Abstracts
Fractal analysis and signal preprocessing of luminol dependent neutrophil photon emission
Michaela Nerudová [1], Michal Cifra [2], Honorata Nawrocka-Bogusz [3]
[1] Department of Circuit Theory, Faculty of Electrical Engineering, Czech Technical University in Prague, Czech Republic
[2] Institute of Photonics and Electronics, Academy of Sciences of the Czech Republic, Czech Republic
[3] Department of Biophysics, University of Medical Science, Poznan, Poland
Neutrophils are highly specialized white blood cells contributing to the immune response. Their basic function is the destruction of phagocytised microorganisms by respiratory burst, one of intercellular destruction mechanisms of pathogens. The mechanism of the respiratory burst consists in a more than ten times increase of the consumption of oxygen, and the production and releasing of large quantities reactive oxygen species (ROS).
Blood from healthy volunteers was used for the purpose of the study. Lithium heparin was used as the anticoagulant. Neutrophils were isolated from full blood by using density gradient method.
The physiotherapy treatment device Viofor JPS was used as a generator of the variable magnetic field. The mean induction of the variable magnetic field was 44.5 µT. The fundamental frequency of pulses of magnetic field was 180-195 Hz. The pulses were administered in the form of packets of pulses (12.5-29 Hz), groups of packages (2.8-7.6 Hz) and series (0.08-0.3 Hz). The form of impulses was close to the peak-shaped. M2P3 program according Viofor JPS device was applied, which means an application with intensity increasing to the chosen value. After reaching the chosen intensity, the value of impulse strength decreases. This process was repeated periodically. The sample with neutrophils was kept in magnetic field for 30 min in the dark at room temperature.
The respiratory burst was induced with PMA (phorbol 12-myristate 13-acetate), in order to produce submaximal stimulation of the respiratory burst.
Luminol is a luminescent probe for multiple types of ROS which are produced in the course of respiratory burst. The higher the concentration of species is (which are able to oxidize the luminol, leading to luminescence) the higher is the detected luminescence. The luminol-amplified photon emission has been measured from neutrophil suspension groups treated or nontreated with pulsed magnetic field with parameters defined above. Photon emission has been also measured from 5 µM luminol in 5 mM sodium bicarbonate buffer with 1.5 mM hydrogen peroxide. Photon emission has been measured using selected low-noise photomultiplier module H7360-01 in a light tight chamber.
We dedicated this study for preprocessing of the photonic signals and basic fractal analysis. We studied normalized fracal dimension (based on the Katz method) and sensitivity of fractal parameters to the preprocessing parameters in the experimental groups: unstimulated neutrophils, PMA-stimulated neutrophils, unstimulated neutrophils+magnetic field, PMA-stimulated neutrophils+magnetic field and luminol in bicarbonate buffer.
Michaela Nerudová [1], Michal Cifra [2], Honorata Nawrocka-Bogusz [3]
[1] Department of Circuit Theory, Faculty of Electrical Engineering, Czech Technical University in Prague, Czech Republic
[2] Institute of Photonics and Electronics, Academy of Sciences of the Czech Republic, Czech Republic
[3] Department of Biophysics, University of Medical Science, Poznan, Poland
Neutrophils are highly specialized white blood cells contributing to the immune response. Their basic function is the destruction of phagocytised microorganisms by respiratory burst, one of intercellular destruction mechanisms of pathogens. The mechanism of the respiratory burst consists in a more than ten times increase of the consumption of oxygen, and the production and releasing of large quantities reactive oxygen species (ROS).
Blood from healthy volunteers was used for the purpose of the study. Lithium heparin was used as the anticoagulant. Neutrophils were isolated from full blood by using density gradient method.
The physiotherapy treatment device Viofor JPS was used as a generator of the variable magnetic field. The mean induction of the variable magnetic field was 44.5 µT. The fundamental frequency of pulses of magnetic field was 180-195 Hz. The pulses were administered in the form of packets of pulses (12.5-29 Hz), groups of packages (2.8-7.6 Hz) and series (0.08-0.3 Hz). The form of impulses was close to the peak-shaped. M2P3 program according Viofor JPS device was applied, which means an application with intensity increasing to the chosen value. After reaching the chosen intensity, the value of impulse strength decreases. This process was repeated periodically. The sample with neutrophils was kept in magnetic field for 30 min in the dark at room temperature.
