Mr. David
R.
Schwoegler
Public Affairs Office
7000 East Avenue, L-797
Livermore, CA 94551
Phone: 925-422-6900
Fax: 925-424-2780
E-mail: newsguy@llnl.gov
Number of Human Subjects projects reported: 58
| LLNL-94-105 | "Investigation of the Relationship between Numerical Chromosomal Aneuploidy in Sperm and Offspring" |
| LLNL-95-126 | "Influence of Heterocyclic Amine Metabolic Polymorphisms on Bioactivation of PhIP in the Human Colon" |
| LLNL-96-109 | "Age Effects on the Incidence of Genetic and Physiological Defects in Human Sperm" |
| LLNL-98-106 | "Melanoma and Other Mortality Rates in LLNL Employees" |
| LLNL-99-121 | "Improved Measurement of Cholinesterase Inhibition" |
| LLNL-00-113 | "Use of Chelating Agents in Radiation Accidents" |
| LLNL-01-101 | "Background Uranium in Urine at LLNL" |
| LLNL-01-116 | "Biomechanics of Human Dentin" |
| LLNL-01-119 | "Changes in Diffraction-limited Spatial Vision Associated with Aging and Retinal Disease" |
| LLNL-02-101 | "Determining the Carcinogenic Significance of Heterocyclic Amines" |
| LLNL-02-106 | "The effect of daily soft-drink consumption on human bone resorption" |
| LLNL-02-114 | "Study of the association of DNA damage with cancer risk" |
| LLNL-02-119 | "Vitamin B12 absorption and metabolism in humans" |
| LLNL-02-124 | "Identification of markers of human exposure to biological agents, LLNL volunteers" |
| LLNL-02-126 | "Identification of markers of human exposure to biological agents. Hemodialysis patients" |
| LLNL-03-103 | "Identification and Development of Biological Markers of Human Exposure to the Insecticide Permethrin" |
| LLNL-03-106 | "Identification of markers of human exposure to biolgoical agents: Vaccinia study" |
| LLNL-03-109 | "Does Tamoxifen Cause DNA Damage in Human Colon?" |
| LLNL-03-113 | "Host Pathogen Interactions: Biomarkers for Early Detection" |
| LLNL-03-114 | "Monitoring Bone Resorption Using 41Ca and Accelerator Mass Spectrometry" |
| LLNL-03-115 | "The effect of dietary protein on calcium metabolism" |
| LLNL-03-118 | "Quantifying the impact of diet on carcinogen exposure" |
| LLNL-03-126 | "Molecular modulation of calcium crystallization" |
| LLNL-04-113 | "Genetic Effects of Benzene Exposure in Human Sperm: A Study of Chinese Factory Workers" |
| LLNL-04-115 | "Monitoring Bone Resorption with Sweat Patches" |
| LLNL-04-116 | "Detecting Chemotherapeutics in Buccal Cells using Time-of-Flight Secondary Ion Mass Spectrometry" |
| LLNL-04-117 | "Determining the Effect of Green Tea on Gene Expression in Buccal, Bladder, and Blood Cells" |
| LLNL-04-118 | "Gene Expression Studies in Human Blood and Saliva" |
| LLNL-04-119 | "Anonymous Semen Donor Program" |
| LLNL-04-121 | "Environmental Epidemiology of Essential Tremor" |
| LLNL-04-123 | "Evaluation and Validation of 41Ca as a Novel Isotopic Technique in Bone Research" |
| LLNL-05-104 | "Prostate cancer screening and dietary HA exposure in African-Americans: Phase II" |
| LLNL-05-109 | "Test Bed Deployment of SIRAD" |
| LLNL-05-113 | "ToF-SIMS analysis of cancer tissue" |
| LLNL-05-115 | "Identifying Metastatic Breast Cells from Peripheral Blood" |
| LLNL-05-118 | "Buccal Mucosa Cell Sampling in Head and Neck Radiotherapy for Molecular Biodosimetry" |
| LLNL-05-121 | "Medical Surveillance for Former Department of Energy Workers" |
| LLNL-05-122 | "41Ca Absorption and Distribution in a Total Hip Replacement Patient" |
| LLNL-05-124 | "The Effects of Nickel (II) on Human Sperm DNA Integrity" |
| LLNL-05-UCM-05-001 | "Systems of Election, Latino Representation, and Student Outcomes in California Schools" |
| LLNL-05-UCM-05-002 | "Implementing Student Excellence - A Unique Opportunity" |
| LLNL-06-101 | "Chlorophylls as Trans-species Blocking Agents" |
| LLNL-06-102 | "Detecting and Monitoring Prostate Cancer Metastasis Using 41Ca AMS" |
| LLNL-06-105 | "A Multiplexed Diagnostic Platform for Point-of-Care Pathogen Detection" |
| LLNL-06-110 | "Metalloproteomics of the Human Blood Cell" |
| LLNL-06-113 | "The Effect of Surveillance on the Thickness of LLNL Melanoma" |
| LLNL-06-117 | "The Clinical Use of 41Ca as a Novel Isotopic Assay for High Precision Non-Invasive Assessment of Metastatic Bone Disease, Therapeutic Response and Disease Progression in Breast Cancer Patients" |
| LLNL-06-121 | "DNA Repair Phenotype and Testicular Cancer Type" |
| LLNL-06-123 | "Leading Innovation: A Case Study of Sustained Innovation" |
| LLNL-06-UCM-05-005 | "Effectiveness of Success Workshops on First-Year Student Academic Performance" |
| LLNL-06-UCM-05-006 | "Evaluation of Adipose Hormones, Serum Cytokine Levels, Serum Oxidative Stress Markers and Presence or Absence of NASH in Morbidly Obese Subjects Undergoing Bariatric Surgery" |
| LLNL-06-UCM-05-008 | "National Survey of Student Engagement (NSSE)" |
| LLNL-06-UCM-06-002 | "Creating a Community College Transfer Culture at UC Merced" |
| LLNL-06-UCM-06-003 | "Engineering Innovation - Measuring the Impacts" |
| LLNL-06-UCM-06-006 | "Genetic Polymorphism of Glutamate Cysteine Ligase in Idiopathic Pulmonary Fibrosis " |
| LLNL-06-UCM-06-007 | "Children's Questions: Insight into Cognitive Development" |
| LLNL-06-UCM-06-008 | "Animals Are Like Humans: How Parents Help Children Understand Biology." |
| LLNL-06-UCM-06-110 | "Animals vs. Inanimate Objects: How Do Parent/Child Conversations Differ?" |
"Investigation of the Relationship between Numerical Chromosomal Aneuploidy in Sperm and Offspring"
Principal Investigator: Dr. Andrew J. Wyrobek, Lawrence Livermore National Laboratory
Project started in: 1994
This project ended in fiscal year 2006.
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 105
Institutional Review Board (IRB) Review:
Type of Review:
Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 07/27/05
IRB approval number: 94-105
Explanation of IRB approval:
PROTOCOL IS CLOSED.
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 0
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
The specific aim of this study has been to determine whether fathers of children with Klinefelter Syndrome (KS; 47, XXY) who are known to have contributed the extra X chromosome have elevated frequencies of sperm bearing abnormal number of chromosomes (aneuploidy) compared to fathers of KS children where the extra X chromosome is known to be of maternal origin. The study population included index cases and the mothers and fathers of the index cases. In this study, the index cases consisted of boys with Klinefelter Syndrome who were six years old or younger at the time of recruitment.
Animal and human evidence indicates that: a) sperm can carry gene mutations or chromosomal abnormalities; b) genetically defective sperm can fertilize eggs; and c) an embryo's ability to survive through development to birth and beyond, depends on the specific chromosomal defect it carries.
Hypothesis:
Fathers of children with KS when the extra X chromosome was paternally-derived have higher levels of sperm with chromosomal abnormalities than fathers of children with KS where the extra X chromosome was maternally derived.
