Delta-Aminolevulinic Acid Dehydratase, Low Blood Lead Levels, Social Factors, and Intellectual Function in an Afro-Brazilian Children Community.

Delta-aminolevulinic acid dehydratase (ALAD) enzyme catalyzes the second phase of the heme biosynthesis and is involved in lead toxicokinetics. This research aimed to evaluate its influence on the relationship between blood lead (PbB) levels and intellectual performance in Afro-Brazilian children. PbB, hemoglobin concentration, ALAD activity, and polymorphism were determined in whole blood. Anthropometric, socioeconomic, and family environment stimuli data were collected with appropriate instruments. The non-verbal intelligence of children and their mothers or guardians was assessed using the correspondent Raven's Progressive Matrix versions. The medians (range) of PbB levels and ALAD activity were 1.0 µg/dL (0.1-21.3) and, 71 U/L (31-113), respectively. ALAD G177C was distributed as follows: 97.9% for ALAD1/1 and 2.1% for ALAD1/2 genotypes. The mean of Raven raw score was 19.3 (± 5.6) points and there were no differences according to sex or environmental Pb exposure. No statistically significant association was observed between PbB level and children's IQ. However, ALAD activity presented an inverse significant association with PbB levels, children's percentile IQ, and children's IQ/Age ratio, suggesting a neuroprotective role of ALAD1 genotype in those with low PbB level.

Mercury Exposure in Munduruku Indigenous Communities from Brazilian Amazon: Methodological Background and an Overview of the Principal Results.

The Amazonian indigenous peoples depend on natural resources to live, but human activities' growing impacts threaten their health and livelihoods. Our objectives were to present the principal results of an integrated and multidisciplinary analysis of the health parameters and assess the mercury (Hg) exposure levels in indigenous populations in the Brazilian Amazon. We carried out a cross-sectional study based on a census of three Munduruku indigenous villages (Sawré Muybu, Poxo Muybu, and Sawré Aboy), located in the Sawré Muybu Indigenous Land, between 29 October and 9 November 2019. The investigation included: (i) sociodemographic characterization of the participants; (ii) health assessment; (iii) genetic polymorphism analysis; (iv) hair mercury determination; and (v) fish mercury determination. We used the logistic regression model with conditional Prevalence Ratio (PR), with the respective 95% confidence intervals (CI95%) to explore factors associated with mercury exposure levels ≥6.0 µg/g. A total of 200 participants were interviewed. Mercury levels (197 hair samples) ranged from 1.4 to 23.9 µg/g, with significant differences between the villages (Kruskal-Wallis test: 19.9; p-value < 0.001). On average, the general prevalence of Hg exposure ≥ 6.0 µg/g was 57.9%. For participants ≥12 years old, the Hg exposure ≥6.0 µg/g showed associated with no regular income (PR: 1.3; CI95%: 1.0-1.8), high blood pressure (PR: 1.6; CI95%: 1.3-2.1) and was more prominent in Sawré Aboy village (PR: 1.8; CI95%: 1.3-2.3). For women of childbearing age, the Hg exposure ≥6.0 µg/g was associated with high blood pressure (PR: 1.9; CI95%: 1.2-2.3), with pregnancy (PR: 1.5; CI95%: 1.0-2.1) and was more prominent among residents in Poxo Muybu (PR: 1.9; CI95%: 1.0-3.4) and Sawré Aboy (PR: 2.5; CI95%: 1.4-4.4) villages. Our findings suggest that chronic mercury exposure causes harmful effects to the studied indigenous communities, especially considering vulnerable groups of the population, such as women of childbearing age. Lastly, we propose to stop the illegal mining in these areas and develop a risk management plan that aims to ensure the health, livelihoods, and human rights of the indigenous people from Amazon Basin.

Genetic Polymorphism of Delta Aminolevulinic Acid Dehydratase (ALAD) Gene and Symptoms of Chronic Mercury Exposure in Munduruku Indigenous Children within the Brazilian Amazon.

