Mr. David R. Schwoegler
Public Affairs Office
7000 East Avenue, L-404
Livermore, CA 94551
Phone: 510-422-6900
Fax: 510-424-2780
Email: newsguy@llnl.gov
Projects are approved by an IRB located at: Lawrence Livermore National Laboratory.
The approving IRB operates under a Multiple Project Assurance (MPA) recognized by DOE or by the Department of Health and Human Services (HHS).
MPA number of the IRB: M-1415-01-XB
Number of Human Subjects Projects reported: 40
Project Identifier: LLNL-84-P-101-03
Project Title:
Human Sperm Chromatin and Chromosomes
Principal Investigator:
Ms. Laurie A. Gordon
Project started in: 1984
This project ended in Fiscal Year 1997.
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
This project is a core function of the Human Genome Center and is funded under A.V. Carrano's Grant. Estimate is based on amount of funding directed for use of this protocol only.
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: May 15, 1996
IRB Approval Number: 84P-101-03
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 5
Type of Human Subjects Involvement:
A sperm donor will be asked at least 5 days in advance to provide a semen sample on a specified day. He will be provided with a sterile container and he will collect a fresh sample and deliver it to the laboratory or an appropriate intermediate contact person. Once the semen sample arrives in the laboratory it will be given time to liquefy, then it will be placed in a special buffer for overnight storage in a refrigerator. The following day, the sample will be washed 3x in another buffer and the sperm will be isolated by centrifugation. Small aliquots of washed sperm cells are then distributed into culture dishes. Specially prepared hamster eggs are added to achieve gamete fusion. After 2 to 3 hours of co-incubation, the eggs are washed free of sperm and cultured overnight. Early the next morning, the eggs are fixed on microscope slides for analysis of pronuclei formed from the sperm heads which penetrated the eggs. Such sperm pronuclei have very extended chromatin that are used for fluorescence in situ hybridization (FISH) mapping to support the Human Genome Project's chromosome mapping efforts. Any remaining sperm cells or seminal constituents are discarded under regulations pertaining to biohazards.
The benefit of this project to society is derived from its contribution to the Human Genome project. The sperm cells (after fusion with hamster eggs) provide a very extended DNA/chromatin target compared with other nuclear targets. This advantage allows us to identify the location of genes and other markers of interest within the human genetic complement with a much greater resolution than with conventional molecular biology techniques. Using fluorescence in situ hybridization (FISH) mapping techniques we can order and estimate distances between markers along the chromosome, taking advantage of the unique, very extended chromatin produced from human sperm using this system. The maps created by using sperm pronuclei support a broader effort to locate, identify and sequence genes, including genes responsible for genetic diseases. A long-term goal of the Human Genome Project is to devise new diagnostic and therapeutic interventions, and improve those currently available.
Donors are NOT subjected to any treatment, chemicals or radiation.
Our donor pool consists of two to five men. We use only one sperm sample per experiment, performing 10-15 experiments per year. Donors are generally asked to donate a sperm sample at intervals of no more than once a month. Only sperm cells will be used for experimental purposes; all other seminal constituents are discarded.
We do not administer a questionnaire in connection with this study. There are no known risks to donating a semen sample.
Project Identifier: LLNL-88-105
Project Title:
Radiation Genotoxicity from Chernobyl Accident
Principal Investigator:
Ms. Irene Jones
Principal Investigator's Institution: LLNL
Project started in: 1988
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: September 17, 1997
IRB Approval Number: 88-105
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 200
Type of Human Subjects Involvement:
a. Objectives
This project applies three somatic mutation assays to a human population exposed to low-dose, whole-body, ionizing radiation for the purpose of defining the exposure and evaluating the dosimetric assays alone and in combination. In addition, the ability of low dose radiation exposure to induce inherited changes in mini-satellite DNA sequence is studied. Over a four-year period, 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. Workers who were exposed during clean up of the site and sarcophagus construction for the reactor are termed "liquidators" by the Soviets were recruited from throughout the Soviet Union. Each person's work was planned to limit their exposure to 25 cGy. 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. Information from each assay will be analyzed independently for the exposed average and variability, alone and in relation to control, and in relationship to variables such as age, smoking and diet. The results of this study should determine: the utility of these biological dosimeters in the study of low-dose human radiation biology; and the advisability of subsequent pursuit of health effects in this population.
b. Methodology
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, 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 mini-satellite DNA sequences are 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; such alterations are inherited mutations.
c. List any agents to which subjects are exposed.
The people studied are not exposed to any agent as a consequence of this study. The liquidators we are studying were exposed to radiation during the clean up of radiation after the Chernobyl nuclear power reaction accident.
d. Involvement of Human Subjects.
Blood samples are collected by our Russian collaborators at St. Petersburg and Tula. All of the contact with the subjects including obtaining informed consent and collection of blood samples is provided by our 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 are bruising at the site of venipuncture and minor infection. Any adverse effects will be treated by the appropriate Russian health service. The volume of blood drawn is 25-45 ml for adults, and 10 ml for children. Informed consent is 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 is required by a parent or legal guardian. The inclusion of children is necessary for the identification of germinal mutations.
In addition, each control and liquidator completes 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.
Project Identifier: LLNL-88-111
Project Title:
Cytogenetic Studies
Principal Investigator:
Dr. James D. Tucker
Project started in: 1988
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: November 13, 1996
IRB Approval Number: 88-111
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 15
Type of Human Subjects Involvement:
Subject involvement in this study will be to provide peripheral blood for cytogenetic analyses, which will include calibration, validation, and quality control for laboratory purposes. In addition, metaphase chromosomes will be utilized for mapping DNA probes, and developing or evaluating potential new cytogenetic methods.
In some cases, we may elect to have donors complete a questionnaire inquiring about lifestyle factors. This is the same questionnaire that we have used for our other studies, and the data may be useful for interpreting the results of our analyses
We obtained blood from 5 subjects since November, 1995. One donor gave 4 times, one gave twice, and the other three people each donated once. The samples were used to provide metaphase cells for hybridization, for evaluating DNA isolation methods, for fluorescence in situ hybridization probe calibration and testing, and media testing. Each of these activities constitutes an essential activity in my laboratory, without which we could not perform our other work.
Society will benefit in several ways. First, samples obtained will be used to verify that our various molecular probes continue to perform at the expected standard. This will directly benefit all of our projects by providing assurance that our procedures are performing appropriately. Normal control samples, like those requested here, are the most appropriate method for accomplishing this task. Second, cross-validation of various techniques continues to play an important role in our evaluation of various molecular methods. Finally, these blood samples provide raw material for testing new procedures on a limited basis. When or if such new procedures become used on a large scale, a separate Human Subjects approval will be sought.
This use of normal human blood is a vital part of our cytogenetic work. Without this supply the rest of our work would be impossible to conduct.
We require human metaphase chromosomes and nuclei on microscope slides for a very large portion of our work. While these slides are needed on a regular basis, the actual number used in any one week or month varies significantly. Thus, our need for fresh peripheral blood can be irregular and sometimes unpredictable. Accordingly, we cannot anticipate with any degree of accuracy how much will be required in any one year. However, the amount requested in previous years has proven to be adequate for our work. Our current request is unchanged, i.e. no more than 8 separate individuals (mostly from BBRP) will provide blood, with no more than 10 venipunctures per individual per year, and no more than 250 mL per person per year. Venipunctures will be performed by the LLNL Health Services Department using normal procedures.
