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: 40
"Radiation Genotoxicity from Chernobyl Accident"
Principal Investigator: Dr. Irene M. Jones, Lawrence Livermore National Laboratory
Project started in: 1988
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Identifier or number: 88-105
Institutional Review Board (IRB) Review:
Type of Review: Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 04/19/01
IRB approval number: 88-105
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 2001
Type(s) of Human Subjects Involvement:
This project applies three somatic mutation assays to a human population exposed to low-dose, whole-body, ionizing radiation to define the exposure and evaluate the dosimetric assays alone and in combination. In addition, the ability of low dose radiation exposure to induce inherited changes in mini-satellite DNA sequences is studied. This study is designed to provide guidance on the use of the biodoismetric assays in future studies of radiation exposed populations.
The project will study approximately 300 Russians exposed to doses in the range of ~5-25 cGy while working on the containment and cleanup of the Chernobyl nuclear power plant accident in Chernobyl, Ukraine after April 1986, and 300 controls for somatic effects. Fifty control and 25 liquidator families will be studied for heritable effects. The somatic cell assays are 1) stable chromosome aberrations in lymphocytes detected by fluorescence in situ hybridization; 2) glycophorin A mutation in red blood cells; and 3) hypoxanthine phosphoribosyltransferase (HPRT) mutation in lymphocytes (both frequency and deletion spectrum). The results of this study should determine the utility of these biological dosimeters in the study of low-dose human radiation exposure; and the advisability of subsequent pursuit of health effects in this population.
All laboratory analyses start with cells in blood samples. The cytogenetic and HPRT studies both require that white blood cells be cultured. Cytogenetic (chromosomal) changes are studied by microscope studies using fluorescent in situ hybridization methods. For HPRT studies, cultured cells are evaluated by microscopic inspection to determine how many cells are mutated; DNA of mutated cells is studied to define any deletions of the HPRT gene. The glycophorin A assay studies red blood cells by treating them first with a preservative, then with fluorescent label for glycophorin A markers; a flow cytometer determines the number of mutated cells. The germinal mutation studies process DNA from peripheral blood so that a large number of special, highly repeated DNA sequences called mini-satellites can be detected in each person. The patterns of these sequences are compared to determine if there are any sequences in a child that are not present in a parent’s somatic cells; such an alteration is the result of a genetic change, mutation, that occurred in a parent’s germ cell that was transmitted to the child.
Blood samples were collected by the PI's Russian collaborators at St. Petersburg and Tula and Moscow. All of the contact with the subjects was provided by the Russian collaborators in conjunction with the Russian Ministry of Health, the agency charged with providing health care to these individuals. The risks to the individual from the drawing of the blood sample were bruising at the site of venipuncture and minor infection. Any adverse effects were treated by the appropriate Russian health service. The volume of blood drawn was 25-45 ml for adults, and 10 ml for children. Informed consent was obtained from both the exposed and the control individual, and spouses, by standard means, using forms and explanations in their native language. Consent on behalf of minor children was required by a parent or legal guardian. The inclusion of children was necessary for the identification of germinal mutations.
In addition, each control and cleanup-worker was asked to complete a brief questionnaire that provides information on their date of birth, current health status, medications they take, medical treatments involving radiation exposure, their current occupation, the dates they were at Chernobyl and the work assignment they had when at Chernobyl. All subject information was coded so that individual identities cannot be associated with results.
The study has been successful in meeting its aims. Final data analyses are in progress but some key conclusions can be stated. Analysis of chromosome translocations in lymphocytes by FISH was the biomarker most sensitive to past, low dose radiation exposure in the Chernobyl clean up workers studied. HPRT mutant frequency in lymphocytes also detected an effect of radiation exposure, but was less sensitive. GPA mutant frequency did not detect an effect of radiation exposure. No increase in the frequency in inherited mutations of mini-satellite sequences were detected.
"Cytogenetic Studies"
Principal Investigator: Dr. James D. Tucker, Lawrence Livermore National Laboratory
Project started in: 1988
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 10/25/00
IRB approval number: 88-111
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 3
Reporting period for number of human subjects:
Fiscal Year 2001
Type(s) of Human Subjects Involvement:
For many years the PI's laboratory has relied on fresh samples of human blood for a variety of purposes, including chromosome and DNA analyses. This continues to be an essential aspect of his laboratory's needs. Approval of this protocol continues to be essential for many aspects of the work in his laboratory, not all of which can be predicted in advance. There is no "experiment" being conducted under this protocol. The present request for access to human subjects is unchanged from prior years.
The objectives of this protocol are to obtain peripheral blood for cytogenetic and DNA analyses, which will include calibration, validation, and quality control for laboratory purposes. In addition, metaphase chromosomes will be utilized for mapping or verifying DNA probes, and developing or evaluating potential new methods.
The laboratory methods vary according to the needs at the time. The consistent factor is the involvement of human subjects, which is limited to phlebotomy and occasionally the use of a questionnaire as noted in the paragraph below.
Human subject involvement in this research activity is limited to a phlebotomy, and occasionally a questionnaire. Subjects are drawn from LLNL employees who are not under the PI's line of supervision. Pregnant women are excluded. During the past fiscal year 3 subjects were recruited, each of whom donated a single sample of 25 to 30 ml. Venipunctures are performed by the LLNL Health Services Department using normal procedures. In some cases donors have been asked to complete a questionnaire inquiring about lifestyle factors. The risks to subjects are those associated with routine phlebotomy. All identifying material is kept in a locked file cabinet in the PI's office to maintain full confidentiality. No identifying information will be published. All subjects are required to complete a consent form.
Since this protocol does not involve an experiment in the usual sense, there is no "conclusion" per se. However, without access to these samples the PI's lab will not be able to conduct their other work in a scientifically valid manner.
"Glycophorin A-based Somatic Cell Mutation Measurements in Blood Samples from Normal Individuals"
Principal Investigator: Dr. Richard G. Langlois, Lawrence Livermore National Laboratory
Project started in: 1991
This project ended in fiscal year 2001.
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 03/09/00
IRB approval number: 91-102
Explanation of IRB approval:
All research was completed and protocol was closed by PI on 3/9/01.
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 2001
Type(s) of Human Subjects Involvement:
This study has been completed. The following information was submitted by the PI as part of the close-out process
The GPA human mutation assay was developed at LLNL, and the assay is now being extensively used for studies of genetic damage in human populations with exposure to potentially mutagenic agents and populations that are potentially unusually susceptible to mutational damage. Thus, the GPA assay provides an important approach for studying the human risk from exposure to genotoxic agents. Blood samples from normal donors were used for both quality control, and for defining the distribution of GPA variant frequencies in unexposed individuals for comparison with individuals with potential mutagen exposure. Blood samples from normal donors were required for quality control for hypothesis driven GPA studies covered by other IRB protocols. IRB Protocol 91-102 covered the collection and use of blood samples (5-30 ml) from volunteers at LLNL that were obtained by standard venipuncture done by members of the LLNL Medical Department.