Genetic polymorphisms involved in mercury toxicokinetics and toxicodynamics may be associated with severe mercury toxicity. This study aimed to investigate the impact of an ALAD polymorphism on chronic mercury exposure and the health situation of indigenous children from the Brazilian Amazon. One-hundred-and-three indigenous children (under 15 years old) were included and genotyped (rs1800435) using a TaqMan validated assay. The mean age was 6.6 ± 4.5 years old, 60% were female, 49% presented with anemia, and the mean hair mercury concentration was 7.0 ± 4.5 (1.4-23.9) µg/g, with 49% exceeding the reference limit (≥6.0 µg/g). Only two children were heterozygous ALAD, while the others were all wild type. Minor allele frequency (ALAD G) and heterozygous genotype (ALAD CG) were 1% and 2%, respectively. The two children (12 and 14 years old) with the ALAD polymorphism had mercury levels above the average as well as had neurological symptoms related to chronic mercury exposure, such as visual field alterations, memory deficit, distal neuropathy, and toe amyotrophy. Both children also reported frequent consumption of fish in the diet, at least three times a week. In conclusion, our data confirm that an ALAD polymorphism can contribute to mercury half-life time, harmful effects, and neuropsychological disorders in indigenous children with chronic mercury exposure to gold mining activity.

Exploring human porphobilinogen synthase metalloprotein by quantum biochemistry and evolutionary methods.

Previous studies have shown the porphobilinogen synthase (PBGS) zinc-binding mechanism and its conservation among the living cells. However, the precise molecular interaction of zinc with the active center of the enzyme is unknown. In particular, quantum chemistry techniques within the density functional theory (DFT) framework have been the key methodology to describe metalloproteins, when one is looking for a compromise between accuracy and computational feasibility. Considering this, we used DFT-based models within the molecular fractionation with conjugate caps scheme to evaluate the binding energy features of zinc interacting with the human PBGS. Besides, phylogenetic and clustering analyses were successfully employed in extracting useful information from protein sequences to identify groups of conserved residues that build the ions-binding site. Our results also report a conservative assessment of the relevant amino acids, as well as the benchmark analysis of the calculation models used. The most relevant intermolecular interactions in Zn2+-PBGS are due to the amino acids CYS0122, CYS0124, CYS0132, ASP0169, SER0168, ARG0221, HIS0131, ASP0120, GLY0133, VAL0121, ARG0209, and ARG0174. Among these residues, we highlighted ASP0120, GLY0133, HIS0131, SER0168, and ARG0209 by co-occurring in all clusters generated by unsupervised clustering analysis. On the other hand, the triple cysteines at 2.5 Å from zinc (CYS0122, CYS0124, and CYS0132) have the highest energy attraction and are absent in the taxa Viridiplantae, Sar, Rhodophyta, and some Bacteria. Additionally, the performance of the DFT-based models shows that the processing time-dependence is more associated with the choice of the basis set than the exchange-correlation functional.

Environmental exposure to lead and hematological parameters in Afro-Brazilian children living near artisanal glazed pottery workshops.

This study aimed to evaluate the exposure to environmental lead (Pb) of children from a traditional community of African descent in Brazil and the effects on hematological parameters. Children (n = 75) aged 5.5-13 years from the exposed areas classified as low (LEx) and moderately (MEx) exposed were compared with children (n = 75) of a control group (CG). Dust Pb loading rates (RtPbs) at children's homes were measured. Peripheral venous blood samples were collected to assess Pb biomarkers of exposure and effects. All Pb determinations were performed by graphite furnace atomic absorption spectrometry. The median (IQR) of RtPb, PbB level, and ALA-D activity were 65 (25-137) µgPb/m2/30 days, 1.0 (0.1-2.8) µg/dL, and 71 (55-86) U/L, respectively. Spearman correlation evidenced the relationship of PbB with RtPb (rho = 0.368, P < 0.001) and ALA-D activity (rho = -0.587; P ˂ 0.001). After adjusting for exposure degree and child's age, a decline of 7.4 U/L in ALA-D activity was associated with a 10-fold increase in PbB. The prevalence ratio of elevated PbB (>5 µg/dL) in LEx and MEx areas were 1.5 and 3.7, respectively. Indoor dust exposure, living near pottery workshops and the secondary exposure were the main determinants of elevated PbB levels, which were associated with hematological effects.

Toxicological effects in children exposed to lead: A cross-sectional study at the Colombian Caribbean coast.