Project Identifier: LLNL-89-106
Project Title:
Protective Breathing Equipment (Respirators) Testing
Principal Investigator:
Mr. Art Biermann
Project started in: 1989
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: May 21, 1997
IRB Approval Number: 89-106
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 12
Type of Human Subjects Involvement:
For Respirator protection testing
The objective of this research is to measure the degree of protection offered by various respirators to a challenge agent while the wearer, a human subject, is performing movements or exercises designed to simulate specific workplace operations. To evaluate the performance of the respirator during the movements, a human subject will don a respirator and enter a test chamber containing the challenge agent. The challenge may be an aerosol, gas, or combination of the two. Leakage of the challenge agent into the respirator is measured to quantify the respirator performance during the simulated operations. In these experiments, testing is performed on the respirator, not the subject; involvement of the subject is required to adequately simulate the workplace activities.
Respirators being tested may include half masks, full face masks, powered air purifying respirators, supplied air line respirators, and self-contained breathing apparatus. For experiments being conducted in fiscal years 1997-98, the research studies focus on several types of powered air purifying and supplied air line respirators supplied by a variety of manufacturers. Also, for these studies, a submicron aerosol of polyethylene glycol (molecular weight of 400) is used as the challenge agent. Concentrations of the aerosol in the test chamber can range from 10 to 80 mg/m3. However, the subject is not exposed to these levels since they enter the test chamber donning the respirator and there is a high degree of protection provided by the respirator. Once in the chamber the subjects execute the planned exercise protocol. Exercises include standing still while breathing normally, bending forward and touching toes, raising arms above head and looking upward, bending knees and squatting, running in place, standing while twisting at the waist, moving the head side-to-side and up and down, hand scooping of oiled pebbles, and the building of a concrete block wall. Subjects are in the test chamber for no longer than 45 minutes. In addition to measurements of aerosol leakage, the pressure difference between the chamber and inside the face piece and the air flow provided by the respirator will be measured.
For the current studies, it is envisioned that 12 to 20 human subjects will eventually be qualified to participate. All subjects will be LLNL employee volunteers and LLNL-respirator qualified. There are no significant risks resulting from using polyethylene glycol as a challenge agent. Many years of human experience in the workplace and in consumer products consisting of polyethylene glycol have not indicated any adverse health effects. No carcinogenic effects have been reported. Further, no specific industrial hygiene inhalation limits have been established for the compound. Subjects may experience fatigue or sore muscles due to the exercise, or respiratory distress if there is a hypersensitivity to the aerosol. Total exposure will not exceed that permitted for an 8-hr workday under current workplace exposure guidelines for a nuisance oil mist. As of the end of fiscal year 1997, 6 subjects have qualified for participation, and, at the most, one subject will have participated in an actual experiment.
Project Identifier: LLNL-90-105
Project Title:
Center for Genome Research
Principal Investigator:
Dr. Anthony V. Carrano
Project started in: 1990
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: March 19, 1997
IRB Approval Number: 90-105
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 3
Type of Human Subjects Involvement:
As part of the research carried out by members of the Human Genome Center, we occasionally need metaphase preparations of human chromosomes to localize probes (small segments of DNA) back on the chromosomes. Our current need for these chromosomes preparations is quite small, so we are obtaining samples less that 5 times per year.
To generate the metaphase chromosomes preparations, a small aliquot of human peripheral blood is obtained from a donor, drawn by a qualified phlebotomist at the LLNL Health Services facility. White blood cells contained therein are cultured for 2-3 days. Chromosomes are harvested from the white blood cells and placed on microscope slides for the analyses. No long term cultured cells are preserved, nor is any personal data collected from the individual providing the blood sample. The donors are not exposed to any hazardous materials as part of this protocol.
There is minimal risk or discomfort to the individual donating the blood sample, but may including temporary pain, bruising, localized infection, and/or fainting. This activity does not involve medical treatment.
Project Identifier: LLNL-90-108
Project Title:
Detection of Aneuploid Human Sperm by In Situ Hybridization and Image Analysis
Principal Investigator:
Dr. Andrew J. Wyrobek
Project started in: 1990
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
This is one third of a $210,000 project.
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: March 19, 1997
IRB Approval Number: 90-108
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 40
Type of Human Subjects Involvement:
Medical Therapy
a. Objectives:
The objectives of this research are to develop new methods for detecting aneuploidy in human sperm using multi-probe FISH and image analyses for automation.
b. Methodology:
A small amount of human semen is smeared onto glass slides and is analyzed for sperm aneuploidy by fluorescence in situ hybridization (FISH) using chromosome specific DNA probes.
c. Ionizing Radiation, Radioactive Substances, or Chemical Substances.
No persons are exposed to these substances for the purpose of our study. Most of the men used in this study are healthy normal men. For validation of our assays, we also utilize a small number of archived semen samples provided by cancer patients before, during and after they have been treated with drug and/or radiation therapies.
d. Involvement of Human Subjects.
1. Men are invited to be volunteers for providing semen samples and some choose to participate in our studies. Samples are delivered to the laboratory and all samples are coded to protect the confidentiality of the donors. Either frozen or fresh samples are used as dictated by the requirements of the specific laboratory methods.
2. There is no know risk to the semen donors.
Project Identifier: LLNL-91-102
Project Title:
Glycophorin A-based Somatic Cell Mutation Measurements in Blood Samples from Normal Individuals
Principal Investigator:
Dr. Richard G. Langlois
Project started in: 1991
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: March 19, 1997
IRB Approval Number: 91-102
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 50
Type of Human Subjects Involvement:
A) Objectives
The glycophorin A (GPA) human mutation assay was developed at LLNL, and this 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 are required for both quality control, and for defining the distribution of GPA variant frequencies in unexposed individuals for comparison with individuals with potential mutagen exposure.
B) Methodology
Blood samples (5-30 ml) from volunteers at LLNL will be obtained by standard venipuncture done by members of the LLNL Medical Department. All samples will first be M,N typed to identify which samples are of blood type MN, as the assay can only be performed on samples of this blood type. Blood samples will be fixed, labeled with monoclonal antibodies and propidium iodide, and analyzed by flow cytometry. The flow cytometer data will be used to calculate the frequency of variant phenotype erythrocytes in each sample.
C) Ionizing Radiation, Radioactive Substances, or Chemical Substances
None
D) Involvement of Human Subjects
Blood samples will be obtained from normal donors in the LLNL employee population for use in the GPA assay for somatic cell mutations in humans. Samples from normal donors will be used for instrument calibration, quality control tests on assay performance, and as test samples for modifications of the GPA assay method. Assay data from normal donors will also be combined with data from other normal donors to provide information on the distribution of variant cell frequencies in unexposed individuals.
Each donor will be assigned a code number by the principal investigator, Dr. Richard G. Langlois. Subject names will be known only to him and appropriate laboratory personnel. Data obtained from individual subjects will be referred to only by the code number so that the identity of donors will be protected.
Blood samples are obtained by standard venipuncture procedures, and it is expected that the subject will be able to function normally immediately. Possible risks and discomforts that may result from the procedure are considered unlikely but include:
a. Temporary pain
b. Bruising and/or soreness of the affected tissue or surrounding tissue
c. Formation of scar tissue
d. Infection
e. Fainting
Project Identifier: LLNL-91-109
Project Title:
Ultratrace Hair Analysis
Principal Investigator:
Dr. Brian D. Andresen
Project started in: 1991
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: May 21, 1997
IRB Approval Number: 91-109
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 30
Type of Human Subjects Involvement:
Objective
Hair can possibly be a biological sample that will reveal chemical exposures in humans. Incidental inhalation and ingestion of particles over extended periods of time may result in the formation of a diagnostic chemical fingerprint in hair samples. Methods will be explored to ascertain if routine chemical exposures in the work environment can be revealed through ultratrace analysis of human hair samples.
