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The objective of the current study was to characterize the generalizability of the oral rinse-based method by analyzing unpublished RNA quality data obtained through a parent study that collected salivary neutrophil samples using the method from a larger sample size and including both men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.

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.

Do Noncoding RNAs Mediate the Efficacy of Energy Psychology?

BACKGROUND: There are over 100 published studies of a therapy called Emotional Freedom Techniques (EFT). This popular form of energy psychology combines elements of established methods like cognitive therapy with acupressure. Our group reported the first evidence of its mechanisms of action at the molecular level, showing that it can influence levels of the stress hormone cortisol. OBJECTIVES: Given recent advances in molecular genomics that have identified noncoding ribonucleic acid (RNA) molecules as important regulators of gene expression, the aim of this study is to explore the possibility that microRNAs play a role in mediating the effects of EFT. METHODS: We measured microRNA levels in stored blood samples from our previous study in which veterans were randomized into an EFT group receiving EFT and treatment as usual throughout a 10-week intervention period, and a control group receiving only treatment as usual during the intervention period and then receiving EFT. A broad panel of 800 microRNAs was probed using a multiplexed, direct hybridization, and detection system. RESULTS: All of the microRNA targets were expressed at low levels and most were below thresholds established by negative control probes. Baseline variability was determined using samples collected from the control group at the start and end of the intervention period, and used to filter out targets that were too noisy under control conditions to be able to distinguish a response to treatment. Analysis of the remaining viable targets found a general trend of reduced expression following EFT, compared to expression levels in samples from the control group during the intervention period. The most notable decreases in expression levels were found for 2 microRNAs: let-7b and let-7c, although no significance was found after adjusting for multiple comparisons. CONCLUSIONS: These preliminary data support the feasibility of measuring microRNA expression level changes that correlate with effective EFT therapy.

Epigenetic Effects of PTSD Remediation in Veterans Using Clinical Emotional Freedom Techniques: A Randomized Controlled Pilot Study.

PURPOSE: To assess the feasibility of measuring changes in gene expression associated with post-traumatic stress disorder (PTSD) treatment using emotional freedom techniques (EFT). DESIGN: Participants were randomized into an EFT group receiving EFT and treatment as usual (TAU) throughout a 10-week intervention period and a group receiving only TAU during the intervention period and then receiving EFT. SETTING: A community clinic and a research institute in California. PARTICIPANTS: Sixteen veterans with clinical levels of PTSD symptoms. INTERVENTION: Ten hour-long sessions of EFT. MEASURES: Messenger RNA levels for a focused panel of 93 genes related to PTSD. The Symptom Assessment 45 questionnaire, Hospital Anxiety and Depression Scale, Insomnia Severity Scale, SF-12v2 for physical impairments, and Rivermead Postconcussion Symptoms Questionnaire. ANALYSIS: Pre-, posttreatment, and follow-up mean scores on questionnaires were assessed using repeated measures 1-way analysis of variance. A Student t test and post hoc analyses were performed on gene expression data. RESULTS: Post-traumatic stress disorder symptoms declined significantly in the EFT group (-53%, P < .0001). Participants maintained their gains on follow-up. Significant differential expression of 6 genes was found ( P < .05) when comparing the expression levels before and after the intervention period in participants receiving EFT. CONCLUSION: Study results identify candidate gene expression correlates of successful PTSD treatment, providing guidelines for the design of further studies aimed at exploring the epigenetic effects of EFT.

Expressive Arts Therapy with Hospitalized Children: A Pilot Study of Co-Creating Healing Sock Creatures©.

A novel form of expressive arts therapy was developed in a pediatric unit and received enthusiastic support from hospital staff and family members because of their impressions that the children were calmer following therapy, as well as throughout the remainder of the hospital stay. A pilot study was conducted to assess the feasibility of quantifying such impressions by measuring changes in the children's mood by self-report. Twenty-five children (mean age 8.34 years, SD 3.77) were recruited for the study, coming from diverse social-economic backgrounds, ethnicities, and an array of medical diagnoses. The results document improvements in mood for children following therapy sessions, compared to children in a wait-list control group. Additionally, a meta-analysis examining external influences and changes in salivary cortisol levels measured before and after therapy sessions illustrates the importance of considering aspects of the clinical setting when assessing the effectiveness of this and other expressive arts therapies for reducing stress during hospitalization.

Expressive arts therapy for hospitalized children: a pilot study measuring cortisol levels.

This pilot study aimed at assessing the feasibility of capturing physiological evidence of reduced stress for hospitalized children following expressive arts therapy. Twenty-five patients were offered a novel form of expressive arts therapy, termed Healing Sock Creatures, during their stay in the hospital. Saliva samples were collected at two times in the afternoon for the purpose of measuring salivary cortisol levels. The patients were randomly assigned to two groups, a treatment group or a wait-list control group. A trend of decreased cortisol levels was apparent following therapy in the treatment group and concurrent steroid treatment, which is common in intensive care units, does not appear to interfere with the ability to measure decreased cortisol levels following therapy. Our results support the design of a formal study to assess physiological biomarkers of stress in hospital settings. To our knowledge, this is the first in-patient study assessing a biomarker of stress following expressive arts therapy for children.

Infrasound sensitizes human glioblastoma cells to cisplatin-induced apoptosis.

