Integrating Ultra-Weak Photon Emission Analysis in Mitochondrial Research.

Once regarded solely as the energy source of the cell, nowadays mitochondria are recognized to perform multiple essential functions in addition to energy production. Since the discovery of pathogenic mitochondrial DNA defects in the 1980s, research advances have revealed an increasing number of common human diseases, which share an underlying pathogenesis involving mitochondrial dysfunction. A major factor in this dysfunction is reactive oxygen species (ROS), which influence the mitochondrial-nuclear crosstalk and the link with the epigenome, an influence that provides explanations for pathogenic mechanisms. Regarding these mechanisms, we should take into account that mitochondria produce the majority of ultra-weak photon emission (UPE), an aspect that is often ignored - this type of emission may serve as assay for ROS, thus providing new opportunities for a non-invasive diagnosis of mitochondrial dysfunction. In this article, we overviewed three relevant areas of mitochondria-related research over the period 1960-2020: (a) respiration and energy production, (b) respiration-related production of free radicals and other ROS species, and (c) ultra-weak photon emission in relation to ROS and stress. First, we have outlined how these research areas initially developed independently of each other - following that, our review aims to show their stepwise integration during later stages of development. It is suggested that a further stimulation of research on UPE may have the potential to enhance the progress of modern mitochondrial research and its integration in medicine.

Pharmacological targeting of ROS reaction network in myeloid leukemia cells monitored by ultra-weak photon emission.

Acute myeloid leukemia (AML) is a blood cancer that is caused by a disorder of the process that normally generates neutrophils. Function and dysfunction of neutrophils are key to physiologic defense against pathogens as well as pathologies including autoimmunity and cancer. A major mechanism through which neutrophils contribute to health and disease is oxidative burst, which involves rapid release of reactive oxygen species (ROS) generated by a chemical reaction network catalyzed by enzymes including NADPH oxidase and myeloperoxidase (MPO). Due to the involvement of neutrophil-derived reactive oxygen species in many diseases and importance of NADPH oxidase and MPO-mediated reactions in progression and treatment of myeloid leukemia, monitoring this process and modulating it by pharmacological interventions is of great interest. In this work, we have evaluated the potential of a label-free method using ultra-weak photon emission (UPE) to monitor ROS production in neutrophil-like HL60 myeloid leukemia cells. Suppression of ROS was achieved by several drug candidates that target different parts of the reaction pathway. Our results show that UPE can report on ROS production as well as suppression by pharmacological inhibitors. We find that UPE is primarily generated by MPO catalyzed reaction and thus will be affected when an upstream reaction is pharmacologically modulated.

Cellular glutathione levels in HL-60 cells during respiratory burst are not correlated with ultra-weak photon emission.

Recently, ultra-weak photon emission (UPE) was developed as a novel tool for measuring oxidative metabolic processes, as its generation is related to reactive oxygen species (ROS). Both an imbalance in ROS or the uncontrolled production of ROS can lead to oxidative stress, which is commonly associated with many diseases. In addition to playing several biological functions, the thiol amino acid glutathione has an important antioxidant function in the body's defense against ROS. Specifically, glutathione is an important endogenous antioxidant that helps maintain oxidant levels. At the cellular level, glutathione is present in its reduced form (GSH) at relatively high concentrations (in the millimolar range) and in its oxidized form (GSSG) at low concentrations (in the micromolar range). Thus, the GSH/GSSG ratio is often used as an indicator of cellular redox state. Here, we used the HL-60 cell line as a model system in order to determine whether UPE is correlated with intracellular GSH and GSSG levels. HL-60 cells were differentiated into neutrophil-like cells and then stimulated to undergo respiratory burst. We then recorded UPE in real time for 9000 seconds and used capillary electrophoresis coupled to mass spectrometry to measure GSH and GSSG levels in cell extracts. We found that although respiratory burst significantly decreased the GSH/GSSG ratio, this change was not significantly correlated with the UPE profile.

Ultra-weak photon emission as a dynamic tool for monitoring oxidative stress metabolism.

