DE102015119990A1 - A method for controlling biological processes in microorganisms - Google Patents

Abstract

The present invention discloses a method for influencing or effecting at least one biological pathway of a microorganism in a substrate. The method comprises steps of: (a) providing a source of non-thermal plasma (NTP) for generating an NTP beam as defined in Table 1; (b) providing a power flow focusing device (EFFM) for transferring the NTP beam into a focused energy flow; (c) providing the microorganism in the substrate; and (d) operating the EFFM having a predetermined pulsed profile as defined in Table 2 to emit the focused energy flow to the at least one microorganism. The present invention further provides a system therefor.

Description

Field of the invention

The invention relates to the field of technology and application of non-thermal plasmas. In particular, the invention relates to means and methods for affecting or effecting biological pathways (or pathways) in microorganisms.

Background of the invention

Thermal plasmas and lasers are widely used in medicine to cut tissue by heat. The effects of such thermal plasmas on tissue are nonselective and difficult to control because they are primarily due to the transfer of intense heat. In contrast, non-thermal plasma does not generate heat, so its effects are more selective. Although electrical discharges producing non-thermal plasma have been known for a long time, their clinical potential has so far been largely ignored, and until recently, applications have been limited to the sterilization of inert surfaces or the modulation of cell attachment. The use of cold plasma for clinical applications requires the mechanistic understanding of the interaction of non-thermal plasma with living tissues.

The patent application US-A-2010/0130911 describes a plasma source device that provides a reduced pressure in the ionization chamber. This device is adapted for the sterilization of wounds. The patent application WO-A-2005/084569 discloses a disposable gas plasma tissue surface renewal device for skin treatment comprising an electromagnetically resonant focusing element. The aforementioned patent applications relate to the treatment of tissues on the surface of wounds.

The patent application KR-A-20140002357 discloses a food surface sterilization process using non-thermal plasma. The sterilization method of the present invention selects a non-thermal plasma suitable for the group of foods and optimizes the treatment conditions so as to increase the sterilizing effect on the microorganisms on the surface of food or food and the remaining agricultural chemical components on the surface Surface of food substances or food to decompose. The patent application US-A-2002/175068 teaches the decontamination of fluids or objects contaminated with chemical or biological agents using a distributed plasma reactor. In this publication, corona discharge can be generated using very short high voltage pulses (pulsed discharge) produced by a Tesla coil. Another pulsed discharge embodiment includes a primary coil surrounding a chamber having a cavity filled with a plurality of secondary coils. In one embodiment, the apparatus includes a ceiling-like structure useful for decontaminating surfaces or a fluid passing between spaced bare electrodes. In another embodiment, the bare electrodes define an internal treatment volume through which contaminated fluid flows.

It has been shown that non-thermal plasma generated by a dielectric barrier discharge (DBD) has dose-dependent effects on mammalian cells in culture, ranging from increasing cell proliferation to inducing apoptosis. It has further been shown that these effects are based primarily on the formation of intracellular reactive oxygen species (ROS), which are known to cause DNA damage ( S. Kalghatgi, C. Kelly, E. Cerchar, A. Fridman, G. Friedman, J. Azizkhan-Clifford, "Effects of Non-Thermal Plasma on Mammalian Cells" PLoS ONE, January 21, 2011, 6 (1) ). The registration US-A-2010/0145253 refers to the application of plasma to living tissue in vitro. Therefore, means and methods for inducing biological and / or biochemical effects on an entire subject in vivo using non-thermal gas plasma treatment are still needed.

The influence of a plasma treatment on seed germination has been investigated ( I. Filatova et al., "The effect of plasma treatment of seeds of some grains and legumes, on their sowing quality and productivity", Rome. Journ. Phys., Vol. 56, pp. 139-143, 2011 ; S. Bozena et al., "Influence of plasma treatment on wheat and oat germination", IEEE Transactions on Plasma Science, Vol. 38, 2010 ). These studies describe a device comprising a vacuum chamber, a centrifugal pump and a microwave resonator. The exposure time to the plasma treatment was over a period of several minutes to more than 40 Minutes. The effects reported in these studies continue to relate primarily to the surface of the seed coat and the sterilization of the seed coat.

The publication of Park et al. (2012) - "Analysis of the biological effects of a non-hepatic plasma on Saccharomyces cerevisiae", Journal of the Korean Physical Society, Vol. 60, pp. 916-920 , teaches the cellular and molecular responses of eukaryotic yeast to a non-thermal plasma at atmospheric pressure. It has been shown that the number of surviving cells is reduced by more than 50% when yeast cells are exposed to DBD plasma. It was further demonstrated in this paper that the protein profile of yeast treated with plasma exposure was affected relative to a control yeast by the plasma treatment.

Several publications teach the improved production of butanol by technically modified Saccharomyces cerevisiae: Steen et al. (2008) - "Metabolic engineering of Saccharomyces cerevisiae for the production of n-butanol", Microbial Cell Factories, Vol. 7, p. 36 discloses that Saccharomyces cerevisiae, which has been engineered with an n-butanol biosynthetic pathway, exhibits a tenfold improved production of n-butanol relative to a control sample. Si et al. (2014) report the improved accumulation of butanol from engineered endogenous 1-butanol pathway Saccharomyces cerevisiae using an endogenous pathway for 1-butanol production in Saccharomyces cerevisiae. Swidah et al. (2015) - "Butanol production in S. cerevisiae via a synthetic ABE pathway is enhanced by specific metabolic engineering and butanol resistance", Biotechnology for Biofuels, Vol. 8, p. 97 report that significant levels of n-butanol in S. cerevisiae can be achieved by transplantation of an n-butanol synthesis pathway from Clostridial sp. into the genome of the S. cerevisiae strain. Sakuragi et al. (2015) - "Enhanced butanol production by eukaryotic Saccharomyces cerevisiae engineered to contain an improved pathway" - Bioscience Biotechnology and Biochemistry, Vol. 79, pp. 314-320 teach the introduction of the butanol production pathway of Clostridium acetobutylicum into the yeast Saccharomyces cerevisiae. It has been reported that the elimination of the glycerol production pathway in yeast contributes to the improvement of 1-butanol production. In addition, the use of trans-enoyl-CoA reductase in the engineered pathway markedly improved 1-butanol production.

There is therefore a long-felt and unmet need to provide systems and methods for inducing biological and biochemical effects on a subject using effective non-thermal gas plasma treatments and protocols.

Summary of the invention

The present invention relates to the field of technology and non-thermal plasma application. In particular, the invention relates to means and methods for affecting or effecting biological pathways in living tissues, microorganisms, fluids, emulsions and gas media.

It is therefore an object of the present invention to disclose a method for affecting or effecting at least one biological pathway of a microorganism in a substrate, the method comprising steps of: (a) providing a source of non-thermal plasma (NTP ) for generating an NTP beam, said NTP beam containing argon, nitrogen and oxygen in a predetermined ratio; (b) providing a power flow focusing device (EFFM) for transferring the NTP beam into a focused energy flow; (c) providing the microorganism in the substrate; and (d) operating the EFFM with a predetermined pulsed profile to emit the focused energy flow to the at least one microorganism.

It is another object of the present invention to disclose a method for influencing or effecting at least one biological pathway of a microorganism in a substrate, the method comprising steps of: (a) providing a source of non-thermal plasma (NTP ) for generating an NTP beam as defined in Table 1; (b) providing a power flow focusing device (EFFM) for transferring the NTP beam into a focused energy flow; (c) providing the microorganism in the substrate; and (d) operating the EFFM having a predetermined pulsed profile as defined in Table 2 to emit the focused energy flow to the at least one microorganism.

It is another object of the present invention to disclose the method as defined above, wherein the EFFM comprises (1) a focusing vector and (2) an energy flow vector selected from the group consisting of an electric field vector, a magnetic field vector, an electromagnetic vector Field vector, a piezoelectric field vector, a piezoelectric field vector, and any combination thereof.

It is another object of the present invention to disclose the method as defined in any of the above, wherein the substrate is selected from the group consisting of a food, a liquid, a beverage, a suspension, a biological culture, a medium, a Growth medium, an emulsion, a biological tissue, a biological organism, a human, an animal, a plant, a fluid, a soil or solid, minerals, media, a gas, mixtures of gas and liquid, gas mixtures, cells, tissue cultures, organs and any combination thereof.

It is another object of the present invention to disclose the method as defined in any of the above, wherein the microorganism is selected from the group consisting of yeast, bacteria, archaea, algae, fungi, protozoa, viruses, spores, hype, prions and any combination thereof.

