Intranasal delivery of shRNA to knockdown the 5HT-2A receptor enhances memory and alleviates anxiety.

Short-hairpin RNAs (shRNA), targeting knockdown of specific genes, hold enormous promise for precision-based therapeutics to treat numerous neurodegenerative disorders. However, whether shRNA constructed molecules can modify neuronal circuits underlying certain behaviors has not been explored. We designed shRNA to knockdown the human HTR2A gene in vitro using iPSC-differentiated neurons. Multi-electrode array (MEA) results showed that the knockdown of the 5HT-2A mRNA and receptor protein led to a decrease in spontaneous electrical activity. In vivo, intranasal delivery of AAV9 vectors containing shRNA resulted in a decrease in anxiety-like behavior in mice and a significant improvement in memory in both mice (104%) and rats (92%) compared to vehicle-treated animals. Our demonstration of a non-invasive shRNA delivery platform that can bypass the blood-brain barrier has broad implications for treating numerous neurological mental disorders. Specifically, targeting the HTR2A gene presents a novel therapeutic approach for treating chronic anxiety and age-related cognitive decline.

Who's calling? Evaluating the accuracy of guessing who is on the phone.

Some people claim to occasionally know who is calling them without using traditional means. Controlled experiments testing these claims report mixed results. We conducted a cross-sectional study of triads examining the accuracy of knowing who was calling using two randomly selected designs: 1) a web server randomly chose the caller before the callee's guess (telepathic/pre-selected trials), and 2) a web server randomly chose the caller after the callee's guess (precognitive/post-selected trials). We also performed exploratory multilevel mixed-effects logistic regressions on the relationship of genetic relationships, emotional closeness, communication frequency, and physical distance data with accuracy. A total of 177 participants completed at least one trial (105 "completers" completed all 12 trials). Accuracy was significantly above chance for the 210 completers telepathic/pre-selected trials (50.0% where the chance expectation was 33.3%, p<.001) but not the 630 completers precognitive/post-selected trials (31.9% where the chance expectation was 33.3%, p = .61). We discuss how these results favor the psi hypothesis, although conventional explanations cannot be completely excluded. Genetic relatedness significantly predicted accuracy in the regression model (Wald χ2 = 53.0, P < .001) for all trials. Compared to 0% genetic relatedness, the odds of accurately identifying the caller was 2.88 times (188%) higher for 25% genetic relatedness (Grandparent/Grandchild or Aunt/Uncle or Niece/Nephew or Half Sibling; ß = 1.06, z = 2.10, P = .04), but the other genetic relatedness levels were not significant. In addition, communication frequency was significant (ß = 0.006, z = 2.19, P = .03) but physical distance (ß = 0.0002, z = 1.56, P = .12) and emotional closeness (ß = 0.005, z = 1.87, P = .06) were not for all trials. To facilitate study recruitment and completion, unavoidable changes to the protocol were made during the study due to persistent recruitment difficulties, including changing inclusion/exclusion criteria, increasing total call attempts to participants, adjusting trial type randomization schema to ensure trial type balance, and participant compensation. Thus, future research will be needed to continue to improve the methodology and examine the mechanism by which people claim to know who is calling, as well as factors that may moderate the effects.

Genetic modulation of the HTR2A gene reduces anxiety-related behavior in mice.

The expanding field of precision gene editing using CRISPR/Cas9 has demonstrated its potential as a transformative technology in the treatment of various diseases. However, whether this genome-editing tool could be used to modify neural circuits in the central nervous system (CNS), which are implicated in complex behavioral traits, remains uncertain. In this study, we demonstrate the feasibility of noninvasive, intranasal delivery of adeno-associated virus serotype 9 (AAV9) vectors containing CRISPR/Cas9 cargo within the CNS resulting in modification of the HTR2A receptor gene. In vitro, exposure to primary mouse cortical neurons to AAV9 vectors targeting the HT2RA gene led to a concentration-dependent decrease in spontaneous electrical activity following multielectrode array (MEA) analysis. In vivo, at 5 weeks postintranasal delivery in mice, analysis of brain samples revealed single base pair deletions and nonsense mutations, leading to an 8.46-fold reduction in mRNA expression and a corresponding 68% decrease in the 5HT-2A receptor staining. Our findings also demonstrate a significant decrease in anxiety-like behavior in treated mice. This study constitutes the first successful demonstration of a noninvasive CRISPR/Cas9 delivery platform, capable of bypassing the blood-brain barrier and enabling modulation of neuronal 5HT-2A receptor pathways. The results of this study targeting the HTR2A gene provide a foundation for the development of innovative therapeutic strategies for a broad range of neurological disorders, including anxiety, depression, attentional deficits, and cognitive dysfunction.