Methodologies
Human hair from individuals known to have been exposed to chemicals in the workplace will be analyzed. New analytical methods will be developed to assay for ultralow levels of and certain organics (e.g., high explosives) and specific elements (e.g., actinides).
The focus of the hair analysis program centers first on reviewing the literature concerning the extraction of target analytes in laboratory-fortified hair samples. Several approaches will be reviewed in detail. For example, enzymatic digestion, organic solvent extractions, and mild acidic extractions. Depending on the harshness of the extraction, solid phase extraction (SPE) may be proposed as a partial method to clean up the extracts. Every extraction technique will be evaluated on its efficiency and ease of use.
The work also includes the development of a laboratory protocol for organic extracts of hair that will either pre-concentrate the sample prior to analysis, or evaporate the sample to dryness and then derive the isolated compounds and elements. It will be proposed to analyze each sample utilizing gas chromatography-mass spectrometry (GC-MS) and inductively coupled plasma mass spectrometry (ICP-MS). We will recommend equipment that is readily available to most all analytical chemistry laboratories.
The use of matrix assisted laser desorption and ionization (MALDI) mass spectrometry will be evaluated for sectional hair analysis. Because hair grows at approximately 1 cm/month it may be used to detect a history of chemical exposures. We plan to review the application of MALDI-MS for hair analysis of HE and actinide incorporation. MALDI-MS will enable us to precisely focus laser power on a selected portion of a hair strand utilizing a miniaturized video camera with newly designed laser optics to monitor the ionization points on the hair shaft. The ions created should allow an ion trap mass spectrometer (with MS/MS option) to detect target species. We will consider in our studies the level of concentration of chemicals in the hair and if at what level the target species must be present in order to be seen with MALDI-MS. Most all of the preliminary work will be performed with laboratory fortified hair samples.
Ionizing Radiation
No radiation experiments are anticipated in these studies. Only hair samples from individuals who work with radioactive materials will be utilized in these studies. No human experiments are anticipated.
Involvement of Human Subjects
1. Hair samples from selected volunteers will be obtained for these studies. Typically hair is obtained during routine hair cuts at the local barber shops. The hair samples are placed in suitable containers and collected for analysis.
2. There is very little risk of injury in obtaining hair samples. The information obtained following the analysis is maintained confidential to all and only available to selected researchers as coded data. No names are associated with the analytical chemistry findings.
Project Identifier: LLNL-92-105
Project Title:
The Effects of Ergonomically Designed Computer Keyboards on the Incidence of Cumulative Trauma Disorders Among VDT Workers
Principal Investigator:
Dr. Stephen R. Burastero
Project started in: 1992
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Total Funding: $259,000
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: May 21, 1997
IRB Approval Number: 105
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 50
Type of Human Subjects Involvement:
Recruited heavy keyboard users (>4 hours per day) involved with data entry and word processing will be eligible for the keyboard trials. Ergonomic keyboard evaluation will be administered to identify various causes of Carpal Tunnel Syndrome (CTS) and/or tendinitis. CTS subjects will be randomized into 3 different groups where they will be assigned to use different keyboards for a 12 week trial period. Selected subjects will also be asked to participate in a laboratory based experiment to measure their wrist deviation patterns using a video analysis system. Subjects will be asked to type from a pangrammic text as their hand and arm movements are monitored using a camera system that relays positional information to a computer for motion analysis.
Standard clinical care will be given regardless of selection into the study.
Project Identifier: LLNL-94-103
Project Title:
Percutaneous Absorption of Radiolabeled Chemicals in Humans
Principal Investigator:
Dr. John S. Vogel
Project started in: 1994
This project ended in Fiscal Year 1997.
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: March 20, 1996
IRB Approval Number: 103
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 10
Type of Human Subjects Involvement:
dermal exposure
Basic research. CS used as tracer.
Objectives: The ultimate objective of this research is to develop and experimentally validate algorithms which predict the absorbed dose following topical application or dermal exposure to chemical substances of therapeutic and toxicological interest. The initial objectives of the experiments are to: (a) measure, in vivo in man, the cutaneous uptake of a series of substituted phenols following exposure to the skin; (b) quantify uptake by application of the chemical 'spiked' with a very small amount of 14C (~nCi) and assess the mass entering the stratum corneum via repeated adhesive tape-stripping and analysis of the strips for 14C; (c) alternatively quantify uptake by selecting phenol derivatives of unique infrared (IR) spectroscopic characteristics and by using attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR) to again determine the quantity of chemical taken up into the stratum corneum (once more, using the adhesive tape-stripping routine); (d) correlate the results from the two methods, and demonstrate that they may be used interchangeably; and (e) compare the experimentally determined levels of chemicals within the stratum corneum at the end of the exposure period with those amounts predicted by dermal uptake models.
Methodology: Radiochemical Methodology: Aqueous solutions of the penetrants, 'spiked' with a very low level of 14C-radiolabel (~10 nCi) will be applied, under occlusion, to the ventral forearms of the human volunteers. Administration will involve saturation of a gauze pad (area = 20 cm2) with approximately 1 mL of solution. After application, the delivery system will be covered with an adhesive, occlusive dressing and left in place for a defined period of time, not to exceed 3-hours. At the end of the exposure time, the gauze is removed and the skin surface is carefully dried with cotton Q-tips. Delivery system and Q-tips will be reserved for analysis. After a period of 10 minutes, sequential layers of stratum corneum at the application site will be removed using preweighed pieces of adhesive tape in the standard way. Up to a maximum of 20 tape-strips will be removed (this is sufficient to excise the entire stratum corneum in most individuals). The tape-strips will then be extracted in methanol and taken to the Center for Accelerator Mass Spectrometry (CAMS) for the analysis of 14C. Background levels will be assessed from tape-strips removed from the contralateral, untreated forearm skin. In this way, the concentration profile of chemicals across the stratum corneum, and the absolute amount taken up into the membrane during the exposure period will be evaluated.
ATR-FTIR Technique: ATR-FTIR is used to obtain spectral measurements of the phenols in the skin of the human volunteers. Prior to administration, a pretreatment spectrum of each subject's stratum corneum will be recorded with ATR-FTIR. After exposure, sequential tape-stripping and spectroscopy will be performed, providing an alternative assay of the distribution of chemical as a function of depth into the stratum corneum (weight of the tape-strips again being used as a marker for position within the membrane). Therefore, the 14C and IR determinations will be performed at the same time, that is, the applied solution of chemical will be both 'spiked' with radioactivity and a spectroscopic marker. Assay of uptake by CAMS can be utilized, then, as a further calibration of the spectroscopic measurements.
Radioactive and Chemical Substances: The chemicals chosen for the study are a series of phenol derivatives, which are commercially available both 14C-radiolabelled and (separately) with significant deuteration [with the exception of cyanophenol, which possesses an inherently unique IR absorbance via the -CºN functionality]. Chemical doses are from 10-50 mg/mL. Radioactive doses are 10 nCi/mL.