In recent years, excessive oxidative metabolism has been reported as a critical determinant of pathogenicity in many diseases. The advent of a simple tool that can provide a physiological readout of oxidative stress would be a major step towards monitoring this dynamic process in biological systems, while also improving our understanding of this process. Ultra-weak photon emission (UPE) has been proposed as a potential tool for measuring oxidative processes due to the association between UPE and reactive oxygen species. Here, we used HL-60 cells as an in vitro model to test the potential of using UPE as readout for dynamically monitoring oxidative stress after inducing respiratory burst. In addition, to probe for possible changes in oxidative metabolism, we performed targeted metabolomics on cell extracts and culture medium. Lastly, we tested the effects of treating cells with the NADPH oxidase inhibitor diphenyleneiodonium chloride (DPI). Our results show that UPE can be used as readout for measuring oxidative stress metabolism and related processes.

Simultaneous and intercontinental tests show synchronism between the local gravimetric tide and the ultra-weak photon emission in seedlings of different plant species.

In order to corroborate the hypothesis that variations in the rate of spontaneous ultra-weak photon emission (UPE) from germinating seedlings are related to local variations of the lunisolar tidal force, a series of simultaneous tests was performed using the time courses of UPE collected from three plant species-corn, wheat and sunflower-and also from wheat samples whose grains were transported between continents, from Brazil to The Netherlands and vice versa. All tests which were run in parallel showed coincident inflections within the UPE time courses not only between seedlings of the same species but also between the different species. In most cases, the UPE inflections were synchronised with the turning points in the local gravimetric tidal variation. Statistical tests using the local Pearson correlation verified these coincidences in the two time series. The results therefore support the hypothesis of a relationship between UPE emissions and, in the oscillations, the local gravimetric tide. This applies to both the emissions from seedlings of different species and to the seedlings raised from transported grain samples of the same species.

Crossing the Boundaries of Our Current Healthcare System by Integrating Ultra-Weak Photon Emissions with Metabolomics.

The current healthcare system is hampered by a reductionist approach in which diagnostics and interventions focus on a specific target, resulting in medicines that center on generic, static phenomena while excluding inherent dynamic nature of biological processes, let alone psychosocial parameters. In this essay, we present some limitations of the current healthcare system and introduce the novel and potential approach of combining ultra-weak photon emission (UPE) with metabolomics technology in order to provide a dynamic readout of higher organizational systems. We argue that the combination of metabolomics and UPE can bring a new, broader, view of health state and can potentially help to shift healthcare toward more personalized approach that improves patient well-being.

Tracking biochemical changes correlated with ultra-weak photon emission using metabolomics.

Ultra-weak photon emission (UPE) is light emitted spontaneously by biological systems without the use of specific luminescent complexes. UPE is emitted in the near-UV/UV-Vis/near-IR spectra during oxidative metabolic reactions; however, the specific pathways involved in UPE remain poorly understood. Here, we used HL-60 cells, a human promyelocytic cell line that is often used to study respiratory burst, as a model system to measure UPE kinetics together with metabolic changes. HL-60 cells were differentiated into neutrophil-like cells by culturing in all-trans-retinoic acid for 7days. We then used a targeted metabolomics approach with capillary electrophoresis-mass spectrometry to profile intracellular metabolites in HL-60 cells and to investigate the biochemical changes based on the measured UPE profile. Our analysis revealed that the levels of specific metabolites, including putrescine, creatine, ß-alanine, methionine, hydroxyproline, serine, and S-adenosylmethionine, were significantly altered in HL-60 cells after inducing respiratory burst. A comparison with recorded UPE data revealed that the changes in putrescine, glutathione, sarcosine, creatine, ß-alanine, methionine, and hydroxyproline levels were inversely correlated with the change in UPE intensity. These results suggest that these metabolic pathways, particular the methionine pathway, may play a role in the observed changes in UPE in HL-60 cells and therefore demonstrate the potential for using UPE to monitor metabolic changes.

Ultraweak photon emission as a non-invasive health assessment: a systematic review.