Quantum aspects of the brain-mind relationship: A hypothesis with supporting evidence.

If all aspects of the mind-brain relationship were adequately explained by classical physics, then there would be no need to propose alternatives. But faced with possibly unresolvable puzzles like qualia and free will, other approaches are required. In alignment with a suggestion by Heisenberg in 1958, we propose a model whereby the world consists of two elements: Ontologically real Possibles that do not obey Aristotle's law of the excluded middle, and ontologically real Actuals that do. Based on this view, which bears resemblance to von Neumann's 1955 proposal (von Neumann, 1955), and more recently by Stapp and others (Stapp, 2007; Rosenblum and Kuttner, 2006), measurement that is registered by an observer's mind converts Possibles into Actuals. This quantum-oriented approach raises the intriguing prospect that some aspects of mind may be quantum, and that mind may play an active role in the physical world. A body of empirical evidence supports these possibilities, strengthening our proposal that the mind-brain relationship may be partially quantum.

What if consciousness is not an emergent property of the brain? Observational and empirical challenges to materialistic models.

The nature of consciousness is considered one of science's most perplexing and persistent mysteries. We all know the subjective experience of consciousness, but where does it arise? What is its purpose? What are its full capacities? The assumption within today's neuroscience is that all aspects of consciousness arise solely from interactions among neurons in the brain. However, the origin and mechanisms of qualia (i.e., subjective or phenomenological experience) are not understood. David Chalmers coined the term "the hard problem" to describe the difficulties in elucidating the origins of subjectivity from the point of view of reductive materialism. We propose that the hard problem arises because one or more assumptions within a materialistic worldview are either wrong or incomplete. If consciousness entails more than the activity of neurons, then we can contemplate new ways of thinking about the hard problem. This review examines phenomena that apparently contradict the notion that consciousness is exclusively dependent on brain activity, including phenomena where consciousness appears to extend beyond the physical brain and body in both space and time. The mechanisms underlying these "non-local" properties are vaguely suggestive of quantum entanglement in physics, but how such effects might manifest remains highly speculative. The existence of these non-local effects appears to support the proposal that post-materialistic models of consciousness may be required to break the conceptual impasse presented by the hard problem of consciousness.

Salivary neutrophil sampling feasibility in general population for gene expression analysis.

OBJECTIVE: Human gene expression studies typically rely on peripheral blood samples as a cellular source, however there are numerous situations in which venipuncture is contraindicated. To this end, an oral rinse-based method for collecting salivary neutrophils as a cellular source for gene expression analyses was previously developed and shown in a pilot study with five male participants to yield mRNA expression results comparable to those obtained from peripheral blood samples. The objective of the current study was to characterize the generalizability of the oral rinse-based method by analyzing unpublished RNA quality data obtained through a parent study that collected salivary neutrophil samples using the method from a larger sample size and including both men and women. RESULTS: The 260/280 nm absorbance ratios of the RNA obtained from 48 participants using the oral rinse-based method were within the expected range (average = 1.88 ± 0.16) for the majority of the samples, and no significant differences in RNA quality were found between participants' health, age group, or gender. Together with published data confirming the integrity of RNA obtained using the same method, these results support the feasibility of using this noninvasive method for obtaining samples for human gene expression analyses.

Exploring Personal Development Workshops' Effect on Well-Being and Interconnectedness.