Involvement of Human Subjects: Human subjects have an occlusive patch (2 X 9 cm) moistened with a 1-ml chemical solution (10-50 mg/ml) applied to their forearm for periods up to 3 h. The application site is then serially tape-stripped and analyzed for chemical penetration with ATR-FTIR. Approximately 10 mg of outer skin tissue is removed from each human subject. Little if no physical risk is posed to the human subject by this procedure. Dose estimates for the application site and whole body are 0.134 rad/cm2 and 0.0075 mr.
Project Identifier: LLNL-94-105
Project Title:
Investigation of the Relationship between Numerical Chromosomal Aneuploidy in Sperm and Offspring: A Feasibility Study
Principal Investigator:
Dr. Andrew J. Wyrobek
Project started in: 1994
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: March 19, 1997
IRB Approval Number: 94-105
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 40
Type of Human Subjects Involvement:
a. Objectives:
The objective of this study is to study the relationship between numerical chromosomal abnormalities in semen and the chance of fathering a child with a chromosomal defect (e.g., Klinefelter syndrome, 47,XXY).
b. Methodology:
Blood from the parents and child is used to determine the parental origin of the abnormal chromosome and sperm from the father is used to determine the frequency of aneuploidy in the sperm.
c. Ionizing Radiation, Radioactive Substances, or Chemical Substances.
None
d. Involvement of Human Subjects.
1. Families are identified who have a child with Klinefelter syndrome. A genetic counselor will approach the family to request participation in our study and will obtain the questionnaire information and arrange for the visit by the nurse. A nurse will go to the family residence to draw the blood and to pick-up the semen sample which was provided by the father. All samples will be coded to protect the identity of the family.
2. There is a small risk to the puncture area associated with drawing blood and a certified nurse will perform this procedure. There are no known risks for the semen donor.
Project Identifier: LLNL-94-116
Project Title:
Personnel Detection and Imaging with Micropower Impulse Radar
Principal Investigator:
Dr. Stephen G. Azevedo
Project started in: 1994
This project ended in Fiscal Year 1997.
Project Funding Information:
Project did not receive funding in Fiscal Year 1997.
Project did not use human subjects in Fiscal Year 1997.
Explanation:
Funding Sources:
The LDRD work included other efforts besides the human subjects work.
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: November 15, 1995
IRB Approval Number: 94-116
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 0
Type of Human Subjects Involvement:
a. Objective:
Micropower radar pulses will be directed at human subjects from behind walls and other structures for the purpose of detecting and imaging the subjects and their motion. This type of radar imaging will provide the information necessary for law enforcement or military personnel to ascertain the location and movement of people in houses or buildings. Safety of law enforcement personnel may depend upon knowledge of a situation behind walls before surreptitious entry or hostage rescue. Such a device could be useful in these types of situations. Initial tests need to be performed to assess its capabilities.
b. Methodology:
1. Define experiment and review with subject, and ascertain the exposure levels. These levels will depend on distance from the radar (no less than 10 cm). Location of the experiments will vary in order to test different wall types and thicknesses, but will be in areas where the radar complies with FCC regulations.
2. Explain that there is no direct benefit to the subject.
3. Explain long term benefit of experiment to subject: development of an imaging array for military and law enforcement.
4. Brief subject on proposed exposure levels, known safety standards and risks, and source of expert information on exposure.
5. Inform subject of maximum possible exposure in the event of a worst case equipment fault, and inform subject should a fault occur. The worst-case fault is described in the consent form.
6. Inform subject that no special exertion is needed and that no discomfort should occur.
7. Obtain signed and dated consent form.
8. Assign code to patient for data logging ID.
9. Conduct experiment. Generally, the experiment will involve placing the subject on the opposite side of a wall from the radar, and using the detection equipment to image his/her location and movement.
c. Ionizing Radiation, Radioactive Substances, or Chemical Substances: None.
d. Involvement of human subjects:
Only the external dimensions of large objects (people) and their motion will be scanned; imaging of internal body structure will not be performed in this experiment. Detection of cardio-pulmonary function will be attempted by measurement of small external body motions. In all cases, the radar transmitter and receiver will not come in contact with the subject. Tests will take place at various locations/buildings on-site at LLNL.
Project Identifier: LLNL-94-117
Project Title:
Direct Measurement of Uptake of Toxic Chemicals from Water, Soil & Dust into Human Skin In Vivo, Phase I: Aqueous Expos
Principal Investigator:
Dr. Kenneth T. Bogen
Project started in: 1994
This project ended in Fiscal Year 1997.
Project Funding Information:
Project did not receive funding in Fiscal Year 1997.
Project did not use human subjects in Fiscal Year 1997.
Explanation:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: January 17, 1996
IRB Approval Number: 94-117
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 0
Type of Human Subjects Involvement:
See abstract.
See Abstract.
In three experiments to be conducted under this protocol, a single (1) human subject will receive an extremely low total dose (<250 nCi and <1µg) involving two (2) 14C-radiolabeled chemicals (chloroform and trichloroethylene) via dermal exposure to the forearm. In each experiment, each of four application sites will be sampled by a superficial shave biopsy that removes 1 to 5 mg (~2 mm2) of tissue from the outer skin layers. The samples will be analyzed for 14C-radiolabel uptake by accelerator mass spectrometry (AMS), a highly sensitive technique allowing measurements of extremely small uptakes that present a negligible health risk to the research subject.
The project will provide critical human in vivo validation for a much larger series of parallel data on dermal uptake of key environmental contaminants (namely, certain chlorinated solvents and pesticides) to be obtained using an in vitro method involving human cadaver skin. These data are needed to accurately assess human exposures to environmental chemicals through the dermal exposure route. The data obtained will be used to establish more detailed procedures to be included in a future protocol applicable to nine additional subjects in a second phase of AMS-based research on human dermal uptake of several environmental pesticides via two additional dermal exposure media (soil and household dust), which media are of critical concern to the funding agency. Chloroform and trichloroethylene were selected for use in Phase I because results from these studies will be directly comparable to previous results we have obtained using similar AMS and other procedures applied to skin in vitro and hairless guinea pig skin in vivo. In parallel with the experiments involving a human subject under this protocol, we will be conducting similar experiments using human cadaver skin in vitro (LLNL Protocol 94-118 [Certification of Exemption], Approved 19 Nov. 1994). The present experiments will allow the first direct comparisons of dermal uptake and its kinetics in vivo using AMS methods applied to human skin tissue, thereby providing a firm basis for extrapolating more easily obtained in vitro AMS results applied to human cadaver skin in setting safety standards for drinking water and hazardous waste sites. Such data cannot be obtained using only experimental animals, due to (1) substantial interspecies variation in dermal permeability for chemicals and (2) very limited data currently available upon which to base valid animal-to-human extrapolations of in vivo dermal uptake of chemicals specifically from soil, dust or water (the media of interest to the sponsoring agency).