Experimental Design:
Research conducted at Lawrence Livermore National Laboratory (LLNL) included (a) determination of the parent of origin of the extra X chromosome and (b) determination of the level of sperm aneuploidy in the father of affected children. At this time no additional laboratory research is being conducted. All samples have been collected, and only data analysis and manuscript preparation are ongoing.
A total of 38 families with a Klinefelter Syndrome (XXY) child were recruited. The blood samples from father, mother, and child plus father's semen were collected and shipped to LLNL. We determined the parental origin of the extra X chromosome of each boy for all the families by polymerase chain reaction (PCR) and sequencing using a polymorphic X-linked microsatellite deoxyribonucleic acid (DNA) marker. Inheritance was paternal in 10 families and maternal in 26. Two families were not informative. The frequencies of aneuploid sperm of each father for all the families were determined by fluorescence in situ hybridization (FISH) using DNA probes specific for chromosomes X, Y, and 21.
For the multiple aneuploidy study, we evaluated the fourth family recruited in the main study. This family had three aneuploid pregnancies prior to the index case. We established that the extra X chromosome of the index child was inherited from the father and that the father produces more aneuploid sperm than we have ever seen in any man including heavy smokers and men who have received cancer chemotherapy. Previously, we obtained tissue blocks from the aneuploid pregnancies that died before birth (trisomy 15 and 22) and determined that the extra chromosomes were also paternal in origin. Using an aliquot of semen, we also determined that the aneuploidy frequency was also elevated for these chromosomes. In addition, we obtained a second semen sample from the father and determined that the aneuploid sperm were elevated persistently over a two-year period. The protocol and reporting procedures for these families was the same as for those in the main study.
Human Subjects Involvement:
According to our protocol, we requested blood samples from the mother, father, and index child. We requested a semen specimen from the father and arranged for telephone interviews. The findings of the family's blood and semen analysis are shared with the family if they are interested and request this in writing. A genetic counselor discussed with each of the parents whether they wanted both individual and study results or just the latter. Findings were conveyed by the study's genetic counselor. If requested in writing by the family, specific findings will be given via mail or telephone to the family's physician. Potential risks to privacy have been addressed.
Anticipated Results:
Children with Klinefelter Syndrome (XXY) and their fathers provide a unique opportunity to characterize the relationship between sperm aneuploidy and aneuploidy at birth. We determined that fathers of children with paternally derived Klinefelter Syndrome produced inherently elevated levels of aneuploid sperm. Data analyses and publication of results are expected to be completed during the next year.
The project has found an age-effect on the frequency of sperm carrying a wrong number of chromosomes (aneuploidy), specifically of the X-Y type.
"Influence of Heterocyclic Amine Metabolic Polymorphisms on Bioactivation of PhIP in the Human Colon"
Principal Investigator: Dr. Paul T. Henderson, Lawrence Livermore National Laboratory
Project started in: 1995
This project ended in fiscal year 2006.
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 126
Institutional Review Board (IRB) Review:
Type of Review:
Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 10/04/05
IRB approval number: 95-126
Explanation of IRB approval:
Continuing review eclipses fiscal year schedule.
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 0
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
Background :
Colon cancer is the second leading cancer killer in the United States. Although the causes remain largely unknown, dietary factors have been strongly implicated as a cause of this disease, and some evidence suggests an increased risk associated with the consumption of well-done cooked meat. The identification of mutagenic and carcinogenic heterocyclic amines (HCAs) in cooked meat has raised the possibility that these compounds may play a role in the development of cancer in humans. Of all the HCAs, 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) is considered to be a significant colon cancer risk factor because it is usually the most mass-abundant in cooked meat and causes colon tumors in rats. Consequently, the cancer risk associated with exposure to PhIP is a major public health issue.
Objectives:
The development of colon tumors in rats administered PhIP has been associated with the formation of large numbers of deoxyribonucleic acid (DNA) adducts in this organ. DNA adducts, which are a type of DNA damage, are the product of reactions between DNA and chemical carcinogens. If these adducts are not repaired, they may lead to changes in DNA and may ultimately result in tumor formation. Therefore, the presence of DNA adducts can give an indication of the carcinogenic potential of a compound. Since adducts are usually formed at very low levels, sensitive techniques must be employed for their detection. So far, the traditional methods have not proven sensitive enough to accurately determine if PhIP forms DNA adducts in the colon of people and the extent of interindividual variation. Our objectives are to: 1) Use PhIP that is labeled with carbon-14 and the sensitive technique of accelerator mass spectrometry (AMS) to measure adducts in human tissue samples and then to compare this with adduct levels detected in rats. This may give an indication of any potential risk of developing PhIP induced tumors in humans. 2) Identify the routes of PhIP metabolism (breakdown) in humans. 3) Determine if interindividual variation in the metabolism of PhIP in the body can influence the amount of PhIP that forms DNA adducts in the colon, which may help find a predictor of individual colon cancer risk.
Methodology:
This study is a collaboration between Lawrence Livermore National Laboratory (LLNL) and the University of Arkansas for Medical Sciences (UAMS), the J. L. McClellan Memorial Veterans Administration Medical Center (VAMC), and the Arkansas Cancer Research Center. In this study, volunteers undergoing colorectal surgery will be administered a single dose of PhIP that is labeled with carbon-14. Tissue samples removed during surgery will then be collected and analyzed by AMS. In addition, blood, urine, and feces will be collected and analyzed to determine how the individuals metabolise PhIP. Approximately six weeks after surgery, a simple urine test will be conducted in which the volunteers will be administered caffeine and their urine analyzed for caffeine breakdown products. Caffeine is used because it is broken down by the enzymes considered to be important in PhIP metabolism.
Involvement of Human Subjects:
Briefly, human subjects who have been previously diagnosed with colon cancer and who are scheduled for surgery at the UAMS University Hospital or the J. L. McClellan Memorial VAMC in Little Rock, Arkansas will be fully informed as to the nature of the study. Subjects will be asked to participate and those willing to take part will provide written informed consent. Volunteers will then be administered [14C]PhIP in a capsule. The dose will not represent a chemical risk to volunteers beyond that presented by a normal diet, since it is roughly equivalent to the exposure resulting from the consumption of up to 2.3 kg of chicken, beef or bacon, depending upon the cooking time and temperature. Furthermore, the radiation exposure represents less than one percent of an individual's annual radiation exposure from natural sources. Blood will be drawn (30 mls by vein puncture) at several time points up until surgery to determine the circulating levels of the compound and its metabolites. Additionally, urine and feces will be collected from approximately 12 hours prior to PhIP administration and for up to 96 hours after administration of the PhIP. Approximately six weeks after surgery, patients who participate in this study will be given 200 mg of caffeine and a urine sample collected. In order to protect the confidentiality of volunteers in the study, records will be maintained at the UAMS and VAMC. LLNL will receive only coded samples for analysis.
Outcome:
This study will: (1) determine if adducts are formed by PhIP in humans at dietary-relevant doses, (2) identify the routes of PhIP metabolism (breakdown) in humans, and (3) determine if interindividual variation in the metabolism of PhIP in the body can influence the amount of PhIP that forms DNA adducts in the colon. This pilot study will be completed once we have collected and analyzed samples from 10 volunteers.
Conclusion:
The results of this study will help determine if PhIP is a human colon cancer risk factor at dietary levels of exposure and may establish a predictor of colon cancer risk.
Subject participation has been completed (or the study does not involve subject contact). There is no ongoing contact with any subjects. Follow-up, including review of private records or mailings, will continue.
"Age Effects on the Incidence of Genetic and Physiological Defects in Human Sperm"
Principal Investigator: Dr. Andrew J. Wyrobek, Lawrence Livermore National Laboratory
Project started in: 1996
This project ended in fiscal year 2006.
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 109
Institutional Review Board (IRB) Review:
Type of Review:
Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 06/27/05
IRB approval number: 96-109
Explanation of IRB approval:
PROTOCOL IS CLOSED.
