[Federal Register Volume 69, Number 168 (Tuesday, August 31, 2004)]
[Notices]
[Pages 53064-53067]
From the Federal Register Online via the Government Publishing Office [www.gpo.gov]
[FR Doc No: 04-19719]



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ENVIRONMENTAL PROTECTION AGENCY

[OPP-2004-0226; FRL-7369-8]


Modified Cry3A Protein (mCry3A) and the Genetic Material 
Necessary for its Production in Corn; Notice of Filing a Pesticide 
Petition to Establish a Temporary Tolerance for a Certain Pesticide 
Chemical in or on Food

AGENCY: Environmental Protection Agency (EPA).

ACTION: Notice.

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SUMMARY: This notice announces the initial filing of a pesticide 
petition proposing the establishment of regulations for residues of a 
certain pesticide chemical in or on various food commodities.

DATES: Comments, identified by docket identification (ID) number OPP-
2004-0226, must be received on or before September 30, 2004.

ADDRESSES: Comments may be submitted electronically, by mail, or 
through hand delivery/courier. Follow the detailed instructions as 
provided in Unit I. of the SUPPLEMENTARY INFORMATION.

FOR FURTHER INFORMATION CONTACT: Mike Mendelsohn, Biopesticies and 
Pollution Prevention Division (7511C), Office of Pesticide Programs, 
Environmental Protection Agency, 1200 Pennsylvania Ave., NW., 
Washington, DC 20460-0001; telephone number: (703) 308-8715; e-mail 
address: [email protected].

SUPPLEMENTARY INFORMATION:

I. General Information

A. Does this Action Apply to Me?

    You may be potentially affected by this action if you are a 
agricultural producer, food manufacturer, or pesticide manufacturer. 
Potentially affected entities may include, but are not limited to:
     Crop production (NAICS 111)
     Animal production (NAICS 112)
     Food manufacturing (NAICS 311)
     Pesticide manufacturing (NAICS 32532)
    This listing is not intended to be exhaustive, but rather provides 
a guide for readers regarding entities likely to be affected by this 
action. Other types of entities not listed in this unit could also be 
affected. The North American Industrial Classification System (NAICS) 
codes have been provided to assist you and others in determining 
whether this action might apply to certain entities. If you have any 
questions regarding the applicability of this action to a particular 
entity, consult the person listed under FOR FURTHER INFORMATION 
CONTACT.

B. How Can I Get Copies of this Document and Other Related Information?

    1. Docket. EPA has established an official public docket for this 
action under docket ID number OPP-2004-0226. The official public docket 
consists of the documents specifically referenced in this action, any 
public comments received, and other information related to this action. 
Although a part of the official docket, the public docket does not 
include Confidential Business Information (CBI) or other information 
whose disclosure is restricted by statute. The official public docket 
is the collection of materials that is available for public viewing at 
the Public Information and Records Integrity Branch (PIRIB), Rm. 119, 
Crystal Mall 2, 1801 S. Bell St., Arlington, VA. This docket 
facility is open from 8:30 a.m. to 4 p.m., Monday through Friday, 
excluding legal holidays. The docket telephone number is (703) 305-
5805.
    2. Electronic access. You may access this Federal Register document 
electronically through the EPA Internet under the ``Federal Register'' 
listings at http://www.epa.gov/fedrgstr/.
    An electronic version of the public docket is available through 
EPA's electronic public docket and comment system, EPA Dockets. You may 
use EPA Dockets at http://www.epa.gov/edocket/ to submit or view public 
comments, access the index listing of the contents of the official 
public docket, and to access those documents in the public docket that 
are available electronically. Although not all docket materials may be 
available electronically, you may still access any of the publicly 
available docket materials through the docket facility identified in 
Unit I.B.1. Once in the system, select ``search,'' then key in the 
appropriate docket ID number.
    Certain types of information will not be placed in EPA's Dockets. 
Information claimed as CBI and other information whose disclosure is 
restricted by statute, which is not included in the official public 
docket, will not be available for public viewing in EPA's electronic 
public docket. EPA's policy is that copyrighted material will not be 
placed in EPA's electronic public docket but will be available only in 
printed, paper form in the official public docket. To the extent 
feasible, publicly available docket materials will be made available in 
EPA's electronic public docket. When a document is selected from the 
index list in EPA Dockets, the system will identify whether the 
document is available for viewing in EPA's electronic public docket. 
Although not all docket materials may be available electronically, you 
may still access any of the publicly available docket materials through 
the docket facility identified in Unit I.B.1. EPA intends to work 
towards providing electronic access to all of the publicly available 
docket materials through EPA's electronic public docket.
    For public commenters, it is important to note that EPA's policy is 
that public comments, whether submitted electronically or in paper, 
will be made available for public viewing in EPA's electronic public 
docket as EPA receives them and without change, unless the comment 
contains copyrighted material, CBI, or other information whose 
disclosure is restricted by statute. When EPA identifies a comment 
containing copyrighted material, EPA will provide a reference to that 
material in the version of the comment that is placed in EPA's 
electronic public docket. The entire printed comment, including the 
copyrighted material, will be available in the public docket.
    Public comments submitted on computer disks that are mailed or 
delivered to the docket will be transferred to EPA's electronic public 
docket. Public comments that are mailed or delivered to the docket will 
be scanned and placed in EPA's electronic public docket. Where 
practical, physical objects will be photographed, and the photograph 
will be placed in EPA's electronic public docket along with a brief 
description written by the docket staff.