The respiratory burst was induced with PMA (phorbol 12-myristate 13-acetate), in order to produce submaximal stimulation of the respiratory burst.
Luminol is a luminescent probe for multiple types of ROS which are produced in the course of respiratory burst. The higher the concentration of species is (which are able to oxidize the luminol, leading to luminescence) the higher is the detected luminescence. The luminol-amplified photon emission has been measured from neutrophil suspension groups treated or nontreated with pulsed magnetic field with parameters defined above. Photon emission has been also measured from 5 µM luminol in 5 mM sodium bicarbonate buffer with 1.5 mM hydrogen peroxide. Photon emission has been measured using selected low-noise photomultiplier module H7360-01 in a light tight chamber.
We dedicated this study for preprocessing of the photonic signals and basic fractal analysis. We studied normalized fracal dimension (based on the Katz method) and sensitivity of fractal parameters to the preprocessing parameters in the experimental groups: unstimulated neutrophils, PMA-stimulated neutrophils, unstimulated neutrophils+magnetic field, PMA-stimulated neutrophils+magnetic field and luminol in bicarbonate buffer.
Patterns of biophoton emission from developing embryos and cell cultures
Lev V. Beloussov
Faculty of Biology, Moscow State University and International Institute of Biophysics (IIB)
The aim of this presentation is to review the main results of the measurements of ultraweak photon emission (UPE) from hen and fish eggs and monolayer cultures of fibroblasts, cardyomyocytes and hyppocamp cells performed in IIB from 1994 to 2005. In the developing hen eggs three UPE emitters of a diminished intensity can be distinguished: egg shell (displaying light-dependent delay luminescence), yolk and embryo proper (light-independent UPE). UPE of fish eggs and cell cultures does not exceed in the average the background level but can be clearly discriminated from the noise by Fourier analysis, revealing a set of periodicities within seconds – dozens of minutes range. In fish eggs UPE Fourier patterns are stage dependent. In cell cultures abrupt changes in Fourier patterns are associated with the administration of cytoskeletal drugs, addition of ligands (FGF in the case of fibroblasts) and changes in physiological conditions. In hyppocamp cells these changes occur spontaneously. Fourier analysis of biophoton emission may be a real time non-invasive tool for estimating physiological state of developing embryos and cell cultures.
Lev V. Beloussov
Faculty of Biology, Moscow State University and International Institute of Biophysics (IIB)
The aim of this presentation is to review the main results of the measurements of ultraweak photon emission (UPE) from hen and fish eggs and monolayer cultures of fibroblasts, cardyomyocytes and hyppocamp cells performed in IIB from 1994 to 2005. In the developing hen eggs three UPE emitters of a diminished intensity can be distinguished: egg shell (displaying light-dependent delay luminescence), yolk and embryo proper (light-independent UPE). UPE of fish eggs and cell cultures does not exceed in the average the background level but can be clearly discriminated from the noise by Fourier analysis, revealing a set of periodicities within seconds – dozens of minutes range. In fish eggs UPE Fourier patterns are stage dependent. In cell cultures abrupt changes in Fourier patterns are associated with the administration of cytoskeletal drugs, addition of ligands (FGF in the case of fibroblasts) and changes in physiological conditions. In hyppocamp cells these changes occur spontaneously. Fourier analysis of biophoton emission may be a real time non-invasive tool for estimating physiological state of developing embryos and cell cultures.
Impact
Daniel Fels
Swiss Tropical Institute (STI), Basel, Switzerland
Science needs funding. While applied research gets huge amounts, basic research gets less and when basic research refers to unknown or innovative topics funding gets close to nil.
The research of cell-internal em-fields is a marginalized topic and the question is how it gets from that position into a funded mainstream position.
