BACTERIAL MUTATION STUDY

• Document Type: CREST
• Collection: STARGATE
• Document Number (FOIA) /ESDN (CREST): CIA-RDP96-00789R003800290001-9
• Release Decision: RIFPUB
• Original Classification: U
• Document Page Count: 42
• Document Creation Date: November 4, 2016
• Document Release Date: October 27, 1998
• Sequence Number: 1
• Publication Date: May 1, 1985
• Content Type: RS

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11
Final Report
Covering the Period October 1983 to October 1984
E~~
~IIJ 1 BACTERIAL MUTATION STUDY
By: BEVERLY S. HUMPHREY EDWIN C. MAY
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Approved by.
ROBERT S. LEONARD, Director
Radio Physics Laboratory
DAVID D. ELLIOTT, Vice President
Research and Analysis Division
d For Re FdAs 2t O'0f087140u-e ClJgRDP916 00 350029001-9
[phone] ? Cable: SRI INTL MPK ? TWX: 910-373-2046
opy No . .............
This document consists of 42 pages.
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CONTENTS
LIST OF ILLUSTRATIONS . . . . .
.
LIST OF TABLES . . . . . . . . . . . .
. iii
EXECUTIVE SUMMARY . . . . . . . . . . . . . . . . . . . . . . 1
I OBJECTIVE . . . . . . . . . . . . . . . . . . . . . . . . 4
II INTRODUCTION . . . . . . . . . . . . . . . . . . . . . . 5
III METHODS OF APPROACH . . . . . . . . . . . . . . . . . . 7
A. Definition of Terms . . . . . . . . . . . . . . . . . . . . 7
1. Biological Terms . . . . . . . . . . . . . . . . . . . 7
2. Procedural Terms . . . . . . . . . . . . . . . . 8
B. Biological Background . . . . . . . . . . . . . . . . . . . 8
C. Experimental Design . . . . . . . . . . . . . . . . . . . . 10
1. Conceptual Replication . . . . . . . . . . . . . . . . . 10
2. Model Testing Criteria . . . . . . . . . . . . . . . . . 11
a. The IDS Model . . . . . . . . . . . . . . . . . . 11
b. The RA or IDSU Model . . . . . . . . . . . . . . 11
D. Protocols . . . . . . . . . . . . . . . . . . . . . . . . 13
1. Preexperiment Protocols . . . . . . . . . . . . . . . . 13
a. Subject Selection . . . . . . . . . . . . . . . . . 13
b. Experiment Site Locations . . . . . . . . . . . 14
c. Hardware Construction . . . . . . . . . . . . . . . 14
2. Presession Protocols . . . . . . . . . . . . . . . . . . 15
3. Session Protocols . . . . . . . . . . . . . . . . . . . 15
4. Postsession Protocols . . . . . . . . . . . . . . . . . 19
IV RESULTS AND DISCUSSION . . . . . . . . . . . . . . . . . 24
V . SUGGESTIONS.FOR FUTURE STUDIES . . . . . . . . . . . . . 36
A. The Role of Feedback . . . . . . . . . . . . . . . . . . . 35
B. Screening Criteria and Presentation of Psychoenergetic Task . . . . 36
C. Biological Protocols . . . . . . . . . . . . . . . . . . . . 37
D. Future Experiments with Other Biological Systems . . . . . . . . 37
1. Single-Cell Systems . . . . . . . . . . . . . . . . . . 37
2. Multicell Systems . . . . . . . . . . . . . . . . . . . 38
Appendix--BIOLOGICAL MATERIALS . . . . . . . . . . . . . . . . ' 39
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ILLUSTRATIONS
1 Sample Bio-PK Form . . . . . . . . . . . . . . . . . . . . . .
2 Sample Bacterial Assay Form . . . . . . . . . . . . . . . . . . .
3 Sample Feedback Form . . . . . . . . . . . . . . . . . . . . .
4 Distribution of Mutation Probability for 187 Baseline Test Tubes.
TABLES
1 Normalized Mutation Rates X
2 Normalized Mutation Rates X
3 Normalized Mutation Rates X
4 Normalized Mutation Rates X
5 Normalized Mutation Rates X
6 Normalized Mutation Rates X
7 Normalized Mutation Rates X
10-6 (Subject
10-6 (Subject
10-6 (Subject
All Subjects . . . . . . . . . .
10-6 (Subject
10-6 (Subject
10-6 (Subject
8 Normalized Mutation Rates x 10-6 (Subject
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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."
