Ultraweak Photon Emission Research Articles

Non-Invasive Label-Free Imaging of Oxidative Processes in Human Skin

Oxidative processes present across all types of organisms, including humans, cause chemical formation of electron excited species with subsequent endogenous ultra-weak photon emission. Thus, imaging of this endogenous chemiluminescence using ultra-sensitive devices potentially enables label-free monitoring of oxidative stress in optically accessible areas of human body, such as human skin. However, no quantified imaging of oxidative processes in human skin has been performed until now using endogenous chemiluminescence under controlled extent of oxidative stress conditions. Furthermore, the mechanisms and dynamics of endogenous chemiluminescence is not fully explored.Here we demonstrate that different degrees of oxidative processes on skin can be spatially resolved through non-invasive label-free endogenous chemiluminescence imaging in a quantitative manner. Additionally, to obtain insight into the underlying mechanisms, we developed and employed a minimal chemical model of skin based on a mixture of lipid (linoleic acid) / melanin / water to show that it reproduces essential features of the response of a real skin to oxidative stress. Our results contribute to novel non-invasive label-free methods for quantitative monitoring of oxidative processes and oxidative stress.We acknowledge support by the Czech Science Foundation, project no. P102/15-17102S. Authors participate in COST Action BM1309 and bilateral exchange project between Czech and Slovak Academies of Sciences, no. SAV-15-22.

Application of Spontaneous Photon Emission in the Growth Ages and Varieties Screening of Fresh Chinese Herbal Medicines.

Ultraweak photon emission emitted by all living organisms has been confirmed to be a noninvasive indicator for their physiological and pathological characteristics. In this study, we investigated the characteristics of spontaneous photon emission (SPE) and the contents of specific active compounds of roots and flowers buds of several fresh Chinese herbal medicines (natural medicines) with different growth ages and varieties. The results revealed that the contents of specific active compounds from same species herbs with different growth ages and varieties were significantly different, and this difference could be reflected by their SPE. Because the contents of specific bioactive constituents in Chinese herbs are closely related to their quality and curative effect, the SPE measurement technique may contribute to the quality control of Chinese herbal medicine in the future.

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.

Spontaneous photon emission: A promising non-invasive diagnostic tool for breast cancer

Ultra-weak photon emission (UPE) has attracted significant scientific attention for its potential to monitor the physiological and pathological characteristics of living systems. In this study, we investigated the strength of spontaneous photon emission (SPE) from the right (R) and left (L) side of body surface of human breast cancer-bearing nude mice models, considering the entire breast cancer growth process, and healthy controls using a photon detector. And then we calculated the ratio of the average SPE strength (ratio (R/L)) between the right and left side of each mouse. Cluster analysis was used to evaluate the accuracy rates of strength-R, strength-L, ratio (R/L) and their combination to identify tumor mice from the controls. Our results revealed that the discriminating powers of different parameters were different in different growth stages of tumor: in tumor incubation period, the accuracy rates of strength-R, strength-L, ratio (R/L) and their combination to identify tumor mice from control mice were 63.6%, 40.9%, 81.8% and 86.4%, respectively; For nude mice with tumor diameter <0.5cm, the accuracies were 72.7%, 45.5%, 86.4% and 90.9%; and for nude mice with tumor diameters larger than 1.5cm, the accuracies were 86.4%, 77.3%, 95.5% and 100%, respectively. These results indicated that the SPE from the body surface of the lesion site could significantly distinguish tumor mice from the controls when tumors were obvious or when there were no obvious morphological changes, although the accuracy was relatively low. The results suggest that SPE, as a sensitive and promising optical method, may contribute to the preliminary screening of breast cancer, especially for early diagnosis, and it may play a critical role in curtailing the effects of breast cancer and improving the survival of patients in the future.

