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Disposition of lead (Pb) in saliva and blood of Sprague-Dawley rats following a single or repeated oral exposure to Pb-acetate.

Biological monitoring for lead (Pb) is usually based upon a determination of blood Pb concentration; however, saliva has been suggested as a non-invasive biological matrix for assessing exposure. To further evaluate the potential utility of saliva for biomonitoring, the disposition of Pb was evaluated in whole blood (WB), red blood cells (RBC), plasma, parotid gland, bone, and saliva following either a single oral dose of 100mg Pb-acetate/kg body weight in rats or approximately 1-week after 5 sequential daily oral gavage doses of 1, 10, or 100mg Pb-acetate/kg/day. Saliva volume, pH, total saliva protein, and alpha-amylase activity were also determined. At specified times post-dosing groups of animals were anesthetized and administered pilocarpine to induce salivation. Saliva was collected, the animals were humanely sacrificed, and tissue samples were likewise collected, weighed, and processed for Pb analysis. Following a single dose exposure to Pb-acetate, Pb was detectable in all samples by 30 min post-dosing. For both the single and repeated dose treatments the concentration of Pb was highest in WB and RBC relative to plasma and saliva. However, the Pb rapidly redistributed (within 5-days post-treatment) from the blood into the bone compartment based on the substantial decrease in WB and RBC Pb concentration, and the concurrent increase in bone Pb following repeated exposure at all dose levels. Although there is clear variability in the observed Pb concentrations in plasma and saliva, there was a reasonable correlation (r(2)=0.922) between the average Pb concentrations in these biological matrices, which was consistent with previous observations. The single oral dose of Pb-acetate resulted in a decrease in salivary pH which recovered by 24h post-dosing and a decrease in alpha-amylase enzyme activity which did recover within 5-days of ceasing exposure. It is currently unclear what impact these slight functional changes may or may not have on Pb salivary clearance rates. These results demonstrate a feasibility to rapidly detect Pb in saliva and suggest that saliva may correlate best with plasma Pb concentration.

Animals↗

Low level lead (Pb) exposure produces learning deficits in young rat pups.

Eleven day old Sprague-Dawley (CD strain) rat pups whose mothers were maintained on a 200 mg/l dosage of lead (Pb) acetate in their drinking water from breeding and through gestation until the pups were weaned exhibited differences in a learning paradigm as compared to controls. No significant differences were noted between the control pups and the experimental pups with respect to acquisition rates but there were significant differences between the two groups with respect to extinction rates. Similar results were obtained in tests of rat pups whose dams had been maintained on a 400 mg/l dosage of lead (Pb) acetate. These data indicate that low level lead (Pb) exposure can induce significant behavioral deficits in young rat pups.

Animals↗

Microanalyzer for biomonitoring lead (Pb) in blood and urine.

Biomonitoring of lead (Pb) in blood and urine enables quantitative evaluation of human occupational and environmental exposures to Pb. State-of-the-art ICP-MS instruments can only analyze metals in laboratories, resulting in lengthy turnaround times, and they are expensive. In response to the growing need for a metal analyzer capable of on-site, real-time monitoring of trace toxic metals in individuals, we developed a portable microanalyzer based on flow-injection/stripping voltammetry (ASV), and validated the system using rat blood and urine spiked with known amounts of Pb. Fouling of electrodes by proteins often prevents the effective use of electrochemical sensors in biological matrices. Minimization of such fouling was accomplished with suitable sample pretreatment and by establishing turbulent flow of blood and urine containing Pb onto the electrode inside the microanalyzer, which resulted in no apparent electrode fouling even when the samples contained 50% urine or 10% blood by volume. No matrix effect was observed for the voltammetric Pb signals, even when the samples contained 10% blood or 10% urine. The microanalyzer offered linear concentration ranges relevant to Pb exposure levels in humans (0-20 ppb in 10% blood samples, 0-50 ppb in 50% urine samples). The device showed excellent sensitivity and reproducibility; Pb detection limits were 0.44 ppb and 0.46 ppb, and % R.S.D. was 4.9 and 2.4 in 50% urine and 10% blood samples, respectively. It gave similar Pb concentrations in blood and urine to those measured by ICP-MS. It offered high throughput (3 min per sample) and economical use of samples (60 microL per measurement) as well as low reagent consumption (1 microg of Hg per measurement), thus minimizing environmental concerns associated with mercury use. Since it is miniaturized, the microanalyzer is portable and field-deployable. Thus, it shows much promise as the next-generation analyzer for the biomonitoring of toxic metals.