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.
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 in
a system psychoenergetically, it is thought to involve causal or energetic processes primarily.
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 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.
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.
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 C
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.
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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robustness of the IDS capability, and to evaluate definitively the efficacy of using Salmonella
as an intrusion detector.
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I OBJECTIVE
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
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.
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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experiments of this type, using a wide variety of physical systems, have been cited in the
parapsychological literature.
One category of candidate target physical systems is biological systems; the
precedent for using these in RA experiments has been well established. Of particular interest
(because of its similarity to the experiment detailed in this document) is Carroll B. Nash's
experiment involving the psychokinetic control of bacterial mutation. * The published abstract
of the Nash experiment is provided here:
Three experimenters each tested 20 subjects not known to be psychically
gifted. Because of procedural errors, results were obtained for only 52
subjects. Each subject was tested in a single run with a separate set of
nine tubes of a mixed culture of lac-negative and lac-positive strains of
Escherichia coli. Mutation of lac-negative to lac positive was mentally
promoted in three of the tubes, mentally inhibited in three, and three of
the tubes served as controls. The mutant ratio of lac positive to total
bacteria was greater in the promoted than in the inhibited tubes, with
two-tailed p < 0.005; less in the inhibited tubes than in the controls, with
two-tailed p < 0.02; and greater in the promoted tubes than in the
controls, although not significantly so. The results are interpreted to
suggest that the rate of bacterial mutation was psychokinetically affected.
The experiment described in this report also undertook to investigate
psychokinetic influence on bacterial mutagenicity, but it differs significantly from the Nash
experiment in certain of its experimental protocols and underlying theoretical assumptions.
The overall objective was also different than that of the Nash experiment in that the SRI
study is concerned with providing a "first order" examination of the existence of psycho-
energetic intrusion detection with biological systems, as a precursor to investigating the
necessity and/or feasibility of countermeasures against such intrusion.
Nash, C. B., "Psychokinetic Control of Bacterial Growth," Journal of the Society for
Psychical Research, Vol. 51, pp. 217-226 (1982).
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III METHODS OF APPROACH
In this chapter, we present a few concepts that are preparatory to a discussion of
the actual protocols used in our experiment. The preliminary or background concepts are
presented in two separate sections: "Definition of Terms," and "Biological Background."
The third section, "Experimental Design," provides an overview of the experiment in terms of
how it was designed to address the proposed theoretical concerns.* The fourth section,
"Protocols," is a detailed summary of all presession, session, and postsession procedures.
In order to provide a framework for discussing the essential components of this
experiment, working definitions of the most salient and most frequently encountered
biological- and protocol-related terminology are provided below.
1. Biological Terms
The following are the most common biological terms:
? Bacteria--Microscopic, unicellular organisms used as the target biological
system in this experiment. The specific species employed was Salmonella
typhimurium, a short, rod-shaped bacterium that is actively motile.
? Histidine--An amino acid essential for the growth of Salmonella
typhimurium.
? Mutation--A genetic change occurring in a small subset of Salmonella that
allows them to grow and divide in the absence of histidine. Such cells are
termed mutant cells or revertants.
? Mutation Frequency--Number of mutant cells per total number of
cells--normalized, for comparison purposes, to number of mutants per
million cells.
? Plating--A process whereby diluted samples of the bacterial cultures in the
experimental test tubes were placed on complete medium plates (i.e., plates
Our theoretical considerations have been fully addressed in the "Executive Summary."
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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. (U) 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.
B. Biological Background
'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 constructed that it can replicate itself in a most exact fashion one
cell generation after another. This is the basis of biological continuity and unity. It is 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 frequency. Such changes include the
presence or absence of certain trace elements (e.g., selenium), plus the presence of physical
or chemical agents (mutagens).
Bacteria 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 vitro tests that employ single microbial and/or mammalian
cells, as well as in vivo tests that employ multicell 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 at SRI International
is using this assay system on a daily basis for Government agencies and commercial clients to
determine the mutagenic potential of chemicals; they have performed such testing over a
period of more than 10 years.
The Salmonella assay employs several tester strains of Salmonella typhimurium,
each with a unique specificity 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 of the essential amino acid histidine because of a mutation in
one of the genes that is needed for 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 in the
absence of 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 is
stably inherited, all progeny of 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.
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 (-v10*).
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 106 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. Experimental Design
1. Conceptual Replication
The experiment undertaken in this study represents a conceptual replication
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.