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 0
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
Although it is well accepted that maternal age can result in adverse consequences to the fetus, it remains unclear whether paternal age also influences fetal viability and pregnancy outcome. Evidence for male-mediated developmental toxicity derives from strong animal data that premating paternal exposures can lead to adverse developmental effects. In addition, there is growing epidemiological evidence that exposures of fathers to environmental toxicants are associated with adverse consequences to the fetus. However, the underlying mechanism for the effects of paternal exposure remain unresolved and is likely to include genetic defects transmitted by sperm.
The hypotheses for this study are to: (1) determine whether there is an effect of a man's age on the types and proportions of genetic damage in sperm measured by various methods and semen quality and (2) examine whether certain diets are associated with higher rates of genetic damage and decreased semen quality.
This research will provide fundamental information on the effects of paternal age and diet on genetic damage to human sperm. These findings will also provide critical data needed for the design and interpretation of studies of paternal effects of exposure to environmental agents.
The sample collection phase is complete. The following laboratory analyses were performed: sperm concentration, visual motility, and computer assisted sperm analysis (CASA) motility; nuclear morphometry assay; sperm chromatin assay, sperm chromosomal aneuploidy, and aberration assay. The following analyses are in progress or will be conducted next year: mutation frequency at various specific genes and micronutrient analyses.
The recruitment phase is completed, the specimens are under laboratory analysis, and the resulting data are in statistical analysis. This research will provide fundamental information on the effects of paternal age and diet on genetic damage to human sperm. These findings will also provide critical data needed for the design and interpretation of studies of paternal effects of exposure to environmental agents.
There are no further procedures involving direct contact with human subjects. Further activity is limited to the analysis of archived samples. All information gathered is treated with respect and confidentiality to protect the privacy of each subject.
"Melanoma and Other Mortality Rates in LLNL Employees"
Principal Investigator: Dr. Mortimer L. Mendelsohn, Lawrence Livermore National Laboratory
Project started in: 1998
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 106
Institutional Review Board (IRB) Review:
Type of Review:
Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 03/13/06
IRB approval number: 98-106
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 0
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
The primary purpose of this study is to follow mortality rates in the Lawrence Livermore National Laboratory (LLNL) population as an indicator of occupational and environmental exposure. Our concern began in the early 1980s when the laboratory experienced a highly significant cluster of melanoma cases. We responded in 1984 with a vigorous melanoma campaign including an aggressively active melanoma clinic, which in turn led to the detection of high levels of early melanomas and zero melanoma mortality from 1984 to 1996. General mortality (i.e., all causes of death) of LLNL employees for this same time interval was 46 percent of U.S. levels, while cancer mortality was 73 percent and cardiovascular mortality was 42 percent. Such data can only be obtained with human studies.
Hypotheses concerned the reality and possible causes of the melanoma cluster and later its disappearance. With regard to mortality in general, we are evaluating what appears to be a very strong and increasing healthy worker effect in association with low tobacco usage, extending well into retirement.
We are using conventional methodology and standard U.S. statistics to evaluate the overall and specific causes of mortality. The relevant census data are collected by laboratory administration and consist of name, social security number (SSN), birth date, and hire and termination dates for all LLNL employees. Periodically (roughly in 10 to 20 year intervals), the data are assembled and submitted to the National Death Index (NDI) where deaths are identified throughout the U.S., and diagnoses are provided to us.
The study is designed so that we have no interaction with the employee, their family, or their medical records. The NDI returns all the identifying information to us soon after relevant deaths are found. We maintain the identifiers in data files kept under lock and key. The identifying data are never transmitted electronically. All our analyses and public data are given as rates, without any identifiers. By definition there is no risk to the dead and a miniscule risk of disclosure to the living due to possible mishandling of the data. The reassurance to the staff about the remarkably high level of their survival has been very beneficial, as might be any possible future detection of unexpected mortality.
Further details on the data collected and its handling:
The research requires periodic mortality studies of the LLNL workforce using the NDI. The crux of this system is the use of sufficient personal identifiers to determine whether an individual is dead or alive. Typically this includes full name, SSN, birth date, state of residence, gender, and race. Death certificates with correspondence in all categories have a 97 percent chance of being correctly attributable to the given individual.
This information was in the past and will be in the future assembled by LLNL's Applied Information System. It covers all laboratory employees in each calendar year of the study (typically five years or more) and is provided on a compact disc (CD) handed to the pricipal investigator (PI). The data are entered into the PI's digital computer, and the CD is kept under lock and key. The computer resides in a locked single-occupant office and is password protected. When necessary, for checking and computational advancements, the data are shared with the collaborator, Dan Moore, who also keeps it on a password protected computer in an adjoining locked office. Periodically, when a mortality study is done, the subset of data representing ex-employees (deaths, transfers, retirees, etc.) are digitally coded and sent by mail to the NDI, U.S. Department of Health and Human Services, Centers for Disease Control and Prevention, National Center for Health Statistics, Hyattsville, Maryland. The NDI searches its registries for matches and mails back positive results with details about the cause of death and the quality of the match. They take scrupulous care of the data and return the files by mail when the study is completed. They also require that we do not use local medical records or family contacts to elaborate or pursue causes of death. We in turn guard the data carefully, as described above, and use it only for general statements about causes of death, i.e., only for rates for a particular disease or category and never for individuals.
We keep the collected identifiers on employees and ex-employees, since the search for death of an employee continues indefinitely with each year of the study, that terminates only when the study ends or the individual dies. The first 13 years of the study have netted 460 deaths among 14,492 persons. If and when the study terminates, the computer files will be erased and the CDs will be destroyed.
We give our full assurance that we have not and will not disclose to the PI any of our subjects, except that required to determine death and cause of death through the NDI.
We have shown a striking absence of melanoma mortality at LLNL for the period from 1984 to 1996, corresponding to the operation of the melanoma clinic. This is associated with a dramatic reduction in invasive melanoma and a brisk increase in early, in situ melanoma. Meanwhile we are in a holding period, waiting for sufficient time to make yet another mortality study a reasonably efficient effort (i.e., until 2008 at the earliest). However, recent budget restrictions have led to the decision to close the melanoma clinic in April 2006. Employees will be encouraged to continue surveillance using their own traditional health caregivers. It is unclear at this time how termination of the clinic will affect either future mortality studies or future melanoma risk to our population. The laboratory has access to two endpoints to study the risk: mortality and cancer incidence. The latter would have more sensitivity but is administratively more difficult to do. In the interim, the prudent thing to do is to preserve the necessary databases. For mortality, this means to continue this IRB-approved project.
"Improved Measurement of Cholinesterase Inhibition"
Principal Investigator: Dr. Garrett A. Keating, Lawrence Livermore National Laboratory
Project started in: 1999
This project ended in fiscal year 2006.
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 121
Institutional Review Board (IRB) Review:
Type of Review:
Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 06/09/05
IRB approval number: 99-121
Explanation of IRB approval:
PROTOCOL IS CLOSED.
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 0
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
Cholinesterase (ChE) inhibition is a consequence of exposure to certain chemical warfare agents and organophosphate (OP) pesticides. Current assessment of ChE inhibition involves the measurement of ChE activity in blood with a colorimetric assay. The percent of ChE inhibition is calculated by subtracting the measured activity from the background or normal level of activity which is usually based on a population average. Interindividual variability in normal ChE levels of 30 percent has been observed in the population so inhibition calculations based on a population average can misrepresent the degree of inhibition in the individual. The purpose of this study is to develop a method to determine the normal level of ChE activity from an inhibited blood sample. Due to differences in the levels and types of ChE between laboratory animals and humans, blood from human subjects is required so that methods developed by this study can be applicable to clinical ChE determinations.
The study is testing whether the ChE activity of an OP-inhibited blood sample can be reactivated so that the normal ChE level can be determined. This involves comparison of ChE activity in the reactivated blood samples with that from normal, untreated blood samples.
