Michael R. Lasarev

Organophosphate Pesticide Exposure and Neurobehavioral Performance in Agricultural and Nonagricultural Hispanic Workers

Our understanding of the health risks of farmworkers exposed to pesticides in their work and home environments is rapidly increasing, although studies designed to examine the possible neurobehavioral effects of low-level chronic pesticide exposure are limited. We measured dialkyl phosphate urinary metabolite levels, collected environmental dust samples from a subset of homes, obtained information on work practices, and conducted neurobehavioral tests on a sample of farmworkers in Oregon. Significant correlations between urinary methyl metabolite levels and total methyl organophosphate (azinphos-methyl, phosmet, malathion) house dust levels were observed. We found the neurobehavioral performance of Hispanic immigrant farmworkers to be lower than that observed in a nonagricultural Hispanic immigrant population, and within the sample of agricultural workers there was a positive correlation between urinary organophosphate metabolite levels and poorer performance on some neurobehavioral tests. These findings add to an increasing body of evidence of the association between low levels of pesticide exposure and deficits in neurobehavioral performance.

Variation in organophosphate pesticide metabolites in urine of children living in agricultural communities

Children of migrant farmworkers are at increased risk of exposure to organophosphate pesticides because of “carry-home” transport processes and residential location. Although this at-risk status is generally recognized, few available reports describe the extent of this exposure among agricultural communities. We quantified dialkyl phosphate (DAP) levels in serial samples of urine from 176 children, 2–6 years of age, in three Oregon communities hosting differing agricultural industries: pears, cherries, and fruit berries. Up to three spot samples of urine were collected from children at the beginning, mid-point, and end of their parents’ work seasons. The median levels of dimethylthiophosphate (DMTP), the most commonly detected metabolite, was significantly higher in urine samples from children in each of the three agricultural communities (17.5, 19.0, and 41.0 ng/mL) relative to a reference group of children who lived in an urban community and whose parents did not work in agriculture (6.5 ng/mL; Kruskal-Wallis, p < 0.001). After controlling for age, sex, and weight, the median level of DMTP in children in the pear community was 1.92 times higher than the level in children of the berry community [95% confidence interval (CI), 1.14–3.23] and 1.75 times higher than the level in children of the cherry community (95% CI, 0.95–3.23). We observed increasing levels of DMTP across the work season only within the berry community. Levels decreased in the cherry community and remained constant in the pear community. Substantial temporal variation within the children followed demonstrates the need for multiple urine samples to most accurately characterize longer term and/or cumulative exposure. The observed variability in urinary DAP levels, between communities and over time, could be attributed to the types and amounts of organophosphate pesticides used, the timing of applications and degradation of residues in the environment, work operations and hygiene practices, the proximity of housing to orchards and fields, or the movement of these working families. Additional studies of variation in pesticide exposure across agricultural regions are needed.

Biomarkers of Chlorpyrifos Exposure and Effect in Egyptian Cotton Field Workers

Background Chlorpyrifos (CPF), a widely used organophosphorus pesticide (OP), is metabolized to CPF-oxon, a potent cholinesterase (ChE) inhibitor, and trichloro-2-pyridinol (TCPy). Urinary TCPy is often used as a biomarker for CPF exposure, whereas blood ChE activity is considered an indicator of CPF toxicity. However, whether these biomarkers are dose related has not been studied extensively in populations with repeated daily OP exposures. Objective We sought to determine the relationship between blood ChE and urinary TCPy during repeated occupational exposures to CPF. Methods Daily urine samples and weekly blood samples were collected from pesticide workers (n = 38) in Menoufia Governorate, Egypt, before, during, and after 9–17 consecutive days of CPF application to cotton fields. We compared blood butyrylcholinesterase (BuChE) and acetylcholinesterase (AChE) activities with the respective urinary TCPy concentrations in each worker. Results Average TCPy levels during the middle of a 1- to 2-week CPF application period were significantly higher in pesticide applicators (6,437 μg/g creatinine) than in technicians (184 μg/g) and engineers (157 μg/g), both of whom are involved in supervising the application process. We observed a statistically significant inverse correlation between urinary TCPy and blood BuChE and AChE activities. The no-effect level (or inflection point) of the exposure–effect relationships has an average urinary TCPy level of 114 μg/g creatinine for BuChE and 3,161 μg/g creatinine for AChE. Conclusions Our findings demonstrate a dose–effect relationship between urinary TCPy and both plasma BuChE and red blood cell AChE in humans exposed occupationally to CPF. These findings will contribute to future risk assessment efforts for CPF exposure.