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. Model Testing Criteria
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. The RA or IDSU Model
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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as inaccessible as possible by the experimental protocols, such that the most direct
explanation of a potentially significant effect is a remote-action mechanism. For the
experiment presented here, the test tubes were fixed and predetermined as to aim for half of
the sessions--that is, Test Tubes 1, 2, and 3 were a priori assigned as those tubes in which
mutation rates were to be inhibited (low aim), Tubes 4, 5, and 6 were designated as controls
(no aim), and Tubes 7, 8, and 9 were assigned as those in which mutation rates were to be
promoted (high aim). In this way, the subject's potential IDS ability to sort tubes psycho-
energetically according to the natural spread of mutation rates was precluded in half of the
sessions.
A second possibility also had to be accounted for--namely, that experi-
menters, who are desirous of a certain outcome to the experiment, might use their own IDS
ability to psychically scan the future. They could thus assign the IDSU sessions to days on
which, for whatever reason (e.g. systematic sequential pipetteing bias in preparation of the
cultures), Tubes 1, 2, and 3 might possess, on average, naturally lower mutation rates than
Tubes 7, 8, and 9. To preclude this possibility, a balanced binary random protocol, with
certain states disallowed, was formulated to determine session sequence. The disallowed
sequences included (1) three RA sessions followed by three IDS sessions ("aaabbb"); (2)
three IDS sessions followed by three RA sessions ("bbbaaa"); and (3) alternating IDS and RA
sessions ("ababab" or "bababa"). The six-session sequence that was actually generated was
RA, RA, IDS, RA, IDS, IDS (i.e., "aababb"), a protocol that was fixed for all subjects prior
to the start of the experiment.
The final possibility that had to be considered was the degree to which the
biological technician should be allowed to know the overall purpose of the experiment, in
general, and to know specific experimental protocols, in particular. It was hypothesized that
the technician's knowledge concerning experimental goals could potentially introduce his (the
technician's own) IDS ability into the experiment as an undesirable variable. For example,
had the technician been cognizant of the aims of the IDSU condition, it is possible that he
could have used his own IDS capabilities either consciously or subconsciously to position the
test tubes in the rack, such that Tubes 7, 8, and 9 possessed the high mutation rate cultures,
and Tubes 1, 2, and 3 the low mutation rate cultures. Rather than precluding or, at the very
least, greatly inhibiting IDS channels (in accordance with the original aim of its design) the
IDSU condition would have created a new, unintended conduit by which IDS could operate
directly. To preclude the potential introduction of this third possible IDS variable into the
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experiment, the technician was kept blind as to all aspects of experimental goals and protocols
until the conclusion of the experiment.
Given the caveats introduced by the experimental design considerations
detailed above, a statistically significant deviation from MCE in the RA or IDSU condition
could be interpreted, theoretically, as follows: (1) the subject somehow mentally "forced"
genetic changes to occur in the bacteria in accordance with his desires to promote mutation in
Test Tubes 7, 8, and 9, and inhibit mutation in Tubes 1, 2, and 3 (the RA hypothesis); or
(2) IDS is able to operate in a goal-oriented mode, such that either the experimenter or the
technician or both were able to adjust their actions in "real time" based on their global scan
of a successful outcome of the experiment in the future (the global IDS hypothesis). To cite
a specific example, if, at the conclusion of the experiment, the technician were finally told the
purpose of the experiment, and that the result had been significant in the IDSU condition,
there exists the following rather circuitous pathway by which his global IDS ability might be
operative: (a) the technician could continually. scan his future and determine that by pipetteing
the cultures a certain way in real time, the IDSU condition would not show overall
significance (that is, future "a" determines action "a" in real time); (b) he might discover,
however, that by adjusting his real-time pipetteing strategy on certain sessions (e.g., by placing
more dilute culture in Tubes 1, 2, and 3 as opposed to Tubes 7, 8, and 9), the IDSU
condition would become significant overall (future "b" determines action "b" in real-time).
It should be noted that the concept of global IDS is predicated on an "alternative futures"
model, and it cannot be determined at present whether such a model possesses any validity.
In any event, given the torturous nature of the pathway that must be postulated for global IDS
to occur, we believe that a statistically significant result in the IDSU condition would be
explained most parsimoniously by the RA hypothesis.