Introduction: Personal development workshops are increasingly popular. This study evaluated the relationships between the measures of well-being, interconnectedness, and extended perception in various workshops and explored which kinds of workshops and individual characteristics predicted changes in these outcomes. Materials and Methods: In a prospective, uncontrolled, within-participant design study, adult participants completed questionnaires and online tasks before and after personal development workshops. Three analyses were conducted: (1) examining the relationships between measures by using only pre-workshop measures using Spearman correlations; (2) exploring change scores pre- to post-workshop and workshop using Wilcoxon signed-rank test; (3) assessing workshop format and content, and individual characteristics as predictors of those change scores multivariate nonparametric regression. The following outcomes were collected: Well-being-Arizona Integrative Outcomes Scale, positive and negative affect, Dispositional Positive Emotions Scale-Compassion subscale, Sleep Quality Scale, Numeric Pain Rating Scale; Interconnectedness-Cloninger Self-Transcendence Scale, Inclusion of Nature in Self and Inclusion of the Other in Self; and Extended perception tasks-Intuition Jar, Quick Remote Viewing, Psychokinesis Bubble, and Time Estimation. The following potential predictor variables were collected: demographic, mental health, psychiatric and meditation history, Single General Self-Rated Health Question, Brief Five-Factor Inventory-10, and the Noetic Experience and Belief Scale. Workshop leaders also selected which format and content characteristics applied to their workshop. Results: Interconnectedness measures were significantly and positively correlated with well-being (ρ: 0.27 to 0.33), positive affect (ρ: 0.20 to 0.27), and compassion (ρ: 0.21 to 0.32), and they were negatively correlated with sleep disturbance (ρ: -0.13 to -0.16) and pain (ρ: -0.11 to -0.16). Extended perception task performance was not correlated with interconnectedness or well-being. General personal development workshops improved subjective interconnectedness, well-being, positive emotion, and compassion, and they reduced sleep disturbances, negative emotion, and pain (all p's < 0.00005). The lecture (p = 0.03), small groups (p = 0.001), pairs (p = 0.01), and discussion (p = 0.03) workshop formats were significant predictors of well-being outcomes. The workshop content categories of meditation (p = 0.0002) and technology tools (p = 0.01) were also predictive of well-being outcomes, with meditation being the most consistent predictor of positive well-being changes. Conscientiousness was the only significant individual characteristic predictor (p = 0.002), although it was associated with increases in some well-being measures and decreases in others. Conclusions: This study provides preliminary evidence for the positive relationship between the subjective sense of interconnectedness and multiple well-being measures and the beneficial effects of some personal development workshops.

Genetics of psychic ability - A pilot case-control exome sequencing study.

INTRODUCTION: It is commonly believed that psychic ability, like many mental and physical traits, runs in families. This suggests the presence of a genetic component. If such a component were found, it would constitute a biological marker of psychic ability and inform environmental or pharmacologic means of enhancing or suppressing this ability. METHODS: A case-control study design was used to evaluate differences between psychic cases and non-psychic controls. Over 3,000 candidates globally were screened through two online surveys to locate people who claimed they and other family members were psychic. Measures of relevance to the claimed abilities (e.g., absorption, empathy, schizotypy) were collected and based on those responses, individuals with indications of psychotic or delusional tendencies were excluded from further consideration. Eligible candidates were then interviewed and completed additional screening tests. Thirteen individuals were selected as the final "psychic cases," and ten age-, sex-, and ethnicity-matched individuals with no claims of psychic ability were selected as controls. DNA from the saliva of these 23 participants was subjected to whole-exome sequencing. Two independent bioinformatics analyses were blindly applied to the sequenced data, one focusing exclusively on protein-coding sequences and another that also included some adjacent noncoding sequences. RESULTS: Sequencing data were obtained for all samples, except for one in the control group that did not pass the quality controls and was not included in further analyses. After unblinding the datasets, none of the protein-coding sequences (i.e., exons) showed any variation that discriminated between cases and controls. However, a difference was observed in the intron (i.e., non-protein-coding region) adjacent to an exon in the TNRC18 gene (Trinucleotide Repeat-Containing Gene 18 Protein) on chromosome 7. This variation, an alteration of GG to GA, was found in 7 of 9 controls and was absent from all psychic cases. DISCUSSION: The most conservative interpretation of these results is that they result from random population sampling. However, when the results are considered in relation to other lines of evidence, the results are more provocative. Further research is justified to replicate and extend these findings.

Effects of intentionally treated water on the growth of mesenchymal stem cells: An exploratory study.