Measuring ultra-weak photon emission as a non-invasive diagnostic tool for detecting early-stage type 2 diabetes: A step toward personalized medicine

The global prevalence of type 2 diabetes is estimated to reach 4.4% by 2030, placing a significant burden on our healthcare system. Therefore, the ability to identify patients in early stages of the disease is essential for both prevention and effective management, and diagnostic methods based on traditional Chinese medicine (TCM) may be suitable for identifying patients with early-stage type 2 diabetes. Here, a panel of three physicians trained in TCM classified 44 pre-diabetic subjects into three syndrome subtypes using TCM-based diagnostics. In addition, ultra-weak photon emission (UPE) was measured at four anatomical sites in each subject. Ten properties encompassing 40 parameters were then extracted from the UPE time series. Statistical analyses, including multinomial logistic regression, were performed using the results of each parameter measured at the four sites. Sixteen UPE parameters were then selected and used to discriminate between the three subtypes of pre-diabetic subjects. Next, Spearman's correlation coefficient was used to quantify the correlation between the 16 UPE parameters and the TCM-based diagnoses. The resulting correlation networks accurately reflected the differences between the three syndrome subtypes. These results suggest that UPE is a suitable tool for detecting subtypes in early-stage type 2 diabetes. In addition, our results provide evidence that TCM may represent an important step toward personalized medicine.

Optically functionalized biomorphism of bean seeds

Cannellini-Lingot beans (Phaseolus vulgaris) have been analyzed as optical collectors for ultra-weak photon emission (UPE) during the very early germination process. The optical path of the internally-generated light was investigated using a custom ray-tracing numerical code. Specific light maps reveal an optical focus in correspondence with the plumula and radical tip regions of the embryo. Endoscopic photon counting of the emission confirmed the numerical prediction. The whole seed shape has been geometrically studied and represented in terms of parabolic surfaces; a specific asymmetric morphology is the consequence of the asymmetric distribution of the embryo inside the seed; it seems to be functional for collecting and focusing UPE on the environmentally sensitive plumula and radical tip.

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, β-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, β-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 from the Seed Coat in Response to Temperature and Humidity—A Potential Mechanism for Environmental Signal Transduction in the Soil Seed Bank

Seeds beneath the soil sense the changing environment to time germination and seedling emergence with the optimum time of year for survival. Environmental signals first impact with the seed at the seed coat. To investigate whether seed coats have a role in environmental sensing we investigated their ultraweak photon emission (UPE) under the variable temperature, relative humidity and oxygen conditions they could experience in the soil seed bank. Using a custom‐built luminometer we measured UPE intensity and spectra (300–700 nm) from Phaseolus vulgaris seeds, seed coats and cotyledons. UPE was greatest from the internal surface of the seed coat. Seed coat UPE increased concomitantly with both increasing temperature and decreasing relative humidity. Emission was oxygen dependent and it was abolished by treatment with dinitrophenylhydrazine, demonstrating the key role of seed coat carbonyls in the phenomenon. We hypothesize that beneath the soil surface the attenuation of light (virtual darkness: low background noise) enables seeds to exploit UPE for transducing key environmental variables in the soil (temperature, humidity and oxygen) to inform them of seasonal and local temperature patterns. Overall, seed coats were found to have potential as effective transducers of key fluctuating environmental variables in the soil.

Ultra-weak photon emission of hands in aging prediction

Aging has been one of the several topics intensely investigated during recent decades. More scientists have been scrutinizing mechanisms behind the human aging process. Ultra-weak photon emission is known as one type of spontaneous photon emission that can be detected with a highly sensitive single photon counting photomultiplier tube (PMT) from the surface of human bodies. It may reflect the body's oxidative damage. Our aim was to examine whether ultra-weak photon emission from a human hand is able to predict one's chronological age. Sixty subjects were recruited and grouped by age. We examined four areas of each hand: palm side of fingers, palm side of hand, dorsum side of fingers, and dorsum side of hand. Left and right hand were measured synchronously with two independent PMTs. Mean strength and Fano factor values of photon counts were utilized to compare the UPE patterns of males and females of different age groups. Subsequently, we utilized UPE data from the most sensitive PMT to develop an age prediction model. We randomly picked 49 subjects to construct the model, whereas the remaining 11 subjects were utilized for validation. The results demonstrated that the model was a good regression compared to the observed values (Pearson's r=0.6, adjusted R square=0.4, p=9.4E−7, accuracy=49/60). Further analysis revealed that the average difference between the chronological age and predicted age was only 7.6±0.8years. It was concluded that this fast and non-invasive photon technology is sufficiently promising to be developed for the estimation of biological aging.

