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.

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 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.

Transcriptional Changes in Cancer Cells Induced by Exposure to a Healing Method.

Energy healing, or healing with intent, is a complementary and alternative medicine therapy reported to be beneficial with a wide variety of conditions. We are developing a delivery technology for a method previously tested in mouse models with solid tumors (the Bengston method) independent of the presence of a healer. The goal of this study was to assess whether stored or recorded energy has an impact on breast cancer cells in vitro, using energy-charged cotton and electromagnetic recording of healers practicing the method. Expression of genes involved in cancer and inflammation pathways was measured by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Treatment of cells using energy-charged cotton resulted in statistically significant changes <1.5-fold. In cells exposed to an electromagnetic recording, 37 genes of 167 tested showed a >1.5-fold change when compared to the control, and 68 genes showing statistically significant fold changes. Two genes, ATP citrate lyase (ACLY) and interleukin 1ß (IL-1ß), were consistently downregulated at 4 and 24 hours of exposure to the recording, respectively, in 3 independent experiments. Both ACLY and IL-1ß were also downregulated in cells exposed to a hands-on delivery of the method, suggesting these 2 genes as potential markers of the healing method.

Distant healing of surgical wounds: an exploratory study.

BACKGROUND: Distant healing intention (DHI) is one of the most common complementary and alternative medicine (CAM) healing modalities, but clinical trials to date have provided ambivalent support for its efficacy. One possible reason is that DHI effects may involve variables that are sensitive to unknown, uncontrolled, or uncontrollable factors. OBJECTIVE: To examine 2 of those potential variables-expectation and belief-we explored the effects of DHI on objective and psychosocial measures associated with surgical wounds in 72 women undergoing plastic surgery. DESIGN: Participants were randomly assigned to 1 of 3 groups: blinded and receiving DHI (DH), blinded and not receiving DHI (control), and knowing that they were receiving DHI (expectancy). Outcome measures included collagen deposition in a surrogate wound and several self-report measures. DHI was provided by experienced distant healers. No differences in the main measures were observed across the three groups. RESULTS: Participants' previous belief in the efficacy of DHI was negatively correlated with the status of their mental health at the end of the study (P = .04, 2-tailed), and healers' perceptions of the quality of their subjective "contact" with the participants were negatively correlated both with change in mood (P = .001) and with collagen deposition (P = .04). A post-hoc analysis found that among participants assigned to receive DHI under blinded conditions, those undergoing reconstructive surgery after breast cancer treatment reported significantly better change in mood than those who were undergoing purely elective cosmetic surgery (P = .004). CONCLUSION: If future DHI experiments confirm the post-hoc observations, then some of the ambiguity observed in earlier DHI studies may be attributable to interactions among participants' and healers' beliefs, their expectations, and their motivations.

Compassionate intention as a therapeutic intervention by partners of cancer patients: effects of distant intention on the patients' autonomic nervous system.

OBJECTIVE: This double-blind study investigated the effects of intention on the autonomic nervous system of a human "sender" and distant "receiver" of those intentions, and it explored the roles that motivation and training might have in modulating these effects. DESIGN: Skin conductance level was measured in each member of a couple, both of whom were asked to feel the presence of the other. While the receiving person relaxed in a distant shielded room for 30 minutes, the sending person directed intention toward the receiver during repeated 10-second epochs separated by random interepoch periods. Thirty-six couples participated in 38 test sessions. In 22 couples, one of the pair was a cancer patient. In 12 of those couples, the healthy person was trained to direct intention toward the patient and asked to practice that intention daily for three months prior to the experiment (trained group). In the other 10 couples, the pair was tested before the partner was trained (wait group). Fourteen healthy couples received no training (control group). OUTCOME MEASURES: Using nonparametric bootstrap procedures, normalized skin conductance means recorded during the intention epochs were compared with the same measures recorded during randomly selected interepoch periods, used as controls. The preplanned difference examined the intention versus control means at the end of the intention epoch. RESULTS: Overall, receivers' skin conductance increased during the intention epochs (z = 3.9; P = .00009, two-tailed). Planned differences in skin conductance among the three groups were not significant, but a post hoc analysis showed that peak deviations were largest and most sustained in the trained group, followed by more moderate effects in the wait group, and still smaller effects in the control group. CONCLUSIONS: Directing intention toward a distant person is correlated with activation of that person's autonomic nervous system. Strong motivation to heal and to be healed, and training on how to cultivate and direct compassionate intention, may further enhance this effect.