It is another object of the present invention to disclose the method as defined in any of the above, wherein the microorganism is further selected from the group consisting of: Candida, corynebacteria, aerobic bacteria, anaerobic bacteria, Listeria monocytogenes, Escherichia coli, Salmonella, Enterobacteriaceae family bacteria, Listeriaceae family bacteria, Gram-positive bacteria, Gram-negative bacteria, Staphylococcus aureus, Pseudomonas aeruginosa, Staphylococcus epidermidis, Staphylococcus haemolyticus, Enterococcus faecalis, Clostridiaceae family, Clostridium, Actinobacteria, and any combination thereof.

It is another object of the present invention to disclose the method as defined in any of the above, additionally comprising steps of affecting or effecting the at least one biological pathway of the microorganism within up to 24 hours from the emission of the focused energy flow to the microorganism, in particular within up to 6 hours from the emission of the focused energy flow to the microorganism.

It is another object of the present invention to disclose the method as defined in any of the above additionally comprising steps of comparing the effect on the at least one biological pathway of the microorganism having the effect in a control organism.

It is another object of the present invention to disclose the method as defined in any of the above, wherein the effect on at least one biological pathway is selected from the group consisting of: activation effect, inactivation effect, metabolic effect, effect on a biochemical pathway, effect on the production of at least one biochemical composition, effect on the production of recombinant proteins, effect on protein mutation, effect on protein expression, effect on protein modification, effect on protein profile, sterilization, infertility, antimicrobial effect, antibacterial effect, deactivating effect, survival effect, Infertility effect, loss of viability effect, killing effect, destruction effect, induction effect, destruction of pathogenic effect, bacterial destruction effect, viral destruction effect, genetic effect, water or gas disinfection effect, effect on water or Ga scavenging parameter, effect on the chemical composition, effect on the water or any other fluid chemistry, bacterial concentration effect, phenol concentration, effect on the chemical composition of crude oil, effect on crude oil components and concentration, effect on the production and / or destruction of biofuel or biodiesel compositions and any combination thereof.

It is another object of the present invention to disclose the method as defined in any of the above, wherein the effect on the at least one biological pathway is an effect on at least one biochemical pathway for the production of a composition selected from the group consisting of biofuel and / or products thereof, biodiesel and / or products thereof, organic compounds, metabolic compounds, metabolites, antibiotics, prodrugs, fermentation products, acid compositions, gas compositions, alcohol compositions, precursors of amino acids, ethanol, 1-propanol, 2-propanol, isopropanol, 2 , 3-butanedione, vicinal diketone, acetoin, acetone, 2,3-butanediol, methanol, isobutanol, butanol, 1-butanol, n-butanol, propanediol, diesel and any isomeric or racemic compound thereof, any intermediate pathway composition or any precursor thereof and any combination thereof.

It is another object of the present invention to disclose the method as defined in any of the above additionally comprising steps of effecting or affecting at least substantially the biological pathway of the microorganism in the substrate while at least substantially the pH, the hydrogen peroxide concentration or a combination thereof of the substrate is maintained.

• a) eine Quelle eines nicht thermischen Plasmas (NTP) zum Erzeugen eines NTP-Strahls wie in Tabelle 1 definiert; und
• b) eine Energieflussfokussierungsvorrichtung (EFFM) zum Überführen des NTP-Strahls in einen fokussierten Energiefluss, wobei die EFFM zumindest eine Öffnung für den Durchgang des NTP-Strahls aufweist und wobei die EFFM zumindest einen Energieflussvektor umfasst ausgewählt aus der Gruppe bestehend aus: i) zumindest einem elektrischen Feldvektor, ii) zumindest einem Magnetfeldvektor, iii) zumindest einem elektromagnetischen Feldvektor, iv) zumindest einem piezoelektrischen Feldvektor und v) zumindest einem piezomagnetischen Feldvektor, wobei das System zusätzlich zumindest einen fokussierenden Vektor umfasst, der dazu konfiguriert ist, den Plasmastrahl zu fokussieren, und wobei die EFFM dazu konfiguriert ist, Pulse des fokussierten Energieflusses mit einem vorbestimmten Profil wie in Tabelle 2 definiert bereitzustellen, um den zumindest einen biologischen Pfad des Mikroorganismus in dem Substrat zu beeinflussen oder zu bewirken, ein Verfahren zu offenbaren wie in irgendeinem des Obigen.

It is another object of the present invention in a system comprising:

• a) a source of non-thermal plasma (NTP) for generating an NTP beam as defined in Table 1; and
• b) an energy flux focusing device (EFFM) for transferring the NTP beam into a focused energy flow, the EFFM having at least one aperture for passage of the NTP beam and wherein the EFFM comprises at least one energy flux vector selected from the group consisting of: i) at least an electric field vector, ii) at least one magnetic field vector, iii) at least one electromagnetic field vector, iv) at least one piezoelectric field vector, and v) at least one piezomagnetic field vector, the system additionally comprising at least one focusing vector configured to focus the plasma beam and wherein the EFFM is configured to provide focused energy flow pulses having a predetermined profile as defined in Table 2 to affect or cause the at least one biological pathway of the microorganism in the substrate to reveal a method as in any of the above strength.

It is another object of the present invention to disclose a method for influencing or effecting at least one biological pathway of a microorganism in a substrate in a system comprising: (a) a non-thermal plasma source (NTP) for generating a NTP beam as defined in Table 1; and (b) an energy flux focusing device (EFFM) for transitioning the NTP beam into a focused energy flow, the EFFM having at least one aperture for passage of the NTP beam and wherein the EFFM comprises at least one energy flux vector selected from the group consisting of: ( i) at least one electric field vector, (ii) at least one magnetic field vector, (iii) at least one electromagnetic field vector, (iv) at least one piezoelectric field vector and (v) at least one piezomagnetic field vector. The system additionally includes at least one focusing vector configured to focus the plasma beam; the EFFM is configured to provide focused energy flux pulses having a predetermined profile as defined in Table 2 to affect or effect the at least one biological pathway of the microorganism in the substrate.

Detailed description of the figures

In order to better understand the invention and its implementation in practice, a plurality of embodiments will now be described by way of non-limiting example with reference to the accompanying drawings, in which: FIG

1A Figure 12C is a schematic illustration of a preferred embodiment of an exemplary Energy Flow Focusing Device (EFFM) for converting an NTP beam into a focused energy flow;

2A and 2 B not in scale schematic views of multiple electric or magnetic or electromagnetic field vectors in an EFFM ( 11a ) and a single electric or magnetic or electromagnetic field vector in an EFFM ( 11b ) as two embodiments of the present invention;

3A not to scale, a schematic view of an atmospheric pressure plasma jet as an embodiment of the present invention illustrated and 3B not to scale, illustrates a schematic view of a corona discharger as an alternative embodiment of the present invention;

4 is a graphic illustration of the production of biochemical compounds by S. cerevisiae, which has a focused energy flux treatment of 5 pulse cycles ( 4A ) and two focused Energy flow treatments of 5 pulse cycles ( 4B ) were compared with a control of untreated S. cerevisiae ( 4C );

5 is a graphic illustration of the production of biochemical compounds by S. cerevisiae, which has a focused energy flux treatment of 7 pulse cycles ( 5A ) and two focused energy flux treatments of 7 pulse cycles ( 5B ) were compared with a control S. cerevisiae ( 5C );

6 is a graphic illustration of the concentration of biochemical compounds generated by S. cerevisiae that undergo a focused energy flux treatment of 5 pulse cycles ( 6A ) and two focused energy flux treatments of 5 pulse cycles ( 6B ) were compared with a control S. cerevisiae ( 6C );

7 is a graphic illustration of the concentration of biochemical compounds generated by S. cerevisiae that undergo a focused energy flux treatment of 7 pulse cycles ( 7A ) and two focused energy flux treatments of 7 pulse cycles ( 7B ) were compared with a control S. cerevisiae ( 7C ).

Detailed Description of the Preferred Embodiments

The following description is provided throughout for all the chapters of the present invention to enable one skilled in the art to utilize the invention, and sets forth the best modes contemplated by the inventor for carrying out the invention. Various modifications, however, are adapted to be apparent to those skilled in the art, as the generic principles of the present invention have been specifically defined to provide means and methods for affecting or effecting biological pathways in living tissues, microorganisms, fluids, emulsions and gas media.

According to one embodiment, the present invention provides a method of affecting or effecting at least one biological pathway of a microorganism in a substrate, the method comprising the steps of: (a) providing a source of non-thermal plasma (NTP) for generating an NTP beam as defined in Table 1; (b) providing a power flow focusing device (EFFM) for transferring the NTP beam into a focused energy flow; (c) providing the microorganism in the substrate; and (d) operating the EFFM having a predetermined pulsed profile as defined in Table 2 to emit the focused energy flow to the at least one microorganism.