OBJECTIVE: This study explored if human primary mesenchymal stem cells (MSCs), derived from two donors and cultivated in a medium made with intentionally treated water, would exhibit more growth and pluripotency than MSCs from the same source but grown in untreated (control) water. DESIGN: To create the treated water, three Buddhist monks directed their attention toward commercially bottled water while holding the intention that the water would enhance the growth of MSCs. Under double-blind conditions, cell culture growth mediums were prepared with the treated and untreated water, which was in turn used to grow the primary MSCs. Primary cells obtained from two donors were designated as Cells #1 and Cells #2. The prediction was that treated water would result in increased cell proliferation, that more cells would enter the cell cycle growth phase, and that there would be increased expression of genes (NANOG, OCT4 and SOX2) associated with improved cell growth and decreased expression of genes (p16, p21, and p53) associated with a decline in cell growth. The improved growth hypothesis was directional, thus one-tailed p-values were used to evaluate the results. RESULTS: Proliferation averaged across Cells #1 and #2 showed overall increased growth in treated as compared to control water (p = 0.0008). Cells #1 and #2 considered separately had differences in the same direction but only Cells #2 showed a significant difference on day 6 (p = 0.01). For cell cycle, there was a significantly greater percentage of Cells #2 in the S interphase with treated vs. control water (p = 0.04). For the gene expression analysis, when considering the average across the two donor cells, only the NANOG gene expression was in the predicted direction (p = 0.01); by contrast, the p16 gene expression was significantly opposite to the predicted direction (p = 0.005, one-tailed, post-hoc). For Cells #1 considered separately, no differences were significant except for p16, which resulted in an effect opposite to the predicted outcome (p = 0.05). For Cells #2, three genes were significantly in the predicted directions: NANOG (p = 0.0008), OCT4 (p = 0.005), and P53 (p = 0.05); p16 was significantly opposite to the prediction (p = 0.001). CONCLUSION: Intentionally treated water appeared to have some biological effects on the growth, pluripotency and senescence of human MSCs. This was especially the case in one of the two donor cells tested, but the effects were not consistently in the predicted direction. As an exploratory study, caution is warranted in interpreting these outcomes, and adjustment for multiple testing would likely reduce some of the weaker effects to nonsignificant. But given the double-blind protocol, as well as several more significant outcomes in the predicted directions, further research is warranted.

Effects of intentionally treated water and seeds on the growth of Arabidopsis thaliana.

OBJECTIVE: . A previously reported experiment indicated that Arabidopsis thaliana seeds with cryptochrome mutation His-CRY2 showed more robust photomorphogenic growth when hydrated with intentionally treated water as compared to untreated water. The present study attempted to replicate that outcome, adding a condition where the seeds were also intentionally treated. Arabidopsis seeds were used because they contain a photosensitive flavoprotein called cryptochrome (CRY). CRY has been proposed as a possible "transducer" of intention in living systems because it is thought to have quantum biological properties, and as such, it might potentially be sensitive to quantum observer effects. DESIGN: Three Buddhist monks directed their attention toward commercially bottled water and Arabidopsis seeds while holding the intention to improve the growth of the plant. As a control condition, no attention was directed at water or seeds from the same sources. Under double-blinded conditions, treated and untreated seeds were placed in an incubator, hydrated with treated or untreated water, and exposed to either continuous blue light or blue plus far-red light. The seed germination process was repeated three times, each time using new seeds. A 2 × 2 × 2 ANOVA, with water, seeds, and light as factors, was used to analyze the results. RESULTS: . Treated water was associated with enhanced photomorphogenic growth, as reflected by a shorter hypocotyl length (p = 0.04) and greater amounts of chlorophyll (p = 0.0005) and anthocyanin (p = 2 × 10-6). Treated seeds resulted in greater amounts of chlorophyll (p = 0.04), but also a longer hypocotyl (p = 0.0004) and less anthocyanin (p = 0.01). Plants exposed to blue plus far-red light were constantly more robust than plants grown under blue light, regardless of the type of water or seed (p < 10-10). CONCLUSION: . Intentionally treated water improved the growth of the His-CRY2 variant of Arabidopsis, confirming results of an earlier experiment. Enhanced growth associated with exposure to blue plus far-red light also confirmed to known effects. A more complex relationship was observed with treated seeds. Further research is required to understand the latter outcome, as it may provide clues about the underlying mechanisms of intentional influences.

Energy Medicine treatments for hand and wrist pain: A pilot study.

