r
r
Final
Report
Covering
the Period October
1983
to October
1984
BACTERIAL MUTATION STUDY
(U)
r
r
May
1985
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EXECUTIVE SUMMARY (U)
'
*
The experiment presented in this document was a conceptual replication of
reported work in the parapsychological literature, claiming positive
statistical evidence for
psychoenergetic interactions with biological systems.
Both the energetic and informational
aspects of human interaction with bacteriological systems were examined, with the ultimate
objective of determining,
to
first order, whether biological systems can be employed as
psychoenergetic “intrusion detectors.”
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There were two principal experimental hypotheses under consideration.
The
first,
which
will be referred to as the
Intuitive Data Sorting (IDS) hypothesis,
posits that individuals
are able to identify or “sort out" locally-deviant subsequences contained within a larger
random sequence using psychoenergetic means.
In our experiment, an IDS hypothesis
predicted that individuals would be able to identify psychically
from, a set of test tubes with a
normal
statistical spread of mutation rate—subsets of test tubes either with slightly higher or
slightly lower average mutation rates than the overall mutation rate for the entire
set.
Because
an IDS mechanism appears to be predicated on an individual’s ability to gain information
about a system psychoenergetically,
it
is thought to involve informational processes primarily.
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The second experimental hypothesis, which
will be referred to as the Remote
Action (RA) or IDS Unfavorable (IDSU)
hypothesis, postulates that certain individuals are
able to effect either a predetermined increase or decrease
in a given samples’s mutation rate
by somehow “mentally” causing physical
(e.g., genetic) changes in the bacteria.
Because an
RA mechanism appears to be predicated on an individual’s ability to effect physical changes
a system psychoenergetically,
it
is thought to involve causal or energetic processes primarily.
|j
. A total of seven subjects contributed six sessions each:
three sessions were
designed to test the IDS hypothesis, and three were designed to test the RA hypothesis.
In
in
all
sessions, the subject was confronted with nine test tubes, which were
visible inside a locked,
environmentally-stable
ice chest.
The tubes contained dilute solutions of the bacterium
Salmonella typhimurium.
The bacteriological preparations were carried out by SRI’s Microbial
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Genetics Department, which routinely uses the Ames Salmonella assay that was adapted for
use
in this study.
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In the IDS sessions, the subjects were able to choose three test tubes in which they
wished to promote the mutation rates psychoenergetically
(high aim), three tubes in which
they wished to inhibit mutation rates (low aim), and three that they wished to leave
“uninfluenced"
as controls (no aim).
In
all of the RA sessions, Tubes
1,
2, and 3 were
predetermined as the low-aim tubes
(the subject would attempt to inhibit mutation rates);
Tubes
4,
5, and 6 were the no aim controls; and Tubes
7,
8, and 9 were the high-aim tubes
(the subject would attempt to promote mutation rates).
The basic premise in comparing the
IDS and RA conditions
is that the subjects were given the opportunity to select high-versus-
low mutation rates from a natural spread of nine in the IDS sessions.
Given the
predetermined tubes of the RA sessions, however, the subjects were required to cause
physical changes in the bacteria,
in order to achieve the desired high-versus-low mutation
rates.
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The overall result of the experiment showed weak
statistical evidence that
individuals are able to sort bacteriological samples according to mutation rate—that
is, a p <
0.05 was obtained overall in the IDS sessions for the mutation rates of the low-aim
test tubes
being lower than the no-aim controls.
Statistical significance was not achieved in any of the
other IDS conditions
(i.e.,
for no-aim mutation rates being less than high aim or for low aim
being
less than high aim).
There were no significant differences for various aims observed in
the RA condition.
It must be concluded, therefore, that while there was some evidence that
subjects are able to gain information psychoenergetically about the mutation rates of
Salmonella, there was no compelling evidence that subjects are able to cause physical
perturbations in these bacteria.
*i According to criteria set forth in the beginning of this study, a physical system
will not be considered a candidate intrusion detector unless there
is clear evidence that
it
is
registering energetic effects
(i.e.. physical perturbations) concomitantly with psychoenergetic
intent.
To
first order, therefore,
it must be concluded on the basis of this one experiment
that the Salmonella bacterium does not appear to be a promising intrusion detector.
~~ Because this
is the only known experiment of
its kind using Salmonella bacteria
as the target biological system, replication
is strongly recommended—both to verify the
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OBJECTIVE (U)
[
The objective of this subtask was to determine the veracity of the claims in the
parapsychological literature regarding psychoenergetic interactions with biological systems.
A
conceptual replication of the most promising of these earlier claims was undertaken,
as a
means to examine whether biological systems register physical effects concomitantly with
psychoenergetic “intent” by an observer.
This
initial experimental effort was an attempt to
determine, to
first order, whether biological systems can eventually be employed as
psychoenergetic “intrusion detectors.”
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II INTRODUCTION (U)
One of the ultimate applications goals of psychoenergetic phenomena
is the
determination of whether psychoenergetic intrusion can be detected, and whether counter-
measures exist against such intrusion.