In another embodiment, the EFFM optionally includes a gas plasma flow regulator or monitoring means configured to control and / or monitor gas plasma flow parameters such as flow rate, flow pressure, mass flow, and gas plasma composition ratios.

In a specific embodiment, the gas plasma discharge comprises a gas mixture combination comprising argon (Ar), N ₂ and O ₂ . It is noted that the gas plasma flow controller or monitoring means is configured to separately control and / or monitor the plasma flow parameters of the gas plasma mixture and each of its components or ingredients.

Reference is now made to Table 1, which sets forth the optional plasma ingredient combinations (Nos. 1 to 10) that are within the scope of the present invention. Table 1: Plasma constituents (percent by weight), R is any material other than Ar, N ₂ or O ₂ ; a, b, c and r refer to the weight percentage of each of the ingredients No. Ar (%) N ₂ (%) O ₂ (%) R (%) 1 80-99 0-10 0-5 0 2 80-95 5-12 0-4 0 3 80-95 0-15 0-5 0 4 80-95 0-12 0-4 0-4 5 90 8th 2 0 6 85 9 6 0 7 90 6 4 0 8th 95 3 2 0 9 100a a = 0.7-1.3 100b b = 0.13-0.06 100c b = 0.01-0.03 0 10 100a a = 0.6-1.2 100b b = 0.075-0.22 100c b = 0.005-0.025 100r r = 0.005-0.075

It is a major aspect of the system and method of the present invention to provide effective focused energy flow treatments and procedures that can be applied to a previously selected substrate or subject to control and / or enhance the biological processes or pathways. The biological processes include effects on metabolite production or production of biochemical compounds and effects on microbial inactivation. It is emphasized that the focused energy flux treatments and methods of the present invention are highly advantageous over the currently conventionally used methods for controlling biological processes in microorganisms. For example, the focused energy flux treatments of the present invention require only a relatively short application or exposure time of the substrate that has taken up the microorganisms (between 8 seconds and up to 30 minutes) over the focused energy flow treatment. Furthermore, the achieved desirable effect is achieved after a relatively short recovery or incubation period from exposure to focused energy flow treatment (up to 6 or 24 or 48 hours). In addition, the focused energy flux treatment of the present invention is applied without requiring any engineering, transformation, or transplantation processes, and / or without a need for additional materials or pretreatments. In addition, the focused energy flux treatments of the present invention may be adjusted for a desirable biological effect and / or predefined microorganism or combination of microorganisms.

It is within the scope of the present invention that by exposing microbes to the focused energy flux treatments of the present invention, advanced generation biofuels or products thereof can be effectively produced.

It is recognized herein that first and second generation biofuels such as ethanol and biodiesel have several inherent limitations and disadvantages that make them significantly less attractive as a long term substitute for petroleum. For example, the primary raw materials of the first generation of ethanol (corn and sugar cane) and biodiesel (rape, soybeans and palm trees) are food-based crops that compete for scarce crops, fresh water and fertilizers. Besides these fuels can not be used in unmodified drives beyond small additions or blends, and they are not applicable to the jet fuel market.

The present invention therefore provides advantageous and useful next generation biofuels (i.e., at least the "third generation" and "fourth generation") based on advanced biochemistry and revolutionary processes. The present invention uses novel metabolic and / or fermentation techniques to produce biofuel compounds such as propanol and butanol derivatives using yeast strains.

A raw material is defined herein as any renewable biological material or biomass that can be used directly or indirectly as a fuel or converted to another form of fuel or energy product.

The advanced biofuels of the present invention encompass the various terms and approaches related to biofuels, including: first generation, second generation, third generation, fourth generation, 1G, 2G, 3G next generation, sustainable, renewable, advanced, green hydrocarbons, etc ,

It is further recognized that the classifications of biofuels are generally based on the type of source, the conversion technology and the properties of the fuel molecules produced.

• a.) Biokraftstoffe, die durch fortgeschrittene Verfahren erzeugt werden, d. h. aus Grundstoffen, die keine Nahrungsmittel sind, wie z. B. Mikroorganismen. Das Endprodukt kann zu den Kraftstoffen äquivalent sein, die durch eine Technologie der ersten Generation (z. B. Ethanol oder FAME) erzeugt wurden, oder es kann ein anderer Typ von fortgeschrittenen Biokraftstoffen sein (wie Propanol oder Butanol). Solche Biokraftstoffe der „nächsten Generation“ werden als nachhaltiger angesehen (sie können ein größeres Ausmaß an Treibhausgas-(„GHG“)-Reduktion bieten und stehen nicht im Wettbewerb mit Nahrungsmittelpflanzen um die Landnutzung).
• b.) Biokraftstoffe mit verbesserten Eigenschaften, die beispielsweise aus nachhaltigen Grundstoffen erzeugt werden, die nicht als mit den Nahrungsmittelproduktionssystemen in nachteiliger Weise im Wettbewerb stehend angesehen werden oder die nicht zu einem Verlust von gespeicherten Kohlenstoff durch Abholzung führen. Diese können mit bestehenden Kraftstoffinfrastrukturen besser kompatibel sein, Treibhausgasersparnisse im Vergleich zu den Biokraftsoffen der ersten Generation ermöglichen oder andere technische Vorteile bieten.

In other aspects, it is further within the scope that the term advanced biofuels is used herein in a general manner to describe:

• a.) Biofuels produced by advanced processes, ie non-food raw materials, such as: B. microorganisms. The final product may be equivalent to the fuels produced by first generation technology (eg, ethanol or FAME), or it may be another type of advanced biofuel (such as propanol or butanol). Such "next generation" biofuels are considered more sustainable (they can provide greater levels of greenhouse gas ("GHG") reduction and are not in competition with food crops for land use).
• b.) biofuels with improved properties, for example, produced from sustainable raw materials that are not considered to be unfairly competitive with food production systems or that do not result in loss of stored carbon through deforestation. These can be more compatible with existing fuel infrastructures, provide greenhouse gas savings compared to first-generation biofuels, or provide other technical benefits.

It is still within the scope that biofuels made from non-food or first-generation technology may also be termed "next generation" or sustainable, or sometimes grouped together with advanced biofuels, even if none advanced process technology is used.

It is further noted that other factors related to land use, competition with food crops, efficiency of production process and overall energy balance, etc. can be considered when referring to advanced biofuels.

The method and system of the present invention is designed and developed to meet and achieve sustainability and fuel quality standards, as well as the need for renewability.

According to one embodiment, the energy flow focusing device (EFFM) of the present invention comprises a controller for controlling the EFFM.

It is further within the scope of the invention that the controller is configured to provide pulses of focused energy flow in a predetermined pulsed manner.

It is further within the scope of the invention that the unique method of the present invention unexpectedly provides an effect below the outer integument of the treated subject. Relevant prior art refers to experiments showing surface modification (ie Feng et al., 2012, Appl. Phys. Lett., 101, 041602 ), in particular the treatment of superficial wounds on the outer surface of the skin (epidermis) or the influencing of the seed coat or the sterilization of the outer layer of the skin or semen. The present invention is further configured to induce a therapeutic or regenerative or beneficial or biological effect on layers or tissues or regions that are not confined to the surface of the subject being treated, for example, affecting bowel disease, providing an effect on cells, or Tissue or cell pathways or providing an effect on wounds that penetrate into deeper layers of the skin and the body.

It is further within the scope of the invention that the system is adapted to provide a synergistic effect with regard to effecting or affecting at least one biological pathway of a microorganism in a substrate as compared to the effect induced by a source or a Non-thermal plasma discharge device (NTP) or through each of the energy flow focusing device (EFFM) for converting the NTP beam into a focused energy flow, administered singly or in partial combination.

In a further embodiment, the method of the present invention is adapted to provide a synergistic effect of at least about 5% with respect to effecting or affecting at least one biological pathway of a microorganism in a substrate having a therapeutic or regenerative or biochemical or beneficial or biological Effect induced on the subject.

The term "non-thermal plasma" or "non-thermal plasma" or "cold plasma" or "NTP" generally refers hereinafter to any plasma that is not in thermodynamic equilibrium, either because the ion temperature is different from the electron temperature, or because the velocity distribution of one of the species does not follow a Maxwell-Boltzmann distribution. It is within the scope of the invention to refer to the NTP by the specific technique used to generate it, e.g. "Gliding arc", "plasma pencil" or "plasma pencil", "plasma needle", "plasma jet", "dielectric interface or barrier discharge", "uniform glow discharge plasma in one atmosphere", "atmospheric plasma", "Non-thermal discharges at ambient pressure", "nonequilibrium atmospheric pressure plasmas", these terms referring to both: non-thermal plasma and plasma operating at or near atmospheric pressure. It is further within the context of the present invention that the term further refers to a "cold" plasma defined as plasma discharges near room temperature at one atmosphere, which is different from other plasmas occurring at hundreds or thousands of degrees above the Ambient temperature work.