Yet, if it gets there it will certainly be due to its interaction with the scientific community and hence, our focus has to be the scientific community. However, the scientific community is just a group within society in general. Therefore, we might focus on both groups.
In the presentation we will look at how we presented us in the past and how we may do so in the future. My major criticism refers to papers that report without hypothesis about correlated results and this in a journal, which nobody reads. We will emphasis, hence, again what good science writing means. I would further like to focus on competitive versus synergistic behaviour in groups and finally discuss public relations.
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Daniel Fels
Swiss Tropical Institute (STI), Basel, Switzerland
Science needs funding. While applied research gets huge amounts, basic research gets less and when basic research refers to unknown or innovative topics funding gets close to nil.
The research of cell-internal em-fields is a marginalized topic and the question is how it gets from that position into a funded mainstream position.
Yet, if it gets there it will certainly be due to its interaction with the scientific community and hence, our focus has to be the scientific community. However, the scientific community is just a group within society in general. Therefore, we might focus on both groups.
In the presentation we will look at how we presented us in the past and how we may do so in the future. My major criticism refers to papers that report without hypothesis about correlated results and this in a journal, which nobody reads. We will emphasis, hence, again what good science writing means. I would further like to focus on competitive versus synergistic behaviour in groups and finally discuss public relations.
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The theoretical scheme reflecting the mechanistic aspects behind UPE
Marek Rác [1], Pavel Pospíšil [1]
[1] Department of Biophysics, Faculty of Science, Palacký University, Třída Svobody 26 1192/12, 771 46 Olomouc, Czech Republic
Since the early 1920s when Gurwitsch firstly described phenomena of UPE countless theories about origin of the UPE were presented. However, in last few years the theory involving ROS became the one most supported by experiments. The schema reflecting the role of ROS in ultra-weak photon emission with the focus given on the mechanistic aspect leading to the formation of electronically-excited species responsible for the photon emission will be presented. While there are reactions in the scheme already proved by experiments still a lot of work has to be done do in order to reach the final understanding of the UPE origin.
This work was supported by the grants no. ED0007/01/01 Centre of the Region Haná for Biotechnological and Agricultural Research and no.CZ.1.07/2.3.00/20.0057 Operational Programme Education for Competitiveness from the Ministry of Education Youth and Sports, Czech Republic
Marek Rác [1], Pavel Pospíšil [1]
[1] Department of Biophysics, Faculty of Science, Palacký University, Třída Svobody 26 1192/12, 771 46 Olomouc, Czech Republic
Since the early 1920s when Gurwitsch firstly described phenomena of UPE countless theories about origin of the UPE were presented. However, in last few years the theory involving ROS became the one most supported by experiments. The schema reflecting the role of ROS in ultra-weak photon emission with the focus given on the mechanistic aspect leading to the formation of electronically-excited species responsible for the photon emission will be presented. While there are reactions in the scheme already proved by experiments still a lot of work has to be done do in order to reach the final understanding of the UPE origin.
This work was supported by the grants no. ED0007/01/01 Centre of the Region Haná for Biotechnological and Agricultural Research and no.CZ.1.07/2.3.00/20.0057 Operational Programme Education for Competitiveness from the Ministry of Education Youth and Sports, Czech Republic
WEMCA: The future within
Frederic Laager
S.U.P.E.R. Lab, Marie Curie Strasse 1, 26129 Oldenburg, Germany
We will show and discuss some of the new results acquired with our Weak Emission Multi-Channel Analyser (WEMCA). Measurements of eggs, cabbage, cress, blood and apples in the different states, will be this year’s topic. We will talk about the fingerprint system that will be available in the software of our device. Further ideas, developments and innovations will also be discussed.
Frederic Laager
S.U.P.E.R. Lab, Marie Curie Strasse 1, 26129 Oldenburg, Germany
We will show and discuss some of the new results acquired with our Weak Emission Multi-Channel Analyser (WEMCA). Measurements of eggs, cabbage, cress, blood and apples in the different states, will be this year’s topic. We will talk about the fingerprint system that will be available in the software of our device. Further ideas, developments and innovations will also be discussed.