Summary of results obtained from this Protocol
The glycophorin A (GPA) assay, developed at LLNL, has been applied to many human studies in the last 10 years. While the samples from this protocol were primarily used for assay development and quality control, these samples were critical to the success of all the GPA studies.
"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
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 08/29/01
IRB approval number: 94-105
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 2001
Type(s) of Human Subjects Involvement:
This is an ongoing study. The specific aim has been to determine whether fathers of children with 47,XXY who are known to have contributed the extra chromosome have elevated frequencies of sperm bearing abnormal number of chromosomes (aneuploidy) compared to fathers of children with 47,XXY where the extra 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 consist of boys with Klinefelter Syndrome (47,XXY) who are six years old or younger. (Men with Klinefelter syndrome have an extra sex chromosome, XXY instead of the usual male arrangement, XY. The symptoms include enlarged breasts, sparse facial and body hair, small testes, and an inability to produce sperm).
Animal and human evidence indicates that: a) both human sperm and animal sperm can carry gene mutations or chromosomal abnormalities; b) genetically defective sperm can fertilize; and c) an embryo’s ability to survive through development to birth and beyond, depends on the specific chromosomal defect it carries. There is no animal model.
The laboratory research conducted at LLNL includes (a) determination of the parent of origin of the extra X chromosome, (b) determination of the level of sperm aneuploidy in the father of affected children.
A total of 38 families with a Klinefelter syndrome(XXY) child have been recruited. The blood samples from father, mother and child plus father’s semen were collected and shipped to LLNL. The team has determined the parental origin of the extra X chromosome of each boy for all the families by polymerase chain reaction (PCR) and sequencing using polymorphic X-linked microsatellite 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 probe specific for chromosomes X, Y and 21.
For the multiple aneuplody study, the team evaluated the fourth family recruited in the main study. This family had three aneuploid pregnancies prior to the index case. The team established that the extra X chromosome of the index child was inherited from the father and that the father produced more aneuploid sperm than this team had ever seen in any man including heavy smokers and men who received cancer chemotherapy. Previously, the team obtained tissue blocks from the aneuploid pregnancies that died before birth (trisomy 15 and 22) to determine whether the extra chromosomes were also paternal in origin. They used an aliquot of semen to determine whether the aneuploidy frequency was also elevated for these chromosomes. In addition, they obtained a second semen sample from the father to determine whether the aneuploid sperm are elevated persistently (two semen samples are two years apart).
According to the protocol, the PI's team requested blood samples from the mother, father, and index child. They requested a semen specimen from the father, and arranged for telephone interviews. Families were compensated $150.00 for their time and participation upon completion of this study ($100 after providing the blood and semen samples and $50 for participating in the 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 want both individual and study results, or just the latter. Findings are conveyed by the study's genetic counselor. The study physician was also available to answer genetics and medical questions. If requested in writing by the family, specific findings were given via mail or telephone to the family's physician. Potential risks to privacy have been addressed.
Children with Klinefelter syndrome (XXY) and their fathers provide a unique opportunity to characterize the relationship between sperm aneuploidy and aneuploidy at birth. The PI's team proposed to determine whether fathers of children with paternally derived Klinefelter syndrome produce inherently elevated levels of aneuploid sperm. This information is critical for understanding the predictive value of aneuploid sperm, for identifying possible underlying paternal host factors for fathering aneuploid offspring, and will serve as one of the first human tests of chromosomally abnormal sperm as a possible mechanism of paternally mediated developmental toxicity. Data analyses and publication of results are expected to be completed during the next year.
"Influence of Heterocyclic Amine Metabolic Polymorphisms on Bioactivation of PhIP in the Human Colon"
Principal Investigator: Dr. Kenneth W. Turteltaub, Lawrence Livermore National Laboratory
Project started in: 1995
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 11/15/00
IRB approval number: 95-126
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 2
Reporting period for number of human subjects:
Fiscal Year 2001
Type(s) of Human Subjects Involvement:
It has been estimated that there will be 93,800 new cases of colon cancer in the United States in 2000. 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.
The development of colon tumors in rats administered PhIP has been associated with the formation of large numbers of 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. The objectives of this protocol 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.
This study is a collaboration between 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 are 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 6 - 8 wks 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. LLNL will receive only coded samples for analysis.
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. 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. Consequently, this study will help determine if PhIP may be a cancer risk factor at dietary levels of exposure and may help find a predictor of individual colon cancer risk. This pilot study will be completed once samples from 10 volunteers have been collected and analyzed.
The results of this study may be published, but the identify of participants will not be released. The confidentiality of all research records will be maintained to the fullest extent possible, and may not be disclosed without the subject's written permission. However, the FDA and/or funding agencies or sponsors may inspect the research records. A signed consent form will be obtained from subjects prior to their participation in the research and in a manner compliant with federal regulations.
"Does Tamoxifen cause DNA Damage in Human Tissues"
Principal Investigator: Dr. Kenneth W. Turteltaub, Lawrence Livermore National Laboratory
Project started in: 1996
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 06/06/01
IRB approval number: 96-103
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 2001
Type(s) of Human Subjects Involvement:
Tamoxifen is a drug used in the treatment of breast cancer and is currently being evaluated for its use as a chemopreventive agent in women at high risk of developing this disease. Tamoxifen is well tolerated and causes relatively few well documented side effects. However, tamoxifen causes liver tumors in rats which is associated with DNA damage. Furthermore, epidemiological evidence suggests that long-term administration of tamoxifen to women leads to an increase in the incidence of endometrial and possibly gastro-intestinal tumors. Consequently, the cancer risk associated with taking tamoxifen as a chemotherapeutic agent is a major public health issue.
The development of liver tumors in rats administered tamoxifen is associated with the formation of large numbers of 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. In women, it is not yet clear if endometrial cancer occurs with tamoxifen as a result of a genotoxic (DNA damaging) or hormonal (estrogenic) mechanism. 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 tamoxifen forms DNA adducts in tissues of women. Our objective is to use [14C]tamoxifen 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 tamoxifen induced tumors in humans.