D. Protocols
1. Preexperiment Protocols
a. Subject Selection
A total of seven subjects were chosen primarily on the basis of their
expressed interest in the field of psychoenergetics. None of the seven participants had ever
taken part in a psychoenergetic experiment involving biological systems, and three of the
group were true neophytes--having never participated in any type of psi experiment. Four of
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the seven had been participants in previous psychoenergetic experiments--i.e., all four had
demonstrated some ability in remote viewing. One of these four subjects also scored
significantly in an earlier SRI random number generator PK experiment, and another had
demonstrated some ability previously in Computer-Assisted Search (CAS) tasks. All of the
participants were SRI employees: one was a statistician, two were secretaries, and the
remainder were research professionals in either physics or computer science.
b. Experiment Site Locations
For reasons stated in Section C.2.a, it was determined that the
biological technician should be kept entirely blind as to all facets of the experiment, and that,
in order to facilitate this situation, the psychoenergetic testing should occur in a location that
was different from the one used for the biological preparations. The Microbial Genetics
Laboratory, therefore, was used for all aspects of preparation of the biological cultures, and a
room in another building at SRI was used for the psychoenergetic sessions.
'Once it had been determined that two separate facilities were
necessary for conducting the experiment, a container had to be constructed that was suitable
for transporting the biological samples from the Microbial Genetics Laboratory to the
psychoenergetics facility. There were three primary factors that dictated the design of the
container: (1) the biological samples had to be protected from extreme variations in
temperature; (2) the samples had to be protected from sunlight; and (3) the container had to
be lockable.
To control against extreme variations in temperature, which can greatly
affect the mutagenicity of Salmonella, a Coleman? ice chest was chosen as the transport
container. Triple-paned insulated glass windows were specially installed in the top and front
side of the ice chest to allow an unobstructed view of the experiment test tubes. Because
sunlight also affects the mutagenicity of the bacteria, a tarpaulin was used to completely cover
the cooler during transport between the biological laboratory and the psychoenergetics facility.
A lock was installed; the key was retained exclusively by the biological technician, to preclude
the possibility of tampering with the biological samples once they had been removed from the
Microbial Genetics Laboratory.
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A series of activities took place prior to the start of every experimental session.
First, the experiment monitor identified the session type from the fixed session sequence,
"aababb," (cf. Section C.2.b), to determine whether the session would be an IDS favorable or
an IDS unfavorable (RA or IDSU) condition.
Second, the technician in the Microbial Genetics Laboratory prepared the
bacterial cultures for the session (see Appendix). Eleven numerically-labelled, sterile,
16-X-150-mm test tubes were aseptically filled with 2.5 ml of glucose minimal broth. Fifty
?l of a 37?C overnight culture of strain TA100 of Salmonella typhimurium was then added to
each tube.* In a standardized manner, the first nine tubes were arranged in a test-tube rack,
which was placed in the specially designed ice chest, and then locked. The remaining two
control cultures were shielded from visible light by a covering of aluminum foil, and were
maintained at room temperature in the Microbial Genetics Department laboratory.
The ice chest, with its enclosed cultures, was placed on a cart and covered with
a tarpaulin to ensure that the mutation rate of the cultures was not affected by sunlight during
transportation from the laboratory to the experimental facility.
Third, the experiment monitor transported the covered ice chest on the cart
from the biological laboratory to another facility at SRI, where the psychoenergetic portion of
the experiment was performed. Prior to the arrival of the subject, the monitor wheeled the
ice chest into a room equipped with a table and two chairs. The ice chest was then
uncovered and positioned in such a way that a seated subject could readily view the nine
numbered test tubes through the glass.
3. Session Protocols
For other than the first session for each subject, a session usually commenced
with feedback to the subject of the previous session's results (to be discussed in "Postsession
* (U) It should be noted that there was no visible evidence of "cloudiness" caused by the
bacterial culture in any of the prepared test tube solutions. The appearance of the liquid
was uniformly that of clear tap water. Thus, there were no visual cues available to the
subject, as to which test tubes might contain greater amounts of the bacterial culture.
15
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Protocols" below). After the presentation of feedback, the subjects were informed whether
they would select the test tubes they wished to influence (IDS favorable), or whether the tubes
had been predetermined as to aim by the protocol (IDS unfavorable) for the current day's
session.
For the IDS favorable session, the subjects were specifically instructed to select
three test tubes that they thought were most susceptible to a decrease in mutation rate, three
most susceptible to an increase in mutation rate, and three that they wished to leave
uninfluenced as controls. It should be emphasized that in all cases where we use the term
"select," we mean that the subjects simply indicated to the monitor the number of the test
tube they were going to attempt to influence. At no time did the subjects have physical
contact with the test tubes, because they were in the locked ice chest--to which the biological
technician, alone, possessed the key.