Potential Hazards
Each experiment will expose the volunteer to a total of 80 nCi plus <1 µg of 14C-radiolabeled test compound. The subject will not be used for more than three experiments, each separated by at least two weeks, and applications will not be repeated at the same location on the subject. The whole-body-equivalent dose corresponding to the 3-experiment maximum 240-nCi exposure is estimated to be approximately 0.5 mrem, or <0.01% of the ~300-mrem U.S. average annual exposure to ionizing radiation from natural background sources (National Council on Radiation Protection and Measurements, 1987, Ionizing Radiation Exposure of the Population of the United States, NCRP No. 93, NCRP, Washington, DC). The corresponding local dose to (a total of ~12 cm2 of) skin is estimated to be <500 mrem. These experimental maximum doses correspond to upper-bound estimates for total increased lifetime risks of 0.1 and 0.01 per million, respectively, for cancer and for transmitting inherited diseases. Acute toxic effects from the chemicals and doses to be used are not expected, and any related cancer risk is considered to be negligible (all chemical doses to be administered are below the corresponding acceptable daily intakes defined by current, applicable California and U.S. Environmental Protection Agency regulations (maximum applied doses = 0.35 micrograms/day for either compound, for a total of 3 days separated by at least 2 wk, which is <10% of the corresponding acceptable levels for daily intake over a lifetime). In no case will a particular application site on a subject be used if it exhibits any sign of cut, abrasion, discoloration, or abnormality of any kind.
Project Identifier: LLNL-95-109
Project Title:
An Investigation of Human Calcium Kinetics Using 41Ca and Stable Isotope Labeled Calcium
Principal Investigator:
Dr. Stewart P. Freeman
Project started in: 1995
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project did not use human subjects in Fiscal Year 1997.
Explanation:
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: March 19, 1997
IRB Approval Number: 95-109
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 0
Type of Human Subjects Involvement:
Objectives
We seek to develop novel methodologies for measuring human calcium kinetics. The development at LLNL of technology capable of measuring the very long lived isotope of calcium, 41Ca (105 yr half-life), at environmental levels, enables this isotope to be added to the cannon of calcium isotopic tracers. 41Ca tracer might be employed where others are precluded for radiological, physiological and/or economical reasons, permitting unique long term studies of resorbing labeled bone and so-call continuous feeding studies. Here, having previously demonstrated the feasibility of the continuous feeding approach, we attempt to do so again with a second subject in order to obtain further data.
Methodology
The concentration of 41Ca in feces and urine obtained before, during and up to 6 months after 15 days of consumption of 41Ca labeled orange juice with each meal will be measured to permit the calculation of various parameters of calcium metabolism. In addition, supporting data and data for comparison will be obtained by performing a conventional stable isotope test beginning on the 10th day of 41Ca consumption. That day, the test will involve the administration of stable calcium isotopes IV and PO and blood draws. Excreta samples obtained that day and subsequently will also be measured for the stable isotopic tracers. After appropriate preparation, the blood and excreta tracer isotope concentrations will be determined by mass spectrometry.
Radioactive and Chemical Substances
The single subject will ingest a total of 40 nCi 41Ca @ 10-6 of total calcium in the form of the carbonate dissolved in orange juice. 2 µg/kg 46Ca and 0.5 mg/kg 44Ca will be administered by IV and PO, respectively, having first been tested for sterility and pyrogenicity.
Human Subject Involvement
The subject will provide urine, fecal and nine 3 mL blood samples. Excreta collections will be spot collections, except for a 3 day complete urine collection and a week long complete fecal collection beginning on the day of the stable isotope test.
The risks to the subject are those associated with the lifetime radiation dose from the subsequent decay of 41Ca absorbed by bone. 40 nCi 41Ca will be ingested of which roughly 10 nCi will be taken up. About 40% of this will be incorporated into the stable bone. The remainder of the radiocalcium will be quickly excreted from the body and is assumed to not contribute to the dose. The radiations as a consequence of decaying 41Ca have such short ranges that essentially only the target organs receive a radiation dose (ADf=0.0059 g . rad/mCi.hr). Assuming that the biological half-life of the stable bone component, and, therefore, the effective radionuclide half-life is 50 years, and that the subject survival post administration is 30 years, the cumulated radioactivity will be 633 mCi.hr and the total average dose to the standard man 4 kg cortical bone mass will be 1 mrad or 0.03 mrem/yr. For comparison, background radiation is approximately a factor of 7000 greater. The essentially negligible dose can also be expressed in terms of cancer risk: the relative risk of a 1 mrad dose to bone is a 0.001% increase and the equivalent absolute risk is about 3 cancer cases per year per 10 billion subjects. (The subject has previously been made surgically menopausal, and, therefore, there is no risk of a radiation dose to any fetus.)
Project Identifier: LLNL-95-110
Project Title:
Molecular Genetics of DNA Repair
Principal Investigator:
Dr. Christine A. Weber
Project started in: 1995
This project ended in Fiscal Year 1997.
Project Funding Information:
Project did not receive funding in Fiscal Year 1997.
Project did not use human subjects in Fiscal Year 1997.
Explanation:
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: March 20, 1996
IRB Approval Number: 95-110
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 0
Type of Human Subjects Involvement:
A. Objectives: Our primary objective is to gain an understanding of the genetic and biochemical bases of the genetic disorders resulting from defects in the DNA repair and transcription factor gene ERCC2 (trichothiodystrophy and xeroderma pigmentosum group D) and why one disorder is cancer-prone and the other is not. To achieve this objective, cells from trichothiodystrophy and xeroderma pigmentosum group D patients and family members are used for genetic analysis of the mutations in the ERCC2 gene and in studies of the DNA repair characteristics of the cells. The data generated provide information that is useful both to our research (greater numbers of patients increases our ability to identify commonly occurring mutations and to correlate specific alterations in the protein with the specific clinical presentation) and to the clinicians and families (confirmation of diagnosis, information on degree of photosensitivity of the cells, knowledge of specific mutations in that family useful for future prenatal diagnosis).
B. Methodology: We receive cultures of archived material established in other laboratories (either clinical or research) along with case histories. We request that no identifying information be provided to us. Patient confidentiality is ensured by blacking out any identifying information received and, whenever possible, duplicating the item and destroying the original. Once received in our laboratory, a code designation is assigned (disorder name, #, and 2 letter city abbreviation) and the samples are grown for experimental use (e.g., survival curves, mutation analysis) as well as frozen for future studies. All experimental notes and materials use only the code designation. The code database (showing clinician names, their sample identification, and our corresponding code designations), patient photographs, and any original documents with blacked out identifying information that must be maintained are kept in a locked file. Experimental results are then provided to the clinician and are published. Publications will typically include case histories (sometimes referencing an already published case history) as well as experimental data. In some cases, recognizable photographs may be included. In such cases, the clinicians are co-authors and obtain consent for publication from the parents, legal guardian, or patient, as appropriate to each case (the patients are typically minors).
C. Human subjects are not exposed to any chemical or radioactive substances or ionizing radiation under this project.
D. Involvement of human subjects
1. Archived cell cultures from trichothiodystrophy and xeroderma pigmentosum group D patients and family members are used for UV-survival studies, DNA repair assays, and preparation of RNA and DNA samples for genetic analysis of the mutations in the ERCC2 gene. No cell samples are collected specifically for this project.
2. Since only archival materials are used, the only risks to which human subjects are exposed under this project involve loss of privacy. Steps for ensuring confidentiality are outlined in B above.
Project Identifier: LLNL-95-111
Project Title:
Detection of Chromosomal Abnormalities in Sperm from Men Carrying Specific Constitutive Chromosomal Genetic Factors
Principal Investigator:
Dr. Andrew J. Wyrobek
Project started in: 1995
This project ended in Fiscal Year 1997.
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project did not use human subjects in Fiscal Year 1997.
Explanation:
Funding Sources:
This is one third of a $210,000 project.
