gene was analyzed by polymerase chain reaction, and the reaction product was used as a target for direct DNA sequencing. Genomic DNA analysis revealed that all 11 workers had the ALAD allele, whereas the entire ALAD gene analysis failed to indicate other variants, except for the Rsa I site. The depletion in erythrocyte ALAD activity was not found to be caused by the ALAD allele.     

Initial occupational exposure to lead: Chromosome and Biochemical findings.

Serial chromosome and biochemical studies were carried out in 11 subjects before and during initial occupational exposure to moderate quantities of lead fumes in a storage battery plant. The rate of abnormal metaphases, mostly with chromatid and one-break chromosome aberrations, was approximately doubled after one month of work; it further increased after two months of work; remained in this range up to seven months of exposure; and then tended to decrease somewhat. Blood lead levels increased progressively in the first few months, then reached a steady state. Urinary lead and coproporphyrin levels increased sharply after one month of work, while urinary delta-aminolevulinic acid (ALA) levels increased moderately. The ALA dehydratase (ALAD) activity of red blood cells (RBCs) was reduced to almost 50% of the initial values after one month, decreased further in subsequent months, and remained decreased through the remainder of the study.     

Lead exposure in highway toll-booth workers

Biomarkers of lead exposure (blood lead, BPb) and effect (erythrocyte protoporhyrin, EP, and activity of delta-aminolevulinic acid dehydratase, ALAD) were measured in 68 male toll-booth operators (aged 22-60 years) on the Zagreb-Karlovac motorway. Average values (arithmetic mean +/- standard deviation) were: 61.8 +/- 29.3 micrograms/L for BPb, 0.70 +/- 0.20 mumol/L erythrocytes for EP, and 50.6 +/- 9.8 U/L erythrocytes for ALAD. All were within the normal range determined for general population (BPb < 150 micrograms/L, EP < 1.62 mumol/L erythrocytes, and ALAD > 35 U/L erythrocytes). A significant positive correlation was found between BPb and EP (r = 0.367, P < 0.01) and an inverse correlation between BPb and ALAD (r = -0.271, P < 0.05) and for EP and ALAD (r = -0.381, P < 0.01). Significant correlations were found between BPb or ALAD and smoking index (r = 0.486, P < 0.01, and r = -0.322, P < 0.01, respectively), whereas BPb also significantly correlated with blood gamma-glutamyl transferase (GGT) activity, which may indicate hepatotoxic effect of alcohol consumption (r = 0.334, P < 0.01). Among standard spirometric tests, BPb inversely correlated with FEV1 (r = -0.251, P < 0.05) and Tiffenau index (r = -0.280, P < 0.05), whereas ALAD positively correlated with FEF75-85 (r = 0.261, P < 0.05) and Tiffenau index (r = 0.314, P < 0.01). Among standard hematologic tests, BPb positively correlated with MCV (r = 0.282, P < 0.05), EP inversely correlated with erythrocyte count (r = -0.253, P < 0.05), and ALAD positively correlated with MCHC (r = 0.306, P < 0.05) and inversely with MCV (r = -0.250, P < 0.05). Although PbB values in these workers are within occupational exposure limits, they are higher than in corresponding occupations in developed countries. This may be explained by greater exposure to lead in ambient air, tobacco (through mainstream and sidestream smoking) and alcohol in this population.     

Changes in urinary δ-aminolevulinic acid at low lead exposure level with special reference to production activity

A 7-year follow-up survey on 53 workers was carried out in a lead storage battery factory to evaluate the significance of urinary delta-aminolevulinic acid (ALA) and blood lead in a work environment where lead in the air was considered to be about or less than the current occupational exposure limit. While lead in the blood and ALA in the urine had a good correlation to each other cross-sectionally as well as longitudinally, geometric means of lead concentrations in the workroom air samples which were collected following grid sampling strategy, did not correlate with ALA in urine significantly. On the contrary, the semiannual production of batteries significantly correlated with changes in mean ALA in urine. The questionnaire survey proved that the prolongation in work hours, lead to an increase in the mean ALA in urine as well as a higher incidence of higher-than-normal urinary ALA. The results clearly demonstrated the importance of the biological indicators, such as lead in blood and urinary ALA, as well as the necessity of paying attention to non-industrial hygiene factors, such as the production rate of batteries and the length of the daily shift, for the protection of the workers' health when lead in the air is moderate.     