Whole blood samples will be serially inhibited with an OP pesticide and inhibition measured with a standard colorimetric assay. Free enzyme will then be inhibited with a radioactive probe and the sample subjected to chemical treatment to reactivate the OP-inhibited enzyme. This activity will then be assayed colorimetrically and with the radioactive probe. The objective of the research is to validate the reactivation methodology so that it can be performed without radioactive probes.
Human blood must be used to perfect the methodology for use with existing clinical protocols for measuring cholinesterase inhibition in patients. Subjects will be solicited by a memo describing the research and involvement of subjects distributed to their mail box. Subjects who agree to participate will meet with the principal investigator and will be provided with a description of the study and the IRB Bill of Rights. Subjects will be asked to schedule an appointment with Health Services to provide the blood sample. Subjects will undergo a standard blood draw to provide 5 ml of blood.
Reactivation of OP-inhibited ChE has been shown after prolonged chemical treatment (several hours) although the level of reactivation has not been complete. This study will combine several reactivation chemistries in an attempt to improve the level of reactivation. Reactivation of enzyme to a level of 90 percent of normal will be required for the new assay to be applicable given the level of variability in the clinical ChE assay. Also, reproducibility of the reactivation assay must exceed 90 percent so that it can be confidently applied to blood samples from different individuals where normal ChE levels can vary by as much as 30 percent. The variability and reproducibility of the activation assay will be tested with normal, untreated blood from the subjects so that accurate estimates of these two parameters can be obtained. Blood from one subject was obtained and stored to test the cholinesterase assay being modified for the reactivation protocol. Results using human blood have been comparable to those obtained with pure cholinesterase and mouse blood. Human subjects will be enrolled and the reactivation protocol performed on human blood samples.
"Use of Chelating Agents in Radiation Accidents"
Principal Investigator: Dr. Richard B. Watts, Lawrence Livermore National Laboratory
Project started in: 2000
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 113
Institutional Review Board (IRB) Review:
Type of Review:
Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 05/26/06
IRB approval number: 00-113
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 0
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
Ca-DTPA and Zn-DTPA are calcium and zinc salts of diethylenetriaminepentaacetate (DTPA) that are administered intravenously and are used as chelating agents for plutonium and other transuranic elements such as americium, californium, and curium. These chelating agents have been in clinical use in major radiation accidents since 1958. They are regulated by the Food and Drug Administration (FDA) and were classified as investigational new drugs (INDs) until recently, when a New Drug Application (NDA) was issued. However, the current stocks of these drugs managed by DOE/Oak Ridge Associated Universities (ORAU) will continue to be managed under IND procedures. There is effectively no alternate to these drugs for chelation of transuranics.
An oral chelating agent, Prussian Blue, is used to treat internal contamination with thallium and cesium-137. Like DTPA, Prussian Blue recently changed status from IND to NDA but current stocks managed by DOE/ORAU will continue to be managed under IND procedures.
There is no hypotheses being tested as this is not a research activity in the strictest sense. Rather, the chelating agents are being maintained on IND status to ensure accurate reporting of possible contamination accidents and to continue to record any side effects. These chelating agents have been used for over 30 years in radiation accidents in the U.S. and worldwide, and it is well documented that the drugs are effective against transuranic contamination.
There is no experimental design associated with the administration of these chelating agents. Rather, the drugs would be administered during significant transuranic, cesium-137, or thallium exposures if there was a significant potential for internal contamination based on evaluation of the patient, the wound counts available, and the elements involved.
In the event of potential internal contamination, exposed workers would be taken to Health Services. If it was determined that a significant transuranic exposure had occurred, the risks and benefits of DTPA treatment would be explained to the worker(s). If the exposure is to cesium-137 or thallium, the risks and benefits of Prussian Blue would be explained to the worker(s). Workers are given the right to refuse treatment although, based on decades of clinical experience, chelation therapy has become a part of widely accepted medical practice in removing heavy metal, cesium, and thallium contamination from humans. If workers agree to being treated with a chelating agent, they are asked to review and sign the appropriate consent form. A baseline medical history, blood and urinalysis are performed prior to treatment. DTPA may be injected intravenously or inhaled. Depending on the level of contamination, treatment may require repeated doses over a period of up to several weeks or months. Initial dose is typically 1 gram, but may be as much as 1 gram 3 times the fist day. Prussian Blue is administered orally with dose ranges of 3 to 20 grams daily in divided doses, with the duration of treatment contingent on exposure dose estimates and excretion rates after initiation of treatment.
The risks involved in DTPA therapy are relatively few and include those inherent in starting intravenous access and drawing blood, electrolyte and blood component problems, and possible exacerbation of kidney or bone marrow disease. Prussian Blue is not absorbed from the gastrointestinal tract and has no known risks other than constipation. Privacy and confidentiality will be maintained to the extent possible. The medical records will be made available to the FDA and to Drs. Albert L. Wiley and Patrick C. Lowry, of the Oak Ridge Institute for Science and Education. Drs. Wiley and Lowry are responsible for reporting all uses of these agents to the FDA. General information relating to a worker's case may be used in professional medical literature, but the individual worker will not be identified in any way. Termination of treatment would be decided on a case by case basis after consultation with the Radiation Emergency Assistance Center/Training Site (REAC/TS) at ORAU and an internal radiation dosimetry expert at LLNL.
Internal dosimetrist:
Although DTPA is not approved by the FDA for uranium neptunium, they may be used for this purpose on an off-label basis after consultation with ORAU.
We expect to continue to monitor the efficacy and safety of these drugs on an as-needed basis.
"Background Uranium in Urine at LLNL"
Principal Investigator: Dr. Lori Johnson, Lawrence Livermore National Laboratory
Project started in: 2001
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 101
Institutional Review Board (IRB) Review:
Type of Review:
Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 08/17/06
IRB approval number: 01-101
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 5
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
Scientific context of the study:
Everyone has some level of naturally occurring uranium in their urine. The amount of uranium present in the urine varies greatly depending on diet, water consumption, where an individual lives, and on many other complex factors. The amount of uranium in an individual's urine can vary significantly over time, as eating habits and environmental uranium levels fluctuate. The Hazards Control Bioassay Laboratory analyzes hundreds of urine samples for uranium from Lawrence Livermore National Laboratory (LLNL) employees that work with uranium on a routine basis each year. This is performed in conjunction with the LLNL Internal Dosimetry Program and is a part of the routine occupational safety surveillance program. The goal of this study is to obtain a representative range of the LLNL population uranium in urine level over time to be used as a control when evaluating the occupational safety surveillance results of LLNL employees that work with uranium on a routine basis. Based on recent demographic information, LLNL uranium workers who participate in the occupational safety surveillance program are mostly male (greater than 85 percent) and more than half of them live in the Central Valley. This study will focus on (but not be restricted to) males who are not occupationally exposed to uranium, with an additional specific request for individuals who reside in the Central Valley.
Human subjects involvement:
It is not possible to construct a simulation model or animal study that incorporates all of the different variables involved in variation of uranium concentration in human urine. Uranium concentrations can vary greatly between individuals; diet, age, gender, metabolism, medication, water and beverage consumption (type and amount), smoking, organic food consumption, vitamin supplements, home and work locations, and many other factors can affect uranium concentration in urine. In particular, several foods and consumer products, such as beets and other root vegetables, shrimp and bottom feeding fish, cigarettes, etc., are known to contain more uranium than meat, poultry, eggs, or fruit. Consumption/use of such items may impact the level of uranium in urine.
Hypothesis:
We are not testing a specific hypothesis. The primary objective of this study is to establish a representative range of uranium in urine for non-occupationally exposed LLNL workers. This range will be used as a control for evaluating the occupational surveillance results (i.e., discriminating between "background" and potentially occupationally enhanced levels of uranium). "Urine Blank" control samples (urine from individuals who do not work with uranium) are routinely collected and analyzed for uranium as part of the Hazards Control Bioassay Laboratory's quality control program. This study will analyze urine and drinking water from a larger population than that provided by the routine "Urine Blank" control samples. Corollary objectives include characterizing how uranium concentration in an individual's urine may vary over time and determining if there is a statistically significant correlation between uranium in urine and uranium in drinking water for the population at LLNL.