Animals↗

Perinatal Lead (Pb) Exposure Increases Mouse Embryonic Weight and Alters Neuronal Gene Expression.

Acute and chronic exposure to lead (Pb) during pregnancy is linked to adverse health outcomes, including delayed neurodevelopment in offspring. However, the pathways by which Pb exposure influences long-term health remain poorly understood. To address this, we measured the effects of perinatal Pb exposure on gene expression including imprinted genes, X-linked genes, and sexually dimorphic genes. Female mice were given control or Pb acetate dosed (32 ppm) drinking water two weeks prior to timed mating until embryonic day (E)10-12, upon which whole embryos were collected, weighed, and sexed at E13-15. From a subset of embryo heads (n&#x2265;9 per sex per group), we extracted and sequenced RNA. We used linear regression to assess Pb impacts on embryonic weight and gene expression across all mice and stratified by sex. Among the differentially expressed genes, we identified significantly enriched pathways. Pb-exposed embryos weighed more than controls (p=0.007), across both sexes. Collectively, we identified 2,920 differentially expressed genes (FDR<0.05), including 31 imprinted genes and 120 X-linked genes upon Pb exposure. Pb exposure altered expression in gene pathways related to neuronal structure and function as well as sexually dimorphic genes (44 for females; 76 for males). These findings highlight perinatal Pb-linked alterations that may drive later-life health outcomes.

DOHaD↗

Incidence of subclinical lead (Pb) exposure in cattle of an industrial area in Greece.

The incidence of subclinical lead (Pb) poisoning was investigated in the cattle population of an industrial area in northern Greece. The cattle were divided into two groups with respect to their alleged exposure to Pb, which was assayed by determining whole blood and hair Pb concentration and erythrocyte delta-aminolevulinic acid dehydratase (ALA-D) activity. The cows which were located closer to potential contamination sources were found to have higher (P < 0.05) blood and hair Pb content, whereas no significant differences were noted in ALA-D activity. Hair Pb increased significantly (P < 0.05) with age, an observation which should lead to further, more extensive research, in order to evaluate the possibility of using hair as indicator of chronic Pb intake.

Animal Feed↗

Effect of lead (Pb) exposure on the activity of superoxide dismutase and catalase in battery manufacturing workers (BMW) of Western Maharashtra (India) with reference to heme biosynthesis.

The aim of this study was to estimate the activity of superoxide dismutase (SOD) and catalase in erythrocytes and malondialdehyde (MDA) in plasma of battery manufacturing workers (BMW) of Western Maharashtra (India) who were occupationally exposed to lead (Pb) over a long period of time (about 15 years). This study was also aimed to determine the Pb intoxication resulted in a disturbance of heme biosynthesis in BMW group. The blood Pb level of BMW group (n = 28) was found to be in the range of 25.8 - 78.0 microg/dL (mean + SD, 53.63 + 16.98) whereas in Pb unexposed control group (n = 35) the range was 2.8 - 22.0 microg/dL (mean + SD, 12.52 + 4.08). The blood level (Pb-B) and urinary lead level (Pb-U) were significantly increased in BMW group as compared to unexposed control. Though activated d- aminolevulinic acid dehydratase (ALAD) activities in BMW group did not show any significant change when compared to control group but activated / non activated erythrocyte - ALAD activities in BMW group showed a significant increase. Erythrocyte- zinc protoporphyrin (ZPP), urinary daminolevulinic acid (ALA-U) and porphobilinogen (PBG-U) of BMW groups elevated significantly as compared to control. A positive correlation (r = 0.66, p < 0.001) between Pb-B and ALA-U were found in BMW group but no such significant correlation (r = 0.02, p> 1.0) were observed in control group. Hematological study revealed a significant decrease of hemoglobin concentration, packed cell volume (%) and other blood indices and a significant increase of total leucocytes count in BMW group in comparison to control group. The serum MDA content was significantly increased (p < 0.001) and the activities of antioxidant enzymes such as erythrocyte- SOD (p < 0.001) and erythrocytecatalase (p < 0.001) were significantly reduced in BMW group as compared to control group. A positive correlation (r = 0.45, p < 0.02) between Pb-B and serum MDA level was observed in BMW group (Pb-B range 25.8 - 78.0 microg / dL) but such significant correlation did not notice in control group (Pb-B range 2.8 - 22.0 microg / dL). The study clearly showed an adverse effect of heme biosynthesis and imbalance of pro-oxidant / antioxidant status in lead exposed battery manufacturing workers resulting in increase in lipid peroxidation associated with decrease in erythrocyte-SOD and erythrocyte-catalase activities.