Preliminary results of luminol photon emission from human neutrophils after pulsed magnetic field treatment in vitro
Honorata Nawrocka-Bogusz [1], Michal Cifra [2]
[1] Department of Biophysics, University of Medical Science, Poznań, Poland. [2] Institute of Photonics and Electronics, Academy of Sciences of the Czech Republic, Czech Republic
Neutrophils are highly specialized white blood cells. Their basic function is executing the phagocytosis and successive killing of phagocytized microorganisms. One of destruction mechanisms of pathogens is the production of reactive oxygen species (ROS), called respiratory burst. The respiratory burst was induced with PMA (phorbol 12-myristate, 13-acetate).
Blood samples from healthy blood donors were used for the study. Lithium heparin was used as the anticoagulant. Neutrophils were isolated from full blood by using density gradient method.
The variable magnetic field in the ELF range with a mean induction of 44.5 µT was applied for 30 minutes. The level of the magnetic induction application was growing at 12 seconds intervals starting from 4.45 µT to the selected value, then the intensity drops to the initial value, the process is repeated cyclically. The fundamental frequency of pulses was 180-195 Hz. The pulses were administered in the form of packets of pulses (12.5-29 Hz), groups of packages (2.8-7.6 Hz) and series (0.08-0.3 Hz).
Luminol is a luminescent probe for multiple types of ROS which are produced in the course of respiratory burst. The higher the concentration of ROS is, which are able to oxidize the luminol (which leads to luminescence), the higher the detected luminescence. The luminol-amplified photon emission has been measured from neutrophil suspension groups treated or nontreated with pulsed magnetic field with parameters defined above. Photon emission has been measured using a selected low-noise photomultiplier module H7360-01 in light tight chamber.
Depending on the age of neutrophils (after blood withdrawal and isolation), the pulsed magnetic field had stimulating or suppressing effect on peak photon emission detected. This indicates an effect of pulsed magnetic field exposure on neutrophil respiratory burst intensity.
Honorata Nawrocka-Bogusz [1], Michal Cifra [2]
[1] Department of Biophysics, University of Medical Science, Poznań, Poland. [2] Institute of Photonics and Electronics, Academy of Sciences of the Czech Republic, Czech Republic
Neutrophils are highly specialized white blood cells. Their basic function is executing the phagocytosis and successive killing of phagocytized microorganisms. One of destruction mechanisms of pathogens is the production of reactive oxygen species (ROS), called respiratory burst. The respiratory burst was induced with PMA (phorbol 12-myristate, 13-acetate).
Blood samples from healthy blood donors were used for the study. Lithium heparin was used as the anticoagulant. Neutrophils were isolated from full blood by using density gradient method.
The variable magnetic field in the ELF range with a mean induction of 44.5 µT was applied for 30 minutes. The level of the magnetic induction application was growing at 12 seconds intervals starting from 4.45 µT to the selected value, then the intensity drops to the initial value, the process is repeated cyclically. The fundamental frequency of pulses was 180-195 Hz. The pulses were administered in the form of packets of pulses (12.5-29 Hz), groups of packages (2.8-7.6 Hz) and series (0.08-0.3 Hz).
Luminol is a luminescent probe for multiple types of ROS which are produced in the course of respiratory burst. The higher the concentration of ROS is, which are able to oxidize the luminol (which leads to luminescence), the higher the detected luminescence. The luminol-amplified photon emission has been measured from neutrophil suspension groups treated or nontreated with pulsed magnetic field with parameters defined above. Photon emission has been measured using a selected low-noise photomultiplier module H7360-01 in light tight chamber.
Depending on the age of neutrophils (after blood withdrawal and isolation), the pulsed magnetic field had stimulating or suppressing effect on peak photon emission detected. This indicates an effect of pulsed magnetic field exposure on neutrophil respiratory burst intensity.