Experimental design:
Human subjects will provide one urine sample and one drinking water sample (of about 100 ml) every quarter for this study. All subjects will be volunteers. Subjects will be encouraged to participate in the study for at least one year and preferably longer. Samples will be assigned a Bioassay Laboratory sample number and processed in the same manner as routine occupational safety surveillance samples. All samples will be analyzed for uranium using inductively coupled plasma-mass spectrometry analysis. Some samples may be analyzed for the uranium-236 isotope by accelerator mass spectrometry. Specific gravity, pH, gross alpha, and gross beta may also be measured periodically to support the uranium measurements. No other constituents will be analyzed.
Anticipated results:
Based on results from the routine occupational safety surveillance samples and this study, we anticipate a wide variety of uranium concentrations in the samples from our study subjects. The study will be successful if we can analyze all urine and drinking water samples from study subjects for uranium and establish a representative background uranium in urine range for the LLNL uranium worker population. The study will be terminated when routine occupational monitoring for uranium in urine is no longer required at LLNL. Excess urine will be disposed of in an appropriate manner. Study sample urine is processed, stored, and disposed of in the same manner as all urine from routine bioassay samples. Fully trained laboratory technicians and associates perform these functions in accordance with Hazards Control Department Radiation Safety Section Bioassay Laboratory procedures and protocols. The urine will not be used for further tests or studies.
"Biomechanics of Human Dentin"
Principal Investigator: Dr. John H. Kinney, Lawrence Livermore National Laboratory
Project started in: 2001
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 116
Institutional Review Board (IRB) Review:
Type of Review:
Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 05/01/06
IRB approval number: 01-116
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 27
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
Scientific context:
The project is designed to study the mechanical properties of human dentin. These properties include, but are not limited to, elastic constants, fracture toughness, and cyclic fatigue behavior. Human subjects are required because we need to perform the mechanical tests on the actual human dentin.
Procedures involving human subjects:
The teeth are to be extracted for clinical reasons. If the clinician decides a tooth should be removed, the patient is asked if they would like to participate in our study by donating the extracted tooth. They are provided with a consent form, a description of the study, and the location of the laboratories where the tooth might be examined. All medical records or identifiers are kept in strictest confidence and are not available to the research team.
Hypotheses:
Specific Aim 1: Biomechanical Properties of Normal Dentin
These studies are designed to measure the elastic and fracture properties of normal human dentin. Specifically, it is hypothesized that the elastic properties, such as Young's modulus, are isotropic (the same in any direction). Other hypotheses consider the fracture properties of dentin: for example, can one measure fracture toughness, and is the fracture toughness isotropic or does it depend on the orientation of the mineralized collagen fibrils?
Specific Aim 2: Biomechanical Properties of Altered Forms of Dentin
In this specific aim, we examine the elastic and fracture properties of altered forms of dentin. Specifically, we will study carious and transparent dentin, as well as what is called reparative dentin (the dentin formed at the pulpal wall in response to trauma).
Specific Aim 3: Biomechanical Properties Affecting Tooth Lifetime
The studies in Aim 3 are intended to explore the nature of failure of the human tooth, and the role that environmental factors might play on tooth strength. In particular, we will explore the fatigue (cyclic stress) behavior of dentin and attempt to determine if there is a limiting stress below which tooth failure cannot occur. Questions that will be addressed include whether there is a critical flaw size in dentin.
Experimental design:
The above specific aims will be accomplished by augmenting the nanomechanical techniques developed in prior years with resonant ultrasound spectroscopy (RUS), fracture mechanics testing, and finite element modeling.
Anticipated results:
We anticipate that the elastic properties will be controlled by the mineralized collagen fibrils. In addition, we believe that failure in the tissue will be dominated by preexisting or induced defects in the microstructure. The study will be terminated in five years at the end of the program.
"Changes in Diffraction-limited Spatial Vision Associated with Aging and Retinal Disease"
Principal Investigator: Dr. Scot S. Olivier, Lawrence Livermore National Laboratory
Project started in: 2001
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 119
Institutional Review Board (IRB) Review:
Type of Review:
Full Board
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 06/05/06
IRB approval number: 01-119
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 36
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
The human eye suffers from significant high-frequency aberrations that can not be corrected by standard corrective eyewear and thus, limit visual acuity. Adaptive Optics (AO), a mature technology used in astronomy to correct for the blurring effect of the atmosphere on astronomical images, can also be used to compensate for ocular aberrations, thereby providing normal eyes with supernormal vision. Lawrence Livermore National Laboratory (LLNL) is developing an adaptive optics system for vision correction based on the LLNL expertise in high-resolution optical control systems to demonstrate, for the first time, diffraction-limited human vision; i.e., vision that is essentially aberration-free. This adaptive optics instrument is an extension of a prototype system developed and in use at the University of Rochester. Using this instrumentation, we will seek to determine both the ultimate resolution and the ideal optical correction for human eyes based on both the optical aberrations and neurological factors.
This project represents a collaborative effort between the Adaptive Optics Group in the Physics and Advanced Technology Directorate at LLNL, led by Dr. Scot Olivier, and the Visual Psychophysics Laboratory in the Department of Ophthalmology at the University of California-Davis Medical Center(UCDMC), led by Dr. John S. Werner. The researchers at LLNL will provide the adaptive optics instrumentation and will collaborate with researchers at UCDMC, who will use the instrument in a clinical setting at the UCDMC.
The broad purpose of our research program is to understand the limits of human visual acuity and the changes in vision associated with aging and retinal disease. The specific purpose of this study is to apply adaptive optics technology to the study of the human eye for two purposes: (1) to measure the visual performance of the eye when virtually all the aberrations of the eye are corrected and (2) to obtain high-resolution images of the retina, thereby allowing correlation of retinal structure with visual performance. Our collaborator's research programs at the UCDMC have been concerned with the optical and neural factors responsible for the normal aging of the human visual system, and cellular mechanisms of age-related macular degeneration(AMD), the leading cause of blindness in the United States. This project brings those two research programs together by allowing correlation of high-resolution images of cells in the retina with visual performance assessed using aberration-free stimuli. The human visual system changes continuously throughout the life span and may result in changes in the quality of life for healthy elderly people that do not come to the attention of health care providers. In one survey, 40 percent of individuals over age 65 years reported that visual changes impair daily life activities. Our collaborator's research has identified some of the optical and neural factors responsible for the degradation of human vision due to aging. However, there are still fundamental questions that have not been addressed about normal aging of the visual pathways that can now be investigated using aberration-free stimulus imaging afforded by adaptive optics. For example, what are the consequences for vision resulting from the losses in photoreceptor numbers beginning in early adulthood? Also, what changes occur in visual resolution as a result of the one-third loss in retinal nerve cells between the ages of 34 and 72 years? By using an adaptive optics instrument, it will be possible to address these questions systematically using established psychophysical (vision science) procedures. With better understanding of normal aging and adaptive optics instrumentation, it will also be possible to study abnormal aging of the human retina, for instance in the presence of retinal disease such as AMD. Changes for year 2005 renewal include a request by the lead group in this study headed, by John S. Werner MD, to the UCDMC IRB board, to lower the study age to children to a minimum age of eight years. As stated in the UCDMC IRB request: "There are certain eye diseases that manifest at a very early age, and by including these young patients, this will allow us to conduct longitudinal studies that will enhance our understanding of disease processes."