This study is a collaboration between LLNL, Leicester Royal Infirmary, UK and the MRC Toxicology Unit, Leicester, UK. In this study, women volunteers undergoing hysterectomy will be asked to fill in a simple questionnaire, which will center particularly on which drugs they are currently taking. They will then be orally administered 50 µCi [1.85 MBq] of [14C]tamoxifen diluted in unlabeled tamoxifen to provide a dose of 20 mg/person, which is the normal daily therapeutic dose. The radiation dose will be 170.9 µSv and is less than the natural background radiation to which people are exposed in daily life during the course of a month. Surgical specimens and a blood sample will be taken at the time of surgery and urine will be collected for 24 hours following [14C]tamoxifen administration. Samples will be taken to the MRC Toxicology Unit for further processing. DNA, protein and metabolites will be extracted from the tissue, urine and blood and sent to LLNL for analysis by AMS. In order to protect the confidentiality of volunteers in the study, records will be maintained at the MRC Toxicology Unit and LLNL will receive only coded samples for analysis.
21 volunteers have been recruited on this protocol since 1996. The first 11 volunteers received a radioactive dose of 10 µCi of [14C]tamoxifen. Although tamoxifen itself was measurable in the tissue, DNA adducts were not detected because the amount of radioactivity administered was too low. Consequently, within the last two years a further 10 volunteers were administered a higher dose, 50 µCi of [14C]tamoxifen. Tamoxifen and/or its metabolites were detected in the uterine tissue at levels of approximately 100ng tamoxifen equivalents /g tissue, demonstrating that the drug actually reaches the uterus. Low numbers of DNA adducts were detected in DNA from both the myometrium (muscle layer) and endometrium (lining) of the uterus. Tamoxifen was also found to covalently bind to myometrial and endometrial protein (7000pg tamoxifen/g tissue) at levels 30 fold higher than to DNA. It was also determined that women have approximately 8 fold greater numbers of DNA adducts in their uterus than similarly dosed 6 week-old rats. However, this level in women is still very low, about 10 fold lower than the number of adducts seen in the livers of rats.
Recruitment to this study is now complete and all samples have been collected and analyzed for the presence of DNA and protein adducts. Over the coming year the research team will determine the amount of 14C which is covalently bound to DNA bases and attempt to identify the DNA adducts formed in human uterine tissue by comparison to adducts of known structure formed in rat liver DNA. The results of this study will provide better information on the risk-benefit ratio for tamoxifen as a chemopreventive drug.
The results of this study may be published, but the identify of participants will not be released. The confidentiality of all research records will be maintained to the fullest extent possible, and may not be disclosed without the subject's written permission. However, the FDA and/or funding agencies or sponsors may inspect the research records. A signed consent form will be obtained from subjects prior to their participation in the research and in a manner compliant with federal regulations.
"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
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 08/02/01
IRB approval number: 96-109
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 2001
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 sperm aneuploidy and semen quality; and (2) examine whether certain diets are associated with higher rates of aneuploidy .
For all laboratory analyses, laboratory technicians were blinded to the identity and the age of the man. The following laboratory analyses were performed: sperm concentration, visual motility, and CASA (computer-assisted sperm analysis) motility; nuclear morphometry assay; sperm chromatin assay, sperm chromosomal aneuploidy and aberration assay.
Study populations were comprised of healthy male volunteers employed or retired from the Lawrence Livermore National Laboratory (LLNL) having no known problems with fertility or subfertility. A total of 97 non-smoking men were recruited for this study. These subjects were between 20 to 80+ years of age. Recruiting steps included: screening for eligibility; obtaining a self-administered questionnaire about dietary habits; obtaining a self-administered questionnaire about medical history and sociodemographic characteristics; obtaining consent for their dosimetry record; and collecting one or two semen specimens. All men involved in the study were appropriately informed of the consent process. All information gathered is treated with respect and confidentiality to protect the privacy of each subject.
The recruitment phase is completed, the specimens are under laboratory analyses, and the resulting data are in statistical analyses. 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 Dynamics of Folate Metabloism in Adults"
Principal Investigator: Dr. John S. Vogel, Lawrence Livermore National Laboratory
Project started in: 1996
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 07/18/01
IRB approval number: 96-113
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 3
Reporting period for number of human subjects:
Fiscal Year 2001
Type(s) of Human Subjects Involvement:
Folate compounds, including folic acid, are important nutritional chemicals found in many foods. Humans require daily intake of small amounts of folates for proper health and development. Marginal to poor intake of folate in approximately 10% of the American public is associated with chronic and developmental diseases that include neural tube defects, cancer, and homocysteinemia (an independent risk factor for coronary heart disease). A recent study related incidence of Alzheimer’s disease to a low folate intake.
The long range goal of this study is to understand the metabolism and dynamics of folate chemicals in humans in terms of known hereditary and environmental factors that might affect the incidence and progression of diseases. The relationships of nutritional intake of folate to such diseases are not fully understood, and the natural processing of folates in healthy humans is not known in detail. Detailed knowledge of normal folate metabolism might illuminate paths to disease that result from modifications of this metabolism.
Studies of these chemicals at levels that are naturally consumed by healthy humans have not been possible because no analytical technique could follow the small daily amounts in easily obtained human samples, such as urine, feces, and blood. The development of accelerator mass spectrometry (AMS) at LLNL for tracing chemicals in small biological samples allows researchers to study folates in humans at relevant intake levels. Volunteers will ingest less than a normal daily dose of folic acid that is labeled at a very low level with radiocarbon. The digestion and distribution of this single dose within their bodies will be studied over a period of 7 months using small samples of their blood, urine, and feces.
The LLNL principal investigator has no contact with the human volunteers at any time. All recruiting, testing, dosing, sampling, and subsequent interactions with the volunteers takes place at the University of California, Davis, either in the Sacramento Medical Center or at the Nutrition Department on the main campus. Volunteers fill out questionnaires about their dietary habits and spend the first full day of the protocol at the University of California Medical Center (Sacramento) during which they take the dose of traceable folate and have a number of blood samples drawn from them over 14 hours through an implanted catheter. Subsequent blood draws are made on a "walk-in" basis at the Medical Center over the next 7 months. Volunteers take stool and urine collections at home during the early phase of the experiment in provided containers. Benefits to volunteers include learning about human nutrition, in general, about their own nutritional status, in particular, and about any important health implications of findings about their samples, if warranted.
Researchers expect to determine an average fate and behavior of nutritionally relevant folate within the adult American population, along with an estimate of the variability in this usual behavior. They expect to determine differences from these data in data obtained from a population that possesses a well known inherited modification of folate metabolism. All data, and any conclusions, will be reported in the open literature, will be provided to the research sponsor (National Institute of Digestive, Diabetic, and Kidney Diseases: grant number R01-DK-45939), and will be useful in establishing optimal guidelines for nutritional or supplemental ingestion of folate.
The results of this study may be published, but the identify of participants will not be released. The confidentiality of all research records will be maintained to the fullest extent possible, and may not be disclosed without the subject's written permission. However, the FDA and/or funding agencies or sponsors may inspect the research records. A signed consent form will be obtained from subjects prior to their participation in the research and in a manner compliant with federal regulations.