Effects of growth altitude on chemical constituents and delayed luminescence properties in medicinal rhubarb

To improve the quality control of herbal drugs, there has been a major shift from evaluating individual chemicals to evaluating multiple-constituent chemicals, given the multi-pharmacology nature of herbal drugs. Therefore, rapid, systematic assays are needed in order to assess the quality of medicinal herbs using a comprehensive, integrated approach. Light-induced delayed luminescence (DL) is used to measure decaying long-term ultra-weak photon emissions following excitation with light. DL is considered to be a sensitive indicator for characterizing the properties of biological systems and herbal medicines with various therapeutic properties. The aim of this study was to examine the feasibility of using DL as a novel quality-assessment tool using rhubarb material as a model system, and to establish the correlation between DL parameters and the chemical constituents of rhubarb. Raw roots and rhizomes were collected from rhubarb (Rheum palmatum L.) at various elevations in western China. HPLC analysis was used to identify fourteen bioactive constituents. Five DL parameters were calculated from the DL decay curves of the rhubarb samples. Statistical tools, including principal component analysis, were used to classify the rhubarb samples using data obtained using two different assays. Finally, Spearman's correlation coefficient was calculated to quantify the correlation between the bioactive compounds and corresponding DL parameters. We found that both the chemical analysis and DL measurements reflect variations in the quality of rhubarb due to environment factor. The DL parameters were correlated significantly with the bioactive chemical constituents. Our results indicate that DL is a promising tool for evaluating multiple constituents and for assessing the therapeutic properties of herbal medicines. Thus, DL may be used as part of a comprehensive system for assessing the quality and/or therapeutic properties of herbal medicines.

Photocount statistics of ultra-weak photon emission from germinating mung bean

Ultra-weak photon emission (UPE) is an endogenous bioluminescence phenomenon present in all biological samples with an active oxidative metabolism, even without an external pre-illumination. To verify the potential of UPE for non-invasive monitoring of metabolism and growth in germinating plants, the aim of this study was to investigate the UPE from a model system - germinating mung bean seedlings (Vigna radiata) - and analyze the statistical properties of UPE during the growth in two different conditions of imbibition (pure water and 1% sucrose). We found that in all days and in both conditions, photocount distributions of UPE time series follow the negative binomial distribution whose parameters changed during the growth due to the increasing ratio of signal-to-detector dark count. Correspondingly for both groups, the mean values of UPE increased during the seedlings growth, while the values of Fano factor show a decreasing trend towards 1 during the 6day period. While our results do not show any significant difference in hypocotyl length and weight gain between the two groups of mung seedlings, there is an indication of a tiny suppressing effect of sucrose on UPE intensity. We believe that UPE can be exploited for a sensitive non-invasive analysis of oxidative metabolism during the plant development and growth with potential applications in agricultural research.

Endogenous spontaneous ultraweak photon emission in the formation of eye-specific retinogeniculate projections before birth.

In 1963, it was suggested [Sperry, R.W. (1963). Chemoaffinity in the orderly growth of nerve fiber patterns and connections. Proc. Natl. Acad. Sci. USA 50, 703-710.] that molecular cues can direct the development of orderly connections between the eye and the brain (the "chemoaffinity hypothesis"). In the same year, the amazing degree of functional accuracy of the visual pathway in the absence of any external light/photon perception prior to birth [Wiesel, T.N and Hubel, D.H. (1963). Single-cell responses in striate cortex of kittens deprived of vision in one eye. J. Neurophysiol. 26, 1003-1017.] was discovered. These recognitions revealed that the wiring of the visual system relies on innate cues. However, how the eye-specific retinogeniculate pathway can be developed before birth without any visual experience is still an unresolved issue. In the present paper, we suggest that Müller cells (functioning as optical fibers), Müller cell cone (i.e. the inner half of the foveola that is created of an inverted cone-shaped zone of Müller cells), discrete retinal noise of rods, and intrinsically photosensitive retinal ganglion cells might have key functions by means of retinal spontaneous ultraweak photon emission in the development of eye-specific retinogeniculate pathways prior to birth.