It is further within the scope of the invention that the generated plasma is selected from a group consisting of positive ions, negative ions, electrons, metastable particles, atoms, free radicals and photons.

According to certain embodiments, the NTP-emitting source is selected from the group consisting of a dielectric interface or barrier discharger, an atmospheric pressure glow discharge, a corona discharge battery, a DC high voltage corona discharger, a DC negative voltage high voltage corona discharger, a positive DC positive voltage corona discharger, a blanked barrier discharger , a moving arc discharge (GD) induced plasma and a plasma jet.

As used herein, the term "plate" generally refers to an article or element or object that preferably has a rounded or circular shape, such as a ring or disc.

The term "about" as used herein refers to ± 25% of the defined amount or measure or value.

The term "substantially" as used herein is defined generally as being something constituent or as part of the nature or being of something. In the context of the present invention, it may refer to a range of ± 25% of the defined amount or measure or value.

The term "subject" as used herein refers to a human, an animal, a plant, flatworms, microorganisms, planarians, fluids, foods, emulsions, suspensions, soil, minerals, media, gas, a liquid, gas mixtures, mixtures of gas and liquid and / or on an object.

The term "substrate" as used herein refers to a food or food product, a liquid, a beverage, a suspension, a biological culture, a medium, a growth medium, an emulsion, a biological tissue, a biological organism, a human, an animal, a plant, a fluid, soil or a solid, minerals, media, a gas, mixtures of gas and liquid, gas mixtures, cells, tissue cultures, organs and any combination thereof.

The term "microorganism" as used herein refers generally to a microscopic living organism, which may be a unicellular or a multicellular. It is within the scope that microorganisms include yeast, bacteria, archaea, algae, corynebacteria, aerobic bacteria, anaerobic bacteria, fungi, protozoa, viruses, spores, hype, candida, prions, and any combination thereof. In specific embodiments, it includes industrial microorganisms that refer to species of microorganisms, such as bacteria and yeast, that are used to produce large scale industrial articles. Industrial microbiology includes the use of microorganisms to produce food or industrial products in large quantities. A variety of microorganisms are used within industrial microbiology including naturally occurring organisms, selected mutants or otherwise treated microorganisms or genetically modified organisms (GMOs).

Non-limiting examples of bacteria included in the scope of the present invention are Listeria monocytogenes, Escherichia coli, Salmonella, bacteria of the Enterobacteriaceae family, Listeriaceae family bacteria, gram-positive bacteria, Gram-negative bacteria, aerobic bacteria, anaerobic bacteria, Staphylococcus aureus, Pseudomonas aeruginosa, Staphylococcus epidermidis, Staphylococcus haemolyticus, Enterococcus faecalis, the Clostridiaceae family, Clostridium, Korynebacterium, Actinobacteria and any combination thereof.

The term "pulse cycle" or "cycle" is defined herein as a predefined pulse sequence profile as shown in Table 2 below. Table 2: Examples of Pulse Cycle Profiles

Reference is now made to Table 2, which presents examples of pulse cycle profiles within the scope of the present invention. The columns define the successive number of pulses in the cycle (1st pulse, 2nd pulse, etc.). Each of the pulses is described by a pulse duration in seconds (the left number) followed by a pause in seconds (the right number). A cycle or pulse cycle is defined as a sequence of pulses (1st pulse, 2nd pulse, etc.), as illustrated in each case in a horizontal row.

In specific embodiments, the number of pulses i in each cycle is at least 4 and up to 30, in particular at least 6 and up to 30 or at least 8 and up to 20 pulses in each cycle.

In some embodiments, the duration of one cycle or pulse cycle is between about 9 seconds and about 120 seconds, and more preferably between about 20 seconds and about 60 seconds.

In other aspects, the number of cycles administered according to the method or plan of the present invention is between 2 and 50, especially between 3 and 40.

It is further within the scope of the present invention that any combination of the pulses defined above may be used and applied in the focused energy flux treatments of the present invention.

The term "energy flow" is generally defined within the context of the present invention as the energy transfer through a surface or towards a surface. The energy flow rate may be defined as the rate of energy transfer per unit area (ie SI units: W * m ^-2 = J * m ^-2 * s ^-1 ) or as the total radiant energy per time. In some aspects, the radiation energy is the electromagnetic radiation. The amount of radiant energy can be calculated by integrating the radiant flux (or radiant power) over time. This is a vector size with its components determined in the normal (or perpendicular) direction to the measurement surface. It is sometimes defined as energy flux density. The energy flow can alternatively be defined as total energy transfer. The rate can be described by SI units: W = J · s ^-1 . It is also referred to as energy flow.

The term energy flow also includes in its scope the Poynting vector aspect, which indicates the directional energy flux density (the rate of energy transfer per unit area) of an electromagnetic field. The SI unit of the poynting vector is watts per square meter (W / m ² ). The Poynting vector is usually denoted by S or N, where
E is the electric field vector; and
H is the magnetic field vector.

An alternative definition of the electric field E and the magnetic flux density B is used. It is also possible to combine the electric displacement field D with the magnetic flux density B to obtain the Minkowski shape of the Poynting vector, or to use D and H to construct another shape (Abraham shape). In some aspects, the poynting vector describes the special case of an energy flow vector for electromagnetic energy. However, any form of energy has its direction of motion in space as well as its density, so that energy flow vectors can also be defined for other types of energy.

The term "focused energy flow" as used herein refers to a plasma or plasma beam that is converted to, coupled to, modified by, transformed by, or generated by at least one of an electric field vector to induce an electric field, a magnetic field vector for inducing a magnetic field and an electromagnetic field vector for inducing an electromagnetic field. Such vectors include ferroelectric agents or elements, ferromagnetic agents or elements, piezoelectric agents or elements or a combination of all elements or any partial combination thereof. According to a main embodiment, the plasma is further converted or adapted or influenced by a focussing vector or focussing element among others as disclosed. It is within the scope of the present invention that focused energy flow refers to plasma oscillations that have been influenced or converted by an energy flux focusing device (EFFM) comprising at least one vector or element selected from the group consisting of at least one ferroelectric element, at least a ferromagnetic element, at least one piezoelectric element, or any combination thereof, as well as a focusing vector or focusing element. The focused energy flow is applied to a subject in a predetermined pulsed manner to induce a biological or renewable or regenerative or beneficial effect.

In one embodiment, the focused energy flow is applied to the subject in a predetermined manner, in particular a pulsed or pulsed mode, which is determined according to the classification (ie taxonomic classification) of the subject being treated or according to the desirable effect (eg, microbial activation or biochemical pathway). For example, a focused energy flow pulse that is designed to induce a regenerative or biochemical effect or beneficial effect in a plant may be different (ie, in terms of pulse duration, pulse rate, pulse intervals, pulse cycle profile, or any combination thereof) from the focused energy flow profile. which is designed to induce a therapeutic or regenerative or biochemical process or beneficial effect in a human and / or the focused energy flow profile designed to induce a beneficial effect on water or biochemical processes in bacteria or yeast or viruses or to produce or control mushrooms. Examples of pulse profiles or cycles or patterns or parameters included in the scope of the present invention are provided in Table 2.

The term "energy flow focusing device" or "EFFM" refers to a device that is remote from or remote from or external to or separate from the NTP source and is configured and configured to communicate with the plasma jet coming from the NTP source NTP source is emitted, too or affect or affect this or to couple with this. It is within the scope of the invention that the EFFM comprise at least one of: an electric field vector, a magnetic field vector, an electromagnetic field vector, a piezoelectric field vector, a piezomagnetic field vector, or any combination thereof, and at least one focusing vector in various configurations and arrangements.

The term "vector" or "energy flow vector" or "energy flux element" as used herein refers to an agent that includes a material or substance or component or structure that is used to provide the focused energy flow. In some embodiments, the vector is configured to provide an electrically and / or magnetically and / or electromagnetically and / or ferroelectrically and / or ferromagnetically induced field. In preferred embodiments, the vector is selected from the group consisting of (1) at least one ferroelectric element, (2) at least one ferromagnetic element, (3) at least one piezoelectric element, (4) at least one piezomagnetic element, and any combination thereof. According to certain embodiments, such vectors or any combination of the aforementioned vectors are used as part of the method and system to effect at least one biological pathway of a microorganism in a substrate. The energy flow vector is configured to convert or affect or affect or couple to the plasma beam emitted from the NTP source.