Biomonitoring Findings for Occupational Lead Exposure in Battery and Ceramic Tile Workers using Biochemical Markers,Alkaline Comet Assay,and Micronucleus Test Coupled with Fluorescence in Situ Hybridisation

Manufacture of lead-containing products has long been associated with various health risks. To get an insight into the related genotoxic risks, we conducted a biomonitoring study in 50 exposed workers and 48 matched controls using a battery of endpoints that sensitively detect the extent of genome instability in peripheral blood lymphocytes. The levels of primary DNA damage were estimated with the alkaline comet assay, while cytogenetic abnormalities were determined with the cytokinesis-block micronucleus (CBMN) cytome assay. Additionally, CBMN slides of 20 exposed and 16 control participants were subjected to fluorescence in situ hybridisation (FISH), coupled with pancentromeric probes to establish the incidence of centromere-positive micronuclei, nuclear buds, and nucleoplasmic bridges. Blood lead levels (B-Pb) were measured with atomic absorption spectrometry. To further characterise cumulative effects of occupational exposure, we measured erythrocyte protoporphyrin (EP) concentrations and delta-aminolevulinic acid dehydratase (ALAD) activity in blood. We also assessed the influence of serum folate (S-folate) and vitamin B₁₂ (S-B12) on genome stability. Compared to controls, occupationally exposed workers demonstrated significantly higher B-Pb (298.36±162.07 vs 41.58±23.02), MN frequency (18.71±11.06 vs 8.98±7.50), centromere positive MN (C+ MN) (8.15±1.8 vs 3.69±0.47), and centromere negative MN (C- MN) (14.55±1.80 vs 4.56±0.89). Exposed women had significantly higher comet tail intensity (TI) and length (TL) than control women. Furthermore, workers showed a positive correlation between age and nuclear buds and MN, between MN and years of exposure, and between S-B₁₂ levels and TI and ALAD activity, while a negative correlation was found between TI and B-Pb. These findings suggest that occupational settings in the manufacture of lead-containing products pose significant genotoxic risks, which calls for developing more effective work safety programmes, including periodical monitoring of B-Pb and genetic endpoints.Key words: blood lead, genetic endpoints, genome damage, human lymphocytes, MN-FISH     

Determination of delta-aminolevulinic acid dehydratase (ALAD) by enzyme immunoassay and its application to evaluation of lead exposure

Enzyme Immunoassay (EIA) technique using a glass bead was established in order to quantify the total amount of delta-aminolevulinic acid dehydratase (ALAD, EC 4, 2, 1, 24) including an inactivated enzyme and to determine the level of ALAD in erythrocytes of workers with moderate lead exposure. EIA was carried out by competitive antibody binding method in which the antigen (ALAD) was linked to a glass bead (O.D. 7 mm). A standard curve was fitted to the function y = A X B/(x-B) + C, where y is the enzymatic reaction product, x is the concentration of ALAD, and A, B and C are parameters. The coefficients of variation in the intra- and inter-assay were 3.7% and 6.5%, respectively. EIA-based amount of ALAD was 136.7 +/- 23 mg/l erythrocyte (mean +/- S.D.) in 66 workers. This ALAD amount was not found to be correlated with the lead level in the blood but correlated with the duration of lead-exposure. An especially good correlation (r = 0.453) was obtained with the inactivated portion of ALAD (EIA-based amount minus activity-based amount). On the other hand, the ratio of restored activity following heat treatment and addition of Zn and dithiothreitol to non-restored activity was well correlated with the lead level (r = 0.874), but not correlated with the duration of lead-exposure (r = 0.218). The inactivated portion of ALAD based on EIA accordingly may be considered a good indicator of the duration of lead exposure, and the activity-based amount of ALAD may be considered a suitable indicator of the extent of the present lead exposure.     

Lack of association of δ-aminolevulinic acid dehydratase genotype with cytogenetic damage in lead workers

Objective The objective of this study was to evaluate the correlations of genetic polymorphism of genotypes -aminolevulinic acid dehydratase (ALAD), blood lead levels (BLLs), zinc protoporphyrin (ZPP), sister chromatid exchanges (SCEs), and high SCE frequency cells (HFCs) in lead workers.Methods Three groups of lead workers were included in the study: high lead exposure group (26 workers), low lead exposure group (31 workers) and control group (30 controls who lived in an area uncontaminated by lead). Blood samples were taken from all subjects and analyzed for lead levels, ALAD genotype and SCE levels.Results Occupationally exposed workers had significantly higher BLLs, ZPP and hemoglobin levels than the controls. There were no differences among the three groups regarding percentages of ALAD 1-1 and ALAD 1-2 genotypes, but the ALAD 2-2 genotype was not detected in any of the three groups. There were no significant differences among the three groups for BLLs, ZPP and hemoglobin levels based on ALAD 1-1 and ALAD 1-2. Average SCE values in the high lead exposure group were significantly greater than those in the control group (6.2 vs 5.2 SCEs/cell, P<0.05). HFC analysis revealed a significantly higher HFC percentage (53.9%) in the high lead exposure group than in the low lead exposure group (16.1%) and the control group (10%). There appeared to be an interaction effect on HFC percentages between smoking and lead exposure. When multiple regression analysis was used, the factors that affected SCE levels were lead exposure and smoking, but ALAD genotypes did not have any significant effect.Conclusions A significant association existed between both SCE and HFC levels and lead exposure. However, different ALAD genotypes were not found to be associated with levels of blood lead and ZPP in the three groups.     