The formal experimental tests will involve presentation of patterns on a computer screen that change over space and time, called sinusoidal gratings (fuzzy bars). We will vary the spatial frequency of the patterns from very coarse (fat bars) to very fine (skinny bars). The gratings will be greenish in color and the overall size will be varied. The computer screen will be viewed through an imaging system containing a combination of optical components (mirrors, lenses, and windows) of similar complexity to a high-end amateur telescope. Tests will be conducted following up to 30 minutes of dark adaptation. Subjects will be asked to press a button to record whether the stimulus was more likely to have been detected in the first or the second interval denoted by beeps. A second set of measurements will present the letter "C" on a computer screen. The subject will be asked to press one of four buttons to denote the orientation of the gap in the letter so that visual acuity can be measured. Images of the subjects' retina may also be recorded using flash photography.
All involvement of human subjects in this project will be under the direction of our collaborators at UC-Davis, following the protocol already approved and subsequently renewed by the IRB at UC-Davis. Prior to any tests or examinations, each subject will be provided and asked to sign the Consent To Participate Form provided by UCDMC. Each subject will then have a standard ophthalmic eye exam prior to joining the study. They will then make about one to three visits to UC-Davis Medical Center, where all of the testing for this project will be conducted. Each visit will be about one to two hours in length, and the subjects will be asked if they would like to take breaks frequently throughout the testing period. During the testing period, the subjects will be seated, have their eyes dilated, and bite down on a molded plastic "bite bar" to stabilize the motion of their head. The dilation requires the use of eye drops. The use of eye drops is standard medical practice and are routinely administered by eye doctors to improve the sharpness of the pictures we take. This plastic bite bar is attached to a steel bar that can be adjusted to a comfortable position for the patient. To measure the aberrations of the eye, it is imperative that the head be still, so once in place the steel bar is tightened down, and the motion of the patient's head is minimized. Every vision group doing similar research has empirically determined that the bite bar set-up is the best method for achieving the desired stability of the eye, as well as providing the most comfort for the patient.
Once comfortably settled and looking into the device, the subject will be asked to look at green and black patterns on a computer screen and press specified buttons depending on what they see. These tasks may seem tedious to the subjects but do not pose a serious risk. A potential risk to the subjects, which is explicitly mentioned in the consent form, is dilation of the pupil of the eye. Subjects are warned about all potential hazards related to pupil dilation before their testing period, and the methods used to mitigate these risks are described. In addition to the computer screen, the subjects will be looking into an infra-red light source [currently a superluminescent diode (SLD)] used for the aberration measurement by the adaptive optics instrumentation. The intensity of this light source was measured by the LLNL Safety Officer and found that it is far below all applicable thresholds for damage or discomfort. The LLNL Safety Officer has calculated that the amount of laser light the subject will be exposed to is about 10 times lower that any ocular damage limits found in the ANSI Z136.1-2000 Safety Handbook. When the subject's retina is photographed, a flash lamp is used for illumination. The light flashes are similar to those routinely experienced by patients in ophthalmic examinations and are well below all applicable damage thresholds but may cause temporary discomfort, and the subjects are notified of this in the consent form. All information obtained in connection with this study will be used in a manner that does not publicly disclose the subject's identity and will be kept confidential. Confidentially is assured by keeping data on each subject on a password-protected computer. Drs. Jack Werner and Thomas Barnes will keep any other records in a locked cabinet behind locked doors. They will view this data for vision research purposes only. A detailed consent form, outlining all potential risks involved in the study, must be reviewed and signed before screening and subsequent testing can begin.
LLNL's involvement in this protocol is the design and support of the hardware used for these tests. Although LLNL personnel may be in the room when the study is being run, they are not involved in subject preparation, application of eye drops, and not involved with subject data collection. These tasks are done by UCDMC personnel exclusively. The LLNL personnel, during alignment of the system, can and may make suggestions regarding placement of subjects, changes to enhance performance, etc., but UC-Davis Medical Center personnel make the final decisions regarding all subject changes.
We expect to discover the limits of normal human visual acuity when optical aberrations in the eye are rendered negligible. The limitations to visual performance will then be due to the physical structure of the retina and to the signal and image processing that takes place in the nerve systems within the eye and in the brain. Precise correction of the aberrations of the eye using the adaptive optics instrumentation is required for success of this project. The quality of this correction will be determined by a series of tests that will include the assessment of retinal images recorded using this instrumentation. We also expect to elucidate the effects of aging and retinal disease on visual performance. This particular study will be complete when the specified subject group has been tested and the results have been interpreted. Subsequent studies may then be initiated to further investigate visual performance issues related to age or disease. The criteria for a successful experiment will be that the data can be interpreted to show a statistically significant relationship between ocular aberrations and visual performance.
"Determining the Carcinogenic Significance of Heterocyclic Amines"
Principal Investigator: Dr. James A. Felton, Lawrence Livermore National Laboratory
Project started in: 2002
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 101
Institutional Review Board (IRB) Review:
Type of Review:
Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 09/01/06
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 8
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
Scientific context of the study:
Recent studies have estimated that many human cancers are caused by modifiable lifestyle factors, including diet. Cooking meat produces compounds that have been shown to cause deoxyribonucleic acid (DNA) mutations in animals and humans and to cause cancer in rats. Although we know that humans are exposed to these compounds, the human health implications of this exposure have not been fully defined. One potential carcinogen found in cooked, well-done meat is 2-amino-1-methyl-6-phenylimidazo[4,5-b] pyridine (PhIP).
There are two objectives in this study: (1) to relate individual differences in PhIP urinary metabolite profiles with metabolic phenotypes to predict risk associated with PhIP exposure and (2) to determine if food items such as wheat bran, green tea, and parsley can affect the absorption and/or metabolism of PhIP, thereby decreasing cancer risk.
Experimental design:
Overall requirements of the study population: The volunteers for this study will be recruited from the local workforce and will include no more than 75 healthy individuals. The study will be conducted in two parts: (1) a metabolism study that will provide a metabolic profile of the individuals every six months for 12 months (a total of three measurements) by completing a 24-hour food diary, collecting urine and saliva, and (2) a preventive intervention study that will measure the effects of one of three different dietary modifications. Participation in the intervention part of the study will entail collecting urine an additional time (a total of four urine collections for each subject). No saliva will be collected for the intervention study. Blood will be collected one time during the study for a phenotyping assay. Participants will be compensated $25 for each completed urine collection and $10 for the blood donation; a total of $110 for completing all aspects of the study.
In both parts of the study, PhIP metabolites are measured by having the volunteers consume approximately 150 grams of cooked chicken (approximately two chicken breasts) that has been cooked to contain a known amount of PhIP. Prior to eating the chicken meal the volunteers will be asked to keep a 24-hour food diary that records a description of the food eaten, how much was consumed, and at what time. Urine samples will be collected before eating the meat and for 24 hours after, in six-hour increments. Volunteers will be asked to refrain from eating grilled meats, including pan-fried or flame-grilled hamburgers, steaks, fish, chicken, and bacon for 24 hours before eating the cooked chicken and again during the urine collection period.
In the metabolism stability study we will perform this basic protocol three times during a 12-month period. Within seven days of completing the urine collection, the individuals will also be asked to provide saliva samples for a caffeine phenotyping assay. Subjects will be asked to abstain from caffeine containing foods for 12 hours prior to the caffeine test dose and during the testing period. Approximately one teaspoon of saliva will be collected prior to being given caffeine in the form of one NoDoz tablet (equivalent to one cup of coffee). Additional saliva samples will be collected 4, 6, and 8 hours after the caffeine dose. Saliva samples will be assayed for caffeine metabolites. Caffeine metabolite ratios correlate with the activity of certain metabolizing enzymes that are important for PhIP metabolism. We will correlate the activity of these enzymes (measured by the caffeine assay) with the amount and type of metabolites excreted in the urine. Blood (no more than 30 ml or two tablespoons) will be collected only one time during the entire study from each subject for sulfotransferase phenotyping. Sulfotransferase is another metabolizing enzyme that is important for the metabolism of PhIP. We will correlate the activity of this enzyme with the amount and type of PhIP metabolites excreted. Trained personnel will collect the blood samples in the Lawrence Livermore National Laboratory (LLNL) Health Services Department under strict antiseptic conditions.