INTRODUCTION: The term "energy medicine" describes healing modalities that manipulate or channel purported subtle energies associated with the body. The objectives of this pilot study were to determine the feasibility of studying energy medicine for people with carpal tunnel pain and gathering relevant preliminary data. METHODS: Following a prospective, within-participant design, participants were recruited to experience a 30 min treatment from one of 17 energy medicine practitioners. Of 374 adults experiencing carpal tunnel pain who were screened for the study, 190 received an energy medicine treatment. Practitioners delivered treatments at close distance, some with and some without light, stationary touch. Outcome measures were collected before, during, and immediately after the treatment, and three weeks later. The primary outcome measure was self-reported pain. Secondary subjective measures included credibility regarding energy medicine and expectancy regarding the efficacy of treatments, pain interference, sleep quality, well-being, mood, and sense of personal transformation. Physiological measures included median nerve conduction velocity, heart rate variability, heart rate synchrony (between the participant and practitioner), and expression levels of neuroinflammation-related genes. RESULTS: On average, self-reported current pain scores decreased 2.0 points post-session and 1.3 points at three weeks compared to baseline values using a 0-10 point scale with 10 denoting worst pain (F(2, 565) = 3.82 p <0.000005). This effect was not influenced by the participants' level of expectancy or credibility regarding the energy medicine modality. Well-being, negative emotion, and sleep quality scores significantly improved at the follow-up visit. Multiple heart rate variability measures significantly changed reflecting increased parasympathetic activity which may indicate decreased stress. No other secondary outcome showed significant change. DISCUSSION: Studying the administration of energy medicine to people with carpal tunnel pain is feasible, although requiring a documented carpal tunnel syndrome diagnosis proved to be prohibitive for recruitment. Our finding of preliminary evidence for positive effects in pain and pain-related outcomes after a single session of energy medicine warrants further controlled investigation.

Effects of the local and geocosmic environment on the efficacy of Energy Medicine treatments: An exploratory study.

INTRODUCTION: Outcomes of medical treatments tend to be highly variable. Some of the underlying variance is due to well-known factors such as age, gender, ethnicity, and effects of local weather. There are also less obvious influences including variations in solar wind, the Earth's geomagnetic field, and the interplanetary magnetic field. This study explored possible effects of these local and solar/geomagnetic variables on the outcomes of energy medicine treatments. The context was a pilot clinical trial involving 17 energy medicine practitioners who treated a total of 190 participants presenting with hand and wrist pain. METHODS: Eighteen environmental variables were correlated against changes in subjective pain and against changes in objective measures of nerve conduction velocity. RESULTS: The results showed that local barometric pressure, interplanetary magnetic field, lunar illumination, proton fluence, electron fluence, and solar radio flux showed statistically significant relationships with these health outcomes (at p < 0.05 or better) before correction for multiple comparison corrections. The variable of barometric pressure had a robust correlation with nerve conduction velocity, surviving adjustment for false discovery rate among the 18 variables at p < 0.05. DISCUSSION: This study lends support for future research into local weather, and potentially also to fluctuations in the solar/geomagnetic environment environmental measures as potential sources of variation in energy medicine sessions.

Possible negentropic effects observed during Energy Medicine sessions.

INTRODUCTION: Previously reported experiments suggest that aspects of the physical environment, in particular measures of negentropy (i.e., order) associated with the statistical outputs of truly random number generators, may be affected during periods of focused mental attention. The present study was designed to conceptually replicate those reports during energy medicine sessions. METHOD: A custom-built "quantum noise generator" (QNG) was used to continuously record and digitize (at 1 KHz) 16 independent channels of random samples (i.e., noise) produced by electron tunneling and avalanche effects in Zener diodes. One metric was developed to quantify temporal dependencies in the noise samples aggregated across the 16 channels, and a second metric was formed that measured spatial dependencies among the 16 channels. The two metrics were combined into a single "spacetime" variable used to measure fluctuations in entropy during 110 half-hour energy medicine sessions. As a control, the same measure was examined in data recorded eight hours after each energy medicine session took place, when no one was in the laboratory. RESULTS: QNG data recorded during the half-hour sessions showed significant deviations from chance expectation, with a peak deviation observed at 24 minutes into the half-hour (z = 4.24, p < 0.00003, two-tail), and with deviations associated with p < 0.05 from 20 to 29 min, after correction for multiple comparisons. By comparison, data recorded eight hours after each session showed uniformly null results. This outcome is consistent with previously reported studies, suggesting that during periods of focused attention negentropic deviations emerge in random physical systems. Counterarguments to this interpretation are discussed, as well as recommendations for future studies.

Spectroscopic analysis of water treated by and in proximity to Energy Medicine practitioners: An exploratory study.