C. How and to Whom Do I Submit Comments?

    You may submit comments electronically, by mail, or through hand 
delivery/courier. To ensure proper receipt by EPA, identify the 
appropriate docket ID number in the subject line on the first page of 
your comment. Please ensure that your comments are submitted within the 
specified comment period. Comments received after the close of the 
comment period will be marked ``late.'' EPA is not required to consider 
these late comments. If you wish to submit CBI or information that is 
otherwise protected by statute, please follow the instructions in Unit 
I.D. Do not use EPA Dockets or e-mail to submit CBI or information 
protected by statute.
    1. Electronically. If you submit an electronic comment as 
prescribed in this unit, EPA recommends that you include your name, 
mailing address, and an e-mail address or other contact information in 
the body of your

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comment. Also include this contact information on the outside of any 
disk or CD ROM you submit, and in any cover letter accompanying the 
disk or CD ROM. This ensures that you can be identified as the 
submitter of the comment and allows EPA to contact you in case EPA 
cannot read your comment due to technical difficulties or needs further 
information on the substance of your comment. EPA's policy is that EPA 
will not edit your comment, and any identifying or contact information 
provided in the body of a comment will be included as part of the 
comment that is placed in the official public docket, and made 
available in EPA's electronic public docket. If EPA cannot read your 
comment due to technical difficulties and cannot contact you for 
clarification, EPA may not be able to consider your comment.
    i. EPA Dockets. Your use of EPA's electronic public docket to 
submit comments to EPA electronically is EPA's preferred method for 
receiving comments. Go directly to EPA Dockets at http://www.epa.gov/edocket/, and follow the online instructions for submitting comments. 
Once in the system, select ``search,'' and then key in docket ID number 
OPP-2004-0226. The system is an ``anonymous access'' system, which 
means EPA will not know your identity, e-mail address, or other contact 
information unless you provide it in the body of your comment.
    ii. E-mail. Comments may be sent by e-mail to [email protected], 
Attention: Docket ID Number OPP-2004-0226. In contrast to EPA's 
electronic public docket, EPA's e-mail system is not an ``anonymous 
access'' system. If you send an e-mail comment directly to the docket 
without going through EPA's electronic public docket, EPA's e-mail 
system automatically captures your e-mail address. E-mail addresses 
that are automatically captured by EPA's e-mail system are included as 
part of the comment that is placed in theofficial public docket, and 
made available in EPA's electronic public docket.
    iii. Disk or CD ROM. You may submit comments on a disk or CD ROM 
that you mail to the mailing address identified in Unit I.C.2. These 
electronic submissions will be accepted in WordPerfect or ASCII file 
format. Avoid the use of special characters and any form of encryption.
    2. By mail. Send your comments to: Public Information and Records 
Integrity Branch (PIRIB) (7502C), Office of Pesticide Programs (OPP), 
Environmental Protection Agency, 1200 Pennsylvania Ave., NW., 
Washington, DC 20460-0001, Attention: Docket ID Number OPP-2004-0226.
    3. By hand delivery or courier. Deliver your comments to: Public 
Information and Records Integrity Branch (PIRIB), Office of Pesticide 
Programs (OPP), Environmental Protection Agency, Rm. 119, Crystal Mall 
2, 1801 S. Bell St., Arlington, VA, Attention: Docket ID 
Number OPP-2004-0226. Such deliveries are only accepted during the 
docket's normal hours of operation as identified in Unit I.B.1.