In the second part of the study, the preventive intervention phase, we will measure the effects of parsley (4 ounces or 1/2 cup daily), bran (10 grams or two bran muffins daily), or green tea (24 ounces or three cups daily) on PhIP urinary metabolites. The subjects will be given their choice of intervention food. Immediately following one of the collection periods for the metabolic phase of the study, the subjects will consume just one of the food supplements daily for at least three, but no more than seven days. At the end of the intervention period, they will again eat cooked chicken and collect urine for 24 hours.
A subject who completes all aspects of the study will provide four 24-hour urine collections, three 24- hour food diaries, three sets of saliva samples, and a blood sample.
This study involves only the minimal risks present in eating cooked chicken and the intervention foods, donating a blood sample, and the discomfort and embarrassment of collecting urine and saliva. The amount of chicken provided will be the minimum amount necessary to provide enough PhIP to be able to detect metabolites. The identity of the subjects will remain entirely confidential. Subjects will be identified by a code, the code will be associated with the subject's name only on the consent form, and the consent forms will be stored in a locked cabinet. Individuals will be allowed to participate in the study only after they feel comfortable with an explanation of the study and have signed the consent form.
This work will increase our understanding of the risks of being exposed to potential carcinogens in cooked meats. The metabolism study will allow us to determine if there are individuals that may be more or less susceptible, based upon their metabolizing enzyme activity. The results from the intervention phase will provide insight into dietary interactions that may reduce risk from exposure.
"The effect of daily soft-drink consumption on human bone resorption"
Principal Investigator: Dr. Darren J. Hillegonds, Lawrence Livermore National Laboratory
Project started in: 2002
This project ended in fiscal year 2006.
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 106
Institutional Review Board (IRB) Review:
Type of Review:
Full Board
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 11/10/04
IRB approval number: 02-106
Explanation of IRB approval:
Most recent approval on record occurred AFTER FY2004 and is noted accordingly.
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 0
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
Scientific context:
Osteoporosis is a debilitating, increasingly widespread disease characterized by compromised bone strength and predisposition to fracture; it can occur at any age, but often follows menopause in women and occurs later in life in men. In general, consistent whole-body calcium equilibrium as well as peak bone strength are maintained between the ages of 30 and 50 in healthy individuals, after which bone mass is consistently lost; rapid postmenopausal bone loss leads to an earlier fracture risk for women versus men. Among the many interrelated factors affecting skeletal health are calcium and vitamin intake, hormone levels, exercise, and many nutritional factors. The interactions of nutrition and the skeletal health are exceedingly complex, requiring the use of human volunteers to assess the pertinent effects and reach conclusions relevant to human biology.
Hypothesis:
We will quantify the effect of phosphorous-containing food (e.g., phosphoric acid added to cola drinks for the purpose of maintaining carbonation) on rate of bone loss, variously reported to be benign and detrimental.
Experimental design:
The following outlines the currently approved protocol:
Up to fifteen postmenopausal volunteers will ingest 10 nCi (80 ng) of 41Ca as CaCO3, a chemical form found in many calcium supplements. Throughout the course of the study, subjects will maintain usual activities and consume their usual diets. To maintain caloric and caffeine intake, diet caffeine-free soft drinks will be used in the study. A day prior to the dose administration (day 1), one blood sample (10 ml, fasted sample), one 24-hour cumulative urine sample, and ten 5 ml saliva samples (fasted) will be collected. Starting from day 14 and continuing to day 90, on the second day of each week, one blood sample (10 ml, fasted) and one 24-hour cumulative urine sample will be collected. One blood sample (10 ml, fasted) and one 24-hour urine sample will be collected on days 120 and 160. Saliva samples (one 5 ml fasted sample for each time point) will be collected on days 4, 8, 15, 22, 29, and 36 of the study. Later in the study, additional fasted saliva samples will be collected on days when a blood sample is being taken (5 ml each).
Description of the procedures involving human subjects:
There are no major risks from participation in this study. The blood drawing is painful only for a few moments and may result in bruising at the puncture site (hematoma), or rarely, an infection. We will be using a licensed nurse and sterile techniques to reduce risk and discomfort, bruising, and infection.
This study involves a small radiation exposure that is less than that commonly experienced through airline travel or medical tests.
Anticipated results:
We anticipate the results will allow quantification of the effect phosphorous-containing soft drinks have on bone resorption. The study is specifically designed to reflect common American diets and soft drink consumption so as to allow utilization of the resulting data for an accurate gauge of whether soft drink consumption has a quantifiable effect. Since the results for our first subject are positive (i.e., we see a quantifiable effect), we plan on recruiting two new subjects for this study, in order to reproduce the results from the first subject.
"Study of the association of DNA damage with cancer risk"
Principal Investigator: Dr. Irene M. Jones, Lawrence Livermore National Laboratory
Project started in: 2002
This project ended in fiscal year 2006.
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 114
Institutional Review Board (IRB) Review:
Type of Review:
Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 01/06/05
Explanation of IRB approval:
PROTOCOL IS CLOSED.
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 0
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
The purpose of this study is to determine whether ability to repair damaged deoxyribonucleic acid (DNA) is related to risk of cancer in radiologic technologists who may have been exposed to radiation during their work. The National Institute of Cancer (NCI) has been following this group for many years, with particular focus on assessing radiation exposure many years ago and occurrence of cancer. Our study is a small part of a much larger study. Human subjects provide the only way to obtain information on actual occupational exposures to carcinogens such as radiation.
The primary hypothesis being tested is that reduced ability to repair damage in DNA is associated with increased risk of cancer in a group of people with occupational exposures to radiation. Reduced repair could lead to increased formation of mutations that contribute to cancer progression. It is also possible that, to the extent that the subjects studied are found to represent a special subset of long-term survivors of treatment, it might be found that poor repair is associated with improved survival from cancer due to greater impact of anti-cancer therapies.
The Livermore studies assess the ability to repair DNA damage by measuring the level of DNA damage in cell lines developed from blood samples by NCI. In this study, we measure the DNA damage (specifically single strand breaks and altered DNA bases) present in cells that have not been treated with radiation. Much of this damage is similar to damage produced by radiation. The level of damage is used to represent the ability of the cells to repair this damage. Higher levels of damage reflect poorer repair when all other factors are kept constant, as we can do in laboratory studies of cells. To address the main question, measurements of DNA damage are made in cells from two groups of people, those who developed cancer and others of the same age and other characteristics who did not. The samples we receive have been assigned a code by our NCI collaborators. We do not know which samples come from people with cancer or which from cancer-free subjects.
NCI scientists have in years past obtained information from the subjects about their work history and medical history. The subjects in this study were, in addition, asked to provide a blood sample so that cell lines could be prepared that would represent them in future studies such as ours. The risks of obtaining the blood sample are minimal. As confidentiality of all information and materials from subjects is protected, providing the information and blood samples carries little risk. The benefits of the study accrue to the general public rather than to individuals in the study. There is potential that studies such as this will contribute to the ability to identify people who are at increased risk of cancer from radiation exposure. This ability could influence policies and practices that relate to both occupational and therapeutic exposures to radiation and to better protection against unwanted effects of radiation on human health.
We will obtain a measure of the ability to repair DNA damage for each subject. The experimental phase of the study will end when data on all the identified subjects are obtained. Our collaborators will then decode the data and relate our findings to the key attributes of the subjects, cancer status, age, history of radiation exposure, and such. Other studies have found that reduced ability to repair DNA damage increases risk of cancer. Those studies addressed other types of DNA damage and were not studies of populations with occupational exposures to radiation. So the outcome of this study will be very interesting and could lead to larger studies in the future.
The preliminary results of the data collected to date based on one measure of DNA damage suggest that there is increased risk of multiple cancers of which one is breast cancer, early onset breast cancer, and thyroid cancer in members of this cohort with elevated level of DNA damage in normal cells. Other measures of DNA damage find elevated risk only in those with multiple cancers of which one is breast cancer. Detailed analysis of the results, incorporating repeat analysis of about 40 samples, is in progress. Based on the preliminary results, the study will be extended in the coming year to include two additional groups of subjects, those with and without cancer of the testes. This extension is the reason for the request for additional samples.