The term "ferromagnetic material" or "ferromagnetic element" as used herein refers to an element comprising a material that exhibits ferromagnetism in a broad sense, including ferrimagnetism. In certain aspects, the ferromagnetic element is selected from the group consisting of a permanent magnet, an electromagnet, a superconducting magnet, and any combination thereof.

It is within the scope of the invention that such materials include elemental metals and in other embodiments include alloys, oxides or other chemical compounds or mixtures thereof. Non-limiting examples of ferromagnetic materials include chromium (IV) oxide, cobalt, dysprosium, ferrite (iron), ferrite (magnet), magnetite (Fe ₃ O ₄ ), α-ferrite (α-Fe), gadolinium, gallium manganese arsenide, iron , Neodymium magnet, nickel, permalloy, a rare earth magnet, a samarium cobalt magnet, suessite, yttrium iron garnet, ferromagnetic alloys, and any combination thereof.

The term "ferroelectric element" as used herein refers generally to a material that has a property of spontaneous electrical polarization that can be reversed or canceled by the application of an external electric field. In other words, ferroelectric materials refer to materials that maintain a permanent electrical polarization that can be reversed or switched in an external electric field. In specific embodiments, ferroelectric materials are pyroelectric and inherently piezoelectric. In some embodiments, ferroelectric capacitors may have the combined properties of memory, piezoelectricity, and pyroelectricity. In some aspects, piezoelectricity refers to the generation of a surface charge in response to the application of an external stress to a material. In other aspects, pyroelectricity generally refers to a change in the spontaneous polarization of a material in response to a temperature change.

It is within the scope of the present invention that ferroelectric materials and / or elements include ferroelectric polymers, particularly polyvinylidene fluoride or polyvinylidene difluoride (PVDF). In alternative embodiments, ferroelectric elements included in the scope of the present invention may at least partially comprise PZT, lead zirconium titanate, ferroelectric oxides, Pb [Zr _(x) Ti _(1-x) ] O ₃ , PbZrO ₃ , barium titanate (BaTiO ₃ ). , Ba (Sr) TiO ₃ , Ba _(1-x) Sr _(x) TiO ₃ and any combination thereof

As used herein, the term "piezoelectric element" generally refers to materials or certain crystals that have the ability to generate electrical voltage in response to an applied mechanical stress. PZT (lead zirconium titanate) is one of the large family of materials whose structure changes with the application of an electric current or which, when loaded, generate electricity. These specific piezoelectric or ferromagnetic effects have the property that when a voltage is applied or a current flows, a volume change occurs in the material.

Therefore, it is recognized herein that according to one embodiment, an important ferroelectric material is lead zirconium titanate (PZT), which is part of the solid state solution formed between ferroelectric lead titanate and anitferroelectric lead zirconate. Various compositions of PZT are used for various applications. For example, lead titanate (PbTiO ₃ ) is preferred for memory applications, whereas piezoelectric applications use the divergent piezoelectric coefficients associated with the morphotropic phase boundary.

It is further within the scope of the present invention that piezoelectric materials such as Pb (ZrTi) O ₃ (PZT) or PbTiO _{3 are used} as generators in which an interactive electro-mechanical energy conversion based on the piezoelectric effect occurs.

It is further recognized herein that PZT (lead zirconium titanate) is an inorganic compound having the chemical formula Pb [Zr _x Ti _1-x ] O ₃ with 0 ≦ x ≦ 1. It is a ceramic perovskite material that exhibits a pronounced piezoelectric effect that has practical applications in the field of electroceramics. It is a perovskite crystal structure in which each unit consists of a small tetravalent metal ion in a lattice of large divalent metal ions. In the case of PZT, the small tetravalent metal ion is usually titanium or zirconium. PZT materials exhibit improved properties such as high sensitivity, high operating temperature, high dielectric constants, and low dielectric loss, ie, in comparison to the metallic oxide-based piezoelectric material barium titanate (BaTiO ₃ ).

The term "piezoelectric element" as used herein refers generally to antiferromagnetic crystals and materials such as piezoelectric ferrite materials, magnetoelectric ceramic materials (e.g., Ba6; xR2x (Nb1; xFe2 + x) O3), nickel, a Ni-Fe alloy, a V-Fe alloy, an Fe-Co-Ni alloy, a Ni-Cr-V alloy, (Fe, Cu system) Monel alloy, nickel ferrite, nickel-copper ferrite, nickel-zinc ferrite, composition systems including magnesium manganese ferrite, nickel cobalt ferrite, etc. Piezo magnetism may be characterized by a linear coupling between the magnetic polarization and the mechanical stress of the system. In a piezo-magnet one can induce a spontaneous magnetic moment by applying a physical tension or a physical deformation or by applying a magnetic field, see IEEE standard 319-1990 (1991) , "IEEE Standard on Magnetostrictive Materials: Piezomagnetic Nomenclature", which is incorporated herein by reference. In addition, it is further within the scope of the invention to use at least one or more of a group comprising magnetostrictive, electromagnetic, piezoelectric, and electrostrictive transducers and elements thereof.

As used herein, the term "focusing vector" refers generally to a component which at least partially comprises a material selected from the group in a non-limiting manner consisting of: a highly reflective coating, glass or an amorphous material, solid or nonporous crystalline materials such as calcium fluoride (CaF ₂ ), polymers, metals such as iron and alloys thereof, stainless steel, aluminum, silver, gold and mixtures thereof, a dielectric coating, an extreme ultraviolet coating, UV-reflective materials, 100-280 nm reflective materials and any combination thereof. Other examples of materials that may be used as the focusing elements within the scope of the present invention may include: titanium, vanadium, chromium, yttrium, zirconium, niobium, molybdenum, technetium, ruthenium, rhodium, palladium, silver, tantalum, tungsten, Rhenium, osmium, iridium, platinum, gold, thallium, lead, diamond-like carbon (DLC), graphite-containing materials, low refractive index materials, blends, blends and / or alloys thereof. The component is arranged to control the flow of energy Bring convergence.

In other embodiments, focusing vector refers to a device, surface, or portion thereof that reflects back at least slightly light or a plurality of wavelengths back to its source, with, for example, a minimum of scatter. In specific embodiments, an electromagnetic wave is reflected back, at least to a minimal extent, along a vector that is parallel in its direction but opposite to the direction from the source of the wave. Reflectors are devices that operate by sending light back to the light source along the same direction of light. The coefficient of luminous intensity RI is the measure of a reflector efficiency which is defined as the ratio of the intensity of the reflected light (luminous intensity) to the amount of light incident on the reflector (normal illuminance).

In one aspect of the invention, aluminum serves as a good reflector (about 92%) of visible light and as an excellent reflector (as much as 98%) of mid and far infrared.

The term "focusing vector" may also refer to an optical coating, which generally means one or more thin layers of material deposited on at least a portion of an optical component, such as on a lens or a mirror, which may be the nature and type How the optics reflect and transmit light changes. In a specific embodiment, a highly reflective coating can be used to create mirrors that reflect more than 99.99% of the light falling on them. More complex optical coatings show high reflection over one wavelength range and antireflection over another range, allowing the generation of dichroic optical thin film filters. The simplest optical coatings are thin layers of metals, such as aluminum, which are deposited on substrates to make mirror surfaces. The metal used determines the reflection characteristics of the mirror. For example, aluminum achieves a reflectivity of around 88% -92% in the visible spectrum. Silver has a reflectivity of 95% -99% even in the far-infrared, but shows a reduced reflectivity (<90%) in the blue and ultraviolet spectral regions. Gold, which has excellent reflectivity (98% -99%) throughout the infrared, has limited reflectivity at wavelengths shorter than 550 nm.

In certain embodiments, an aluminum mirror coating has the highest reflectance of any metal in the 200-400 nm (UV) and 3000-10000 nm (far IR) regions. In the visible range of 400-700 nm it is somewhat surpassed by tin and silver and in the range 700-3000 nm (near IR) by silver gold and copper. Nonetheless, as defined in any of the above embodiments, the focusing elements are low reflecting (LR) elements, and structural elements of the device are adapted to reflect UV, visible light and / or IR only to a very minor extent (e.g. less than 7.5%).

In alternative embodiments, high reflectance (HR) coatings are normally based on a periodic layer system composed of two materials, one with a high index such as zinc sulfide (n = 2.32) or titanium dioxide (n = 2.4) and one with a low-index material such as magnesium fluoride (n = 1.38) or silica (n = 1.49). This periodic system significantly improves the reflectivity of the surface in the certain wavelength range called bandstop filter, the width of which is determined by the ratio of the two indices used, while the maximum reflectivity increases to almost 100% with the number of layers in the stack. The thicknesses of the layers are generally designed as a quarter of a wavelength, so that reflected rays interfere constructively with one another to maximize reflection and minimize transmission. Coatings composed of deposited lossless dielectric materials can achieve reflectivities greater than 99.999% over a relatively narrow wavelength range. Conventional HR coatings can achieve 99.9% reflectivity over a wide range of wavelengths (tens of nanometers in the visible spectral range).