Adult↗

Effects of lead (Pb) exposure during gestation and lactation on female pubertal development in the rat.

Lead (Pb) can delay sexual maturation; however, the mechanism and critical time of insult are not clearly defined. Therefore, we assessed maternal Pb levels during low-level gestational and/or lactational exposure, as well as blood and tissue Pb in developing fetuses in relation to the subsequent detrimental effects of Pb on puberty-related hormones and the onset of female puberty. Adult Fisher 344 female rats were gavaged daily with either a 1-ml solution of PbAc containing 12 mg/ml Pb or an equal volume of sodium acetate (NaCl), for the controls, from 30 days prior to breeding until their pups were weaned at 21 days. By cross-fostering at the time of birth, the pups were either exposed to PbAc or NaAc during gestation only, lactation only, or during both gestation and lactation. Pb delayed the timing of puberty and this delay was associated with suppressed serum levels of insulin-like growth factor-1 (IGF-1), luteinizing hormone (LH), and estradiol (E(2)). Liver IGF-1 mRNA was not affected, suggesting that Pb altered translation and/or secretion of IGF-1. We reported previously that peripherally derived IGF-1 acts at the hypothalamic level to facilitate LH release at puberty; hence, we suggest that the action of Pb in decreasing circulating IGF-1 contributes to the delayed puberty. The detrimental effects occurred regardless of the developmental time of exposure, although gestational exposure appeared more sensitive to the effects of Pb. Also, the effects noted were with blood Pb levels less than previously reported and these levels are relevant to human health concerns.

Administration, Oral↗

Lead (Pb+2) promotes apoptosis in newborn rat cerebellar neurons: pathological implications.

Despite mandated reduction in environmental lead (Pb++), such exposure still poses a public health hazard for children, with devastating effects on CNS development. To replicate aspects of this neurotoxicity, we used cultured granule cells from newborn rat cerebella to study whether apoptotic or necrotic death is the major consequence of exposure to low micromolar concentrations of Pb++. At a low dose, 1 microM (EC50 approximately equal to 10 microM), Pb++ does not affect glutamate-induced neuronal necrosis but promotes neuronal apoptosis, as characterized morphologically by cell shrinkage and chromatin condensation, biochemically by the typical internucleosomal DNA fragmentation and functionally by dependence on new synthesis of macromolecules (cycloheximide- and actinomycin D-sensitive). The low micromolar doses of Pb++ that promote apoptosis are well within the blood level range reported to impair CNS function in children and to alter synaptogenesis in the neonatal rat brain. Thus these in-vitro results suggest that the highly neurotoxic action of Pb++ in the developing CNS of children might depend on a facilitation of apoptosis. The Pb+2-elicited potentiation of neuronal apoptosis is attenuated by treatment with the voltage-sensitive Ca+2 channel agonist Bay K8644, which suggests the possible use of this agonist for treatment of the neurotoxic effects of Pb++.

Animals↗

Effect of magnitude and timing of maternal pregnancy blood lead (Pb) levels on birth outcomes.