Oxidative Stress and DNA Damage in Agricultural Workers

ABSTRACT Oxidative stress and DNA damage have been proposed as mechanisms linking pesticide exposure to health effects such as cancer and neurological diseases. A pilot study of pesticide applicators and farm workers working in the fruit orchards of Oregon (i.e., apples, pears) was conducted to examine the relationship between organophosphate (OP) pesticide exposure and oxidative stress and DNA damage. Urine samples were analyzed for OP metabolites and 8-hydroxy-2′-deoxyguanosine (8-OH-dG). Lymphocytes were analyzed for oxidative DNA repair activity and DNA damage (Comet assay) and serum analyzed for lipid peroxides (i.e., malondialdehyde [MDA]). Cellular DNA damage in agricultural workers was validated using lymphocyte cell cultures. Urinary OP metabolites were significantly higher in farm workers and applicators (p < .001) when compared to controls. 8-OH-dG levels were 8.5 times and 2.3 times higher in farm workers and applicators, respectively, than in controls. Serum MDA levels were 4.9 times and 24 times higher in farm workers and applicators, respectively, than in controls. DNA damage and oxidative DNA repair were significantly greater in lymphocytes from applicators and farm workers when compared with controls. A separate field study showed that DNA damage was also significantly greater (p < .001) in buccal cells (i.e., leukocytes) collected from migrant farm workers working with fungicides in the berry crops in Oregon. Markers of oxidative stress (i.e., reactive oxygen species, reduced levels of glutathione) and oxidative DNA damage were also observed in lymphocyte cell cultures treated with an OP. The findings from these in vivo and in vitro studies indicate that pesticides induce oxidative stress and DNA damage in agricultural workers. These biomarkers may be useful for increasing our understanding of the link between pesticides and cancer.

Symptoms of Gulf War Veterans Possibly Exposed to Organophosphate Chemical Warfare Agents at Khamisiyah, Iraq

Abstract During the 1991 Gulf War, some Allied troops were potentially exposed to satin/cyclosarin as the result the destruction of the destruction of Iraqi munitions at Khamisiyah. To evaluate the prevalence of past and current symptoms known to be associated with exposure to these chemical warfare agents; the authors conducted a computer-assisted telephone survey of 2,918 U.S, Gulf War veterans, Veterans who had participated in or witnessed the demolition in 1991 were more likely to report historical or extant symptoms than were veterans from other military units. These results should be viewed cautiously because they are based on symptoms recalled nine years after the event without precise characterization of exposure. Nonetheless, the findings suggest that symptoms consistent with low-level sarin exposure may have initially occurred, and health effects may have per-sisted in the veterans who were nearest to the demolition activity. Further research is warranted.

Illness experience of Gulf War veterans possibly exposed to chemical warfare agents.

BACKGROUND During the 1991 Gulf War, some Allied troops were potentially exposed to chemical warfare agents as the result of the detonation of Iraqi munitions at Khamisiyah. METHODS In 1999, we conducted a computer-assisted telephone survey of 2918 Gulf War veterans from Oregon, Washington, California, North Carolina, and Georgia to evaluate the prevalence of self-reported medical diagnoses and hospitalizations among this potentially exposed population and among comparison groups of veterans deployed and nondeployed to the Southwest Asia theater of operations. RESULTS Troops reported to be within 50 kilometers of the Khamisiyah site did not differ from other deployed troops on reports of any medical conditions or hospitalizations in the 9 years following the Gulf War. Hospitalization rates among deployed and nondeployed troops did not differ. Deployed troops were significantly more likely to report diagnoses of high blood pressure (odds ratio [OR]=1.7); heart disease (OR=2.5); slipped disk or pinched nerve (OR=1.5); post-traumatic stress disorder (OR=14.9); hospitalization for depression (OR=5.1); and periodontal disease (OR=1.8) when compared to nondeployed troops. There was a trend for deployed veterans to report more diagnoses of any cancer (OR=3.0). CONCLUSIONS These findings do not provide evidence of any long-term health effect associated with exposure to the detonation of chemical warfare agents, but support findings from other investigations of increased morbidity among deployed troops. The prevalence of cancer among this population of deployed troops merits ongoing attention.

Two resting potential levels regulated by the inward- rectifier potassium channel in the guinea-pig spiral modiolar artery

1 Intracellular in vitro recordings were made from 771 cells from the spiral modiolar artery (SMA). The initial resting potentials (RPs) displayed a bimodal distribution that was well modelled as a mixture of two Gaussian distributions. About half of the cells had an average RP of −74 mV, and were termed high‐RP cells, whereas the other half had an average RP around −41 mV, and were termed low‐RP cells. Preparations that were incubated for longer than 24 h contained significantly more high‐RP cells than those incubated for less than 8 h. 2 When labelled with the fluorescent dye propidium iodide, 68 and 36 cells were identified as smooth muscle cells (SMC) and endothelial cells (EC), respectively. The RP and input resistance were not significantly different between these two types of cell. Dye coupling was observed only in ECs. Dual cell recordings with 0.2–1.0 mm separation demonstrated the simultaneous existence of high‐ and low‐RP cells and a heterogeneous low‐strength electrical coupling. 3 The high‐RP cells were depolarized by ACh and by high extracellular potassium concentration (high K+). The low‐RP cells were usually hyperpolarized by moderately high K+ (7.5–20 mm) and by ACh. The high K+‐induced hyperpolarization was suppressed by barium (Ba2+, 10–50 μm). The putative gap junction blocker 18β‐glycyrrhetinic acid suppressed the ACh‐induced responses in SMCs, but not in ECs. 4 Low‐RP cells could rapidly shift the membrane potential to a permanent high‐RP state spontaneously or, more often, after a brief application of hyperpolarizing agents including high K+, ACh, nitric oxide and pinacidil. Once shifted to a high‐RP state, the responses of these cells to high K+ and ACh became similar to those of the original high‐RP cells. 5 High‐RP cells occasionally shifted their potentials to a low‐RP state either spontaneously or after a brief application of 10‐50 μm Ba2+ or 100 μm ouabain. Once shifted to the low‐RP state, the response of these cells to high K+ and ACh became a hyperpolarization. The shift between high‐ and low‐RP states was largely mimicked by wash‐in and wash‐out of low concentrations of Ba2+. The shift often showed a regenerative process as a fast phase in its middle course. 6 It is concluded that the cochlear SMA in vitro is composed of poorly and heterogeneously coupled SMCs and ECs, simultaneously resting in one of two distinct states, one a high‐RP state and the other a low‐RP state. The two RP states are exchangeable mainly due to all‐or‐none‐like conductance changes of the inward‐rectifier K+ channel.