From a phenomenological perspective, the term
psychoenergetic intrusion can entail what appears to be either energetic or informational
processes, or both, as indicated by the following set of operative definitions:
•
The direct perturbation of physical systems that appear to be well shielded
against, or otherwise inaccessible
to, human influence
(energetic).
•
The psychoenergetic acquisition of information thought to be secure against
access (informational).
•
The perturbation of a physical system that occurs indirectly as a result of an
individual’s attempts to acquire information through psychoenergetic means
(energetic and informational).
Only those intrusions that entail causal interactions with physical systems are likely to be
detected.
A physical system
will not be considered a candidate intrusion detector, therefore,
unless
it registers energetic effects directly
(as a
result of intentional perturbation), or
indirectly
(as a result of concomitant acquisition of information).
• In the parapsychological literature, the energetic manifestations of psycho-
energetic intrusion are variously referred to as remote action (RA), remote perturbation (RP),
psychokinesis (PK),
telekinesis (TK), and so forth; informational processes are most often
referred to as remote viewing (RV), clairvoyance, precognition, and the
like.
The term
countermeasures may be defined as the shielding or jamming of psychoenergetic intrusion by
either physical or mental processes.
i
Before the higher-order problem of countermeasures can be addressed,
experimental verification of the existence of psychoenergetic intrusion must
first be obtained.
Detection of the putative energetic aspects of psychoenergetic intrusion can be accomplished
most directly by designing experiments in which an individual’s primary task
is to actively
attempt to cause perturbations in various types of physical systems.
Numerous RA
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with medium containing histidine), and selective minimal glucose plates
(i.e., plates with medium lacking histidine)
to allow for the growth and
appearance of bacterial colonies.
2.
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Procedural Terms
\
(
The following are the most common procedural terms:
•
Subject—One of seven volunteers who undertook to psychoenergetically
influence the mutagenicity of the bacterial samples.
•
Monitor—The individual recording the events that transpired during an
experimental session, and supervised the subject’s activities.
•
Technician—The microbiologist who was responsible for
all aspects of the
pre- and post-session preparation of the biological samples.
•
Session—A single sitting in which the subject attempted to
(1) increase the
mutation rate of bacteria placed in three
test tubes,
(2) decrease the
mutation rate of those placed in three different test tubes, and
(3) leave yet
a different group of three uninfluenced as “controls."
Each of the seven
subjects contributed six such sessions.
•
Trial—An attempt by a subject to psychoenergetically influence
(or not
influence, as in the case of control test tubes) the bacterial culture
in a
single test tube.
There were nine such
trials in each experimental session.
•
Controls—Two types of controls were employed in this experiment:
intrasession and extrasession.
Intrasession control test tubes consisted of
three bacterial test tubes, which the subject was instructed not to attempt to
actively influence, from among the set of nine session test tubes.
Extra-
session controls consisted of two tubes per session that were prepared by the
technician in exactly the same manner as the session test tubes, but were
not used as part of the experimental session set of nine tubes.
The
extrasession controls remained at
all times in the Microbial Genetics
Laboratory, and provided the requisite data for establishing an independent
measure of mutation rate.
•
Feedback—A drawing presented to the subject that indicated his/her
performance on a given session.
Feedback for a given session was typically
administered prior to the start of the subject’s next session.
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B.
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Biological Background
V
In this section, we
will give a general overview of the Ames Salmonella assay
that
is used routinely by SRI’s Microbial Genetics Department, and that was adapted
for use
in
this experiment to study psychoenergetic effects on mutation frequency.
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The heritable material of living organisms
is contained in the DNA (RNA in some
viruses)
, a large molecule so constated that
1, can replicate
itseif In a most exact fashion one
cell generation after another.
This
is the basis of biological continuity and unity.
It
,s also,
however, the basis for biological diversity, which occurs through mutations.
Each mutation
alters the action of a specific gene, which
is a genetic entity with
its own specific end product,
or protein.
Genes are very stable structures, but each has
its own spontaneous mutation
frequency.
The probability that a spontaneous mutant cell will be obtained every time a cell
divides
is constant, provided the environmental conditions are unchanged.
Changes in the
environment are known to influence the mutation Irequency.
Such changes include the
presence or absence of certain trace elements
(e.g.. selenium), plus the presence of physical
or chemical agents (mutagens).
V
Bacterla provide a convenient way to study mutations because millions of cells
can be"grown in a very short period of time.
Over the past few years, several bacterial assays
have been developed to screen chemicals for their ability to induce mutation.
Because there
is a close correlation between mutagenesis and carcinogenesis, such mutagenicity assays are
very often used together with the in vilro tests that employ single microbial andlor mammalian
cells, as well as in vivo tests that employ mullicell organisms from insects
(fruit
fly)
to
mammals (rodents).
One of the best known bacterial mutagenesis assays
is the Salmonella!
mammalian microsome histidine reverse mutation assay developed by Dr. Bruce Ames at the
University of California in Berkeley.
The Microbial Genetics Department a. SRI International
is using this assay system on a daily basis lor Government agencies and commercial clients to
determine the mutagenic potential o[ chemicals; they have performed such testing over a
period of more than 10 years.
