Grantee Research Project Results
2001 Progress Report: Biomarkers and Neurobehavioral Effects of Perinatal Exposure to Chlorpyrifos and Other Organophosphate Insecticides
EPA Grant Number: R828611Title: Biomarkers and Neurobehavioral Effects of Perinatal Exposure to Chlorpyrifos and Other Organophosphate Insecticides
Investigators: Wilkins, John R. , Moeschberger, Melvin L. , Lindsay, Ronald L. , Weghorst, Christopher M. , Dietrich, Kim
Current Investigators: Wilkins, John R. , Moeschberger, Melvin L. , Weghorst, Christopher M. , Dietrich, Kim , Nishioka, M.
Institution: The Ohio State University , University of Cincinnati
Current Institution: The Ohio State University , Battelle Memorial Institute , University of Cincinnati
EPA Project Officer: Hahn, Intaek
Project Period: February 12, 2001 through February 11, 2004 (Extended to February 11, 2006)
Project Period Covered by this Report: February 12, 2001 through February 11, 2002
Project Amount: $1,126,463
RFA: Biomarkers for the Assessment of Exposure and Toxicity in Children (2000) RFA Text | Recipients Lists
Research Category: Children's Health , Human Health
Objective:
The objective of this research project is to employ biomarkers of exposure and susceptibility in a longitudinal design. The proposed research will evaluate the putative relationship between perinatal exposure to chlorpyrifos (CP) and other organophosphate (OP) insecticides, and adverse neurobehavioral effects among infants and young children. Cohort ascertainment will involve recruitment of 176 women in their second trimester of low-risk pregnancy into the longitudinal study, which will follow only healthy full-term newborns for 2 years.
Progress Summary:
To date, study protocols and questionnaires have been written and piloted, and 23 women have been recruited into the study; all have been willing to provide the necessary blood and urine specimens. Fourteen of these women were approached in one of the Neighborhood Health Clinics/Oregon State University Clinic; five were referrals from a local OB/GYN practitioner, three from flyers that were posted in different daycare centers around the Columbus metro area, and one woman was a referral by word of mouth. No results are available for the urine specimens, but they have been aliquotted and frozen for future batch analysis. All blood samples have been analyzed for Pb and ZnPP. Blood specimens have been centrifuged, aliquotted, and frozen. Five of the recruited mothers have delivered their infants.
Data Collection Protocols: Maternal Biological Specimens
Urine (needed for Cotinine, TCP/IMP, DAP). Two overnight and morning voids are required of participants (overnight and morning samples go into the same container). Subjects will be given the collection equipment (collection "hats" and two 500 mL containers) after signing the consent form at the time of recruitment. Subjects will collect the sample during the night and the next morning, after being recruited. The participant will be asked to store the urine in her own refrigerator in biohazard bags until the specimen can be picked up by project personnel. The default plan is for project personnel to call the subject and then pick up specimens the next morning by 9:00 a.m. If the participant cannot be reached (or she cannot accommodate the protocol), other arrangements will be made. Project personnel will have a cooler with two blue ice bags for keeping specimens cool. Urines will be taken to Marcia Nishioka's laboratory at Battelle, where her staff will aliquot, measure, and freeze the specimens for subsequent analyses.
Cotinine Testing Protocol. After bringing urine samples to room temperature, the following is performed: gloves are put on, specimen container is opened, NicoMeter strip package is opened, held by the handle, and dipped up to maximum sample line for 5 seconds, then the strip is placed horizontally over the specimen container for 15 minutes. Results are read as lowest number with reddish purple color (of any shade or intensity). The color must appear in at least one of the zones or the test is nonfunctional and should be repeated with a new strip. The container is labeled "Wilkins" and the participant's ID number, then the container is placed back in the cooler and taken to Room 1120 in the James Cancer Hospital, and placed in a freezer with other samples. Cotinine testing on maternal urine will be performed by personnel in Marcia Nishioka's laboratory at Battelle. The same person should perform all cotinine tests to minimize inter-rater variability.
Blood (Pb/ZnPP, PON1). Two 3 mL tubes are required: Lithium heparin tube for Pb/ZnPP and sodium heparin tube for paraoxonase (PON1). Sodium heparin tubes will be kept in the cooler after drawing and then will be taken to Chris Weghorst's laboratory on the 11th floor in the James Cancer Hospital.