OBJECTIVE: Associations between magnitude and timing of maternal pregnancy blood lead (Pb) levels (BLLs), birth weight, and total days of gestation were examined, as well as associations with related clinical diagnoses of low birth weight (LBW), preterm, and small-for-gestational-age (SGA) birth. STUDY DESIGN: Among a sample of 262 mother-infant pairs studied retrospectively, one-way analysis of variance and regression statistics were used to measure the relationship between level of maternal pregnancy BLLs and birth outcomes while controlling for key maternal and newborn factors. RESULTS: Women with maximum pregnancy BLLs (max-PBLLs) > or =10 microg/dl tended to give birth earlier and their babies were at substantially increased risk for preterm and SGA birth. By holding other explanatory factors constant, each unit increase in max-PBLL above 10 mug/dl was found to be associated with a decrease of -0.3 in total days of gestation. Compared to women with lower levels, women with max-PBLLs > or =10 microg/dl were at a threefold increased risk for preterm birth (adjusted OR=3.2, 95% CI 1.2-7.4) and more than a fourfold increased risk for having an SGA infant (adjusted OR=4.2, 1.3-13.9). Second trimester maximum BLLs > or =10 microg/dl were associated with a steep decrease in total days of gestation (a decrease of -1.0 days per each unit increase above 10 microg/dl). CONCLUSIONS: These data provide evidence of the adverse effects of maternal pregnancy BLLs, particularly when levels are > or =10 microg/dl. Prenatal Pb exposure at these levels was associated with significant decreases in total days of gestation and an increased risk of preterm and SGA birth.

Adult↗

Lead (Pb) accumulation and genotoxic responses in Ludwigia repens J.R. Forst.: a physiological and molecular approach.

In this study, the potential genotoxic effects and phytoremediation capacity of Ludwigia repens J.R. Forst. were evaluated under lead (Pb) stress in contaminated aquatic environments. To achieve this, clonal L. repens plants were used to investigate their ability to remove Pb from freshwater systems and the experimental setup was established in controlled aquarium conditions. The plants were exposed to 0, 10, 25, 50 and 100 micromolar (&#x3bc;M) concentrations of Pb(NO3)2 in a 0.2% Hoagland nutrient solution for a period of ten days. Experimental results showed that Pb accumulated in both stems and leaves of L. repens. Although Pb levels did not meet hyperaccumulator criteria, the bioaccumulation index (BAI), bioconcentration factor (BCF) and translocation factor (TF) values revealed that the plant was capable of accumulating noteworthy amounts of Pb. In parallel, band profile analysis revealed new band appearances only with the UBC 812 primer, while no band loss or new band formation was detected with the other primers (UBC 808, UBC 826, UBC 833 and UBC 834). Instead, only changes in band intensities were observed, indicating a low polymorphism rate and a high level of genomic template stability (GTS). The findings also indicate that L. repens exhibits notable tolerance to Pb stress, as supported by high BAI, BCF and TF values in the absence of visible phytotoxic symptoms. Additionally, the consistent and progressive decline in mineral nutrient levels across Pb treatments, together with the moderate decline in total chlorophyll content, further supports the existence of a coordinated physiological adjustment, potentially reflecting a tolerance mechanism aimed at maintaining ionic balance under heavy metal stress. Moreover, low polymorphism rates and high genomic template stability (GTS) values derived from molecular analyses suggest that this species may serve as a genetically stable and physiologically resilient aquatic plant. These combined traits highlight its potential to contribute effectively to phytoremediation applications, particularly when co-cultivated with established Pb hyperaccumulator species.

Lead↗

Low level lead (Pb) exposure during gestation and lactation: assessment of effects on pubertal development in Fisher 344 and Sprague-Dawley female rats.

Studies using both Fisher 344 and Sprague-Dawley (SD) rat lines have shown that gestational and/or lactational maternal lead (Pb) exposure causes delayed reproductive maturation in their respective female offspring. Because these studies utilized different experimental regimens for dosing and for monitoring Pb levels, it has not been possible to determine which rat line provides the best model for low level Pb toxicity studies. This study was designed to address this issue. Adult Fisher and SD female rats were dosed with either a solution of PbAc containing 12 mg of Pb/ml or sodium acetate (NaAc) for controls. Dosing began 30 days prior to breeding and continued until their pups were weaned at 21 days of age. At the time of breeding and through weaning the blood lead (BPb) levels in the Fisher dams averaged 37.3 microg/dl and the SD dams averaged 29.9 microg/dl. Pb delayed the timing of puberty (p < 0.01) in Fisher offspring, and suppressed serum levels of luteinizing hormone (LH, p < 0.001) and estradiol (E2, p < 0.01). These effects did not occur in the SD offspring. Doubling the dose given to the SD rats increased their BPb levels to 62.6 microg/dl, yet there were still no effects noted. These results indicate that Fisher offspring are more sensitive to maternal Pb exposure with regard to puberty related insults than are SD rats, suggesting that the Fisher line may be a more reliable rodent model to study the effects of low level Pb toxicity.