We conducted a systematic review (SR) of the peer reviewed scientific literature on ultraweak photon emissions (UPE) from humans. The question was: Can ultraweak photon emissions from humans be used as a non-invasive health assessment? A systematic search was conducted across eight relevant databases: PubMed/MEDLINE, BIOSIS, CINAHL, PSYCHINFO, All of Cochrane EBM databases, GIDEON, DoD Biomedical Research, and clinicaltrials.gov from database inception to October 2011. Of the 1315 studies captured by the search strategy, 56 met the inclusion criteria, out of which 1 was a RCT, 27 were CCT, and 28 were observational and descriptive studies. There were no systematic reviews/meta-analyses that fit the inclusion criteria. In this report, the authors provide an assessment of the quality of the RCT included; describe the characteristics of all the included studies, the outcomes assessed, and the effectiveness of photon emission as a potential health assessment tool. This report demonstrates that the peer reviewed literature on UPE and human UPE measurement in particular is surprisingly large. Most of the human UPE literature is of good to high quality based on our systematic evaluation. However, an evaluation tool for systematically evaluating this type of "bio-evaluation" methodology is not currently available and would be worth developing. Publications in the peer reviewed literature over the last 50 years demonstrate that the use of "off-the-shelf" technologies and well described methodologies for the detection of human photon emissions are being used on a regular basis in medical and research settings. The overall quality of this literature is good and the use of this approach for determining inflammatory and oxidative states of patients indicate the growing use and value of this approach as both a medical and research tool.

Towards whole-body ultra-weak photon counting and imaging with a focus on human beings: a review.

For decades, the relationship between ultra-weak photon emission (UPE) and the health state of the body is being studied. With the advent of systems biology, attention shifted from the association between UPE and reactive oxygen species towards UPE as a reflection of changed metabolic networks. Essential for this shift in thinking is the development of novel photon count statistical methods that more reflect the dynamics of the systems organization. Additionally, efforts to combine and correlate UPE data with other types of measurements such as metabolomics be key to understand the complexity of the human body. This review describes the history and developments in the area of human UPE research from a technical - methodological perspective, an experimental perspective and a theoretical perspective. There is ample evidence that human UPE research will allow a better understanding of the body as a complex dynamical system. The future lies in the further development of an integrated UPE and metabolomics platform for a personalized monitoring of changes of the system towards health or disease.

Attributes characterizing spontaneous ultra-weak photon signals of human subjects.

Sixty visible range photon signals spontaneously emitted from the dorsal side of both hands of fifteen human subjects are analyzed with the aim of finding their attributes. The signals are of 30 min duration and detected in bins of 50 ms by two synchronized photo multipliers sensitive in the range (290-630 nm). Each signal is a time series of 36,000 elements. The attributes of its signal are determined from the statistical properties of time series. The mean and variance of time series determine the attributes signal strength and intercept (p0) and slope (p1) of the Fano Factor curve. The photon count distribution of the time series determines squeezed state parameters |α|, r, θ and ϕ, squeezed state index (SSI), and sum of the squares of residue (SSR). The correlation between simultaneously detected signals determines intercept (c0) and slope (c1) of their correlation curve. The variability of attributes is studied by calculating them in smaller intervals covering the entire signal. The profile of attribute at 12 sites in a subject is more informative and biologically relevant.

Statistical analysis of the spontaneously emitted photon signals from palm and dorsal sides of both hands in human subjects.

Photon signals emitted spontaneously from dorsal and palm sides of both hands were recorded using 6000 time windows of size T=50 ms in 50 healthy human subjects. These photon signals demonstrated universal behaviour by variance and mean. The data sets for larger time windows up to T=50s were obtained by merging the data recorded with T=50 ms. The behaviour of Fano factor regarding different window sizes was investigated. The Fano factor hovered around one in signals up to T=3s and increased slowly with the increase in window size. This indicated super-Poissonian distribution of photo counts. The Fano factor curve F(T) obtained by averaging all subjects and locations had a characteristic shape. Data suggest that the shape is essentially a combination of a smaller sub-population of individuals with sub-Poissonian and a larger sub-population with predominantly super-Poissonian photo count distribution. Averaging the data obtained by randomly shuffling observed data sets was flat and did not show any structure with T. The same applied both to the observed background data sets and the data sets obtained by randomly shuffling background in 50 measurements. The Fano factor was also flat in 50 measurements documented both by a standard LED as well as its shuffled data sets. The structure in the shape F(T) is characteristic of human signals. It may contain valuable information about metabolic processes and may have diagnostic relevance.

Using ultra-weak photon emission to determine the effect of oligomeric proanthocyanidins on oxidative stress of human skin.