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: March 20, 1996
IRB Approval Number: 95-111
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 0
Type of Human Subjects Involvement:
a. Objectives:
The objective of this study is to determine whether men who carry chromosomal abnormalities produce higher rates of chromosomally abnormal sperm than do chromosomally normal men.
b. Methodology:
A small amount of sperm is smeared onto glass slides and is analyzed for sperm aneuploidy by fluorescence in situ hybridization (FISH) using chromosome specific DNA probes.
c. Ionizing Radiation, Radioactive Substances, or Chemical Substances.
None
d. Involvement of Human Subjects.
1. Men who carry chromosomal abnormalities are identified by a collaborating physician in a medical genetics clinic and the doctor arranges for the sperm samples. Samples are delivered to the laboratory and frozen. All samples are coded to protect the confidentiality of the donors.
2. There is no know risk to the semen donors.
Project Identifier: LLNL-95-115
Project Title:
Determining Metabolism Differences by Urine Analysis
Principal Investigator:
Dr. James S. Felton
Project started in: 1995
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: May 21, 1997
IRB Approval Number: 95-115
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 12
Type of Human Subjects Involvement:
subjects will eat cooked meat that naturally contains up to 400 parts-per-billion of PhIP.
The objectives of this work are to understand the human susceptibility to the cooked food containing carcinogenic heterocyclic amines. Individuals will eat normally cooked meat products, primarily chicken, and their urine will be collected over a 10 h time period. Subjects will be exposed to food. Unfortunately, cooked food contains 1-500 ppb of very potent carcinogens and the subjects will be exposed up to 200 ug of compounds similar to PhIP (2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine) at a single meal. Individuals will undergo the risk of eating carcinogens, but no more than they do if they are not vegetarians.
Project Identifier: LLNL-95-119
Project Title:
Chromosome Painting
Principal Investigator:
Dr. Joe N. Lucas
Project started in: 1995
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: July 16, 1997
IRB Approval Number: 95-119
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 5
Type of Human Subjects Involvement:
The goal of this project is to develop and validate improved procedures for the assessment of radiation-induced genetic damage. These studies may lead to procedures: (a) for a more accurate assessment of exposure in accidentally or occupational exposed individuals to guide medical treatment or to establish improved correlation between exposure and untoward medical consequences; (b) to improve our understanding of the genetic consequences of exposure to low levels of ionizing radiation; and (c) to increase the sensitivity with which low level radiation exposure can be assessed. An essential requirement for the use of biomarkers in exposure assessment and dosimetry is quantification of the parameters that drive the overall uncertainty. In the simplest case (i.e., under conditions where ad defines the dose-response relationship), only three parameters must be known for biodosimetric reconstruction of radiation dose to an individual. These parameters are the measured frequency in the individual (Yi), the background frequency (Yb), and the slope (a) of the calibration curve (Eqn. 1). Under such conditions, uncertainties in these three parameters will determine the overall uncertainty in the dose estimate.
D = (Yi - Yb) / a (1)
We have already measured a for some radiations. We are now measuring the Yb in equation (1) for unexposed people.
Methodology: Peripheral blood samples (~5ml) will be taken from volunteers or during routine medical examination, cultured, and the lymphocytes spread onto glass slides for the analysis of chromosome aberration frequencies. The principal method for measuring chromosome aberrations will involve chromosome painting technology.
Project Identifier: LLNL-95-126
Project Title:
Influence of Heterocyclic Amine Metabolic Polymorphisms on Bioactivation of PhIP in the Human Colon
Principal Investigator:
Dr. Kenneth Turteltaub
Project started in: 1995
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: November 13, 1996
IRB Approval Number: 95-126
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 10
Type of Human Subjects Involvement:
This study has three objectives:
(1) To establish whether radiolabeled PhIP-DNA adducts can be detected in humans exposed to a quantity of PhIP comparable to a level of exposure that might occur from the diet;
(2) To determine whether a lower dose of 14C-labeled PhIP can be used that will still allow detection of the adduct in the resected specimen; and
(3) To establish the minimal amount of 14C-labeled compound that can be used to study DNA adducts and metabolites using accelerator mass spectrometry.
Ten human subjects who have been previously diagnosed with colon cancer and who are scheduled for colon resection at the UAMS University Hospital and the VAMC will be informed as to the nature of the study and then asked to participate. They will be asked to read and sign the consent form. Patients may be of either sex and must be >= 18 yrs old. Twenty-four (24) hours prior to surgery patients who consent to participate will be administered [14C]-PhIP in a gelatin capsule, per os (0.07 - 1 µg/kg; maximum activity 15.6 µCi/person). Lactose powder will be used as a bulking agent in the capsule. Two patients each will be given doses of 1 µg/kg, 0.25 µg/kg, and 0.070 µg/kg of 14C-PhIP. Blood will be drawn (50 ml by vein puncture) 12 hr, 24 hr, and 36 hr after PhIP administration to determine the circulating levels of the compound and its metabolites. Additionally, urine (50 - 100 ml) will be collected just prior to PhIP administration and for 24 hr after administration of the PhIP containing capsule to determine clearance rates. Approximately 24 hrs after the capsule is given, the patients will undergo colon surgery and the colon tissue removed during surgery will be placed on dry ice. Specimens will then be divided into four portions, labeled, quick frozen in liquid nitrogen, and then stored at -70 deg. C until sufficient quantities are collected for overnight shipment to LLNL.
Approximately 6 - 8 wks after surgery, patients who participate in this study will be phenotyped for caffeine metabolism. Caffeine is used because it is an indicator of CYP1A2 phenotype and N-acetyltransferase phenotype which are the predominant enzymes involved in PhIP metabolism. Results from this test will be used to assess human variation in metabolism capability. At this time, the participants will be asked to take 200 mg caffeine (2 No-Doz tablets). The patients will be asked to empty his/her bladder 4 hours later. This specimen will be discarded. The bladder will again be emptied one hour later (a total of 5 hrs after taking the caffeine) and this urine will be treated with HCL to preserve metabolites and frozen at -20 deg. C until extraction and analysis can be performed. High performance liquid chromatography (HPLC) will be used to quantitatively measure caffeine urinary metabolites. Phenotype will be assigned based on characteristic peak ratio calculations which have been previously described.
To avoid contamination of the sample with exogenous carbon-14, only disposable, one-time use supplies will contact the sample. The location and amount of tissue removed will be determined by the surgeon but will principally be tumor tissue. The frozen tissue will be homogenized and the DNA will be extracted using phenol:chloroform followed by anion exchange chromatography. The purified DNA will be analyzed for 14C-content by accelerator mass spectrometry and 32P-postlabeling to determine adduct levels.
Patients with uncontrolled cardiovascular disease (eg, hypertension, angina, etc.); patients with hepatic dysfunction as determined by bilirubin >1.2 mg/dl, SGOT >40 U/L, and alkaline phosphatase >120 U/L; and patients with abnormal renal function as determined by BUN >20 mg/dl will not be eligible. In addition, patients who have undergone previous radiotherapy to the abdomen or pelvis or previous chemotherapy within the last 6 months will also be excluded from the study.
Project Identifier: LLNL-96-102
Project Title:
Cytogenetic Blood Analyses of NCI-sponsored Studies
Principal Investigator:
Dr. James Tucker
Project started in: 1996
This project ended in Fiscal Year 1997.