"Vitamin B12 absorption and metabolism in humans"
Principal Investigator: Dr. Bruce A. Buchholz, Lawrence Livermore National Laboratory
Project started in: 2002
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 119
Institutional Review Board (IRB) Review:
Type of Review:
Full Board
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 08/02/06
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 0
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
Vitamin B12 (B12 or cobalamin) deficiency is caused primarily by malabsorption. Malabsorption of B12 is diagnosed as pernicious anemia. The gold standard for clinical assessment of B12 absorptive capacity is the Schilling test. Schilling tests are, however, rarely used currently to diagnose B12 malabsorption because the test is cumbersome, unreliable, and expensive, and the material required for its performance, 57Co-cyanocobalamin, is both difficult to procure and dispose. The ability of accelerator mass spectrometry (AMS) to precisely measure attomolar (10 to 18) fluctuations in carbon-14 (14C) in biological samples presents an opportunity to develop an improved and reliable test of B12 absorptive capacity.
Freshly absorbed B12 enters the circulation bound to the serum transport protein, transcobalamin. We have obtained preliminary evidence indicating that a recently characterized common polymorphism (G775C) in transcobalamin may significantly influence B12 absorption, as well as cellular delivery, metabolism, and turnover of the vitamin. AMS will be used to conduct in vivo mass-balance studies of B12 distribution, metabolism, and elimination. The relation between kinetics of B12 absorption and turnover and several genetic variations in enzymes related to B12 will be explored. Human subjects are required to accurately develop a suitable clinical tool for assessing B12 absorption and kinetics.
Objectives
The purpose of the study is to assess the absorption and turnover of a tracer dose of [14C]B12 in individuals with normal and impaired B12 absorptive capacity and to determine the influence of transcobalamin genotype on B12 absorption and turnover in humans.
Methodology
Individual subjects will be studied over a one-year period. On the day of dosing, they will be given a single, oral dose of [14C]B12 (100 nCi, 2.25 microgram, taken in water) followed by a light breakfast (a donut and a cup of coffee or apple juice). Blood will be drawn at baseline, and at 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, and 24 hours; and at 2, 3, 4, 5, 6, 7, 10, 14, 21, 28, 35, 42, 49, 100, 150, 200, 250, 300, and 350 days postdosing (35 total blood draws). Blood collected over the first day of the study will be from an indwelling catheter. Subsequent blood draws will be made by needle stick. Mean volume of blood taken at each time point will be ~16 mL (3 teaspoons). Subjects will collect complete (cumulative) 24-hour urine samples and all stool samples on day 0 (starting 24 hours before taking the oral dose of labeled B12) and on each day for 10 days post-dosing (11 samples total). All samples will be processed for 14C measurement at the University of California (UC)-Davis and analyzed by AMS at LLNL. Bioavailabilty and pharmacokinetics will be determined using the data provided by AMS.
Involvement of Human Subjects
Dosing and sampling are described above. Potential risks to the study participants are minimal. Slight discomfort, bruising, and rarely infection may result from the blood draws. To minimize discomfort, bruising, and infection, a licensed phlebotomist using sterile techniques will perform all blood draws. An experienced physician or registered nurse will insert the in-dwelling catheter required for the first day blood collections. The subjects might become anemic due to the multiple blood draws. A 60-mg iron supplement will be given to each volunteer after the 24-hour blood draw to replace the iron removed by the blood draws performed during the first 24 hours. Individual hematocrits will be monitored throughout the study. At-risk individuals will be released from the study. The study will not affect the treatment regimen for the pernicious anemia patients. Although the administered B12 is radioactive, subjects will be exposed to a very small amount of radiation (total effective dose equivalent = 8.16 mrem). For contrast, one receives 3 mrem during a coast-to-coast airline flight. Subjects will complete a dietary questionnaire before beginning the study and be screened for genetic variations in enzymes related to the use of B12, and specifically the G775C genotype in transcobalamin. All subject information will be kept confidential in locked file cabinets by the UC-Davis principal investigator (PI). Subjects will only be identified by number (e.g., S1, S2) in all communication with LLNL. Informed consent agreements will be obtained from all participants and stored at UC-Davis by the PI.
Outcome
The study will provide new information on the absorption and dynamics of B12 in healthy and pernicious anemia subjects under treatment. The study will be terminated at the end of the sampling time.
Conclusion
The study results will constitute a major advance toward understanding the dynamics of B12 absorption and metabolism.
"Identification of markers of human exposure to biological agents, LLNL volunteers"
Principal Investigator: Dr. Kenneth W. Turteltaub, Lawrence Livermore National Laboratory
Project started in: 2002
This project ended in fiscal year 2006.
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 124
Institutional Review Board (IRB) Review:
Type of Review:
Full Board
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 06/30/05
IRB approval number: 02-124
Explanation of IRB approval:
PROTOCOL IS CLOSED.
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 0
Reporting period for number of human subjects:
Fiscal Year 2006
Type(s) of Human Subjects Involvement:
This study is one component of a broad, long-term Grand Challenge program to elucidate the components and mechanisms of host-pathogen interactions that occur in infectious diseases. Tools of genomics, proteomics, biochemistry, engineering, and computer modeling will be applied to study mechanisms of pathogen virulence and host responses in animals and humans. The purpose of this study is to determine if human blood samples can be used to define the spectrum of molecular changes that occur in infection and the time course of these changes.
The long-term goals of this study are to address the following three hypotheses:
1) Determine if specific biochemical components in blood can be used to detect infection before clinical symptoms are present. These components could be used for pre-symptomatic detection of a natural emerging epidemic or a biological attack.
2) Determine if the spectrum of host biological changes varies among infectious disease types. The spectra of host responses could be used for early diagnosis of infection type or initiation of therapy.
3) Determine if biological agents with similar virulence mechanisms produce similar patterns of host marker responses. This would allow classification of unknown genetically modified organisms or newly emerging diseases into virulence classes, suggesting likely candidates for therapeutic intervention.
The experimental design is to use the following analytical approaches to identify potential biochemical markers for human infection with vaccinia virus. Broad screens of many candidate markers will be used in this initial discovery phase. We propose studies of gene expression in blood cells, antigen profiles of serum, and protein profiles of serum. Blood samples will be collected into PAXgene tubes to lyse the cells and stabilize the messenger ribonucleic acid (mRNA). Purification of mRNA, reverse transcription, and quantitative polymerase chain reaction (PCR) are used for gene expression analysis. We are collaborating with a company, Source Precision Medicine, to perform PCR analyses for 30 to 40 gene loci such as inflammatory cytokines, chemokines, proteinases, proteinase inhibitors, adhesion molecules, receptors, and intracellular signaling systems. Blood samples collected into a serum separation tube are aliquoted for use in protein and antigen studies. The technique of 2-D gel electrophoresis is being used to identify protein spots with differential expression between samples. Surface enhanced laser desorption/ionization-mass spectrometry (SELDI-MS) and liquid chromatography-mass spectrometry (LC-MS) will also be used to screen for protein differences. A collaboration with the company, Rules-Based Medicine, has been established to perform multiplex immunoassays for 150 serum components including cytokines, signaling factors, autoimmune antigens, and infectious agent serum antibodies.
This protocol covers the use of blood samples from Lawrence Livermore National Laboratory (LLNL) volunteers as part of this program. LNLL samples will be used for methods development and for defining the biochemical characteristics of control subjects. Volunteer subjects will be recruited from LLNL staff. Up to 19 additional volunteer subjects will be enrolled in this, the second year of the project. Participation will involve providing a blood sample not to exceed 15 ml, and up to five additional blood samples could be requested per person per year. The medical procedure for obtaining this material is standard (not of an experimental nature) venipuncture, and it is expected that the subject will be able to function normal