Exposure of skin to ultraviolet (UV) radiation triggers oxidative stress in skin tissue that can lead to erythema, early skin aging or even cancer. It is suggested that oligomeric proanthocyanidins (OPCs), phytonutrients that belong to the polyphenol family have an anti-oxidant/anti-inflammatory activity on the skin. Measuring ultra-weak photon emission (UPE) is a non-invasive, fairly-sensitive and convenient technique for continuously monitoring oxidative stress. The present study was undertaken to confirm anti-oxidant activity of the specific OPCs cream formulation in human skin by measuring UPE of skin. In the present study 25 healthy female subjects participated. As a baseline measurement of skin, UPE was recorded from the dorsal surface of the subjects' hands before (spontaneous UPE) and after exposure to UV (UV-induced UPE). The effects of the OPCs cream on spontaneous and UV-induced UPE were measured using a fractionated UV exposure protocol. UV exposure resulted in an increase in UPE from both hands. Repeat UV exposure also resulted in a long-term increase of spontaneous UPE. This is likely due to depletion of anti-oxidant capacity of skin resulting in sensitization of skin to UV. It was assessed by measuring spontaneous UPE at 80 min after each UV exposure. Application of the OPCs cream immediately after UV exposure resulted in a significant (approx. 30%) decrease in UV-induced UPE. Topical OPCs cream application also reduced sensitization of skin to UV following repeated UV exposure (i.e., reduced long-term increase in spontaneous UPE). This study indicates that the specific OPCs cream formulation significantly decreases UV-induced oxidative stress in human skin based on UPE measurement. It therefore suggests that regular use of this OPCs cream might protect skin from harmful effects of UV.

Biophoton detection and low-intensity light therapy: a potential clinical partnership.

Low-intensity light therapy (LILT) is showing promise in the treatment of a wide variety of medical conditions. Concurrently, our knowledge of LILT mechanisms continues to expand. We are now aware of LILT's potential to induce cellular effects through, for example, accelerated ATP production and the mitigation of oxidative stress. In clinical use, however, it is often difficult to predict patient response to LILT. It appears that cellular reduction/oxidation (redox) state may play a central role in determining sensitivity to LILT and may help explain variability in patient responsiveness. In LILT, conditions associated with elevated reactive oxygen species (ROS) production, e.g. diabetic hyperglycemia, demonstrate increased sensitivity to LILT. Consequently, assessment of tissue redox conditions in vivo may prove helpful in identifying responsive tissues. A noninvasive redox measure may be useful in advancing investigation in LILT and may one day be helpful in better identifying responsive patients. The detection of biophotons, the production of which is associated with cellular redox state and the generation of ROS, represents just such an opportunity. In this review, we will present the case for pursuing further investigation into the potential clinical partnership between biophoton detection and LILT.

Free radicals and low-level photon emission in human pathogenesis: state of the art.

Convincing evidence supports a role for oxidative stress in the pathogenesis of many chronic diseases. The model includes the formation of radical oxygen species (ROS) and the misassembly and aggregation of proteins when three tiers of cellular defence are insufficient: (a) direct antioxidative systems, (b) molecular damage repairing systems, and (c) compensatory chaperone synthesis. The aim of the present overview is to introduce (a) the basics of free radical and antioxidant metabolism, (b) the role of the protein quality control system in protecting cells from free radical damage and its relation to chronic diseases, (c) the basics of the ultraweak luminescence as marker of the oxidant status of biological systems, and (d) the research in human photon emission as a non-invasive marker of oxidant status in relation to chronic diseases. In considering the role of free radicals in disease, both their generation and their control by the antioxidant system are part of the story. Excessive free radical production leads to the production of heat shock proteins and chaperone proteins as a second line of protection against damage. Chaperones at the molecular level facilitate stress regulation vis-à-vis protein quali y control mechanisms. The manifestation of misfolded proteins and aggregates is a hallmark of a range of neurodegenerative disorders including Alzheimer's disease, Parkinson's disease, amylotrophic lateral sclerosis, polyglutamine (polyQ) diseases, diabetes and many others. Each of these disorders exhibits aging-dependent onset and a progressive, usually fatal clinical course. The second part reviews the current status of human photon emission techniques and protocols for recording the human oxidative status. Sensitive photomultiplier tubes may provide a tool for non-invasive and continuous monitoring of oxidative metabolism. In that respect, recording ultraweak luminescence has been favored compared to other indirect assays. Several biological models have been used to illustrate the technique in cell cultures and organs in vivo. This initiated practical applications addressing specific human pathological issues. Systematic studies on human emission have presented information on: (a) procedures for reliable measurements, and spectral analysis, (b) anatomic intensity of emission and left-right symmetries, (c) biological rhythms in emission, (d) physical and psychological influences on emission, (e) novel physical characteristics of emission, and (f) the identification of ultraweak photon emission with the staging of ROS-related damage and disease. It is concluded that both patterns and physical properties of ultraweak photon emission hold considerable promise as measure for the oxidative status.