Spontaneous Photon Emissions in Photoreceptors: Potential Convergence of Arrhenius Reactions and the Latency for Rest Mass Photons to Accelerate to Planck Unit Energies

         Spontaneous or phasic voltage shifts normally occur across the plasma membranes of photoreceptors without impingement from external photons on average once every ~38 s (26 mHz). There have been arguments that all living cells spontaneously exhibit Bokkon-type ~10-12 W·m-2 ultraweak photon emissions (UPE) independent of temperature-based Arrhenius reactions. However the non-Gaussian distribution of the phasic voltage-photon events can be accommodated by the results of accelerating mass equivalents of resting photons into the time frame of single electron orbits which would require ~38 s. This condition is strongly dependent upon the presence of free protons within Pollack’s interfacial water states. The duration associated with the energy from the magnetic moment of a proton in a 1 nT magnetic field generated by 2 pA currents through channels across the membrane when divided into Planck’s constant is ~38 s. Second shell activation energies (~10-20 J) that are correlated with proton movements within a human photoreceptor’s volume of water result in a rate constant that is equivalent to energy within the range of Cosmic Microwave Background. One solution to accommodate the median emission of ~2·10-12 W·m-2 as a mass volume within a cell, the equivalent magnetic field strength of 1 nT, the interconnected 10-20 J and the diffusivity coupled to the hydrogen line suggests UPE may be reverse reactions from all living mass originating as Popp’s primordial solar photon energy. If these convergence solutions are valid then non-local photon formation would be the source of these spontaneous shifts in photoreceptors that would be mediated by a plethora of local physical chemical manifestations.

Polychromatic spectral pattern analysis of ultra-weak photon emissions from a human body

Ultra-weak photon emission (UPE), often designated as biophoton emission, is generally observed in a wide range of living organisms, including human beings. This phenomenon is closely associated with reactive oxygen species (ROS) generated during normal metabolic processes and pathological states induced by oxidative stress. Application of UPE extracting the pathophysiological information has long been anticipated because of its potential non-invasiveness, facilitating its diagnostic use. Nevertheless, its weak intensity and UPE mechanism complexity hinder its use for practical applications. Spectroscopy is crucially important for UPE analysis. However, filter-type spectroscopy technique, used as a conventional method for UPE analysis, intrinsically limits its performance because of its monochromatic scheme. To overcome the shortcomings of conventional methods, the authors developed a polychromatic spectroscopy system for UPE spectral pattern analysis. It is based on a highly efficient lens systems and a transmission-type diffraction grating with a highly sensitive, cooled, charge-coupled-device (CCD) camera. Spectral pattern analysis of the human body was done for a fingertip using the developed system. The UPE spectrum covers the spectral range of 450–750nm, with a dominant emission region of 570–670nm. The primary peak is located in the 600–650nm region. Furthermore, application of UPE source exploration was demonstrated with the chemiluminescence spectrum of melanin and coexistence with oxidized linoleic acid.

Phosphene perception is due to the ultra-weak photon emission produced in various parts of the visual system: glutamate in the focus.

Phosphenes are experienced sensations of light, when there is no light causing them. The physiological processes underlying this phenomenon are still not well understood. Previously, we proposed a novel biopsychophysical approach concerning the cause of phosphenes based on the assumption that cellular endogenous ultra-weak photon emission (UPE) is the biophysical cause leading to the sensation of phosphenes. Briefly summarized, the visual sensation of light (phosphenes) is likely to be due to the inherent perception of UPE of cells in the visual system. If the intensity of spontaneous or induced photon emission of cells in the visual system exceeds a distinct threshold, it is hypothesized that it can become a conscious light sensation. Discussing several new and previous experiments, we point out that the UPE theory of phosphenes should be really considered as a scientifically appropriate and provable mechanism to explain the physiological basis of phosphenes. In the present paper, we also present our idea that some experiments may support that the cortical phosphene lights are due to the glutamate-related excess UPE in the occipital cortex.

The Physical Mechanism for Retinal Discrete Dark Noise: Thermal Activation or Cellular Ultraweak Photon Emission?

For several decades the physical mechanism underlying discrete dark noise of photoreceptors in the eye has remained highly controversial and poorly understood. It is known that the Arrhenius equation, which is based on the Boltzmann distribution for thermal activation, can model only a part (e.g. half of the activation energy) of the retinal dark noise experimentally observed for vertebrate rod and cone pigments. Using the Hinshelwood distribution instead of the Boltzmann distribution in the Arrhenius equation has been proposed as a solution to the problem. Here, we show that the using the Hinshelwood distribution does not solve the problem completely. As the discrete components of noise are indistinguishable in shape and duration from those produced by real photon induced photo-isomerization, the retinal discrete dark noise is most likely due to ‘internal photons’ inside cells and not due to thermal activation of visual pigments. Indeed, all living cells exhibit spontaneous ultraweak photon emission (UPE), mainly in the optical wavelength range, i.e., 350–700 nm. We show here that the retinal discrete dark noise has a similar rate as UPE and therefore dark noise is most likely due to spontaneous cellular UPE and not due to thermal activation.