"Elucidating Dynamics of Beta-Carotene Metabolism in Adults"
Principal Investigator: Dr. John S. Vogel, Lawrence Livermore National Laboratory
Project started in: 1998
This project ended in fiscal year 2001.
Funding for Human Subjects Research:
This project involves the use of multiple protocols/subprojects.
Number of protocols/subprojects associated with this project: 1
Identifier or number: 98-104
Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 07/19/00
IRB approval number: 98-104
Explanation of IRB approval:
This protocol expired and was re-opened under protocol #01-110
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 2
Reporting period for number of human subjects:
Fiscal Year 2001
Type(s) of Human Subjects Involvement:
Deficiencies in vitamin A are correlated with ill health and disease. For example, low vitamin A levels in the eyes leads to poor night vision or lack of color vision. Vitamin A is needed for normal growth of epithelial tissues that protect us from infections in our eyes, mouth, nasal passages, gut, and urinary tracts, as well as being needed for the production of lubricating fluids at body orifices. Vitamin A plays other essential roles in reproduction, cell differentiation, and bone development. High circulating levels of compounds, such as beta-carotene that break down to vitamin A, correlate strongly with low chronic disease conditions. Since vitamin A is harmful to tissues at high concentration and since it is not stored within the body to be called upon when needed, compounds like beta-carotene that break down to vitamin A may be the body’s primary store of this vitamin. However, the ability to absorb beta-carotene from digested foods and to convert it to vitamin A was recently shown to vary widely from person to person. The reasons for this variability are not yet known. A person who does not absorb beta-carotene and subsequently convert it to vitamin A may need a modified diet or supplements to reduce chronic disease and other health risks, including cancers. However, an understanding of the distribution and conversion of dietary beta-carotene will be needed before such diets and supplements are designed.
The objective of this protocol is to determine the uptake of dietary beta-carotene, its conversion to retinoids, and quantify its eventual excretion. Investigators hope to determine the source of the variation among people that leads to different levels of nutrient absorption and utilization.
Two normal daily doses of beta-carotene (1 milligram) containing a very low level of radiocarbon (100 nCi) will be ingested 60 days apart by up to 20 volunteers in a high fat mixture of yogurt and olive oil. Additional doses of vitamin A (10 mg) and beta-carotene (20 mg) labeled with stable isotopes will be co-administered with the radiocarbon labeled dose. Small (5 ml) serial blood samples are drawn over a 60-day period after dosing to monitor the uptake and conversion of the labeled carotene to vitamins. Additional blood samples are taken at 74, 88, 102, 116, and 140 days after the second dosing. Urine and fecal collections are made for the first 30 days after each dosing to find the elimination route of the ingested beta-carotene. The chemical identity of the radiocarbon-labeled vitamins in plasma, urine and feces is quantified by AMS.
The LLNL principal investigator has no contact with human volunteers at any time. All recruiting, testing, dosing, sampling, and subsequent interactions with the volunteers takes place through the University of California at Davis, either at the Sacramento Medical Center or at the Nutrition Department on the main campus. Up to a total of 20 healthy subjects, 10 men and 10 women, will be recruited. Because the beta-carotene is radiolabeled, the subjects will be exposed to a very small amount of radiation (total effective dose is comparable to a coast-to-coast airline flight, 3 mrem). The stable isotope forms contain deuterium in place of hydrogen and have no added toxicity associated with the label. The amounts of vitamin A and beta-carotene are routinely consumed in over-the-counter supplements and pose no additional risk to the participants. Samples of blood, urine, and feces will be collected from them for up to 7 months.
The first three volunteers absorbed and utilized beta-carotene well. Investigators were able to identify and quantify the various retinoid chemicals derived from beta-carotene in the blood samples over the entire time span. They expect to find some volunteers who will not absorb carotene and utilize it as a source of vitamin A, since up to 40% of the subjects in a previous study showed such incomplete use of dietary beta-carotene. The detailed kinetic and metabolic profiles should indicate the causes of any malabsorption.
Investigators are exploring the relationship between diet and disease initiation and progression. Improved dietary and supplement recommendations arising from this study could minimize the human and economic cost of diseases due to nutrient imbalances.
The results of this study may be published, but the identify of participants will not be released. The confidentiality of all research records will be maintained to the fullest extent possible, and may not be disclosed without the subject's written permission. However, the FDA and/or funding agencies or sponsors may inspect the research records. A signed consent form will be obtained from subjects prior to their participation in the research and in a manner compliant with federal regulations.
"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.
Institutional Review Board (IRB) Review:
Type of Review: Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 04/06/01
IRB approval number: 98-106
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 660
Reporting period for number of human subjects:
Fiscal Year 2001
Type(s) of Human Subjects Involvement:
The melanoma phase of the study is to search in the National Death Index (NDI) for melanoma deaths among employees of the Lawrence Livermore National Laboratory who worked at least 6 months during the period July 1, 1984 to Dec 31, 1996. These dates coincide with the operation of the Melanoma Clinic at LLNL, and researchers wish to ascertain the effectiveness of the clinic in preventing melanoma mortality. The general mortality phase of the study is to use the same search to review all sources of death beginning Jan 1 1984 but otherwise as above. Both studies can only be done in the relevant human subjects.
For melanoma researchers are testing the hypothesis that the operation of the melanoma clinic reduced LLNL mortality in relation to national rates. For general mortality, the hypothesis is that LLNL employees are healthy compared to the overall US population.
The design of both phases is that of a classical mortality study using US rates as a basis for comparison. The NDI will provide coded causes of death based on name, birth-date, social security number, race, and sex, with the requirement that the research team not explore mortality status further with families, local institutions, or medical records.
There will be no direct contact of families or employees regarding health status. The results will be shared widely with employees and the public, but only in the form of overall rates and with no divulgence of private information. The researchers will address the significance of the results as fairly and honestly as possible. The files are being carefully protected.
The expected melanoma-specific outcome of this study will be updated melanoma mortality rates for men and women, and their comparison to US rates. The overall study outcome will be updated disease-specific standardized mortality ratios for all coded diseases. Researchers anticipate that in both cases the rates and confidence limits will reflect favorably on the Laboratory’s health status. The final polishing and dissemination of results should be completed within the next year.