Quantum squeezed state analysis of spontaneous ultra weak light photon emission of practitioners of meditation and control subjects.

Research on human ultra-weak photon emission (UPE) has suggested a typical human emission anatomic percentage distribution pattern. It was demonstrated that emission intensities are lower in long-term practitioners of meditation as compared to control subjects. The percent contribution of emission from different anatomic locations was not significantly different for meditation practitioners and control subjects. Recently, a procedure was developed to analyze the fluctuations in the signals by measuring probabilities of detecting different numbers of photons in a bin and correct these for background noise. The procedure was tested utilizing the signal from three different body locations of a single subject, demonstrating that probabilities have non-classical features and are well described by the signal in a coherent state from the three body sites. The values indicate that the quantum state of photon emitted by the subject could be a coherent state in the subject being investigated. The objective in the present study was to systematically quantify, in subjects with long-term meditation experience and subjects without this experience, the photon count distribution of 12 different locations. Data show a variation in quantum state parameters within each individual subject as well as variation in quantum state parameters between the groups.

Differential effects of relaxation techniques on ultraweak photon emission.

BACKGROUND: Evidence has accumulated favoring the possible role of oxidative stress in the pathogenesis of many chronic diseases. Meditation is utilized as an adjunct to conventional medical treatment for several clinical conditions. A few studies suggest a role of long-term meditation in the control of the free-radical metabolism. Many techniques for recording reactive oxygen species (ROS) have been made available. However, most are invasive and none are applicable to all conditions. Attention has recently been drawn to spontaneous ultraweak photon emission (UPE). However, the application of this method in meditation studies is very limited. OBJECTIVE: The present study recorded spontaneous UPE at multiple anatomic locations of subjects with long-term experience in transcendental meditation (TM) and compared this with a group that practiced other meditation techniques (OMT) and with subjects having no meditation experience. METHODS: The study examined the anatomic pattern of UPE of 20 subjects practicing TM, compared to 20 subjects practicing OMT, and 20 control subjects with no experience in meditation. Subjects were men who were reported to be healthy and nonsmokers. Meditation was not practiced on the day prior to recording. UPE was recorded in a dark room, using a highly sensitive, cooled photomultiplier system designed for manipulation in three directions. The protocol for the multisite registration of UPE included recording 12 anatomic locations, including the anterior torso, head, neck, and hands. RESULTS: Data demonstrated emission intensities in the TM and OMT groups that were 27% and 17% lower, respectively, compared to the control group. The decrease was recorded at all anatomic locations. The percent emission contribution of each location to total emission was very similar for the three groups. CONCLUSIONS: Data supported the hypothesis that persistent meditation resulted in decreased UPE. However, the determination of oxidation levels as the source of group differences needs to be verified further to confirm our hypothesis.

Thermoluminescence in ultra-high dilution research.

OBJECTIVE: Thermoluminescence of homeopathically prepared ultra-high dilutions of lithium chloride in deuterium oxide (D2O) (10 (-30) g cm (-3); C15 LiCl in D2O) and similarly prepared D2O (C15 D2O) was compared in three experiments varying (1) time between preparation of substance and time of experimentation, and (2) time between irradiation and thermoluminescence recording. Data were also compared with a normal solution 0.1 M LiCl in D2O and with pure D2O. METHODS: To record thermoluminescence, the solutions were frozen in liquid nitrogen (77 K, -196 degrees C), irradiated with 1 kGy, and after a succession of definite storage periods, were progressively rewarmed to 238 K (-35 degrees C). RESULTS: Thermoluminescence patterns of C15 LiCl in D2O, C15 D2O, and D2O were similar. The average thermoluminescence intensity of C15 LiCl in D2O tended to be less than C15 D2O when tests were performed shortly after preparation of the substances and with short storage time between irradiation and recording of thermoluminescence. Thermoluminescence of D2O was significantly lower than from C15 D2O, particularly when the time between irradiation and recording was increased. CONCLUSION: The nature of the phenomena here described still remains unexplained. Nevertheless, data suggest that thermoluminescence might be developed into a promising tool to study homeopathically prepared ultra- high dilutions.