Animals↗

Lead (Pb) alters the norepinephrine-induced secretion of luteinizing hormone releasing hormone from the median eminence of adult male rats in vitro.

In the present study, we evaluated the in vitro effects of lead (Pb) on basal and stimulated luteinizing hormone releasing hormone (LHRH) and Prostaglandin E2 (PGE2) secretion. Median eminences (ME) were removed from brains of adult male rats and preincubated for 15 minutes in Krebs-Ringer bicarbonate glucose buffer in an atmosphere of 95% O2-5% CO2. These media were discarded and all MEs were subjected to one of the following experiments. In Experiment 1, all MEs were incubated for 30 minutes in medium only. These media were collected and replaced with medium only (controls) or with medium containing Pb doses ranging from 5 to 20 microM. After this 60-minute incubation, media were collected, then replaced with new medium containing 60 microM norepinephrine (NE), or NE plus each dose of Pb, then incubated for a final 30-minute period. Experiment 2 was conducted as above, except PGE2 (2.8 microM) replaced the NE. In both experiments, the amounts of LHRH released was measured by RIA. In experiment 3, NE was again used for the challenge; however, this time, the amount of PGE2 released was measured by RIA. Results indicate that Pb did not alter basal LHRH release, but compared with controls, significantly blocked NE-induced LHRH release in a dose-related manner. Conversely, Pb had no effect on the PGE2-induced release of LHRH. Additionally, Pb did not alter basal PGE2 release; however, it significantly blocked the NE-induced release of PGE2. Since NE-induced LHRH release is mediated by PGE2, these results support the hypothesis that Pb is capable of altering the hypothalamus and suggest that this effect is due, at least in part, to the diminished PGE2 synthesis/release within the ME, resulting in diminished LHRH secretion.

Animals↗

Lead (Pb(+2)) impairs long-term memory and blocks learning-induced increases in hippocampal protein kinase C activity.

The long-term storage of information in the brain known as long-term memory (LTM) depends on a variety of intracellular signaling cascades utilizing calcium (Ca2+) and cyclic adenosine monophosphate as second messengers. In particular, Ca(+2)/phospholipid-dependent protein kinase C (PKC) activity has been proposed to be necessary for the transition from short-term memory to LTM. Because the neurobehavioral toxicity of lead (Pb(+2)) has been associated to its interference with normal Ca(+2) signaling in neurons, we studied its effects on spatial learning and memory using a hippocampal-dependent discrimination task. Adult rats received microinfusions of either Na+ or Pb(+2) acetate in the CA1 hippocampal subregion before each one of four training sessions. A retention test was given 7 days later to examine LTM. Results suggest that intrahippocampal Pb(+2) did not affect learning of the task, but significantly impaired retention. The effects of Pb(+2) selectively impaired reference memory measured in the retention test, but had no effect on the general performance because it did not affect the latency to complete the task during the test. Finally, we examined the effects of Pb(+2) on the induction of hippocampal Ca(+2)/phospholipid-dependent PKC activity during acquisition training. The results showed that Pb(+2) interfered with the learning-induced activation of Ca(+2)/phospholipid-dependent PKC on day 3 of acquisition. Overall, our results indicate that Pb(+2) causes cognitive impairments in adult rats and that such effects might be subserved by interference with Ca(+2)-related signaling mechanisms required for normal LTM.

Animals↗

Size distribution of aerosols (PM10) and lead (Pb) near traffic intersections in Mumbai (India).