Lead (Pb) is an environmental pollutant with a toxicity that is a serious public health problem. The aim of this research was to evaluate the associations between Pb exposure and morphometric, hematological and biochemical parameters, mRNA expression of the P53, SOD1, ALAD, TNF and INF-γ genes, ALAD polymorphisms (db SNP ID: rs1800435) and Intelligence Quotient (IQ) in children from the Colombian Caribbean. Blood lead levels (BLL) were determined in 554 participants between the ages of 5-16 years old, from different places of the Colombian Caribbean. A health survey was given to assess risk factors. Whole blood was used for hematology and plasma employed to analyze markers of hepatic toxicity. Gene expression was quantified from blood mRNA by RT-PCR. The ALAD polymorphism was characterized by PCR-RFLP, and the Kaufman's brief intelligence test was employed to estimate the IQ. The mean BLL was 3.5 ±â€¯0.2 µg/dL. The site of greatest exposure to Pb was Tasajera, a poor fishing community, with an average of 8.9 ±â€¯0.8 µg/dL. Breastfeeding was associated with high BLL. Morphometric characteristics and IQ were negatively correlated with BLL. The blood platelet count and the mean corpuscular hemoglobin concentration showed positive and negative correlations with BLL, respectively. Negative relationships with BLL were observed with the ratios Neutrophils/Eosinophils and Neutrophils/Basophils, whereas for BLL and Neutrophils/Monocytes the association was positive. The associations between morphometric and some hematological parameters with BLL were age- and gender-related. The expression of ALAD, SOD1, INF-γ and P53 mRNA was down-regulated according to the BLL, whereas TNF showed an opposite trend. In short, fishing communities are at a high risk of Pb exposure. This xenobiotic can affect physical development and IQ, as well as hematological parameters, even at low concentrations. In addition, it can regulate the transcription of genes associated with inflammation, apoptosis, cell cycle, heme synthesis, and oxidative stress.

Effects of ALAD genotype on the relationship between lead exposure and anthropometry in a Cohort of Mexican children.

OBJECTIVES: Lead exposure is associated with children's growth, but this relationship may depend on the presence of susceptibility factors, including genetic variation. Blood lead levels (BLL) differ by ALAD (aminolevulinic acid dehydratase) genotype. We investigated the association between BLL and growth in Mexican first-graders with different ALAD genotypes. METHODS: Children between the ages of 6-8 years (n = 602) attending first grade in schools within the vicinity of a metal foundry in Torreón, Mexico were enrolled into a randomized controlled trial (RCT) testing the efficacy of iron and/or zinc supplementation on blood lead levels (BLL) and cognition. BLL and anthropometry were assessed at baseline (height, height-for-age z-score (HAZ), knee height, head circumference), after 6 (head circumference) and 12 months (height, HAZ, knee height). Children with ALAD1-1 and ALAD1-2/2-2 were compared. The study sample included 538 and 470 participants who had complete data at baseline and follow-up, respectively. Separate multivariable linear regression models adjusted for covariates were used to test the association between BLL at baseline and each anthropometric measure. Covariates included age, sex, hemoglobin, crowding, and maternal education. BLL x ALAD genotype interaction term was tested. RESULTS: Median BLL (10.1 µg/dL) did not differ by ALAD genotype. After covariate adjustment, baseline BLL was inversely associated with baseline height, HAZ, and knee height. The association (ß [95% CI]) between BLL and baseline height (-0.38[-0.68, -0.09]), HAZ (-0.07[-0.12, -0.02]) and knee height (-0.14[-0.25, -0.02]), was somewhat stronger in children with ALAD1-2/2-2 than ALAD1-1 (-0.09[-0.16, -0.02], -0.02[-0.03, -0.004] and -0.04[-0.06, -0.01], respectively). No associations between BLL and growth at 6 or 12 months were detected irrespective of ALAD genotype. CONCLUSIONS: BLL was adversely associated with anthropometric measures among Mexican children. ALAD genotype may be a susceptibility factor for the effects of lead on child growth.

House sparrow biomarkers as lead pollution bioindicators. Evaluation of dose and exposition length on hematological and oxidative stress parameters.

House sparrows (Passer domesticus) have been proposed as a key ecological indicator of urban pollution. Remarkably, we lack knowledge about the physiological effects of lead on this bird species. Therefore, this study was aimed to evaluate the effect of Pb on several physiological parameters in house sparrows exposed to environmental Pb concentrations. In a first experiment, birds were exposed to Pb sub-lethal doses (from 1.3 to 14.0 µg of Pb/g animal/day) during 5 days, which resulted in a dose response increase of blood Pb levels and decrease of blood ALAD activity. However, at the higher doses tested (> 7 µg of Pb/g animal/day) the blood ALAD activity inhibition (~82%) remained constant. Hematocrit and hemoglobin were significantly reduced only at the highest-doses, and the stress indicator, heterophils to lymphocyte (H/L) ratio, did not show apparent changes. In a second experiment, house sparrows were exposed to Pb in drinking water (12.3 ppm) during either 15 or 30 days. Pb concentration used in this study was enough to produce blood lead levels equivalents to those found recently in house sparrows inhabiting urban areas, reduced blood ALAD activity and inversion of the H/L ratio. Decreasing blood ALAD activities were correlated with increasing blood Pb levels. In addition, Pb exposure produced modification in the levels of hepatic antioxidant enzymes, increased GST activity and decreased CAT activity, without lipid peroxidation. In conclusion, our results suggest that blood ALAD activity is a reliable and sensitive biomarker for environmental Pb exposure in house sparrows, additionally chronic exposure produce physiological stress (H/L inversion) and small changes in antioxidant enzyme activity. Finally, this specie could be considered a bioindicator for monitoring the urban Pb contamination.