The development of nontoxic agents that can selectively enhance the cytotoxicity of chemotherapy is an important aim in oncology. This study evaluates the ability of infrasound exposure to sensitize glioblastoma cells to cisplatin-induced apoptosis. The infrasound was delivered using a device designed to replicate the unique infrasound emissions measured during external Qigong treatments. Human glioblastoma cell lines harboring wild-type p53 (U87) or mutant p53 (U251, SF210, and SF188) were treated in culture with cisplatin, infrasound emissions, or the combination of the 2 agents. Induction of apoptosis was quantified after 24 hours by flow cytometry following annexin V/propidium iodide staining. Infrasound emissions alone, delivered at moderate levels (~10 mPa) with dynamic frequency content (7-13 Hz), did not induce apoptosis, yet combining infrasound with cisplatin augmented the induction of apoptosis by cisplatin in all the 4 cell lines (P < .05). Increased cellular uptake of the fluorophore calcein associated with infrasound exposure was quantified by fluorescence microscopy as well as flow cytometry, demonstrating increased cell membrane permeability. The 4 cell lines differed in the degree to which infrasound exposure increased calcein uptake, and these differences were predictive of the extent to which infrasound enhanced cisplatin-induced apoptosis. When exposed to specific frequencies, membrane permeabilization also appeared to be differentially responsive for each cell line, suggesting the potential for selective targeting of tissue types using isolated infrasonic frequencies. Additionally, the pressure amplitudes used in this study were several orders of magnitude less than those used in similar studies involving ultrasound and shock waves. The results of this study provide support for using infrasound to enhance the chemotherapeutic effects of cisplatin in a clinical setting.

Evaluation of biofield treatment dose and distance in a model of cancer cell death.

OBJECTIVE: This study assessed the potential influence of biofield treatment on cultured human cancer cells and whether such influence was affected by varying the duration of the treatment (dose) or the distance between the biofield practitioner and the target cells. DESIGN: Biofield treatment dosage was assessed from a short distance (0.25 meters) in three independent experiments involving 1, 2, or 5 treatments, along with another set of three independent and comparable mock experiments. Biofield treatment distance was assessed at 0.25, 25, and ∼ 2000 meters involving two treatments in three independent experiments along with another set of three mock experiments. INTERVENTION: Biofield treatments were delivered by a highly acclaimed biofield practitioner with the intention of diminishing growth of the cells or inducing cancer-cell death. OUTCOME MEASURE: Cell viability was quantified 20 hours after treatments, using a spectrophotometric assay for live-cell counting. The dependent measure for each experiment was the log ratio of the cell viability values of treated samples (biofield or mock) over the values of untreated control samples. RESULTS: A trend of decreasing cell viability with increasing biofield dose was evident in the first set of experiments assessing dose-response; however, no such effect was evident in the second set of experiments evaluating biofield treatment distance. Mock experiments yielded relatively stable viability ratios in both sets of experiments. Linear regression analysis and hypothesis testing of the data taken as a whole did not yield statistical significance at p<0.05. CONCLUSIONS: These results represent the first indication of a biofield treatment dose-response in a controlled laboratory setting. The data are inconclusive because of the inability of reproduce the cellular response in a replicate experiment.

Evaluating biofield treatments in a cell culture model of oxidative stress.

OBJECTIVE: To test whether healing treatments by biofield practitioners can protect normal human brain cells against cell death induced by oxidative stress. DESIGN: Cultured human brain cells were exposed to increasing concentrations of hydrogen peroxide and cell death was quantified by computerized time-lapse microscopy. Biofield treatments were delivered to cells from a short distance in 24 independent experiments. Six highly experienced biofield practitioners each participated, all with exceptional reputations within their respective communities (4 independent experiments each). An equal number of control experiments involving no healing intervention were conducted to provide a measure of intrinsic variability of the experimental system. Experiments were conducted with blinding applied to each of the scientists and randomized sample assignment. INTERVENTION: Healing treatments were delivered to cells from a short distance by a single practitioner, before and after exposure to hydrogen peroxide, for a total of 30 minutes. OUTCOME MEASURE: Cell death was quantified over a 4-hour period following experimental treatments. RESULTS: We found no significant difference in cell death rates between treatment and control groups.

Independent motile microplast formation correlates with glioma cell invasiveness.

Diffuse brain invasion contributes to the poor prognosis for patients with gliomas. Analyzing glioma cell migration in vitro, we have demonstrated the spontaneous shedding of anucleate cell fragments that separate from glioma cell bodies and maintain viability from hours to days. Unlike previously described cell fragments that are released from cells as diffusible vectors, glioma cell fragments are independently motile. We used computerized time-lapse microscopy to characterize the formation of these independent motile microplasts (IMMPs) in human cell cultures derived from the most highly invasive glial tumor, glioblastoma. IMMPs were larger than previously described cell fragments, ranging in size from approximately 2% to nearly half of the area of their parent cells. Complex cell-like behaviors-including establishment of polarity, extension of lamellipodia and filopodia, and change in direction of movement-remained intact in IMMPs. The average direction and velocity of the IMMPs were indistinguishable from those of their parent cells. IMMPs formed at a significantly higher rate in glioma cell lines rendered more invasive by overexpression of invasion-related genes than in vector-transfected controls. The correlation with cell invasiveness indicates that IMMP formation may be related to the cell-invasive phenotype. Further investigation will determine whether IMMPs represent a novel addition to the growing list of viable cell fragments with biological relevance.