The subjects were informed by the monitor that the set of nine test tubes, as
with all biological systems, represented a natural 'spread of mutation rates, and that their task
would most likely be facilitated if they tried to mentally influence the tubes in the direction
they were already naturally inclined. A typical experimental session proceeded as follows:
? The subject chose whether he/she wished to begin the session with either
the high-aim or low-aim condition.
? Having decided upon the condition, the subject was directed first to "select"
(i.e., psychoenergetically identify) a test tube that already exhibited a
mutation rate in the direction of the chosen condition, and then to mentally
"promote" it (i.e., via some type of Remote Action mechanism) in that
direction. For example, in the low-aim condition, a subject would be
encouraged to "psychically scan" the session set of nine test tubes, and to
select one that was predisposed toward producing a low mutation rate. The
subject then attempted to mentally influence the biological culture in the
tube, in an effort to further inhibit the mutation rate.
? For a given condition, a subject selected a tube and attempted to influence
it for as long as he/she deemed necessary (usually 30 to 60 seconds),
selected a second tube to concentrate on, then selected a third tube and
concentrated on it. The strategies employed in the attempts at influencing
the cultures were left to the subject's discretion. In the low-aim condition,
for example, several subjects envisioned or "willed" that the Salmonella
were dying, or would starve to death once they were plated in a histidine-
free medium. In the high-aim condition, several subjects reported that they
envisaged the bacteria dividing and multiplying at an extremely rapid rate,
or that they were changing somehow into a new strain of bacteria that grew
well in the absence of histidine.
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? The subjects were encouraged to complete their efforts with all three test
tubes in a given condition before moving on to the other condition, so as to
avoid dividing their attention between the selection of high- and low-aim
test tubes, and to maintain continuity of imagery strategies during the
"influence" phases. The subjects were also admonished to avoid excessive
strategy searching during the influence phase, in order to ensure that their
attention would be focused on the actual task at hand rather than on the
strategy employed.
? After having finished with three tubes in one condition, the subject was
encouraged to take a brief rest. The same selection and concentration
process was then carried out with three test tubes of the subject's choice in
the other condition.
For every IDS unfavorable session, the protocol dictated a priori that Test
Tubes 1, 2, and 3 (the first three tubes on the left as the subject faced the ice chest) were
"low aim," Tubes 4, 5, and 6 were "no aim" or controls, and Tubes 7, 8, and 9 (the last
three tubes on the right) were "high aim." The subject was specifically instructed to
"mentally cause" a decreased mutation rate in. Tubes 1, 2, and 3 and an increased mutation
rate in Tubes 7, 8, and 9, and not to attempt to actively influence the mutation rates of
Tubes 4, 5, and 6. A typical experimental session proceeded as follows:
? As in the case of the IDS favorable session, the subject chose whether
he/she wished to begin the session with either the high-aim or low-aim
condition.
? In the low-aim condition, the subject attempted to inhibit the mutation rates
in Test Tubes 1, 2, and 3, one at a time and in the order of his choosing.
Again, the subject determined the time of "concentration effort" per tube,
which lasted from approximately 30 to 60 seconds. In the high-aim
condition, the subject endeavored to increase the mutation rates in Tubes 7,
8, and 9, one at a time and in the order of his/her choosing. The
strategies employed in these attempts at mentally influencing mutation rates
were the same as those discussed for the IDS favorable session above.
? As in the IDS favorable condition, the subject was directed to complete the
effort with all three test tubes in one condition before shifting attention to
the other condition; a rest period between the two conditions was also
encouraged.
The monitor's principal task during both IDS favorable and IDS unfavorable
sessions was to fill out a Bio-PK Form (see Figure 1) . Prior to start of the session, the
"Session I.D.," "Viewer I.D.," and "Date" portions of the form were completed. Once the
subject began his process of selection, a session "Start Time" was noted, and the test-tube
selections were recorded by filling in the boxes (provided on the form's test-tube icons) with
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FIGURE 1
SAMPLE BIO-PK FORM
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the appropriate test-tube-selection numbers. The order of selection and an estimate of the
duration of effort per each test tube were noted in the "Comments" section. Upon debriefing
the subject at the end of a session, the monitor also recorded any comments the subject
wished to make regarding possible strategies employed in the performance of the task, and
any personal statements the subjects wished to volunteer pertaining to their state-of-mind,
health, and so forth.