"Chromosome Aberration Persistence Study"
Principal Investigator: Dr. James D. Tucker, Lawrence Livermore National Laboratory
Project started in: 1998
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 04/23/01
IRB approval number: 98-111
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 2001
Type(s) of Human Subjects Involvement:
This effort was a 1-year Supplement to the Chernobyl Program Project designed to quantify the persistence of chromosome translocations induced by ionizing radiation. The major emphasis was upon doses of 0.2 to 0.5 Gy, which are similar to those received by most of the Chernobyl clean-up workers. Most exposures to ionizing radiation are below 0.5 Gy, which are lower than those used in previous in vitro and in vivo experiments. One possible mechanism for translocation loss is cell lethality due to partly coincidental and partly correlated types of radiation damage other than translocations. This suggests that the magnitude of loss will decrease as dose decreases, and that it may become vanishingly small at doses such as those encountered by the clean-up workers. The purpose of performing this work is to improve the dosimetry for the Chernobyl clean-up workers by assessing the magnitude of the loss of translocations in vitro after low-dose exposure. All contact with the human subjects has been completed, and the laboratory phase of the work is also done. Only the final data analyses remain.
The aim of the research team was to evaluate the persistence of chromosome aberrations induced in vitro in human peripheral blood lymphocytes by acute exposure to ionizing radiation. This work was needed to refine the biological dose estimates of the Chernobyl clean up workers. The hypothesis was that translocation frequencies decline following acute exposure to ionizing radiation, reaching dose-dependent plateaus. The study has been successful in that researchers have documented the decline of translocation frequencies following acute in vitro exposure to ionizing radiation in peripheral blood lymphocytes. The study will be completed as soon as the final data analyses have been performed and the papers have been published. These tasks should be completed in the coming year.
"Molecular Genetic Analysis of Infertility in Men"
Principal Investigator: Dr. Andrew J. Wyrobek, Lawrence Livermore National Laboratory
Project started in: 1999
This project ended in fiscal year 2001.
Funding for Human Subjects Research: No Funding Sources Reported
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 01/03/00
IRB approval number: 99-104
Explanation of IRB approval:
This protocol was closed by the PI on 11/15/00
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 2001
Type(s) of Human Subjects Involvement:
Infertility affects 20% of couples in the US who are in their reproductive age. In 40% of these cases, the problem can be identified in the male partner. A significant portion of male factor causes of infertility are due to defects in spermatogenesis. It is unfortunate that even with the significant advances in the field of assisted reproduction, we know very little about sperm and egg production in human beings. Recent studies have shown that loss of function of genes located on the Y chromosome can cause spermatogenic defects in men. It was our endeavor in this project to further characterize the molecular basis of infertility in several well-defined populations of infertile men. We studied quantitative defects in gene expression of genes located on X and Y chromosomes, as well as autosomes.
The specific objectives of this project were to: 1) identify genes that map to autosomes or sex chromosomes that are necessary for men to make sperm, 2) explore the function of each gene by using cDNA microarrays, a powerful tool of modern human genetics, to identify alterations in the expression of specific genes in infertile men with low or no sperm counts, and 3) determine the order of function of each gene and identifying mutations which disrupt their function.
Men with various types of infertility were selected for this study including men with decreased sperm density, men with no sperm in the semen and no sperm due to obstructive condition. In addition, normal fertile men and men who were the biological parents and/or siblings of men with various problems as mentioned above were also selected for this study. A letter was sent to each potential participant asking for their consent to join this study. Blood samples and semen samples were collected from all men. Fine-needle aspirates or test is biopsy were done on certain infertility phenotypes.
The two major accomplishments of this projects were 1) significant progress towards the development of enabling technology for the study of expression microarrays for RNA samples isolated from small amounts of tissue, such as biopsies, and 2) the identification of genes that were enriched in human meiosis.
"Absorbed Beta-Carotene: A Retinoid Source in Humans"
Principal Investigator: Dr. John Vogel, Lawrence Livermore National Laboratory
Project started in: 1999
Funding for Human Subjects Research: No Funding Sources Reported
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 07/18/01
IRB approval number: 99-109
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 2001
Type(s) of Human Subjects Involvement:
Localized tissue deficiencies in vitamin A are correlated with cancer development. The local generation of vitamin A, independent of alimentary absorption, may explain the apparent ability of the nutrient beta-carotene to reduce risks of cancer in these tissues. This ability to convert beta-carotene to vitamin A was recently shown to vary from person to person. A person who depends on primary absorption or digestive production of vitamin A from beta-carotene requires a different diet and supplements to reduce cancer risks than a person whose body responds to localized vitamin deficiencies by making the vitamin in situ.
The level of vitamin A production from circulating beta-carotene, and its variability among people, is not known for humans. Under this protocol, researchers hope to determine the rate and form of conversion of circulating beta-carotene to retinoids and quantify its excretion. They will determine the chemical form of the hydrophobic retinoids and the modes of their distribution.
Since any beta-carotene that is eaten passes through the intestine and liver to produce retinoids like vitamin A, the retinoids derived directly from circulating beta-carotene cannot be distinguished through experiments involving ingested beta-carotene. Therefore, researchers will inject the beta-carotene directly into the blood stream after mixing a small amount of [14C]-beta-carotene directly with blood withdrawn from the volunteer. Serial blood samples are drawn over a 60-day period after dosing. Urine and fecal collections are taken for the first 18 days after dosing to find the elimination route of the injected beta-carotene. The quantity of the isotope-labeled vitamin and pro-vitamin in retrieved plasma is quantified by AMS.
Because the administered beta-carotene is radiolabeled, the subject will be exposed to a very small amount of radiation (total effective dose equivalent * 0.5 mrem). By comparison, the total effective dose (TED) equivalent of a chest radiograph, lateral view is 8 mrem and the TED of a coast to coast airline flight is 3 mrem. Blood draws could cause momentary discomfort and/or bruising. A licensed phlebotomist, registered nurse, or physician will be responsible for the blood draws to minimize these risks.
The researchers anticipate highly quantitative conclusions about the value of beta-carotene circulating on lipoproteins as a source of vitamin A in humans. Success is defined as quantitating any such conversion of carotene to vitamin A. They have already succeeded under that criteria. The study will be done when they have enough data from several volunteers to determine the variability of this process in the general population.
The results of this study may be published, but the identify of participants will not be released. The confidentiality of all research records will be maintained to the fullest extent possible, and may not be disclosed without the subject's written permission. However, the FDA and/or funding agencies or sponsors may inspect the research records. A signed consent form will be obtained from subjects prior to their participation in the research and in a manner compliant with federal regulations.