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
Protocol/Subproject Identifier: 96-102
IRB Review:
Type of Review: Full Board
Most Recent Approval: March 20, 1996
IRB Approval Number: 96-102
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 300
Type of Human Subjects Involvement:
Initial Study: We will perform cytogenetic analyses on peripheral blood from up to 300 human subjects over a period of three years. This is part of a large, multi-center study funded by the Radiation Epidemiology Branch of the National Cancer Institute to examine the effects of chronic occupational exposure to ionizing radiation by X-ray technologists. Blood volumes will be approximately 3 ml per donation. The total number of samples we receive will not exceed 300 in the three years of this study. We will perform chromosome painting to enumerate structural chromosome aberrations from each of these samples.
Second study: We will perform cytogenetic analyses on up to 300 human subjects over a period of two years. This also is part of a large, multi-center study funded by the Radiation Epidemiology Branch of the National Cancer Institute. The purpose is to examine the effects of exposure to ionizing radiation produced by weapons testing at the Semipalatinsk facility in Siberia, Russia. The subjects will come from the Altai region of southern Siberia, which is generally downwind from Semipalatinsk. The involvement of the NCI was requested by the Russians and the study is collaborative in nature.
The objectives of the study are as follows:
(1) To determine, by uniform and sensitive thyroid screening, the occurrence of nodular disease and cancer of the thyroid gland in two Russian populations exposed to substantial levels of radioactive fallout from atomic weapons tests.
(2) To relate disease occurrence to the dose level and type of radiation affecting the gland. To accomplish these objectives, thyroid examinations will be offered to approximately 2000 exposed subjects living in Altai as children, 1000 of them in southwestern Altai during 1949 and another 1000 in the northeastern Altai during 1962, and to another 1000 nonexposed or minimally exposed subjects (controls). The subjects to be screened will be drawn from rosters already developed or that are now being developed. Control subjects will be matched by gender, year of birth, and ethnicity. Average age of the people in 1949 or 1962 will be ~5, and in all cases no greater than 15 years. Thus, all subjects are now adults. No blood will be obtained from children. No radiation will be administered to these people as part of this project.
Of these ~3000 subjects, ~10% will be selected for cytogenetic evaluation. According to the NCI documentation, this will involve a single phlebotomy of ~30 ml. All subjects will complete a series of informed consent forms, one or more for the thyroid screening effort (which does not involve LLNL) and one for the phlebotomy.
All samples will be analyzed for structural chromosome aberrations by hybridization with chromosome-specific composite DNA probes ('chromosome painting'). The data will be used to estimate radiation doses, which will be related to adverse health effects.
Project Identifier: LLNL-96-103
Project Title:
Does Tamoxifen Cause DNA Damage in Human Tissues
Principal Investigator:
Dr. Kenneth W. Turteltaub
Project started in: 1996
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: May 21, 1997
IRB Approval Number: 96-103
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 16
Type of Human Subjects Involvement:
Objective:
The question of an increased cancer risk to women taking tamoxifen as a chemotherapeutic agent is a major public health issue. This study addresses whether or not there are DNA adducts present in human tissues after tamoxifen administration. The results will provide better information on the risk-benefit ratio for tamoxifen as a chemopreventive drug.
It is of concern that 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. Analysis of human tissues for the presence of tamoxifen induced DNA damage following the administration of 14C labelled tamoxifen may give an indication of any potential risk of developing tamoxifen induced tumors. This is of concern as tamoxifen may be used as long-term adjuvant treatment in women at high risk of developing breast cancer.
Human Subject involvement:
Women admitted to York District Hospital for breast, endometrial, colon or liver surgery will be fully informed of the purpose of the study and will be asked to participate. Those willing to take part will be asked to fill in a simple questionnaire which will center particularly on which drugs they are currently taking. They will be administered 10 µCi [370 KBq] of 14C labelled tamoxifen diluted in unlabelled tamoxifen to provide a dose of 20 mg/person, which is the normal therapeutic dose. The tamoxifen will be taken orally taken orally 12-18 hours prior to surgery. The radiation dose based upon a 4 day [96 hour] half-life clearance time will be 20.9 µSv which is approximately 20% of the radiation dose of a chest X-ray. This is a conservative [worst case] estimate based upon the clearance time of a 20 mg dose of tamoxifen in men [Adam et al. {1980} Cancer Treatment Reports 64:761-764].
Surgical specimens and a blood sample will be taken at the time of surgery. Urine will be collected up until 48 hours after capsule administration. Samples will be taken to the MRC Toxicology Unit for further processing. DNA will be extracted from the tissue and sent to LLNL for analysis by AMS. Blood plasma and urine will be analyzed for the presence of tamoxifen and metabolites by HPLC.
Project Identifier: LLNL-96-104
Project Title:
Do Trace Levels of Aflatoxin B1 Cause DNA Damage in Human Tissures
Principal Investigator:
Dr. Kenneth W. Turteltaub
Project started in: 1996
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: May 21, 1997
IRB Approval Number: 96-104
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 24
Type of Human Subjects Involvement:
Objectives:
It is the aim of this study to investigate if trace levels of aflatoxin B1 which may be present in the United Kingdom diet as a food contaminant will cause DNA damage in the human liver and colon. This project will provide quantitative data for species extrapolation, and will therefore determine whether the levels of aflatoxin B1 permitted in the human diet in the United Kingdom may pose a cancer risk.
Aflatoxin B1 is a secondary fungal metabolite, which is a naturally occurring dietary contaminant. Animal studies and human epidemiological studies have strongly linked aflatoxin B1 exposure with an increased incidence of hepatocellular carcinoma, however, human studies have frequently been hampered by presumptive intake information and poor cancer incidence data. In addition, the issue of aflatoxin B1 and liver cancer has been the focus of a long-standing debate concerning causative contributions of factors such as hepatitis B virus infection in countries where liver cancer is common. In addition, aflatoxin B1 has been implicated as a human colon carcinogen.
Aflatoxin B1 exposure is not limited to tropical countries due to the importation of potentially contaminated foods to the Western World. For this reason, there are regulations in the United Kingdom for commonly contaminated foods, for example 4 µg/kg for nuts and nut products. However, not all potentially contaminated foods are monitored for contamination, hence, human exposure may be occurring.
Briefly, human subjects will be fully informed of the nature of the study. Subjects will be asked to participate and those willing to take part will provide written informed consent. Patients will be asked to fill in a simple questionnaire which will center particularly on which drugs they are currently taking and diet. They will then be administered 14C aflatoxin B1 in a capsule, taken orally prior to surgery. Surgical specimens and a blood sample will be taken at the time of surgery. Samples will be taken to the Jack Birch Unit, University of York for further processing. DNA will be extracted from the tissue and sent to LLNL for analysis by AMS. Urine samples will be analyzed at LLNL or the Jack Birch Unit. Blood will be separated to obtain lymphocytes, hemoglobin and albumin. These purified fractions will be sent to LLNL for analysis.
Project Identifier: LLNL-96-105
Project Title:
A Comparison of the Use of 41Ca Tracer and Conventional Biomarkers to Assess Bone Turnover in Healthy Adults.
Principal Investigator:
Dr. Stewart Freeman
Project started in: 1996
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: March 19, 1997
IRB Approval Number: 105
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 16
Type of Human Subjects Involvement:
Objectives
Novel methodologies for measuring human calcium kinetics are being pursued. The development at LLNL of technology capable of measuring the very long-lived isotope of calcium 41Ca (105 yr halflife) at environmental levels, enables this isotope to be added to the cannon of calcium isotopic tracers. 41Ca tracer might be employed where others are precluded for radiological, physiological and/or economical reasons, permitting unique long-term studies of resorbing labeled bone and so-called continuous feeding studies. This experiment tests 41Ca protocols against conventional calcium isotope tracers.