The size distribution of aerosols was measured near traffic intersections of Marol link road in air quality control region (AQCR1), which is a moderately industrial area and Dadar Khodad circle in AQCR2, which is a heavily commercial core of the Mumbai City. The reason behind selecting the two unidentical regions was to study the contribution from vehicles to the size separated PM10 and that of Pb. It is recognised that particulates in urban air are responsible for serious health effects. As very small particles are assumed to be important for the adverse health effects, the particle size distribution is thus an important factor that needs to be addressed whenever the particulates pollution is concerned. The size measurements were done with a cascade impactor of eight stages with a back-up filter. It effectively separates the particulate matter into nine-sizes ranging from 0.0-0.4 to 9.0-10.0 microm. Samples were analysed in nine-particle size fractions with special reference to a toxic metal - lead (Pb) by atomic absorption spectrophotometry (AAS). It was found that PM10 and Pb at both the intersections could easily be classified by the size distribution. The fractions of the PM10 and that of Pb showing a tendency of trimodal distributions with the first peak at coarse mode approximately 9.0-10.0 microm, second at approximately 5.8 microm and the third at coarse mode approximately1.1 microm. The significant percentage of Pb was found in the range below 2.5 microm at both the intersections. However, Pb in AQCR1 is found in the coarser range as well, which could probably be the influence of various industrial activities in the area. PM10 concentration values in the coarser range in AQCR2 are associated with the resuspension of dust particles and mechanical erosions.

Aerosols↗

Effects of chronic lead (Pb) exposure on neurobehavioral function and dopaminergic neurotransmitter receptors in rats.

Sprague-Dawley rats were maternally and permanently exposed to Pb (1000 ppm in their drinking water as lead acetate). Behavioral functions were examined starting at post-natal day (PN) 84. Lead exposure did not change spatial learning in the radial arm maze, but induced higher locomotor activity as observed in the open-field and in the radial arm maze. Lead treatment did not impact motor coordination. Autoradiographic analysis of brain sections indicated that Pb-exposure did produce a decrease in [125I]sulpride (D2 receptor antagonist) binding in the cerebral cortex, but not in the striatum and thalamus nucleus. No change was found in [125I]SCH-23982 (D1 receptor antagonist) binding. Since the cortical dopaminergic system is critical for cognitive processes and motor behavior, it is possible that Pb-related change in D2 receptors may mediate to it induced hyperlocomotor activity.

Animals↗

Intrahippocampal administration of lead (Pb) impairs performance of rats in the Morris water maze.

We examined spatial learning in the Morris water maze after daily acute bilateral micro-injection of 13.9 ng sodium acetate (NaAc) or 37.9 ng lead acetate (PbAc) in 1 microliter volumes into the dorsal hippocampus of normal adult rats. After six days of injections and water maze training, rats injected with NaAc were able to find a hidden platform in 8.3 s, and those injected with PbAc were significantly slower (15.2 s; p < 0.02). In a second experiment, rats were trained to find a hidden platform before injections began and then tested in order to determine if intrahippocampal injections of Pb affected the recall of a previously learned task. The escape latency on the first day after injections began was increased slightly when compared to the last day of training before injections, however the NaAc and PbAc groups were not significantly different over three days of injections. Both treatment groups performed as well as they did before injections began by the second day of injections. These results suggest that the direct injection of Pb into the hippocampus impairs the acquisition but not the recall of the spatial learning task in the Morris water maze.

Analysis of Variance↗

The transfer of lead (Pb) from earthworms to shrews (Myosorex varius).

Shrews (Myosorex varius) were bait-trapped close to Stellenbosch in the Western Cape, South Africa, and analyzed to determine the Pb content of the liver, kidneys, muscle tissue, brain, skull, and bone. Shrews were also fed in the laboratory on mince meat and Pb-contaminated live earthworms for 12-17 days and the Pb content compared with control shrews from the field. The Pb levels in the control shrews from the field were exceptionally high and exceeded levels generally considered to be critical for the onset of pathological symptoms in target organs. The shrews that were fed with Pb-contaminated earthworms had significantly higher Pb concentrations in their livers and kidneys than the control shrews. Given that Pb is bioaccumulated in earthworms, this study showed that predation on earthworms could constitute a major pathway for the entry of Pb into the food chain. M. varius could, similar to other species of shrews in other parts of the world, be a suitable biological monitor to study the environmental hazard of Pb pollution.

Animals↗