Photocount distribution of photons emitted from three sites of a human body.

Spontaneous photon emission from 30 sites on the skin of a live human subject is measured at different times and on different days. Signals from three representative sites of low, intermediate and high intensities are selected for further analysis. Fluctuations in these signals are measured by the probabilities of detecting different numbers of photons in a bin. The probabilities have non-classical features and are well described by the signal in a quantum squeezed state of photons. Measurements with bins of three sizes yield same values of three parameters of the squeezed state. A procedure for making correction due to background noise is developed. The correction changes the parameters of the quantum state. The new state appears more like a coherent state of photons.

Anatomic characterization of human ultra-weak photon emission in practitioners of transcendental meditation(TM) and control subjects.

BACKGROUND: Research on human ultra-weak photon emission (UPE, biophoton emission) has raised the question whether a typical human emission anatomic percentage distribution pattern exists in addition to individual subject overall anatomic summation intensity differences. The lowest UPE intensities were observed in two subjects who regularly meditate. Spectral analysis of human UPE has suggested that ultra-weak emission is probably, at least in part, a reflection of free radical reactions in a living system. It has been documented that various physiologic and biochemical shifts follow the long-term practice of meditation and it is inferred that meditation may impact free radical activity. OBJECTIVE: To systematically quantify, in subjects with long-term transcendental meditation (TM) experience and subjects without this experience, the UPE emission of the anterior torso, head and neck plus the hands in an attempt to document the differences by the two groups. SUBJECTS: Subjects were 20 men reported to be healthy and nonsmokers. Each of the subjects in the meditation group had practiced TM twice daily for at least the past 10 years. METHODS: UPE in 20 subjects was recorded in a dark room using a highly sensitive, cooled photomultiplier system designed for manipulation in three directions. The protocol for multisite registration of spontaneous emission includes recording of 12 anatomic locations of anterior torso, head, and hands. RESULTS: Data demonstrate emission intensities that are lower in TM practitioners as compared to control subjects. The percent contribution of emission from most anatomic locations was not significantly different for TM practitioners and control subjects. Exceptions are the contributions of throat and palm. CONCLUSION: In subjects with long-term TM experience, the UPE emission is different from control subjects. Data support the hypothesis that free radical reactions can be influenced by TM.

Anatomic characterization of human ultra-weak photon emission with a moveable photomultiplier and CCD imaging.

Ultra-weak photon emission of a living system has received scientific attention because of its potential for monitoring oxidative metabolism and oxidative damage to tissues. Heretofore, most studies have focused only on the emission from hands. The data regarding emission from other anatomic locations are limited. A previous multi-anatomic site recording of four subjects quantitatively demonstrated that the emission from several corresponding anatomic locations could differ by as much as a factor of 4. The data also suggested a "common" anatomic emission percentage distribution pattern. This information raised the question whether such a typical anatomic percentage emission exists. The objective of the present paper is to systematically replicate the emission from identical anatomic locations to document whether the anatomic percentage distribution pattern is generic. Part 1 includes the recording of ultra-weak photon emission from one sample subject over the torso, head and upper extremities with a highly sensitive charge-coupled device (CCD). Part 2 includes the analysis of that data to select a series of anatomic locations that were subsequently studied with a group of 20 subjects utilizing a highly sensitive, cooled and moveable (in three directions) photomultiplier system. Total sum emission of all recorded anatomic locations per subject fluctuates in this study almost 5-fold between subjects. However, the contribution of each anatomic location to the total emission from each subject was approximately the same percentage for each subject and similar to the sample CCD subject. The deviation of the anatomic percentage contribution for each subject was also established. The study presents evidence that there is a "common" anatomic percentage distribution pattern of ultra-weak photon emission for corresponding locations from each subject.