D. How Should I Submit CBI to the Agency?

    Do not submit information that you consider to be CBI 
electronically through EPA's electronic public docket or by e-mail. You 
may claim information that you submit to EPA as CBI by marking any part 
or all of that information as CBI (if you submit CBI on disk or CD ROM, 
mark the outside of the disk or CD ROM as CBI and then identify 
electronically within the disk or CD ROM the specific information that 
is CBI). Information so marked will not be disclosed except in 
accordance with procedures set forth in 40 CFR part 2.
    In addition to one complete version of the comment that includes 
any information claimed as CBI, a copy of the comment that does not 
contain the information claimed as CBI must be submitted for inclusion 
in the public docket and EPA's electronic public docket. If you submit 
the copy that does not contain CBI on disk or CD ROM, mark the outside 
of the disk or CD ROM clearly that it does not contain CBI. Information 
not marked as CBI will be included in the public docket and EPA's 
electronic public docket without prior notice. If you have any 
questions about CBI or the procedures for claiming CBI, please consult 
the person listed under FOR FURTHER INFORMATION CONTACT.

E. What Should I Consider as I Prepare My Comments for EPA?

    You may find the following suggestions helpful for preparing your 
comments:
    1. Explain your views as clearly as possible.
    2. Describe any assumptions that you used.
    3. Provide copies of any technical information and/or data you used 
that support your views.
    4. If you estimate potential burden or costs, explain how you 
arrived at the estimate that you provide.
    5. Provide specific examples to illustrate your concerns.
    6. Make sure to submit your comments by the deadline in this 
notice.
    7. To ensure proper receipt by EPA, be sure to identify the docket 
ID number assigned to this action in the subject line on the first page 
of your response. You may also provide the name, date, and Federal 
Register citation.

II. What Action is the Agency Taking?

    EPA has received a pesticide petition as follows proposing the 
establishment and/or amendment of regulations for residues of a certain 
pesticide chemical in or on various food commodities under section 408 
of the Federal Food, Drug, and Cosmetic Act (FFDCA), 21 U.S.C. 346a. 
EPA has determined that this pesticide petition contains data or 
information regarding the elements set forth in FFDCA section 
408(d)(2); however, EPA has not fully evaluated the sufficiency of the 
submitted data at this time or whether the data support granting of the 
pesticide petition. Additional data may be needed before EPA rules on 
the pesticide petition.

List of Subjects

    Environmental protection, Agricultural commodities, Feed additives, 
Food additives, Pesticides and pests, Reporting and recordkeeping 
requirements.

    Dated: August 23, 2004.
Janet L. Andersen,
Director, Biopesticides and Pollution Prevention Division, Office of 
Pesticide Programs.

Summary of Petition

    The petitioner's summary of the pesticide petition (PP) is printed 
below as required by FFDCA section 408(d)(3). The summary of the 
petition was prepared by the petitioner and represents the view of the 
petitioner. The petition summary announces the availability of a 
description of the analytical methods available to EPA for the 
detection and measurement of the pesticide chemical residues or an 
explanation of why no such method is needed.

Syngenta Seeds, Inc.

PP 4G6808

    EPA has received a pesticide petition (4G6808) from Syngenta Seeds, 
Inc., P.O. Box 12257, 3054 Cornwallis Road, Research Triangle Park, NC 
27709-2257, proposing pursuant to section 408(d) of the FFDCA, 21 
U.S.C. 346(d), to amend 40 CFR part 174 to establish an exemption from 
the requirement of a tolerance for the plant-pesticide modified Cry3A 
protein (mCry3A) and the genetic material necessary for its production 
in or on all corn.