Methodology
In all cases, 41Ca is administered to experimental subjects managed at a collaborating institution: UC Davis, the Palo Alto VA, 96-105. The tracer is either administered as a single bolos dissolved in food or as many smaller amounts with meals for up to a month for the continuous feeding studies. Excreta samples from the period of tracer administration and up to six months later are analyzed for 41Ca and the kinetics are calculated. The conventional isotope tests involve a day of IV and PO tracer administration, and urine collection and a blood draw for tracer analysis. Subjects are maintained in a metabolic ward for a week for the mass balance determinations, or to stabilize their kinetics prior to providing urine and blood samples for measurement of the biomarkers in that test.
Radioactive and Chemical Substances
Each subject consumes approximately 5 nCi 41Ca @ 10-6 of total calcium in the form of the carbonate dissolved in orange juice. The standard isotope tests involve the administration of 5 µCi 45Ca or stable isotopes tested for sterility and pyrogenicity.
Human Subject Involvement
A total of 21 adult male and post-menopausal female subjects have been or will be recruited into these studies. Eight subjects will consume a single bolos of 41Ca and another eight will continuously consume the tracer for a month. Subjects will provide occasional urine samples during and after tracer administration until the urine tracer level has stabilized. Then, about 100 days after beginning the experiment, the subjects will be admitted to a metabolic ward where they will provide further urine samples and blood samples while on a measured diet.
The risks to the subjects as a consequence of these studies are those associated with the lifetime radiation dose from the subsequent decay of 41Ca absorbed by bone. Normalized to the 5 nCi oral dose, the International Commission on Radiological Protection calculates the 50 year integrated committed effective dose to be 6 µrem, with an associated risk for all stochastic effects (fatal cancers, non-fatal cancers and severe hereditary effects) of 3´10-9. The 0.1 µrem per year dose is 1 three-millionth of the average background radiation dose of 360 mrem per year. A typical chest X-ray is 10-15 mrem.
Project Identifier: LLNL-96-106
Project Title:
Chernobyl Dosimetry
Principal Investigator:
Dr. Lynn R. Anspaugh
Project started in: 1996
This project ended in Fiscal Year 1997.
Project Funding Information:
Project did not receive funding in Fiscal Year 1997.
Project did not use human subjects in Fiscal Year 1997.
Explanation:
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: May 15, 1996
IRB Approval Number: 96-106
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 0
Type of Human Subjects Involvement:
No radiation will be administered, but the level of radiation that was received during cleanup at Chernobyl will be monitored/researched
Objectives:
To assess the health and environmental consequences of the Chernobyl accident. The US DOE has agreed to provide scientific support for these assessments. The present project is part of the DOE/EH program to help reconstruct radiation doses for populations that participate in the epidemiology studies. Current biodosimetry technologies do not permit the reconstruction of doses for everyone in these studies. However, the approach taken here is to perform biodosimetry using the frequency of chromosome aberrations in blood lymphocytes detected by "chromosome painting" (i.e., FISH) on selected individuals to validate and refine the dose-reconstruction methods used at Chernobyl. The initial objective is to calibrate other less well established methods of dosimetry and to intercalibrate with other methods of similar reliability.
Individuals will be selected to participate in this study on the basis of expected dose range that they have experienced (we are most interested in individuals within the dose range of 0.5 to 2 Gy) and on the basis of other dosimetric information. Up to half of the individuals selected will be controls; a few (<5) individuals selected may have untreated leukemia and some (<15) may have eye cataracts.
In the Ukraine, 5 to 10 cc of peripheral blood will be drawn from each individual by qualified medical personnel at IRP. The blood will be drawn into a heparinized vacutainer by standard venipuncture. The investigators at LLNL will receive coded samples to assure donor anonymity. At LLNL, metaphase chromosomes will be painted using our FISH technique and the radiation dose evaluated for each individual sampled. The biodosimetry results will be provided to IRP. Parallel FISH analysis will also be performed at IRP and intercomparisons made with LLNL.
Immediate benefits include improved doses for the epidemiology studies planned at Chernobyl. Longer-term benefits include improved risk coefficients for radiation-induced effects.
Project Identifier: LLNL-96-107
Project Title:
A Synergy of Science and Art
Principal Investigator:
Stephen C. Sesko
Project started in: 1996
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: May 21, 1997
IRB Approval Number: 107
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 12
Type of Human Subjects Involvement:
The objectives of the research project are twofold:
1) to examine the cognitive processes that non-scientists go through in coming to understand something about science, and
2) to examine the processes of how this understanding becomes translated into a visual, graphic product.
The involvement of human subjects:
The involvement of the participants will be limited to the gathering of qualitative data from; 1) both the textual and the annotated graphical journals that the students normally keep of the work they do on the class project; 2) one-on-one audio-taped interviews with the student-participants, based upon, but not exclusive to these journals; and 3) the oral presentation -- with question and answer session -- that the students normally go through at the end of the class. The only addition that the researchers will request is that the students provide textual material to explain the progress of their graphical journals.
No chemical or radiation exposure will occur.
The number of participants who will be involved in the coming year is at present unknown. We will recruit the participants from a group with an upper limit of approximately 40 students, both present and past.
Other information that will be solicited in the form of copies of the two different journals are an: a) annotated graphics journal that the students normally keep to document the development of their art or graphics project; b) affective journal in which they record their thoughts, "a-ha's", the research sources and results that lead to the art or graphic project, and other cognitive indications of how they have worked their way through the research and the development of the final art or graphics project. These journals are referred to in the answer to the first part of question 1 in this document.
The protocol consists of the review of personal journals and the emergence of themes and codes which show "threads" of thought that can be drawn together into commonalties. From these themes and codes, interview questions are developed to investigate these themes in greater depth and breadth. Once the data are collected it is possible to develop a theory of what has occurred in the process being investigated, according to Glaser and Strauss (1967).
The use of journal review and audio-taped interview are also standard methodologies in anthropology, educational anthropology, and often in sociology.
None of these methodologies are experimental.
The research participants elucidate the issues in the study from their own observations, using their own words. The research is conducted from a phenomenological perspective: what the structure and essence of the science art experience phenomenon are for these participants. The data come from the audio-taped interviews and the journals.
Stories, narratives, and anecdotes have always been a way of creating meaning; they are one way of knowing. The research participants will explain their experiences with science and art in the stories they will tell during their audio-taped interviews. They will select details of their experiences, reflect upon them, make personal sense of them, then narrate them to the researchers. Taken together, the stories coalesce into the researcher's understanding of the participant's experiences. The researcher then attempts to discover what is both consistent and different among these stories. This analysis becomes the researcher's understanding of how these research participants think about and use art and science.
Project Identifier: LLNL-96-108
Project Title:
Genetic and Molecular Analysis of a Normal Human Population
Principal Investigator:
Dr. Janice Pluth
Project started in: 1996
Project Funding Information:
Project received funding in Fiscal Year 1997.
Project used human subjects in Fiscal Year 1997.
Funding Sources:
Project does not involve use of multiple protocols/subprojects.
IRB Review:
Type of Review: Full Board
Most Recent Approval: May 21, 1997
IRB Approval Number: 96-108
Number of Human Subjects who participated in this project/protocol during
FY 1997 (10/1/96 - 9/30/97): 225
Type of Human Subjects Involvement: