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Federal Register / VoL 66. 'No. 138 / Thursday. July 18, 19^ / Notices 
/Appendix K. Footnotes 
1. This table is derived from the text in 
Appendices G and K and is not tn be oaed in 
lieu of Appendices G and K. 
2. The criteria in this grid address only the 
biological hazard assented with organisms 
,! containing recombinwt DNA. Other hazards 
j accompanying the large scale ciiltlvation of 
I such organisms (e^J, toxic properties of 
products; physic^ mechanical and chemical 
I aspects of downstream processing) are not 
I addressed and must be considered, 
j separately, 'albeit in conjunction with this 
'll 
l| Appendix K. Definitions to Accompany 
1 Containment Grid and Proposed 
j Modification of ^pen^ K 
I Accidental release-^^^e . 
, pnintentioiial discharge of a 
j microbiological agent (Le,' 
I microoiganism or virus) or eukaryotic 
cell due to a failure in the containment 
,1 system. \ 
Biological barrier — An impediment, 
(naturally occurring or introduced] to 
the infectivlty and/or 'survival of a" 
microbiological agent or eukaryotic 'cell 
I once it has been i^eased into the 
j environment 
‘j Closed system — system which by 
■ij its design and proper operation. 
^ I prevents release of a microbiological 
I agent or eukaryotic cell contained 
li therein. 
Containment — The confinement of a 
microbiological agent or eukaryotic cell 
that is being cultured, stored, 
manipulated, transported or destroyed 
I in order to prevent or limit its contact 
i with people and/or the environmenL 
I Methods used to achieve this include: 
physical and biological barriers and 
inactivation using physical or chemical 
means. 
de minimis release — A release of 
viable microbiological agents or 
eukaryotic cells that do not result in the 
establishment of disease in healthy 
> people, plants or animals or in 
uncontrolled proliferation of any 
microbiological agents or eukaryotic 
cells. 
Disinfection — A process by which 
viable microbiological agents or 
eukaryotic cells are reduced to a level 
unlikely to produce disease in healthy 
people, plants or animals. 
I Good Large Scale Practice {GLSP) 
Organism — For an organism to qualify 
for GLSP consideration, it must meet the 
following criteria: (Reference: 
I Organization for Economic Cooperation 
and Development, Recombinant DNA 
Safety Considerations, 1987, p. 34-35). 
a. The host organism should be non- 
pathogenic, should not contain 
adventitious agents and should have en 
I extended history of safe large-scale use 
: or have built-in environmental 
limitations that permit optimum growth 
in the large-scale setting but limited 
survival without adverse consequences 
in the environment. - . . 
b. The recombinant DNA-engineered 
organism should be non-pathogenic, 
should be as safe in the large-scale 
setting as the host organism, end 
Without adverse consequences in the 
environment . 
c. The vector/insert should be well 
characterized and free from known 
harmfiil sequences; should be limited in 
size as mu(± as possible to the DNA 
-required to perform the intended 
function; should not increase the 
stabib^ of the construct in the 
environm^t unless that Is a 
requirement of the intended function; 
should be poorly mdbilizable; and 
should not transfer any resistance 
niarkers to microorganisms not known 
to acquire them naturally if such 
acquisition could comproi^e the use of 
a drug to control disease agents in 
human or veterinary medibine or 
agriculture. 
Inactivation — Any process .that 
destroys the ability of a specific 
microbiological agent dr eukaryotic cell 
'to self-replicate. 
Incidental release — The discharge of 
a microbiological agent or eukaryotic 
cell from a containment system that is 
expected when the system is 
appropriately designed and properly 
operated and maintained. 
Minimization — The design £ind 
operation of containment systems in 
order that any incidental release is a de 
minimis release. 
Pathogen — ^Any microbiological agent 
or eukaryotic cell containing sufficient 
genetic information, which upon 
expression of such information is 
capable of producing disease in healthy 
people, plants or animals. 
Physical barrier — Equipment, 
facilities and devices (e.g., fermentors, 
factories, filters, thermal oxidizers) 
designed to achieve containment 
Release — ^The discharge of a 
microbiological agent or eukaryotic cell 
from a containment system. Discharges 
can be incidental or accidental. 
Incidental releases are de minimis in 
nature; accidental releases may be de 
minimis in nature. 
B. Addition of Appendix D-XVII to the 
"NIH Guidelines " 
The foUov/ing section is added to 
Appendix D: 
Appendix D-XVII 
Dr. Malcolm K, Brenner of St Jude. 
Children's Research Hospital of 
Memphis, Tennessee, can conduct 
experiments on patients with acute 
myelogenous leukemia (AML). Using the 
LNL6 retroviral vector, the autologous 
bone marrow cells will be transduced 
with the gene coding for neomycin 
resistance. The purpose of this gene 
marking experiment is to determine 
whether the source of relapse after 
autologo.us bone marrow transplantation 
for acute myelogenous leukemia is 
residual malignant cells in the harvested 
marrow or reoccurrence of tumor in the 
patient Determining the source of 
relapse should indicate whether or not 
purging of the bone marrow is a 
necessary procedure. 
C Addition of Appendix D-XVIIl to the 
•^NIH Guidelines- 
The following section is. added to 
Appendix D. 
Appendix D-XVIII 
Dr. Malcom IC Breimer of St Jude 
Children’s Research Hospital of 
Memphis, Tennessee, can conduct 
experiments on pediatric patients with 
Stage D (disseminated] neuroblastoma 
who are being treated with high-dose 
carboplatin and etoposide in either 
phase I/n or phase II trials. All the 
patients in these studies will be 
subjected to bone marrow . 
transplantation since it will allow them 
to he exposed to chemoradiation that 
would be lethal were it not for the 
availability of stored autologous marrow 
for rescue. 
The bone marrow cells of these 
patients will be transduced with the 
gene coding for neomycin resistance 
using the LNL6 vector. The purpose of 
this gene marking study is to determine 
whether the source of relapse after 
autologous bone marrow transplantation 
is residual malignant cells in the 
harvested marrow or residual disease in 
the patient Secondly, it is hoped to 
determine the contribution of marrow 
autographs to autologous reconstitution. 
D. Addition of Appendix D-XDC to the 
-NIH Guidelines” 
The following section is added to 
Appendix D; 
Appendix D-XIX 
Dr. Albert B. Deisseroth of the MD - 
Anderson Cancer Center of Houston, 
Texas, can conduct experiments on 
patients with chronic myelogenous 
leukemia who have been reinduced into 
a second chronic phase or cytogenetic 
remission after accelerated phase or 
blast crisis. The patients in these studies 
will receive autologous bone marrow 
transplantation. Using the LNL6 vector, 
the bone marrow cells will be 
transduced with the gene coding for 
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Recombinant DNA Research, Volume 14 