"Methods Development for Studies of Genetic Damage and DNA Repair Function"
Principal Investigator: Dr. Irene Jones, Lawrence Livermore National Laboratory
Project started in: 1999
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 04/24/01
IRB approval number: 99-117
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 11
Reporting period for number of human subjects:
Fiscal Year 2001
Type(s) of Human Subjects Involvement:
The goal of this work is to increase understanding of the factors that determine the amount of genetic damage in cells, both as a result of exposure to radiation or chemicals and as a result of normal life processes. It is believed that increased genetic damage in cells of the body is associated with increased risk of cancer. People differ in the health consequences of exposure to chemicals and radiation, but the biological reason for these differences is not known. One reason that individuals may differ in their response to exposures is that they may differ in their ability to repair damage in their cells. This project is developing sensitive tests to measure the ability of cells to remove DNA damage by DNA repair functions. To be applicable to studies of human populations, the methods use cells from peripheral blood.
The long-term goal is to test the hypothesis that the risk of cancer, or other health outcomes, following radiation exposure is increased in people with reduced DNA repair function. Underlying this hypothesis is the expectation that there is variation in DNA repair function in healthy people, that this variation is largely determined by the forms of DNA repair genes that they inherited from their parents, and that this genetic endowment defines a susceptibility that is only revealed after exposure to radiation.
This project works to develop sensitive measures of DNA repair function using cells in human peripheral blood. The methods developed will ultimately be employed to study the relationship between repair function, genetic determinants of DNA repair, and risk of radiation exposure.
The tests use cells obtained from a blood sample. In the laboratory, the cells are exposed either to radiation or to chemicals that damage the genetic material. The amount of damage produced and how quickly this damage is repaired are studied. Alternatively, extracts may be made of the cells and the ability of the extracts to repair damage in a synthetic DNA studied.
This year the goal was to recruit a small (10-15) multi-ethnic group of individuals whose age range was ~40 to 60 years, to match subjects in a proposed case control study currently under review at NIH. The majority of subjects are BBRP staff who had participated in earlier studies, who were subsequently approached for potential participation in the DNA repair capacity studies. This group was supplemented by a few LLNL employees from other directorates known to the researchers, or referred to them by other participants, who met the criteria outlined above.
Prior to participation in the study, each potential subject reviews and signs a consent form that presents the purpose, procedure, alternatives, risks and benefits of participation. Each subject provides one or more blood samples and is paid $10 for each sample. Each sample is given a code and only that code is used in experimental work, data analysis, and data presentation. A subject when first contacted may be asked to complete a questionnaire to obtain information needed to interpret the experimental results, e.g., their age, smoking history, diet, and exposure to radiation and chemicals. The questionnaire is given a code and only coded information is communicated to others. For evaluating the reliability and reproducibility of methods, it may be desirable to request that some donors provide samples at multiple times, as much as weekly. To avoid the implication of requesting a long term commitment, each subject will be asked to complete the informed consent form each time he/she agrees to provide a sample. For most people this will be once or twice, but for a few others this might be as many as four times in a month. Pregnant women will not be excluded from the study, but there will be a preference to not ask for repeat samples from them.
The next set of results will include the development of statistical methods for analysis for data from the rate of repair of single strand break and base damage assay that is being developed by the researchers. These analyses will define the sensitivity of the assay to detect differences between groups, such as in case control studies. These results will be submitted for publication Spring 2002.
Results of these studies will be included as preliminary data for grants to DOE and NIH. The first stage of success will be funding to conduct studies of radiation exposed populations. The second stage of success will be finding an association between reduced DNA repair function and increased risk of cancer after radiation exposure.
As long as studies continue using repair function assays, this protocol will be kept active. It provides the basis for method development and quality control.
"Endoscopic Sub-Surface Optical Imaging for Cancer Detection"
Principal Investigator: Dr. Stavros Demos, Lawrence Livermore National Laboratory
Project started in: 1999
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 08/13/01
IRB approval number: 99-119
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 15
Reporting period for number of human subjects:
Fiscal Year 2001
Type(s) of Human Subjects Involvement:
There would be tremendous advantages for the development of imaging techniques that could provide sensitive and accurate real time histo-pathologic diagnosis to guide therapy. If, for example, a physician could examine patient tissues with an optical device that could provide the same information as a tissue biopsy, then a biopsy may not need to be performed. Extending the application to a fibro-optic scope device such as a cystoscope to examine the bladder or colonoscope, any tissue sampling will be more directed and precise with the elimination of unnecessary biopsies. Currently, many benign tissue samples are submitted from these and other locations resulting in an enormous cost to the health care system and anxiety and uncertainty for patients. An in-vivo sensitive and accurate imaging system that can provide histo-pathologic information would be an enormous advancement in clinical medicine both for diagnosis and monitoring of on-going therapy. In the field of surgery and interventional radiology, such imaging systems could assist with more accurate and complete removal of cancerous tissue by determining whether extra tissue margins are necessary. Many internal benign lesions would no longer require surgical removal to confirm their benign nature.
The goal of this research effort is to develop an endoscopic subsurface optical imaging system that has the capability to image different tissue components located underneath the surface. The images obtained using this subsurface technique delineate the differences in depolarized backscattering light between normal and cancer tissues arising from differences in the cellular level. These differences include size and concentration of cells and light scattering centers within and between cells, and their absorption properties. In addition, tissue autofluorescence is explored as a secondary method for the detection of superficial cancer lesions. This second method aims in probing differences in the biochemical constituents of cancer tissues as a result of its different metabolism.
In this research effort, the PI is testing the optical imaging modality's ability to address a number of clinical situations associated with bladder, colon, prostate and kidney cancers. They first need to know if in a variety of tumors the system is able to: 1) identify the depth of penetration of the tumor (this would be especially applicable in bladder and colon cancers); 2) identify if the tumor was unifocal or multifocal (renal cancer would be such a situation since it may be either unifocal or multifocal); 3) whether it would act as an aid in identifying cancer from non-cancer tissue in order to aid research.
The research approach is the following: 1) Utilize a prototype imaging system to establish methodology and explore limits of detection using isolated animal (obtained from the local market) and human tissues; 2) Conduct studies in human tissues to establish methodology for its use in vivo and to demonstrate its applicability in the analysis of different types of lesions; 3) Design and build endoscopic subsurface imaging systems that can reach different parts of the human body; 4) Conduct pilot studies to explore its applicability in clinical research and clinical diagnostics; 5) Investigate the possibility of combining this technique with other optical imaging approaches which can serve as a verification method once a "suspicious" lesion is detected.
Human tissue samples obtained from the surgeon immediately after the operation will be used in this investigation. There are no risks for the patient from this study.
The outcome of this work is to provide the basic understanding as well as a proof of principle that will allow the researchers to move to a Phase II where the investigation will be performed in vivo. In the long term, the endoscopic imaging technology to be developed will utilize inexpensive technology and will be easy to operate once the imaging methodology is established. As a result, this technology may become an easily accessible screening and diagnostic tool to the medical community due to the anticipated low cost of acquisition and operation and the limited expertise and training required. This imaging technology may find application in the early detection/screening of cancer, in providing minimally invasive monitoring of the tumor’s response to various stages of treatments and in assisting during surgery by providing information on the depth of penetration of the tumor.