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    Pursuant to section 408(d)(2)(A)(i) of the FFDCA, as amended, 
Syngenta Seeds, Inc., has submitted the following summary of 
information, data, and arguments in support of their pesticide 
petition. This summary was prepared by Syngenta Seeds, Inc., and EPA 
has not fully evaluated the merits of the pesticide petition. The 
summary may have been edited by EPA if the terminology used was 
unclear, the summary contained extraneous material, or the summary 
untentionally made the reader conclude that the findings reflected 
EPA's position and not the position of the petitioner.

A. Product Name and Proposed Use Practices

    A modified Cry3A Bacillus thuringiensis insect control protein and 
the genetic material necessary for its production in Event MIR604-
derived corn is proposed for use as a plant-incorporated protectant 
active ingredient. Production of the mCry3A protein within corn plants 
confers resistance to damage caused by the western corn rootworm and 
northern corn rootworm, which are major corn pests in the United 
States. A temporary exemption from tolerances is being requested in 
conjunction with a proposed Experimental Use Permit to allow large-
scale field testing of field corn plants derived from Syngenta Seeds' 
transformation event MIR604.

B. Product Identity/Chemistry

    1. Identity of the pesticide and corresponding residues. A modified 
Cry3A (mCry3A) Bacillus thuringiensis insect control protein is 
produced in transgenic corn plants derived from transformation Event 
MIR604. A cry3A gene from Bacillus thuringiensis subspecies (subsp.) 
tenebrionis was recreated synthetically to optimize for expression in 
corn. Additional changes in this corn-optimized gene were made, such 
that the encoded mCry3A protein has enhanced activity against larvae of 
the western corn rootworm (diabrotica virgifera virgifera) and northern 
corn rootworm (diabrotica longicornis barberi). Event MIR604-derived 
corn plants express the synthetic modified cry3A gene, introduced via 
transformation vector pZM26, and display resistance to these pests. The 
native Cry3A protein of Bacillus thuringiensis subsp. tenebrionis is a 
carbon absorber (ca) 73 kDa polypeptide of 644 amino acids. By 
comparison, the mCry3A protein expressed in Event MIR604 corn is a ca 
67 kDa polypeptide of 598 amino acids. Its amino acid sequence 
corresponds to that of the native Cry3A protein, except that (1) its N-
terminus corresponds to methionine-48 of the native protein and (2) a 
cathepsin G protease recognition site has been introduced into the 
protein, conferring enhanced activity toward western and northern corn 
rootworms. Residues of the mCry3A protein, and/or breakdown products 
thereof, are present in corn grain and other tissues of Event MIR604-
derived plants.
    2. Magnitude of residue at the time of harvest and method used to 
determine the residue. A determination of the magnitude of residue at 
harvest is not required for residues exempt from tolerances. However, 
the petitioner has provided data on the quantity of mCry3A protein 
measured in various plant parts. Average mCry3A levels in grain from 
Event MIR604-derived hybrid field corn plants were less than 1 part per 
million (ppm) on a dry-weight or fresh-weight basis, as measured by 
enzyme linked immunosorbent assay (ELISA). Average mCry3A levels 
measured in chopped whole Event MIR604-derived hybrid corn plants were 
less than or equal to ca. 20 ppm on a dry-weight basis and less than or 
equal to ca. 8 ppm on a fresh-weight basis.
    3. A statement of why an analytical method for detecting and 
measuring the levels of the pesticide residue are not needed. An 
analytical method is not required because this petition requests an 
exemption from tolerances. However, the petitioner has submitted an 
analytical method for detection of the mCry3A protein by ELISA.