At the conclusion of the session, and after the departure of the subject
from the psychoenergetics facility, the monitor once again covered the ice chest, then
transported it on the cart back to the Microbial Genetics Laboratory.
The microbiologist removed the nine bacterial cultures from the ice chest and
placed them, together with the additional two extrasession control cultures, in an incubator
(G24 Environmental Incubator Shaker, New Diunswick Scientific Company, Inc., Edison,
New Jersey). The bacterial cultures were shielded from visible light by aluminum foil, and
grown with gentle shaking (100 rpm) for about 24 hours.
Following the incubation period, testing of the eleven bacterial cultures to
determine the extent of mutation induction was initiated. The testing was divided into two
parts:
? Quantitation of number of cells plated, which measures the number of
plated cells that are able to form colonies (CFU) on medium containing
histidine (yeast complete medium).
? Quantitation of mutant cells, which measures the number of cells that are
able to grow in the absence of histidine.
The quantitation of CFU was accomplished according to a standardized set of
laboratory procedures. First, each of the eleven bacterial cultures (i.e., the cultures contained
in the nine-session test tubes plus the two controls) was serially diluted by combining 0.20 ml
of the culture with 1.80 ml of sterile saline until an overall million-fold dilution was obtained
(10-6). Complete medium plates were then divided into three sections with a marking pen,
and a 10-?1 aliquot of the 10-4, 10-5, and 10-6 dilutions were then delivered in triplicate to
the appropriate sections on the plate. The 10-?l spots were allowed to dry on the surface of
the'solid medium in the plates. The plates were then incubated at 37?C for up to 24 hours
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to allow for growth and appearance of colonies. The colonies were then counted manually
using a New Brunswick Scientific Bactronic Colony Counter, Model C110, and the data were
recorded for each dilution. In all cases, the 10-5 bacterial dilution provided an acceptable
number of nonoverlapping individual colonies to back-calculate the total number of bacteria
in 1 ml of the overnight culture.
The quantitation of the number of mutants (histidine independent cells) present
in the overnight cultures was obtained according to the following set of procedures. First, a
100-41 aliquot (approximately 108 cells) of the undiluted culture was combined with two ml of
molten (43?C) top agar in a 13-X-100-mm test tube. Vortexing of the cells in the top agar
ensured even distribution of the cells when poured on the selective minimal glucose plates.
Each culture was then plated in triplicate, and the plates were incubated at 37?C for 48
hours. When the incubation process was complete, revertant colonies were counted manually,
and individual plate counts were recorded.
For each of the eleven bacterial cultures, an average CFU for 10 Al of the 10-5
dilution was obtained and adjusted in terms of CFU per ml of the original culture. The
mutation frequency was then calculated as
mutation frequency/10 6 cells = number mutant cells/ml
X 106
When all of the calculations for a given session were completed, the micro-
biologist provided the monitor with an experimental data sheet detailing the growth rates of
the biological samples and the pertinent experimental parameters involved in the biological
procedures (see Figure 2). The most salient column to note on this form is the
"Mutants/IO6Cells," which is the number of cells per million cells per test tube that were able
to grow in the absence of histidine. The most desirable outcome of a session was obtained if
the subject's "high-aim" test tubes exhibited the highest values from this column, and "low-
aim" test tubes exhibited the lowest values.
After obtaining the form for a given session, the monitor completed a session
feedback sheet (see Figure 3). The test tube numbers of the subject's choices for the IDS
favorable condition (or the predetermined Numbers 1, 2, 3, 7, 8, and 9 for the IDS
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Experiment: t
Innoculation Date: Time: Q4 Broth Batch Date:
Bacteria Stock Incubation Date: ! Start Time: End Time: Tot 4l Time:14.2Arc
Survival (YC Plates) Incubation Start Time:/0'4a A? 4
OEnd Time:d: Total Tiae: Temp:r
Mutation (Bio Plates) In bation Start Time: Op 4 End Time: 41'%m M f1t Total Time; o,)- Teap: ~: ?c
Bio Plat Batch Date: YC Plate Batch Date: 4 ;$ Saline Batch Date: __
1 -7- 1294
colonies 10 1 average auganta/
Tube / Mutants/.100 ml Aver. Mutant/.1 ml dil: 10' 10-j I 10- cells/ml 10 cells
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