Singlet oxygen production in Chlamydomonas reinhardtii under heat stress.

In the current study, singlet oxygen formation by lipid peroxidation induced by heat stress (40 °C) was studied in vivo in unicellular green alga Chlamydomonas reinhardtii. Primary and secondary oxidation products of lipid peroxidation, hydroperoxide and malondialdehyde, were generated under heat stress as detected using swallow-tailed perylene derivative fluorescence monitored by confocal laser scanning microscopy and high performance liquid chromatography, respectively. Lipid peroxidation was initiated by enzymatic reaction as inhibition of lipoxygenase by catechol and caffeic acid prevented hydroperoxide formation. Ultra-weak photon emission showed formation of electronically excited species such as triplet excited carbonyl, which, upon transfer of excitation energy, leads to the formation of either singlet excited chlorophyll or singlet oxygen. Alternatively, singlet oxygen is formed by direct decomposition of hydroperoxide via Russell mechanisms. Formation of singlet oxygen was evidenced by the nitroxyl radical 2,2,6,6-tetramethylpiperidine-1-oxyl detected by electron paramagnetic resonance spin-trapping spectroscopy and the imaging of green fluorescence of singlet oxygen sensor green detected by confocal laser scanning microscopy. Suppression of singlet oxygen formation by lipoxygenase inhibitors indicates that singlet oxygen may be formed via enzymatic lipid peroxidation initiated by lipoxygenase.

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.

Delayed luminescence: an experimental protocol for Chinese herbal medicines.

In Chinese medicine, raw herbal materials are used in processed and unprocessed forms aiming to meet the different requirements of clinical practice. To assure the chemical quality and therapeutic properties of the herbs, fast and integrated systematic assays are required. So far, such assays have not been established. Delayed luminescence (DL) refers to a decaying long-term ultraweak photon emission after exposure to light. Its decay kinetics under certain conditions may be a sensitive indicator reflecting the internal structural and chemical/physiological state of a biological system. DL measurements have been used in many applications for quality control. However, relatively little research has been reported on dried plant material such as Chinese herbs. The objective of the present study is to establish a protocol for direct and rapid DL measurements of dried Chinese herbal materials, including the determination of the dependence on: (a) the optimal excitation time utilizing a white light source; (b) the optimal size of the grinded herbal particle; and (c) the humidity conditions before and during measurement. Results indicate that stable and reproducible curves of DL photon emission depend mainly on the water content of herbal materials. To investigate the application of the established DL measurement protocol, non-processed and processed Aconitum (Aconitum carmichaelii Debx.), wild and cultivated rhubarb (Rheum palmatum L.) and ginseng (Panax ginseng C.A.Mey) of different ages were measured using DL. The results suggest that DL technology is a potential tool for assessment of dried Chinese herb qualities. The results warrant a further exploration of this technique in relation to therapeutic properties of the herbs. Copyright © 2016 John Wiley & Sons, Ltd.

Ultra-weak Photon Emissions Differentiate Malignant Cells from Non- Malignant Cells In Vitro

A fast, inexpensive, and accurate method for differentiating normal (non-malignant) cells from malignant cells could facilitate diagnosis and subsequently treatment. Although blood constituents are the current dominant indicators, we have found that Spectral Power Densities (SPD) obtained from only 100 s of measurements of spontaneous ultra-weak photon emissions (UPE) from cell cultures significantly differentiated malignant from non-malignant states. Breast cells were particularly differentiable from nonbreast cells according to their SPD profiles. More critically the combination of only 3 discrete frequency increment changes in SPD profiles accurately classified 85% of malignant breast cells from normal breast cells in culture. These results confirm results from our mouse experiments and preliminary observations from our human measurements that appropriately analyzed and interpreted SPD from very brief samples of UPE may be a viable tool for early detection of malignancy.