Multi-layer mirrors constructed from hundreds of alternating layers of high mass metal such as molybdenum or tungsten and a low mass spacer such as silicon vacuum deposited on a substrate such as glass cause the mirror to have EUV light (wavelengths shorter than about 30 nm) of the desired wavelength is stronger than what a normal metal mirror would do in visible light. By using multilayer optics, it is possible to reflect up to 70% of the incident EUV light (at a particular wavelength selected when the mirror was constructed).

It is also within the scope of the invention that the method and system for effecting or affecting at least one biological pathway of a microorganism in a substrate may be operated and configured as a holographic prism affecting the subject being treated. In this embodiment, the focusing vector or element in combination of the electromagnetic vectors is configured to simultaneously satisfy the requirements of high discretization and accuracy. It is recognized herein that holographic prismatic elements or holograms have the advantages of high temperature stability, high efficiency, relatively small size, and light weight, as well as the ability to tolerate sharp changes in optical and geometric characteristics (eg, devitrification). In other aspects, the method and system for effecting or affecting at least one biological pathway of a microorganism in a substrate is designed and operated as a diffraction grating device.

In certain aspects, the at least one focussing vector component is configured to focus and amplify the plasma energy flow. In other aspects, the at least one focusing vector is configured to combine and / or collect the generated focused energy flow at a center and / or reduce its loss. According to yet another aspect According to the invention, the at least one focusing element is configured to increase the efficiency of the discharged plasma energy flux. Furthermore, without wishing to be bound by theory, the method and system of the present invention is further configured to adjust and match the energetic resonance of the focused energy flow that interacts with the subject or subject being treated.

Now, magneticohydrodynamics (MHD) (magnetic fluid dynamics or hydromagnetics) as one aspect of the present invention will be referred to. This embodiment relates to the studies on the dynamics of electrically conductive fluids. Examples of such fluids include plasmas, liquid metals and salt water or electrolytes. It is recognized herein that the fundamental concept behind MHD is that magnetic fields can induce currents in a moving conductive fluid, which in turn creates forces on the fluid and also alters the magnetic field itself. The set of equations describing MHD are a combination of the Navier-Stokes equations of fluid dynamics and the Maxwell equations of electromagnetism. Magneto-hydrodynamics (MHD) is the physical-mathematical framework that governs the dynamics of magnetic fields in electrically conductive fluids, eg. In plasmas and liquid metals.

The term "fluid" as used herein generally refers to a substance that flows or continuously deforms. In certain aspects, such a substance deforms continuously under an applied shear stress. The term "fluid" includes liquids, gases, liquid-gas mixtures, emulsions, suspensions, plasma, and to a degree, also plastic solids. Non-limiting examples of "fluids" included within the scope of the present invention include any liquid or fluid or suspension such as water, oil, diesel, naphtha, a beverage, crude oil, milk, honey, ketchup, blood, others Media, water-in-oil mixtures or oil-in-water mixtures, petroleum, fuels, petroleum, LPG, also known as LPG, GPL or LP gas, liquid propane gas or propane or butane or mixtures thereof, biofuels and Products thereof, biodiesel and products thereof, hydrocarbon gas, gas, liquids and mixtures thereof, gas mixtures and any combination thereof.

The term "crude oil" as used herein is intended to include petroleum, petroleum, refined petroleum, fossil fuels, naturally occurring and unprocessed petroleum, and any product or fraction thereof. In some aspects, the above terms refer to a naturally occurring yellow to black liquid found in geological formations near the surface of the earth. Such a liquid is generally refined into various types of fuels. It can consist of hydrocarbons of different molecular weight and other organic components.

The term "unit discharge" as used herein refers to voltage oscillation at one time.

It is in accordance with one embodiment of the invention that a system and method for influencing or effecting at least one biological pathway of a microorganism in a substrate be provided. The method includes steps of: providing a source of non-thermal plasma (NTP) for generating an NTP beam as defined in Table 1; Providing an energy flow focusing device (EFFM) for transferring the NTP beam into a focused energy flow; Providing the microorganism in the substrate; and operating the EFFM with a predetermined pulsed profile as defined in Table 2 to emit the focused energy flow to the at least one microorganism.

This method may be operated in a system comprising, among other modules, a non-thermal gas plasma emitting source for emitting a plasma jet at a predetermined composition rate or with predetermined flow parameters (eg, as defined in Table 1) and an energy flux focusing device (EFFm). The EFFM includes, among others, at least one of an electric field vector, a magnetic field vector, an electromagnetic field vector or a combination thereof, and a focusing vector.

In a specific embodiment, the EFFM comprises a plasma jet plate having at least one aperture for the passage of the plasma jet, this plasma jet plate having a first surface and a second, opposite surface, the first surface of the plasma jet plate being equipped with a energy flux vector selected from the group consisting of: (1) at least one ferroelectric element for providing the field, (2) at least one ferromagnetic element for Providing the array, (3) at least one piezoelectric element for providing the field, (4) at least one piezomagnetic element for providing the field, and (5) a combination thereof, and wherein at least one focusing vector is configured to focus the plasma jet. The EFFM may further include at least one controller to control the EFFM. In this way, the EFFM and the controller are configured to adjust any of the at least one vector in a predetermined manner so as to provide the focused energy flow in a predetermined pulsed manner to effect or augment at least one biological pathway of a microorganism in a substrate influence.

It is in accordance with a primary embodiment of the invention that the EFFM is adapted to couple the plasma jet to a focused energy flow or to convert the plasma jet into a focused energy flow. Thus, the present invention uniquely provides a device for emitting a focused energy flow in a predetermined pulsed manner to induce therapeutic and / or regenerative and / or biochemical or biological effects and / or beneficial or renewable effects on a subject.

It is according to another embodiment of the invention that the system and the method for an indirect treatment of the object or a treatment be designed and operated remotely. Gadri et al., 2000, Surface Coatings Technology 131: 528-542 and Laroussi and Lu, 2005, Appl. Phys. Lett. 87: 113902 and Montie et al., 2000, IEEE Trans Plasma Sci 28: 41-50 and Topala and Nastuta, 2012, Plasma for Bio-decontamination, Medicine and Food Security, NATO Science for Peace and Security Series A: Chemistry and Biology. ISBN 978-94-007-2851-6, Springer Science + Business Media BV, p. 335 and Middelkoop et al., "Burn wound healing: a role for plasma medicine" and Vasile Nastuta et al., 2011, Journal of Physics D: Applied Physics 44 (10): 105204 are publications incorporated herein by reference and nonlimiting examples of NTP. This type of NTP in use is z. B. A decaying plasma (afterglow) - from longer-lived chemical species. The NTP density and energy is z. Of moderate density - the subject is placed away from the electrodes. However, a larger NTP volume can be generated by using multiple electrodes. The distance of the target from the NTP generating electrode is about 5 to 20 cm, especially 15 cm arcing (filamentous discharge) unlikely to contact the subject at any power setting. In this system, there is no electrical conduction through the target. The suitability for irregular surfaces is high - the remote nature of NTP generation means maximum flexibility of using NTP afterglow flow.

In other embodiments, the NTP used is an atmospheric plasma jet (APPJ). The plasma can be generated using principles of corona discharge, DBD and micro discharges. Examples of applications of the NTP plasma or modified plasma may include the treatment of living cells or living tissue, wound healing, cancer cell apoptosis, blood clotting, d. H. in wounds, bone tissue modification, sterilization and decontamination. In such a case, the low temperature plasma jet is driven by high voltage pulses. In a specific embodiment, the plasma jet works in helium. In accordance with a primary object, the system for applying a modified plasma to a subject is applied to produce positive medical results relating to the recovery process of wounds, i. H. burnt wounds, skin regeneration and re-epithelization.

It is according to another embodiment of the invention, wherein the system is designed and operated for direct treatment of the object; Lee et al., 2005, Surface Coatings Technol 193: 35-38 ; Sladek and Stoffels, 2005, J. Phys. D: Appl. Phys. 38: 1716-1721 and Stoffels et al., 2002, Plasma Sources Sci. Technol. 11: 383-388 are publications incorporated herein by reference and as non-limiting examples of systems designed and operated for direct handling. This type of NTP in use is z. Active plasma - short and long lived species. The NTP density and energy is z. B. higher density target in the direct path of a flow of active NTP. The distance of the target from the NTP generating electrode is about 1-5 cm, arcing may occur at higher power settings, may contact the target. In this system, electrical conduction through the target is provided during normal operation, but possible during arcing. The suitability for irregular surfaces is moderately high - NTP is transmitted directly into the target, requiring either rotation of the target or multiple NTP emitters.