C. Mammalian Toxicological Profile

    Syngenta Seeds is providing the results of a mammalian toxicology 
study, in vitro digestibility study, heat stability study and 
bioinformatics evaluations conducted on the mCry3A protein. These 
studies, summarized herein, demonstrate the lack of toxicity of the 
mCry3A protein following acute oral high-dose exposure to mice, rapid 
degradation of mCry3A upon exposure to simulated mammalian gastric 
fluid, instability of the mCry3A protein upon heating, and the lack of 
significant amino acid sequence homology of the mCry3A protein to 
proteins known to be mammalian toxins or human allergens.
    When proteins are toxic, they are known to act via acute mechanisms 
and at very low doses, Sjoblad, R.D., J.T. McClintock and R. Engler 
(1992) Toxicological Considerations for Protein Components of 
Biological Pesticide Products. Regulatory Toxicolology and 
Pharmacology. 15: 3-9. Therefore, when a protein demonstrates no acute 
oral toxicity in high-dose testing using a standard laboratory 
mammalian test species, this supports the determination that the 
protein will be non-toxic to humans and other mammals, and will not 
present a hazard under any realistic exposure scenario, including long-
term exposures.
    Because it is not feasible to extract sufficient mCry3A protein 
from transformed plants for toxicology studies, mCry3A protein was 
produced in recombinant E. coli by over-expressing the same modified 
cry3A gene that was introduced into Event MIR604 corn plants. Following 
purification from E. coli, dialysis and lyophilization, the resulting 
sample, designated test substance MCRY3A-0102, was estimated by ELISA 
to contain ca. 90.3% mCry3A protein by weight. Side-by-side comparisons 
of mCry3A in test substance MCRY3A-0102 with mCry3A extracted from 
Event MIR604-derived corn plants indicated that mCry3A from both 
sources is biologically active against the same target pest species, 
has the same apparent molecular weight by SDS-PAGE, immunoreacts with 
the same anti-Cry3A antibody, and is not apparently post-translation. 
Additionally, peptide mapping of 40% of the mCry3A polypeptide by mass-
spectral analysis confirmed the identity and intended amino sequence of 
mCry3A in test substance MCRY3A-0102. Nucleotide sequencing of the 
entire DNA insert in Event MIR604-derived plants also confirmed that 
the mCry3A protein produced in the plants has the intended amino acid 
sequence. These data justify the use of test substance MCRY3A-0102 in 
safety studies as a surrogate for mCry3A as produced in Event MIR604-
derived plants.
    An acute toxicity study was conducted in mice according to EPA's 
Test Harmonized Guideline OPPTS 870.1100. Test substance MCRY3A-0102 
was administered orally by gavage to five male and five female mice at 
a dose of 2,632 milligrams/kilogram (mg/kg) body weight, representing 
ca. 2,377 mg of pure mCry3A protein/kg body weight. A negative control 
group (five males and five females) concurrently received the dosing 
vehicle alone, an aqueous suspension of 1% methylcellulose, at the same 
dosing volume used for the test substance mixture. No test substance-
related mortalities or clinical signs of toxicity occurred during the 
14-day study. One female mouse in the test group was euthanized the day 
following dosing due to adverse clinical signs resulting from a dosing 
injury (confirmed by post-mortem examination). At study termination, 
macroscopic and

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microscopic examination of all major organs of the surviving mice 
revealed no treatment-related abnormalities. Body weight, body weight 
gain and organ weights (brain, liver, kidneys, and spleen) were 
comparable in the control and test groups. There was no evidence of 
toxicity. Accordingly, the LD50 value for MCRY3A-0102 in 
male and female mice is greater than 2,632 mg/kg body weight, and the 
LD50 value for pure mCry3A protein is greater than 2,377 mg/
kg body weight, the single dose tested.
    Extensive bioinformatics searches of public protein data bases 
revealed that the mCry3A protein shows no significant amino acid 
homology to proteins known to be mammalian toxins or known or suspected 
to be human allergens. Additional information and testing indicate that 
the mCry3A protein does not have properties that would suggest it has 
the potential to become a food allergen. The source of native Cry3A 
protein (Bacillus thuringiensis) is not known to produce food 
allergens. Unlike allergenic proteins, which typically are present at 
1-80% of the total protein in an offending food, the average mCry3A 
concentration measured in raw grain derived from Event MIR604 corn 
represents less than 0.0001% of the total protein. This calculation is 
based on corn grain containing 10% total protein by weight, and assumes 
less than 1 ppm mCry3A in the grain. Additionally, due to degradation 
via food processing methods, mCry3A will not likely be present in 
processed food products, or will be present in only trace quantities. 
The mCry3A protein produced in transformed corn plants is not targeted 
to a cellular pathway for glycosylation, and shows no evidence of post-
translational glycosylation. Bioactivity of mCry3A is lost upon heating 
at 95 [deg]C for 30 minutes. Upon exposure to simulated mammalian 
gastric fluid containing pepsin, mCry3A rapidly degrades.
    The native Cry3A protein has had a history of safe use as a 
component of spore preparations of the microbial insecticide Bacillus 
thuringiensis subsp. tenebrionis, as an encapsulated component of a 
microbial insecticide derived from Bacillus thuringiensis subsp. san 
diego, and as a plant-incorporated protectant in Bacillus thuringiensis 
potato.
    The genetic material occurring in the subject plant-incorporated 
protectant active ingredient has been adequately characterized. This 
genetic material (i.e., the nucleic acids DNA and RNA), including 
regulatory regions, necessary for the production of mCry3A in all corn 
will not present a dietary safety concern. ``Regulatory regions'' are 
the DNA sequences such as promoters, terminators, and enhancers that 
control the expression of the genetic material encoding the protein. 
Based on the ubiquitous occurrence and established safety of nucleic 
acids in the food supply, a tolerance exemption under the FFDCA 
regulations has been established for residues of nucleic acids that are 
part of plant-incorporated protectants in 40 CFR 174.475 (66 FR 37817 
July 19, 2001) (FRL-6057-5). Therefore, no mammalian toxicity is 
anticipated from dietary exposure to the genetic material necessary for 
the production of mCry3A protein in all corn.