In Silico Studies of Mammalian δ-ALAD Interactions with Selenides and Selenoxides.

Previous studies have shown that the mammalian δ-aminolevulinic acid dehydratase (δ-ALAD) is inhibited by selenides and selenoxides, which can involve thiol oxidation. However, the precise molecular interaction of selenides and selenoxides with the active center of the enzyme is unknown. Here, we try to explain the interaction of selenides and the respective selenoxides with human δ-ALAD by in silico molecular docking. The in silico data indicated that Se atoms of selenoxides have higher electrophilic character than their respective selenides. Further, the presence of oxygen increased the interaction of selenoxides with the δ-ALAD active site by O…Zn coordination. The interaction of S atom from Cys124 with the Se atom indicated the importance of the nucleophilic attack of the enzyme thiolate to the organoselenium molecules. These observations help us to understand the interaction of target proteins with organoselenium compounds.

Low blood lead levels impair intellectual and hematological function in children from Cartagena, Caribbean coast of Colombia.

Lead produces numerous biochemical and physiological changes in humans, including hematological disorders, toxic effects on the central nervous system and in the function of several organs. The aim of this study was to determine blood lead levels (BLL) in children from Cartagena, Colombia, associating those with hematological and liver damage markers, the intelligence quotient (IQ), as well as with gene expression of the aminolevulinate dehydratase (ALAD), superoxide dismutase 1 (SOD1), gamma interferon (INF-γ), tumor necrosis factor (TNF) and tumor protein (p53). To achieve this purpose, 118 blood samples were collected from children 5-16 years old, with their respective informed consent from their parents. BLL was measured by atomic absorption; hematological parameters were obtained with automated systems; plasma was utilized to analyze hepatic toxicity markers, alanine aminotransferase (ALT), gamma-glutamyltransferase (γ-GT) and alkaline phosphatase (ALP); the Kaufman Brief Intelligence Test (K-BIT) was administered to measure the IQ; and gene expression was quantified from blood RNA. The mean BLL was 1.7±0.3µg/dL. A low proportion of the children (3.4%) had BLL above the CDC recommended limit (5µg/dL). BLL were correlated weakly, but negatively with child age, weight, height, body mass index, platelets wide distribution, mean platelet volume, γ-GT and IQ. There were not significant changes in the expression of evaluated genes. These results support the hypothesis that BLL below 5µg/dL may still be a detrimental factor on children's cognitive abilities, development and hematology, in line with recent concerns that there is no safe level of pediatric lead exposure.

Association between Blood Lead Levels and Delta-Aminolevulinic Acid Dehydratase in Pregnant Women.

Blood lead levels (BLLs) and delta-aminolevulinic acid dehydratase (ALAD) activity are considered biomarkers of lead exposure and lead toxicity, respectively. The present study was designed to investigate the association between BLLs and ALAD activity in pregnant women from Durango, Mexico. A total of 633 pregnant women aged 13-43 years participated in this study. Blood lead was measured by a graphite furnace atomic absorption spectrometer. ALAD activity was measured spectrophotometrically. Mean blood lead was 2.09 ± 2.34 µg/dL; and 26 women (4.1%) crossed the Centers for Disease Control (CDC) recommended level of 5 µg/dL. ALAD activity was significantly lower in women with levels of lead ≥5 µg/dL compared to those with BLLs < 5 µg/dL (p = 0.002). To reduce the influence of extreme values on the statistical analysis, BLLs were analyzed by quartiles. A significant negative correlation between blood lead and ALAD activity was observed in the fourth quartile of BLLs (r = -0.113; p < 0.01). Among women with blood lead concentrations ≥2.2 µg/dL ALAD activity was negatively correlated with BLLs (r = -0.413; p < 0.01). Multiple linear regression demonstrated that inhibition of ALAD in pregnant women may occur at levels of lead in blood above 2.2 µg/dL.

Early hematological and immunological alterations in gasoline station attendants exposed to benzene.

INTRODUCTION: Elucidation of effective biomarkers may provide tools for the early detection of biological alterations caused by benzene exposure and may contribute to the reduction of occupational diseases. This study aimed to assess early alterations on hematological and immunological systems of workers exposed to benzene. METHODS: Sixty gasoline station attendants (GSA group) and 28 control subjects were evaluated. Environmental and biological monitoring of benzene exposure was performed in blood and urine. The potential effect biomarkers evaluated were δ-aminolevulinate dehydratase (ALA-D) activity, CD80 and CD86 expression in lymphocytes and monocytes, and serum interleukin-8 (IL-8). The influence of confounding factors and toluene co-exposure were considered. RESULTS: Although exposures were below ACGIH (American Conference of Governmental Industrial Hygienists) limits, reduced ALA-D activity, decreased CD80 and CD86 expression in monocytes and increased IL-8 levels were found in the GSA group compared to the control subjects. Furthermore, according to multiple linear regression analysis, benzene exposure was associated to a decrease in CD80 and CD86 expression in monocytes. CONCLUSIONS: These findings suggest, for the first time, a potential effect of benzene exposure on ALA-D activity, CD80 and CD86 expression, IL-8 levels, which could be suggested as potential markers for the early detection of benzene-induced alterations.

Possíveis efeitos do cobre sanguíneo sobre parâmetros hematológicos em idosas / Possible effects of blood copper on hematological parameters in elderly

INTRODUÇÃO: O cobre é um elemento traço essencial e sua homeostase é importante, principalmente em idosos, uma vez que seu metabolismo está associado a doenças neurodegenerativas e distúrbios na eritropoiese, entre outros. OBJETIVO: O presente estudo avaliou a associação entre cupremia, parâmetros hematológicos e estresse oxidativo. MATERIAIS E MÉTODOS: Amostras de sangue de 39 mulheres idosas (grupo de estudo) e de 20 indivíduos adultos saudáveis (grupo-controle) foram coletadas. As concentrações de cobre sérico foram quantificadas por espectrometria de massa por plasma indutivamente acoplado (ICP-MS), a atividade e o índice de reativação da enzima δ-aminolevulinato desidratase (ALA-D) foram determinados por espectrofotometria e parâmetros sanguíneos foram analisados em sistema automatizado. Os resultados foram expressos em média ± desvio padrão (DP). RESULTADOS: As concentrações de cobre, parâmetros hematológicos e índice de reativação da ALA-D para ambos os grupos encontraram-se dentro dos valores de referência. Porém, a atividade da ALA-D (11,47 ± 2,81 U.L-1) foi significativamente inferior no grupo de estudo em comparação com o grupo-controle. Correlações de Spearman (observadas somente nas mulheres idosas) entre as concentrações de cobre versus hemoglobina, hematócrito e atividade da ALA-D foram -0,384; -0,408 e -0,395, respectivamente (p < 0,05). No entanto, o índice de reativação da ALA não apresentou correlação com a cupremia. DISCUSSÃO E CONCLUSÃO: Os resultados mostraram que o cobre, mesmo estando nos limites considerados valores de referência, pode estar envolvido na inibição da ALA-D, o que pode alterar parâmetros hematológicos, como a síntese de hemoglobina. Neste sentido, sugere-se que os níveis de referência para o cobre em idosos sejam reavaliados.

INTRODUCTION: Copper is an essential trace element, and its homeostasis is important, mainly among the elderly, since their metabolism is associated with neurodegenerative diseases and erythropoiesis disorders, among others. OBJECTIVE: This study evaluated the association among cupremia, hematological parameters and oxidative stress. MATERIAL AND METHODS: Blood samples from 39 elderly women (study group) and 20 health individuals (control group) were collected. The concentrations of serum copper were quantified by ICP-MS. The activity and enzyme ALA-D reactivation index were determined by spectrophotometry and blood parameters were analyzed in the automated system. The results were expressed as mean ± standard deviation. RESULTS: Concentrations of copper, hematological parameters and ALA-D reactivation were within the reference values in both groups. However, ALA-D activity (11.47 ± 2.81 UL-1) was significantly lower in the study group compared to the control group. Spearman correlations (observed only in elderly women) between copper concentration versus hemoglobin, hematocrit and ALA-D activity were -0.384, -0.408 and -0.395, respectively (p < 0.05). Nonetheless, ALA reactivation index was not related to cupremia. DISCUSSION AND CONCLUSION: The results showed that copper, although it is within accepted reference values, may be involved in ALA-D inhibition, which may affect hematological parameters such as hemoglobin synthesis. Thus, the reference levels for copper in the elderly should be reviewed.