It is according to another embodiment of the invention, wherein the system is designed and operated in a method of electrode contact; Kelly-Wintenberg et al., 1999, J. Vac. Sci. Technol. A 17 (4): 1539-44 ; Laroussi et al., 2003, New J. Phys. 5: 41.1-41.10 ; and Montenegro et al., 2002, J. Food Sci. 67: 646-648 are publications incorporated herein by reference and nonlimiting examples of embodiments included in the scope of the present invention. This type of NTP in use is z. Active Plasma - all chemical species including the shortest lived and ion bombardment. The NTP density and energy is z. Highest density target within the NTP generation field. The distance of the target from the NTP generating electrode is approximately ≤ 1 cm, arcing between electrodes and target may occur at higher power settings. In this system, the system is operable with respect to the electrical line z. When the target is used as an electrode or when the target is electrically conductive between mounted electrodes. The suitability for irregular surfaces is moderately low - close spacing is required to maintain NTP uniformity. Electrodes can be shaped to match a defined consistent surface.

It is according to another embodiment of the invention that the system is designed and operated to emit a plasma that is affected by piezoelectric materials.

Examples of piezoelectric materials used in the system of the present invention include the material Pb (ZrTi) O ₃ or PbTiO ₃ . Such materials in combination with a focussing vector material are used to convert the NTP beam into a focused energy flow.

It is according to another embodiment of the invention that the system is designed and operated to dispense a non-thermal plasma generated from a plasma source (ie, a plasma jet) and exposed to the EFFM configured to generate one emit focused energy flow in a predetermined pulsed manner.

It is according to another embodiment of the invention, wherein the system is designed and operated to dispense a non-thermal plasma coupled to a magnetic field. Liu Jingjing et al., 2005, Plasma Science & Technology, Vol. 7, No. 5, pp. 3073-3077 and Zongbao Feng et al., 2012, App. Phys. Lett. 101, 041602 are publications incorporated herein by reference and provided as non-limiting examples of embodiments included within the scope of the present invention.

It is according to another embodiment of the invention, wherein the system and method are designed and operated to deploy a ferroelectrically coupled non-thermal plasma field. Dunaevsky A. et al., 2001, Journal of Applied Physics 90: 8 4108-4114 and Holzer F. et al., 2005, Plasma Chemistry and Plasma Processing, 25: 6 595-611 are publications which are incorporated herein by reference and which are provided as nonlimiting examples of embodiments included within the scope of the present invention.

It is according to another embodiment of the invention that the method and system are designed and operated to emit a non-thermal plasma coupled to at least one of the following: a magnetic field vector, an electric field vector, or a combination thereof. Such a converting or coupling device (ie, EFFM) is shown herein to have a significantly enhanced effect or, in other embodiments, a synergistic effect in terms of efficacy and improved properties and results (ie, effecting at least one biological pathway a microorganism in a substrate) relative to the application of a non-thermal plasma without the EFFM device.

It will be up now 1 In the drawing, which is not to scale a schematic illustration of a cross-section of preferred embodiments of the EFFM as an exemplary embodiment of the present invention. The figure shows elements that the EFFM is equipped with or mounted to the EFFM. According to certain embodiments, a high voltage electrode (HV) is used. 360 , in the 1 shown powered by a high frequency high voltage power source with energy. The electrode 360 stands by a plasma jet plate 1 which preferably comprises at least partially a polymeric material. Non-limiting examples of such polymeric materials may include polycarbonates, polystyrene (PS), polyesters, polyphenylene oxide, acrylonitrile butadiene styrene (ABS), polyoxymethylene (POM), also called acetal, polyacetal, and polyformaldehyde, styrene-acrylonitrile, polyimide, and blends and polymeric combinations thereof. In this embodiment, the HV electrode is 360 positioned in the center of the plasma jet plate.

According to a specific embodiment, no stress is applied to the treated object or subject 400 created. The object 400 , which may be a microorganism or a human or animal body or a vegetable or liquid or gaseous sample, acts as a blind, and the discharge of the non-thermal plasma may be referred to as a dummy barrier dielectric barrier (FE-DBD).

In an alternative embodiment, the plasma exhauster is referred to as an HV negative DC corona.

Still in the in the 1A to 1C featured embodiment is the plasma jet plate 1 equipped with an Energy Flow Focusing Device (EFFM) comprising vectors or elements configured to convert or affect HV-NTP application. According to one embodiment, at least one vector 4 , in particular a single vector 4 arranged in the first surface of the plasma jet plate, ie in the middle of the plasma jet plate. The at least one vector or the at least one coupling element 4 together with the focusing vector 6 is configured to alter or modify or affect the plasma output, ie, by improving the effectiveness or efficiency of the modification obtained over larger ranges or to have more intense or pronounced effects not limited to surface modification of the treated subject, but also to affect layers, areas or tissues or paths under the outer integument of the subject being treated. The at least one vector 4 may be selected from the group consisting of: at least one ferroelectric element, at least one ferromagnetic element, at least one piezoelectric element and any combination thereof. The resulting focused energy flow 8th is discharged in a predetermined pulsed manner. The plasma jet plate is further with at least one focusing vector or element 6 equipped. In this figure, the focusing vectors 6 arranged on the outer edge of the plasma plate, ie in an opposite configuration. It is argued that the focusing vectors or elements have a significant and highly important effect on the focusing and / or enhancement of the modification region or the effectiveness or efficiency that the focused energy flow 8th on the subject 400 exercises. It is further argued that the method described herein is configured to adjust or adjust or tune the energetic resonance of the focused energy flow that interacts with, or is delivered to, the subject.

In the 1B and 1C are the focusing vectors 6 on a surface of the EFFM in both sides of the plasma jet opening 8th in opposite directions attached. The focusing vectors are disposed opposite each other so as to reflect the emitted plasma ions and focus them toward the subject being treated, thus increasing the penetration depth of the focused energy flow. In one embodiment, the focusing vectors are 6 characterized by an amorphous configuration. In another embodiment, the opposing focusing vectors are 6 through a curved ( 1B ) or even ( 1C ) Cross section on a diagonal between the x and y axes of the plasma jet plate 1 characterized.

With regard to the material of the focusing vectors, in one embodiment they comprise solid materials such as CaF ₂ . The 1B and 1C further show embodiments where the electromagnetic field vectors (ie magnets) 4 in both sides of the plasma jet opening 8th under the focusing vectors 6 are arranged without contacting them. In a specific embodiment, the electromagnetic vectors are arranged in an opposite orientation in both sides of the plasma jet aperture to affect the reflected plasma beam. It is within the scope of the invention that the system of 1 can be adapted to cause at least one biological pathway of a microorganism in a substrate.

In other embodiments, electromagnetic field vectors, e.g. B. ferromagnetic elements in pairs, arranged along the inner circumference of the EFFM. In specific embodiments, six pairs of ferromagnetic elements are positioned radially in the outer edge of the EFFM. In some aspects, the vectors are in pairs of segments 3a and 3b arranged to configure an outer and inner ring-like structure. In a further aspect of the invention, the at least one pair of ferromagnetic elements is a ferromagnetic material, in particular magnetite, ferrite, cobalt or a cobalt alloy, iron or iron oxides such as Fe ₃ O ₄ , a samarium cobalt magnet (eg SmCo ₅ or SmCo series 1: 5, Sm ₂ Co ₁₇ or SmCo series 2:17), neodymium magnets, ferromagnetic, ferroelectric or ferrimagnetic materials such as Fe ₃ O ₄ , magnetic ferrites, alfa-ferrite (α-Fe) or beta-iron ( β-iron) or other paramagnetic substances or any combination thereof.

Thus, the system and method is designed to provide a unique source of focused energy flow driven by the synergistic effect of combining the NTP with the EFFM, the at least one ferroelectric and / or piezoelectric element, and / or comprises at least one magnetic field vector results, these fields having at least one focusing vector (in 1 shown).

According to one embodiment, the at least one electromagnetic field vector or the at least one ferromagnetic element is arranged radially around a plasma jet opening in the EFFM.

According to a further embodiment, the at least one electromagnetic field vector or the at least one ferromagnetic element is arranged in groups of pairs or triples around the plasma jet opening in the EFFM.

According to another embodiment, the EFFM comprises at least one pair of electromagnetic field vectors or ferromagnetic elements.