Combining ultra-weak photon emission with biochemical parameters having a direct link to REDOX processes in a collagen induced arthritis animal model
Eduard van Wijk [1,4], Masaki Kobayashi [2], Masutaka Kumasaka [2], Thomas Hankemeier [1], Jan van der Greef [1,3], Roeland van Wijk [4]
[1] Leiden University, Leiden, The Netherlands; [2] Tohoku Institute of Technology, Sendai, Japan; [3] TNO Netherlands Organization for Applied Scientific Research, Zeist, The Netherlands; [4] Meluna Research, Amersfoort, The Netherlands
This research is based on the notion that there is convincing evidence for a role of oxidative stress in the pathogenesis of many chronic diseases including Rheumatoid Arthritis (RA). It has stimulated the utilization of a non-invasive technique in human research based on spontaneous ultra-weak photon emission (UPE). UPE can be used as an effective biological marker of ROS inhibition and/or promotion of healing. Moreover recent developments in Metabolomics enable the measurement of biological molecules with a direct link to oxidation/reduction processes by using electrochemical detection. This so-called CoulArray technology enables the profiling of bodyfluids of biochemical components of this class. To combine both measurement techniques, arthritis is induced in mice by a combined injection of lipopolysaccharide (LPS) together with the joint cartilage component type II collagen. This injection of CII and LPS was continued every 2 weeks up to day 56.
Mice were imaged at day 70. Imaging was made with the highly sensitive CCD equipment at Prof Masaki Kobayashi’s laboratory. A Spectral Instruments 600 series CCD camera system (Spectral Instruments, Inc., AZ, USA) was used. Mounted CCD42-40 (e2v technologies Ltd., Essex, UK), which is a back-illuminated, fullframe operation CCD with a 2048 x 2048 pixel resolution and 13.5 x 13.5 mm pixel size. The camera system is equipped with a cooling head to maintain the CCD at -120°C using a closed-cycle mechanical cryogenic unit.
Imaging was made without and with using luminol. After imaging blood was collected for metabolomics measurements.
Data analysis is in process and first results will be discussed at the Seminar.
The combination of UPE measurement and Metabolomics allows both a system level and a wide biological level to be correlated. It could lead to a better understanding of the processes and the diagnostic value of differences.
Eduard van Wijk [1,4], Masaki Kobayashi [2], Masutaka Kumasaka [2], Thomas Hankemeier [1], Jan van der Greef [1,3], Roeland van Wijk [4]
[1] Leiden University, Leiden, The Netherlands; [2] Tohoku Institute of Technology, Sendai, Japan; [3] TNO Netherlands Organization for Applied Scientific Research, Zeist, The Netherlands; [4] Meluna Research, Amersfoort, The Netherlands
This research is based on the notion that there is convincing evidence for a role of oxidative stress in the pathogenesis of many chronic diseases including Rheumatoid Arthritis (RA). It has stimulated the utilization of a non-invasive technique in human research based on spontaneous ultra-weak photon emission (UPE). UPE can be used as an effective biological marker of ROS inhibition and/or promotion of healing. Moreover recent developments in Metabolomics enable the measurement of biological molecules with a direct link to oxidation/reduction processes by using electrochemical detection. This so-called CoulArray technology enables the profiling of bodyfluids of biochemical components of this class. To combine both measurement techniques, arthritis is induced in mice by a combined injection of lipopolysaccharide (LPS) together with the joint cartilage component type II collagen. This injection of CII and LPS was continued every 2 weeks up to day 56.
Mice were imaged at day 70. Imaging was made with the highly sensitive CCD equipment at Prof Masaki Kobayashi’s laboratory. A Spectral Instruments 600 series CCD camera system (Spectral Instruments, Inc., AZ, USA) was used. Mounted CCD42-40 (e2v technologies Ltd., Essex, UK), which is a back-illuminated, fullframe operation CCD with a 2048 x 2048 pixel resolution and 13.5 x 13.5 mm pixel size. The camera system is equipped with a cooling head to maintain the CCD at -120°C using a closed-cycle mechanical cryogenic unit.
Imaging was made without and with using luminol. After imaging blood was collected for metabolomics measurements.