Biomarkers of lead exposure

Biomarkers of exposure, effect, and susceptibility are reviewed in relation to lead exposure. Of the biomarkers of lead exposure, blood lead (Pb-B), mainly red cell lead, is a representative of soft tissue lead, and most widely used as measures of body burden and absorbed (internal) doses of lead. Urine lead (Pb-U) as well as plasma lead (Pb-P) increases exponentially with increasing Pb-B under a steady-state situation and is a reflection of recent exposure. The amount of lead in plasma and urine (MPb-P and MPb-U) after administration of a chelating agent (e.g. CaEDTA) can be useful for biomarkers of internal exposure of lead, reflecting the mobilizable pool of lead which consists of mainly blood and soft tissue lead with only a small fraction derived from bones. The critical effects in bone marrow arise mainly from the interaction of lead with some enzymatic process responsible for heme synthesis. The effects can be used for the biomarkers of effects. They are the inhibition of delta-aminolevulinic acid dehydratase (ALAD) and the variation in some metabolite concentrations (e.g. delta-aminolevulinic acid in urine (ALA-U), blood (ALA-B) or plasma (ALA-P), coproporphyrin in urine (CP), zinc protoporphyrin (ZP) in blood). The activities of pyrimidine nucleotidase (P5'N) and nicotinamide adenine dinucleotide synthetase (NADS) in blood are also decreased in lead exposure, and nucleotide contents in blood is altered in lead exposure. These effects of lead on human can be also useful biomarkers of effect. The differences in levels of heme precursors between two types of ALAD genotypes might be attributable to those in the affinity of different ALAD isozymes to lead. ALAD1 homozygotes have higher levels of ZP and ALA in comparison with ALAD2 carriers at the high lead exposure, suggesting that ALAD1 homozygotes might be more susceptible for disturbance in heme biosynthesis by lead than ALAD2 carriers.     

The protective effect of delta-aminolevulinic acid dehydratase 1-2 and 2-2 isozymes against blood lead with higher hematologic parameters

Previous studies have suggested that delta-aminolevulinic acid dehydratase (ALAD) types 1-2 or 2-2 are protective against the toxicity of blood lead (PbB) when zinc protoporphyrin (ZPP) levels are low because of differential binding of lead in erythrocytes. The hypothesis is that subjects with the ALAD 1-1 genotype are more susceptible to lead exposure with impaired hematologic synthesis and therefore that iron nutrition is more important in those with the ALAD 1-1 genotype. The purpose of this study was to prove the protective effect of ALAD 1-2/2-2 against PbB with higher hematologic parameters. Data on 1,219 male workers from eight lead-using factories in the Republic of Korea were examined in this cross-sectional study. Blood samples were evaluated for PbB, ZPP, hemoglobin (Hb), and serum iron (SFe) concentrations and ALAD genotypes. The overall prevalence of the ALAD 1-2/2-2 genotype was 9.3%, which was associated with lower log ZPP (p < 0.001) and higher Hb (p = 0.014) levels. For the subjects with normal iron status (SFe levels > 60 micro g/dL), those with the ALAD 1-1 genotype were more likely to be anemic (adjusted odds ratio of 5.2; 95% confidence interval, 1.2-22.6) than those with ALAD 1-2/2-2. The study confirms the protective effects of ALAD 1-2/2-2 polymorphisms against PbB on hematologic pathways. In order to promote health and to minimize the toxicity of lead exposure more effectively, the nutritional management of iron in Korean workers should take both their ALAD genotypes and occupational lead exposures into account.     

Improved method for the adjustment of urinary delta-aminolevulinic acid concentration

The work was aimed at finding whether the ratio of delta-aminolevulinic acid (ALA) and creatinine (Cn) concentration (ALA/Cn), presently used in occupational health practice for evaluation of lead exposure gives a better assessment of ALA excretion than uncorrected ALA concentration itself, as well as at finding a better, but not complicated method for adjustment. ALA and Cn concentrations were measured in untimed urine samples of altogether 390 men and women (age: 18-60 years) not occupationally exposed to lead. In agreement with others, ALA/Cn was found to be an unsuitable method of adjustment for the differences in ALA concentration due to the different concentrations of samples. This can be explained by the different renal handling of ALA and Cn, proved by the literature data. The exponential relationship between ALA/Cn and Cn concentration raised the possibility of adjustment to the logarithm of Cn concentration (ALA/log.Cn). This simple method provided a more reliable index, the value of which was independent of the actual Cn concentration of urines within a wide range (2-32 mmol/liter). The recommended biological limit value (70 mumol/log.Cn mmol) separates equally well from normal values, both in dilute and concentrated urines. The evaluation of occupational lead exposure might be more reliable using this index, instead of uncorrected ALA concentration or the ALA/Cn ratio.