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The Salmonella assay employs several tester strains ol Salmonella iyphimurium,
each with a unique sped!, city for detecting chemical mutagens.
The Salmonella strains, under
optimum conditions, have a generation time of less than 30 minutes.
The bacterial strains are
unable to grow in the absence ol the essential amino acid histidine because of a mutation ,n
one o! the genes that
is needed
lor histidine synthesis.
When these bacteria are plated on
defined selective medium having
little or no histidine,
little or no growth occurs except those
few bacteria that spontaneously mutate back to histidine independence
(ability to grow ,n the
absence ol histidine).
In this case,
a nonfunctional gene product
is reverted back to a
functional one.
This event allows the mutant cells to grow and divide.
Because a mutation
,s
stably inherited,
all progeny ol the mutated cells retain the
ability to grow in the absence of
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histidine.
Distinct individual colonies
will appear on the solid selective growth medium, with
each colony containing billions of progeny of the spontaneously mutated
cells.
Exposure of
the bacteria to a chemical mutagen
will result in an increased number of colonies appearing
on the solid selective growth medium, due to an increase in mutation induction.
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In the Ames Salmonella assay,
a small amount of histidine
is added to the growth
medium to allow for a few cell divisions of
all the plated histidine requiring mutants (~10*).
Such growth
is often necessary for chemical mutagenesis to occur.
The results of the
Salmonella assay are usually expressed in terms of the number of revertant colonies per
amount of chemical added to the selective growth medium, which
is usually delivered to the
plate
in 25-ml volumes.
Because of the presence of limited histidine in the selective medium,
the
results of the Ames Salmonella are considered "semiquantitative," since residual growth
on
all plates
(control as well as chemical treated) does not allow for quantitative survival
determination.
A quantitative mutation frequency, however, can be determined.
It
is more
labor intensive than the standard Ames assay, because survival determination requires diluting
of the
cell cultures, and a different growth medium
is needed for determining
(1) the mutant
fraction and survivors for each of the controls, and
(2)
the different exposure concentrations
of the test chemical.
The mutation frequency
is defined
in terms of number of mutants per
given number of surviving cells, usually per
10e
cells.
Because of
its simplicity and the rapid response time of about two days, the
Ames Salmonella assay can readily be adapted to study the effect of RA on the mutation
frequency.
Such an adaptation was established by SRI’s Microbial Genetics Department
for
use
in this experiment; a detailed discussion of the specific biological procedures that were
followed can be found in Section D, Protocols.
C.
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Experimental Design
I.
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Conceptual Replication
The experiment undertaken in
this study represents a conceptual replication
'is
:m
of the Nash experiment described in our Introduction chapter.
The replication presented here
is termed conceptual, because several of the experimental details of the Nash experiment
have been changed and improved.
First, two potential mechanisms have been postulated that
could account for the acquisition of a statistically significant effect—that
is, an IDS hypothesis
has been advanced,
in addition to the more established RA hypothesis.
Second, Salmonella
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typhimurium rather than Escherichia coli were used as the target bacterial cultures.
Because
this particular species of Salmonella
is used most frequently by SRI’s Microbial Genetics
Department in toxicity studies,
its behavior
is particularly well understood in terms of assay
conditions and experimental protocols.
Finally, the Nash analysis was extended to include
multiple analyses of variance.
f As in the Nash experiment, nine test tubes
filled with dilute bacterial
culture were used per session.
Mutation from histidine dependence to histidine independence
was mentally promoted by the subject in three of the tubes, mentally inhibited in three, and
the remaining three tubes served as controls.
For the purposes of obtaining baseline data, two
additional control
test tubes
(for a
total of eleven altogether per session) were prepared in the
same manner as the session test tubes, but were kept in the Microbial Genetics Laboratory.
2.
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Model Testing Criteria
a.
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The IDS Model
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As mentioned previously, there were two primary models under
investigation in this experiment.
A pivotal concept to the
first, or IDS favorable model,
is
freedom of choice: namely, that by using some type of psi-mediated informational processes,
subjects have the opportunity to select out locally-deviant subsequences from a larger random
sequence.
For example,
in half of the sessions, the subjects were allowed to select the three
test tubes in which they wished to promote mutation, and the three test tubes in which they
wished to inhibit mutation.
A
statistically significant deviation from mean chance expectation
(MCE)
in this condition, therefore, could be interpreted theoretically in two ways:
(1)
the
subjects somehow mentally “forced” genetic changes to occur in the bacteria
in accordance
with their desires to either promote or inhibit mutation rates
(the RA hypothesis); or
(2) given
the natural spread of mutation rates in a biological system, the subject was able to psycho-
energetically sort those test tubes containing bacteria with high mutation rates from those
tubes containing bacteria with low mutation rates
(a session-by-session IDS hypothesis).
b.
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The RA or IDSU Model
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The second model under investigation has been termed the Remote
Action (RA) or Intuitive Data Sorting Unfavorable (IDSU) model.
In this condition, the
conduits by which either the subject or experimenter are able to select test tubes are rendered
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