Previously reported experiments suggest that healing intention focused toward water, or merely taking place in the vicinity of water, affects the hydrogen-oxygen (HO) covalent bonds. This claim was explored in the context of a clinical energy medicine pilot study involving 17 practitioners and 190 participants. In a "direct" test, samples of water were directly treated by the practitioners; in an "indirect" test, aliquots attached to lanyards were worn by practitioners and participants as they were engaged in healing sessions. Samples of laboratory-grade distilled water and Fiji brand water were used in the tests, and the water was analyzed using an Attenuated Total Reflection (ATR) Fourier Transform Infrared (FTIR) spectrometer equipped with a liquid nitrogen-cooled detector. The comparison of interest was the ensemble average spectrum recorded during pre- vs. post-intentional healing periods in the primary infrared absorption portion of the water spectrum. The analyses indicated that distilled water directly treated by the practitioners resulted in a change in the HO bond at the wavenumber 3200 cm-1 (p < 0.03, two-tailed). No effect was observed with the Fiji water. The distilled water in aliquots worn by practitioners also resulted in a significant change at the same wavenumber (p = 0.0004, two-tailed). No effects were observed in Fiji water aliquots worn by practitioners or participants, or in distilled water worn by participants. This study contributes to previously reported observations suggesting that the structure of water reacts in an anomalous way to healing intentions. Such effects appear to involve some form of energetic influence, but that is not yet well established. Nor is it certain that the observed effect can only be due to intention; it is conceivable, for example, that an unidentified environmental factor may have been responsible for the observed comparisons. However, given similar results observed in several experiments so far, including the present study, further research seems warranted.

A case study of extended human capacity perception during Energy Medicine treatments using mixed methods analysis.

INTRODUCTION: Some "energy medicine" modalities, such as Reiki, continue to show evidence for efficacy, but the underlying mechanisms of action are still uncertain. To help illuminate possible mechanisms, this exploratory mixed-method study using qualitative and quantitative analyses investigated: 1) How do energy medicine sessions function from the perspective of a seer, i.e., an individual trained in techniques to enable perception of purported subtle energies, and 2) Do any of these observations correlate with changes in participants' subjective pain? METHOD: One seer reported observations during 30-minute energy medicine sessions delivered by 17 practitioners to participants experiencing chronic hand or wrist pain. Data were coded using an inductive approach, followed by thematic and descriptive analyses. Content analysis was used to assign whether each code was mentioned by the seer for each participant, generating 119 binary variables, one for each code. The relationship between these binary variables and change in participant pain scores after the energy medicine session and three weeks later were explored using linear regression models. RESULTS: The qualitative thematic analysis identified six major themes from the seer's notes: "Experience of the Practitioner," "Experience of the Participant," "Space and Other Beings" (referring to the qualities of the physical space the energy medicine session occurred in and ostensible non-physical beings present during session), "Participant-Practitioner Relationship," "Healing Process," and "Attributes of Energy." The energy medicine methods used varied for each participant, according to the practitioners' perceptions of each individual's needs. The linear regression models yielded significant associations between changes in pain scores and various codes, including energy color, practitioner touch and the presence of ostensible non-physical beings supporting the session, although their significance did not persist after correction for multiple comparisons. DISCUSSION: Future studies investigating the mechanisms of energy medicine may benefit from including perceptions by seers in their outcome measures.

Effects Induced In Vivo by Exposure to Magnetic Signals Derived From a Healing Technique.

Energy healing is a therapy said to manipulate and balance the flow of "energies" in the body. One such technique, the Bengston Healing Method (BHM), has shown some success in healing malignant tumors in animals and humans, but the mechanism of action and factors influencing therapeutic success of this method are poorly understood. In this study, we tested in vivo the antitumor potential of magnetic signals recorded during BHM healing. Balb/c mice engrafted with 4T1 breast cancer cells were exposed to this recording for 4 h/d on a weekly or daily basis for 28 days; control mice were not exposed at all. Tumors showed a trend to grow slower in the treatment versus control group during the fourth week of treatment. Elevated leukocyte counts, associated with an increase in blood levels of granulocyte-macrophage colony stimulating factor and interleukin-6, were observed in tumor-bearing mice exposed to the BHM recording but not in healthy animals exposed to the recording. This suggests that exposure to a recording of BHM may induce a biological response in tumor-bearing mice, but limited effects on tumor growth when observed within the predefined end point of 28 days. Studies involving longer end points are recommended to observe the progression of tumor growth.