D. Aggregate Exposure

    1. Dietary exposure--i. Food. Average mCry3A levels measured in 
grain from Event MIR604-derived hybrid field corn plants were less than 
1 ppm on a dry-weight or fresh-weight basis. Processed corn products or 
by-products used in food are unlikely to have measurable mCry3A 
protein, or will have only trace amounts. Oral exposure is not expected 
to result in adverse health effects, because of a demonstrated lack of 
toxicity to mammals and the rapid digestibility of the mCry3A protein. 
It is expected that any mCry3A protein consumed will be digested as 
conventional dietary protein.
    ii. Drinking water. Little to no exposure via drinking water is 
anticipated. Due to the demonstrated mammalian safety profile of 
mCry3A, such exposure would not present a risk.
    2. Non-dietary exposure. Non-dietary exposure is not anticipated, 
due to the proposed use pattern of the product. Exposure via dermal or 
inhalation routes is unlikely because the active ingredient is 
contained within plant cells. However, if exposure were to occur by 
non-dietary routes, no risk would be expected because the mCry3A 
protein is not toxic to mammals.

E. Cumulative Exposure

    Because there is no indication of mammalian toxicity of the mCry3A 
protein or the genetic material necessary for its production, it is 
reasonable to conclude that there will be no cumulative effects for 
this active ingredient.

F. Safety Determination

    1. U.S. population. The lack of mammalian toxicity at high levels 
of exposure to the mCry3A protein demonstrates the safety of the 
product at levels well above possible maximum exposure levels 
anticipated via consumption of all food commodities produced from corn 
plants that produce mCry3A. Moreover, little to no human dietary 
exposure to mCry3A protein is expected to occur via transformed corn. 
Due to the digestibility and lack of toxicity of the mCry3A protein, 
and its very low potential to become an allergen in food, dietary 
exposure is not anticipated to pose any harm for the U.S. population. 
No special safety provisions are applicable for consumption patterns or 
for any population sub-groups.
    2. Infants and children. Based on the mammalian safety profile of 
the active ingredient and the proposed use pattern, there is ample 
evidence to conclude with a reasonable certainty that no harm will 
result to infants and children.

G. Effects on the Immune and Endocrine Systems

    The active ingredient is derived from sources that are not known to 
exert an influence on the endocrine or immune systems.

H. Existing Tolerances

    The registrant is not aware of any existing tolerances or tolerance 
exemptions for mCry3A protein and the genetic material necessary for 
its production as an active ingredient.
    However, exemptions from tolerances exist for use of the native 
form of Cry3A protein as a plant-incorporated protectant in Bt potato 
(40 CFR 180.1147) and as a component of an encapsulated Bacillus 
thuringiensis microbial insecticide (40 CFR 180.1108).

I. International Tolerances

    No Codex maximum residue levels exists for the plant-incorporated 
protectant modified Cry3A Bacillus thuringiensis protein and the 
genetic material necessary for its production in corn.
[FR Doc. 04-19719 Filed 8-30-04; 8:45 am]
BILLING CODE 6560-50-S