Associations between race and ethnicity and treatment for chronic pain in the VA.

UNLABELLED The purpose of this study was to identify racial and ethnic differences in patient-reported rates of treatment for chronic pain and ratings of pain-treatment effectiveness among veterans treated in Veterans Affairs (VA) facilities. This was a cross-sectional analysis of data from 255,522 veterans who participated in the VA Survey of the Healthcare Experiences of Patients (SHEP) in Fiscal Year 2005. Measures included demographics, the Veterans Rand Health Survey-12, a single item inquiring if the patient received treatment for chronic pain in the VA within the prior 12 months, and a single item asking the patient to rate the effectiveness of chronic pain care. In a logistic model adjusting for demographics, pain interference, and mental health status, male and female veterans who were Hispanic (OR 1.39 [95%CI 1.26-1.53] and OR 1.57 [1.02-2.43], respectively) or non Hispanic black (OR 1.43 [1.33-1.54] and OR 1.35 [1.02-1.78], respectively) were more likely to report receiving treatment for chronic pain in the prior 12 months compared to non Hispanic white veterans. Among veterans who reported receiving treatment for chronic pain, non Hispanic black men were less likely to rate pain-treatment effectiveness as very good or excellent, compared to non Hispanic white men (OR .809 [.720-.910]). PERSPECTIVE In our study, Hispanic and non Hispanic black veterans reported receiving chronic pain treatment more frequently than white veterans. Among veterans reporting pain treatment, non Hispanic black men were somewhat less likely to report receiving highly effective treatment than white men. Further research is needed to understand the reasons for these differences and their potential clinical implications.

Self-reported exposures and their association with unexplained illness in a population-based case-control study of Gulf War veterans.

Many factors have been considered as possible causes of the unexplained illness reported by veterans of the Gulf War (GW). In this study, we report an analysis of risk factors and unexplained illness in a population-based sample of GW veterans who underwent clinical evaluation. Multiple risk factors were compared in 241 veterans who met criteria for unexplained illness and 113 healthy controls. Results suggest that GW unexplained illness is most highly associated with combat conditions, heat stress, and having sought medical attention during the GW. When controlling for multiple simultaneous exposures during the GW, interactions around pyridostigmine bromide, insecticides and repellents, and stress were not significant. These results indicate that most unexplained illness in GW veterans cannot be explained by neurotoxic effects of exposures to chemicals that inhibit cholinesterase activity.

The Cycad Genotoxin MAM Modulates Brain Cellular Pathways Involved in Neurodegenerative Disease and Cancer in a DNA Damage-Linked Manner

Methylazoxymethanol (MAM), the genotoxic metabolite of the cycad azoxyglucoside cycasin, induces genetic alterations in bacteria, yeast, plants, insects and mammalian cells, but adult nerve cells are thought to be unaffected. We show that the brains of adult C57BL6 wild-type mice treated with a single systemic dose of MAM acetate display DNA damage (O 6-methyldeoxyguanosine lesions, O 6-mG) that remains constant up to 7 days post-treatment. By contrast, MAM-treated mice lacking a functional gene encoding the DNA repair enzyme O 6-mG DNA methyltransferase (MGMT) showed elevated O 6-mG DNA damage starting at 48 hours post-treatment. The DNA damage was linked to changes in the expression of genes in cell-signaling pathways associated with cancer, human neurodegenerative disease, and neurodevelopmental disorders. These data are consistent with the established developmental neurotoxic and carcinogenic properties of MAM in rodents. They also support the hypothesis that early-life exposure to MAM-glucoside (cycasin) has an etiological association with a declining, prototypical neurodegenerative disease seen in Guam, Japan, and New Guinea populations that formerly used the neurotoxic cycad plant for food or medicine, or both. These findings suggest environmental genotoxins, specifically MAM, target common pathways involved in neurodegeneration and cancer, the outcome depending on whether the cell can divide (cancer) or not (neurodegeneration). Exposure to MAM-related environmental genotoxins may have relevance to the etiology of related tauopathies, notably, Alzheimers disease.