Data analysis is in process and first results will be discussed at the Seminar.
The combination of UPE measurement and Metabolomics allows both a system level and a wide biological level to be correlated. It could lead to a better understanding of the processes and the diagnostic value of differences.
A new
method for removal of cosmic-ray-induced artifacts in ultra-weak photon
emission images
Felix Scholkmann [1], Everine van de Kraats [2]
[1] Biomedical Optics Research Laboratory, Division of Neonatology, University Hospital Zurich, 8091 Zurich, Switzerland
[2] Sino‐Dutch Centre for Preventive and Personalized Medicine, Division of Analytical Biosciences, LACDR, Leiden University, 2300 RA Leiden, The Netherlands
When performing a two dimensional measurement of ultra-weak photon emission (UPE) (UPE imaging) with long exposure durations (i.e. 30 min), artifacts in the UPE images are caused as a result of cosmic-rays hitting the charge-coupled device (CCD) camera. These cosmic-ray-induced artifacts (CRAs) impair the image quality and impede proper image analysis. We present a method for UPE image improvement that (i) detects CRAs by thresholding and analyzing the individual decay properties of CRAs, (ii) removes the detected CRAs by inpainting, i.e. restoration of the deteriorated image parts based on a discrete cosine transform-based penalized least square (DCT-PLS) regression, and (iii) denoises the image by adaptive Wiener filtering with noise estimation based on local image statistics. The processing steps of the proposed method will be discussed in detail. Examples will be given of how to use the method for biomedical and biological UPE imaging applications, e.g. for improving the quality of UPE images of human hands.
Felix Scholkmann [1], Everine van de Kraats [2]
[1] Biomedical Optics Research Laboratory, Division of Neonatology, University Hospital Zurich, 8091 Zurich, Switzerland
[2] Sino‐Dutch Centre for Preventive and Personalized Medicine, Division of Analytical Biosciences, LACDR, Leiden University, 2300 RA Leiden, The Netherlands
When performing a two dimensional measurement of ultra-weak photon emission (UPE) (UPE imaging) with long exposure durations (i.e. 30 min), artifacts in the UPE images are caused as a result of cosmic-rays hitting the charge-coupled device (CCD) camera. These cosmic-ray-induced artifacts (CRAs) impair the image quality and impede proper image analysis. We present a method for UPE image improvement that (i) detects CRAs by thresholding and analyzing the individual decay properties of CRAs, (ii) removes the detected CRAs by inpainting, i.e. restoration of the deteriorated image parts based on a discrete cosine transform-based penalized least square (DCT-PLS) regression, and (iii) denoises the image by adaptive Wiener filtering with noise estimation based on local image statistics. The processing steps of the proposed method will be discussed in detail. Examples will be given of how to use the method for biomedical and biological UPE imaging applications, e.g. for improving the quality of UPE images of human hands.
Contribution of LED-photobiomodulation to wound healing management
Michele Pelletier Aouizérate [1]
[1] Led academy President
Photobiomodulation is the process by which specific wavelengths are absorbed by chromophores in a selective way. This triggers signaling processes in biological cells. This process occurs naturally (natural photobiomodulation (PBM)), when walking outside for example. It is also a natural dynamic phototherapy. The process is modulated by many parameters that includes irradiance, the optimal combination of wavelengths, their mode of continuous or pulsed emission and in this case its transmission frequency, moment of day, etc. We briefly discuss some clinical cases for which the PBM was used alone or in combination. We discuss the prospects that are very promising.
Michele Pelletier Aouizérate [1]
[1] Led academy President
Photobiomodulation is the process by which specific wavelengths are absorbed by chromophores in a selective way. This triggers signaling processes in biological cells. This process occurs naturally (natural photobiomodulation (PBM)), when walking outside for example. It is also a natural dynamic phototherapy. The process is modulated by many parameters that includes irradiance, the optimal combination of wavelengths, their mode of continuous or pulsed emission and in this case its transmission frequency, moment of day, etc. We briefly discuss some clinical cases for which the PBM was used alone or in combination. We discuss the prospects that are very promising.