The results of this study may be published, but the identify of participants will not be released. The confidentiality of all research records will be maintained to the fullest extent possible, and may not be disclosed without the subject's written permission. However, the FDA and/or funding agencies or sponsors may inspect the research records. A signed consent form will be obtained from subjects prior to their participation in the research and in a manner compliant with federal regulations.
"Improved Measurement of Cholinesterase Inhibition"
Principal Investigator: Dr. Garrett Keating, Lawrence Livermore National Laboratory
Project started in: 1999
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 07/03/01
IRB approval number: 99-121
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 2001
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% 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.
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 distributed to their mail box which describes the research and subject involvement. 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% 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% so that it can be confidently applied to blood samples from different individuals where normal ChE levels can vary by as much as 30%. 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.
"Botulinum Toxiod Vaccine"
Principal Investigator: Dr. Stephen Burastero, Lawrence Livermore National Laboratory
Project started in: 1999
Funding for Human Subjects Research: No Funding Sources Reported
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 09/28/01
IRB approval number: 99-130
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 2001
Type(s) of Human Subjects Involvement:
LLNL programs are underway which may pose a risk of exposure to LLNL employees to the bacterium Clostridium botulinum. LLNL Health Services Department (HSD) in its role of providing occupational medical services to LLNL employees has decided that to best protect high risk employees, the vaccine Pentavalent (ABCDE) Botulinum Toxoid be provided as an additional measure of protection from this disease. The vaccine is only available as an Investigational New Drug (IND) with the Centers for Disease Control (CDC) being the primary Principle Investigator. HSD will be a collaborator in this research with the co-investigator being Steve Burastero, MD.
The objective of this protocol is to provide immunological protection against the botulinum toxin for the LLNL employees identified by HSD as at risk to exposure by administration of the Pentavalent (ABCDE) Botulinum Toxoid (vaccine).
Botulinum toxoid is not available as a licensed product in the United States. Therefore, the Food and Drug Administration (FDA) has agreed to allow the CDC to supply botulinum toxoid under rules that apply to INDs. Dr. Burastero has registered with the CDC as a co- investigator and meets all of the requirements of the CDC. The immunizations will be given in the LLNL Health Services Department under a written protocol approved by Dr. Burastero (the physician co-investigator). Records will be maintained, and results reported according to the CDC protocol. The toxoid will be administered only to healthy men and women, between the ages of 18 and 65 years. The effects of administration of the toxoid during pregnancy have not been studied, therefore pregnant woman will be excluded. The toxoid will only be given to those high risk individuals actively working or who expect to be working with known cultures and purified botulinum toxins. No one will be administered a subsequent booster immunization unless laboratory tests have shown the level of antitoxin type B and/or E to be below a satisfactory level.
Only LLNL employees at high risk of exposure to botulinum toxins will be vaccinated. There are currently three and could be as many as ten identified employees at LLNL. (None have been vaccinated to date). LLNL will not administer this vaccine to any pregnant women. A standard pregnancy test will be administered to women who are or would become pregnant prior to vaccination. The initial series is given at 0-2-12 weeks and 1 year. A booster may be recommended two years following the last if antibody levels are insufficient. This follow up test involves standard venipuncture to obtain the small amount of blood for analysis of levels of antibodies to the toxoid. Identified information regarding the administration of the toxoid and subsequent symptoms associated with the administration, and followup laboratory test results will be provided to the CDC. Each recipient must sign a CDC consent form. One copy is given to the recipient; one copy is maintained by the clinical investigator; and the third copy is returned to the CDC Drug Service. An LLNL addendum to the CDC consent form will be completed and maintained by LLNL. HSD maintains medical records according to state and federal laws. Form CDC 519.7, "Response to Investigational New Drug", must be completed for each recipient and returned to the CDC Drug Service.
It is expected that vaccinated individuals will develop satisfactory levels of antibodies for protection against botulism. The antibody response is individual, 90% of persons immunized with the initial series and a subsequent booster show effective antibody levels. There are no other products or drugs which give protection against botulism. The potential health risk reduction (no development of the disease) for these individuals vs the usually minor local and very infrequent systemic effects warrant participation in this program.
"PEREGRINE 3-D Monte Carlo Dose Calculations"
Principal Investigator: Dr. Marie-anne Descalle, Lawrence Livermore National Laboratory
Project started in: 1999
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 09/24/01
IRB approval number: 99-131
Number of human subjects who participated in this project/protocol/subproject in the last reporting period: 37
Reporting period for number of human subjects:
Fiscal Year 2001
Type(s) of Human Subjects Involvement:
External beam radiation therapy is one of the standard modalities in cancer treatment. It is non-invasive and takes advantage of the susceptibility of cancer tissue to ionizing radiation to destroy the tumor while leaving normal tissues intact. The success of radiation therapy depends on diagnosis and tumor localization, treatment planning, and dose delivery. Treatment planning is the determination of optimal treatment parameters for the management of a patient's disease with radiotherapy. Because it is not feasible to directly measure radiation dose in a patient, the dose distribution resulting from treatment planning usually serves as the best estimate of the dose delivered to the patient. Today, even the most sophisticated three-dimensional radiation treatment planning systems use a variety of simplifications for actual dose calculations. The PEREGRINE dose calculation system, developed at LLNL, provides highly accurate dose calculations, based on a 3D computer simulation of radiation transport in both the beam delivery system and the patient. Because PEREGRINE calculations are substantially more accurate than conventional dose calculations, it is believed that their implementation in the clinic will result in improvements in the safety and effectiveness of radiation therapy.
The primary objective of this study is to evaluate the clinical utility of the PEREGRINE dose calculation system in radiation oncology treatment planning. Secondary objectives from the National Institutes of Health (NIH) point of view are to provide NIH trainees with experience in advanced dose calculation methodology, establish the Radiation Consultation Workstation (a data/video link between NIH and LLNL) as a feasible interactive facility for complex, online treatment planning, implement a technology transfer collaboration with LLNL, and establish an NIH presence at the forefront of radiotherapy treatment planning. Additional secondary objectives from the LLNL point of view are to develop an ongoing relationship with the NIH/National Cancer Institute with the goal of drawing on their substantial research and clinical expertise in the area of radiation therapy.