PON1 Blood Handling. Five tubes will be assembled as described below and labeled "Wilkins," subject ID, and date of draw: gloves are put on, hood light is turned on, surface under hood is wiped down with 70 percent ethanol, sodium heparin tube is placed in a rack under the hood, extra green tube (2 mL) is moved from the refrigerator (contains solution for an additional analysis) to the rack under hood, place two empty green tubes (2 mL) in the rack under hood, place two empty clear nalgene tubes (2 mL) to rack under hood, remove green top of tube, use 1 mL hand pipette to aliquot 500 µL into extra tube (discard pipette in "glass disposal" container to left of hood), use 2 mL pipette to aliquot 1 mL into each of the two empty green tubes. Place the two empty green tubes in the Elementary-Body Agglutination (EBA) 12 centrifuge so that the tubes are oriented symmetrically (cap hinges are both oriented in an upward direction), set centrifuge for RCF of 12,500 (G force as opposed to RPM), set timer for 10 minutes, and press start. After centrifuge stops, place tubes in rack under hood, use 1 mL hand pipette to aliquot approximately 400 µL into clear nalgene tubes. Inspect for turbidity, especially for red cells. If it appears red cells were pipetted, recombine plasma and cells, centrifuge again and aliquot plasma again with fresh nalgene tubes. Place extra tube back in rack in refrigerator and place four remaining tubes in appropriate rack in freezer. Wipe down hood surface with germicidal solution and then 70 percent ethanol.
Lead Blood Handling. Lithium heparin tubes will be taken to the Children's Hospital Outpatient Laboratory. Dr. Thornton is coordinating this effort. Pre-printed requisition forms (including name, address, age, sex, and race items) will be filled out and specimens will be dropped off at the desk. The Lithium heparin blood must be well mixed (gently turn upside down and back upright 10-15 times) and cooled. We will avoid freezing and vigorous shaking of the sample. Attempts will be made to ensure samples will be drawn coincident with other laboratory tests required during pregnancy. If necessary, project personnel will draw the specimens.
Data Collection Protocols: Infant Biological Specimens
(1) Urine (0-3 months for TCP/IMP, DAP).
(2) Urine (every 7 months from 3 to 24 months for TCP/IMP, DAP). Mothers will collect urine by using protocols and collection kits supplied by us. Collection kits contain simple directions, a form or sticker on which to note what time the child was put to bed and when diaper was changed, and a prelabeled zip-seal bag with the child's ID number for the wet diaper. Mothers will send in samples by using protocols and mailing kits supplied by us. Mailing kits consist of a plastic zip-seal biohazard bag to hold the zip-seal bag containing the diaper, a bubble wrap padded self-seal envelope to serve as a tertiary container, and a pre-addressed postage-paid fiberboard outer box. The box will be marked with a biohazard label with "Urine Sample" clearly marked. Mothers will send diapers directly to Battelle Memorial Institute.
(3) Blood (at 12 months for Pb and PON1), same protocol as above.
(4) Blood (at 24 months for Pb) ), same protocol as above.
Maternal Covariate Factors
(1) Socioeconomic status (income, education)-Hollingshead Four-Factor Index of Social Status.
(2) Family structure.
(3) Parental intelligence-WASI.
(4) Quality of care taking environment-HSQ.
(5) Parental stress-PSI.
(6) Clinical birth data (birth weight, gestational maturity, evidence of fetal distress)
(7) Other co-factors-ethnicity, gender, birth order, number of children in home, alcohol, tobacco and other drugs.
Infant Outcome Variables
(1) BSID-II (Mental Development Index, Psychomotor Development Index, Behavior Rating Scale).
(2) Child Development Index-CDI.
Future Activities:
The next 6 months will be devoted to aggressively recruiting women into the study and obtaining necessary biological specimens and questionnaire information. As infants are born, the effort will shift to receipt of biological specimens from infants, as well as neurobehavioral assessments of infants. Analyses for insecticide metabolites will proceed as more samples are obtained.
Journal Articles:
No journal articles submitted with this report: View all 2 publications for this projectSupplemental Keywords:
children, biomarkers, exposure, susceptibility, organophosphate, chloropyrifos, insecticide, neurobehavioral effects, perinatal exposure., RFA, Scientific Discipline, Health, Toxicology, Genetics, Health Risk Assessment, Susceptibility/Sensitive Population/Genetic Susceptibility, Biochemistry, Children's Health, Molecular Biology/Genetics, genetic susceptability, health effects, sensitive populations, infants, vulnerability, biomarkers, adolescence, health risks, chlorpyrifos, measuring childhood exposure, exposure, neurotoxicity, longitudinal study, endocrine disruptors, children, children's vulnerablity, assessment of exposure, insecticides, neurobehavioral effects, perinatanl exposure, biological markers, developmental disorders, genetic susceptibility, organophosphate pesticides, environmental hazard exposuresRelevant Websites:
Progress and Final Reports:
Original AbstractThe perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Conclusions drawn by the principal investigators have not been reviewed by the Agency.