Patients will be screened in the NIH Radiation Oncology Branch (ROB) Clinic. Patient suitability for treatment will be evaluated in light of the existing and anticipated research needs of the ROB. Treatment planning for each individual will follow the schema normally used in the development of a three dimensional conformal radiotherapy treatment plan. The plan actually used for the implementation of the patient's treatment will be generated by the NIH treatment planning system. After the patient has been treated, the treatment parameters used on the patients will serve as input to the PEREGRINE dose calculation system. PEREGRINE will not be used to develop a radiation treatment plan. It is used only to provide accurate dose calculations based on the beam and patient parameters established in the NIH treatment plan. Differences between the NIH- and PEREGRINE-generated dose distributions will be assessed by three experienced radiation oncologists for clinical acceptability. Three radiation oncologists will score each NIH treatment plan based on the more-accurate PEREGRINE dose calculation. A score of YES means that the NIH plan would have benefited from further optimization, and is less acceptable for patient treatment, based on PEREGRINE findings. A score of NO means that the Treatment Planning System (TPS) plan is still considered acceptable and needs no additional optimization.
There are no additional medical risks to patients as a result of participation in this study. Each patient is treated using the best standard therapy for his or her disease. No experimental treatments will be applied to patients on this protocol. All patients who meet the inclusion criteria are eligible for this protocol. A patient's rights representative is available to patients on this protocol. Patients may ask any questions about the study, and may withdraw their consent at any time without compromising their ability to receive protocol related medical care from the NIH. LLNL’s role in this study is to provide 3D Monte Carlo dose calculation of dose delivered to each subject. In support of this, a patient’s treatment plan will be provided to LLNL. In discussions with NCI staff on results of calculations, complications or tumor recurrence may also be discussed for individual patients. However, patient identification is protected from LLNL researchers providing PEREGRINE calculations. This is accomplished through a patient coding scheme, in which each patient case is assigned a number, and his/her identity is never shared with LLNL personnel. No research results will be returned to the patient.
By comparing dose distributions calculated by the NIH treatment planning system with more accurate dose calculations provided by the PEREGRINE system, we will be able to better establish the role and importance of accurate, 3D Monte Carlo calculations in radiation therapy.
"Analyses of Spermatozoa from Testicular Cancer Patients Exposed to Radiotherapy"
Principal Investigator: Dr. Andrew Wyrobek, Lawrence Livermore National Laboratory
Project started in: 2000
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Full Board
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 11/15/00
IRB approval number: 00-101
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 2001
Type(s) of Human Subjects Involvement:
Some environmental agents have been shown to cause mutations in somatic cells, but for many agents, as well as ionizing radiation, little information is available on the effects on human gametes and thus on the genetic risk to future generations. Fluorescence In Situ Hybridization (FISH) represents a valuable methodology to detect chromosomal abnormalities in germ cells which are known to be major contributors to infertility, pregnancy loss, mental retardation, infant death and behavioral abnormalities.
The goal of this research is to investigate the effects of ionizing radiation on semen quality and genetic damage in sperm of Stage I Seminoma patients (testicular cancer) who are treated with radiotherapy. The research questions include: (1) What are the effects of radiotherapy on genetic quality of human male germ cells? (2) Do the effects persist with time after the end of treatment?
Stage I Seminoma patients are invited to participate in our study by providing semen samples before, during and after radiotherapy treatment. All samples are coded to protect the confidentiality of the patients. Fresh and frozen samples are used as dictated by the requirements of the specific laboratory methods. There is no known risk to the semen donors. The samples are collected and conventional semen parameters are analyzed. A small proportion of human semen is smeared onto glass slides and is analyzed for sperm aneuploidy and structural aberrations by Fluorescence In Situ hybridization (FISH) using specific-DNA probes.
This study will evaluate and characterize effects of exposure to ionizing radiation on the production of various categories of chromosomally abnormal sperm.
Several newly developed sperm FISH methods provide an efficient approach to study structural and numerical chromosomal abnormalities produced by environmental mutagenic and cytotoxic agents and thus to evaluate the potential genetic risk to future generations.
"A Model for Genetic Susceptibility: Melanoma"
Principal Investigator: Dr. Harvey Mohrenweiser, Lawrence Livermore National Laboratory
Project started in: 2000
Funding for Human Subjects Research:
This project does not involve the use of multiple protocols/subprojects.
Institutional Review Board (IRB) Review:
Type of Review: Expedited
Approving Institution: Lawrence Livermore National Laboratory
Most recent approval: 12/23/00
IRB approval number: 00-102
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 2001
Type(s) of Human Subjects Involvement:
Melanoma is a paradigm for gene-environment interactions in the development of cancer. Sun exposure, the major environmental risk factor, although responsible for 65-90 percent of melanoma, has a small relative risk for the development of melanoma when genetic factors are measured: phenotype (skin color, hair color, eye color), nevus number, and possibly DNA repair. No other cancer has both such a well-identified genetic component and a well-substantiated environmental factor. We hypothesize that there are wide variations among individuals in terms of susceptibility to melanoma. This variation is probably influenced in a heterogeneous manner by multiple susceptibility genes, and sun exposure, the major known exogenous factor, may exert its influence interacting with these genes.
The objective of this large, international population-based case control study is determination of the relative risk for developing melanoma due to polymorphisms in the nucleotide excision repair genes, the pathway for repair of DNA damage induced by sunlight. The LLNL component of the proposal will be the resequencing of DNA repair genes to identify common genetic variation in a population of individuals with melanoma.
To identify the common (polymorphic) variants, we will resequence 18 genes in 50 individuals with multiple primary melanomas. The DNA samples to be screened for DNA sequence variation will be randomly selected by Dr. Marianne Berwick from the repository of more than 400 samples obtained from patients with multiple primary melanomas being collected for this study at Memorial Sloan Kettering Cancer Institute. The 50 DNA samples sent to LLNL for the resequencing will be randomly assigned numbers 1-50 and thus the specific donors will be anonymous.
The results from the LLNL effort will only identify variants and will not address questions related to functional relevance. The results of the complete study will provide quantitative data on the contribution of variation in a specific group of DNA repair genes in melanoma risk, data that will form the basis for credible public health recommendations regarding solar exposure and screening.
Our proposed study will permit evaluation of the public health impact of genetic mutations and polymorphisms and their interaction with sun exposure, via estimation of relevant population parameters in a novel study design.
The results of this study may be published, but the identify of participants will not be released. The confidentiality of all research records will be maintained to the fullest extent possible, and may not be disclosed without the subject's written permission. However, the FDA and/or funding agencies or sponsors may inspect the research records. A signed consent form will be obtained from subjects prior to their participation in the research and in a manner compliant with federal regulations.
"Quantifying the Effects of Preventive Foods on the Metabolism of a Prostate Carcinogen in Humans and in Prostate Cells Grown in Culture"
Principal Investigator: Dr. James Felton, Lawrence Livermore National Laboratory
Project started in: 2000
Funding for Huma