According to another embodiment, the EFFM comprises at least one pair of mutually-oriented electromagnetic field vectors or ferromagnetic elements.

According to another embodiment, the EFFM comprises at least a pair of mutually-oriented pairs of electromagnetic field vectors or ferromagnetic elements.

According to another embodiment, the at least one pair of electromagnetic field vectors or ferromagnetic elements is arranged in a parallel orientation.

According to another embodiment, the at least one pair of electromagnetic field vectors is arranged in the same plane and in other embodiments these electromagnetic field vectors are at different levels successively on opposite axes.

According to another embodiment, the at least one pair of electromagnetic field vectors or ferromagnetic elements is positioned such that the poles of the electromagnetic field vectors or the ferromagnetic elements have attractive polarities or repulsive polarities.

In another embodiment, the one or more electromagnetic field vectors or coupling elements are configured as a disk having a diameter of about 0.5 to about 2.25 cm. In other embodiments, the one or more electromagnetic field vectors or couplers are made of or otherwise contain APC 841 material, or any suitable material, e.g. As defined in Table 3 below. The NTP plasma emitted from the NTP source to the EFFM comprising a plasma jet plate is converted by the EFFM comprising at least one electromagnetic field vector and at least one focusing vector and fed out.

It will now be on the 2A and 2 B Not in scale, two sets of schematic views of the EFFM ( 11a respectively. 11b ) illustrate. The EFFM 11a comprises a plurality of magnets, which are arranged here on the circumference of the plate, while the EFFM 11b includes at least one (here: exactly one) magnet, the z. B. is arranged at the central inner part of the EFFM. The EFFM 11a includes a rounded textured plate 1 with at least one central opening 2 which is adapted in size and shape to accommodate therein at least one plasma-emitting source. Along the inner periphery of the plate, a plurality of electromagnetic field vectors or coupling elements are mounted to convert the plasma jet with an electromagnetic and / or ferroelectric and / or piezoelectric element. The plasma jet plate further comprises at least one focusing vector (not shown). In another aspect, the plasma jet is used herein to focus the at least one field generated by the at least one vector to provide a focused energy flow: see, for example, vectors 3a and 3b , The EFFM as defined above is by means of a device 20 with defined arms 3D-oriented and spatially arranged.

It is within the scope of the invention that the EFFM and especially the plate, and more particularly the second surface of the plate, is made of or contains polymers such as 70% acrylonitrile butadiene styrene (ABS) and polystyrene (PS). It is further within the scope of the invention that the plate be made from its Mixture of polymers or contains such as 70% PS and 30% ABS (wt.% / Wt.% Or mol / mol).

In some embodiments, the angle θ is between the electromagnetic field vectors or coupling elements 3a and 3b varied between 0.5 ° and 270 °.

It is within the scope of the invention that the electromagnetic field vectors or coupling elements are placed in pairs, each of which is arranged as its first coupling element and a second coupling element. This first coupling element is arranged opposite to the second coupling element, d. H. the angle θ between the first and second coupling elements is 180 °.

It is further within the scope of the invention that at least one of the electromagnetic field vectors or coupling elements, ie, the cobalt or cobalt alloy magnet is made, such as a samarium cobalt magnet (eg, SmCo ₅ or SmCo Series 1: 5, Sm ₂ Co ₁₇ or SmCo series 2:17), neodymium magnets, ferromagnetic, ferroelectric or ferrimagnetic materials such as Fe ₃ O ₄ , magnetic ferrite, alfa-ferrite (α-Fe) and beta-iron (β-iron); paramagnetic substances such as platinum, aluminum and oxygen; diamagnetic agents that are repelled by both poles. Compared with paramagnetic and ferromagnetic substances, diamagnetic substances such as carbon, copper, water, plastic and any mixtures and combinations thereof.

It is further within the scope of the invention that the EFFM plate has six coupling elements positioned at 12 o'clock, 2 o'clock, 4 o'clock, 6 o'clock, 8 o'clock and 10 o'clock. It is further within the scope of the invention, wherein the coupling element at 12 o'clock and / or 10 o'clock is a cobalt-containing coupling element, d. H. a magnet. It is further within the scope of the invention, wherein the coupling element at 2 o'clock and / or 4 o'clock is a magnetic-type coupling element, i. H. a magnet. It is further within the scope of the invention, wherein the coupling element at 6 o'clock and / or 8 o'clock is an alpha-ferrite coupling element, i. H. a magnet.

It is further within the scope of the invention that the pairs of poles of the magnets or poles of the coupling elements have an S-S, N-N or S-N orientation.

It is further within the scope of the invention that the NTP plasma discharger be provided in a plasma discharge technology. Non-limiting examples of NTP discharge technology within the scope of the present invention include glow, corona, atmospheric plasma jet (APPJ), dielectric barrier discharge, micro hollow cathode discharge (MHCD) uniform glow discharge plasma in one atmosphere (OAUGDP), plasma needle, atmospheric pressure glow discharge, a DC high voltage corona discharger, a DC negative voltage high voltage corona discharger, a positive DC positive voltage corona discharger, a blanked dielectric barrier discharger, a plasma induced discharge arc (GD), and a plasma jet.

It will be up now 3A In the drawing, which is not to scale, illustrates a schematic view of an atmospheric plasma jet as an alternative embodiment of the present invention. Like Nehra et al. (see Nehra V., Kumar A., Dwivedi H. (2008) - "Atmospheric Non-Thermal Plasma Sources", Int. J. Eng. 2 (1): 53-68 , which is incorporated herein by reference), is a discharge capable of generating non-thermal plasmas at atmospheric pressure, the atmospheric pressure plasma. This type of APPJ consists of two concentric electrodes through which a mixture of helium, oxygen or other gases flows. In this arrangement, the inner electrode is coupled to a 13.56 MHz radio frequency power at a voltage between 100-250V, and the outer electrode is grounded. By applying the RF power, the discharge is ignited and works with a base gas that flows between an outer grounded cylindrical electrode and a central electrode and produces a high velocity effluent of highly reactive chemical species. The high frequency power driven central electrodes accelerate free electrons. These energetic electrons undergo inelastic collisions with the base gas to produce molecules, atoms, free radicals, and additional excited-state ion-electron pairs. Once the gas leaves the discharge volume, ions and electrons are rapidly lost through recombination, but the fast-flowing outflow still contains neutral metastable species and radicals. The key operational features of the APPJ are as follows: (1) It produces a stable, homogeneous and uniform discharge at atmospheric pressure; (2) it operates at high frequency (RF) power of 250 W and a frequency of 13.56 MHz; (3) the ionized gas from the plasma jet exits through the nozzle where it is directed to the substrate and therefore used in downstream processing; (4) it works without a dielectric cover over the electrode and is free of filaments, streamers and arcing; (5) the gas temperature of the discharge is as low as 50 ° C, which allows to treat sensitive surfaces without damage, or as high as 300 ° C, which makes it possible to treat rugged surfaces more aggressively. (6) It shows a great similarity to the low-pressure Gleichspannungsglimmentladung.

It will be up now 3B Still referring to a not-to-scale schematic view of a corona discharger (see details in FIG Nehra et al. ) as alternative embodiments of the present invention. Corona discharge technology involves the generation of a non-thermal plasma. It directly influences the generation of free radicals. The corona discharge exists in various forms, depending on the polarity of the field and the geometrical configuration of the electrode. Corona discharge arrays comprising asymmetric electrode pairs and resulting from the electric field surrounding the inhomogeneous electrode assemblies driven by a continuous or pulsed DC voltage. The development of a coronary discharge preferably comprises the following steps: an asymmetric electrode configuration is made; a high voltage is applied and a free electric charge is made available; and generating a multiplication of an electric current and avalanche breakdown.

In other alternative embodiments involving DBD plasma technology, the plasma discharges between two electrodes, wherein at least one electrode insulated with a dielectric layer can operate in a wide range of geometrical configurations, such as classical bulk discharge, surface discharge, and the coplanar discharge. Volume unloaders may also have either planar or coaxial arrangements. In planar electrode arrays, the two electrodes are parallel to each other and one or both of the dielectric barriers are always located either (i) on the powered or grounded electrode or (ii) on both electrodes or (iii) between the two metal electrodes. The electrodes at DBD may also be arranged in a coaxial manner with one electrode within the other, with at least one or both of the dielectric barriers either (i) on the outer side of the inner electrode / on the inner side of the outer electrode, or (ii) on both electrodes are arranged opposite one another or (iii) between the two cylindrical electrodes. Apart from volume discharges, other designs also exist which use either a surface discharge geometry or a co-planar charge geometry. Surface discharge devices have a thin and long electrode on a dielectric surface and an extended back electrode on the back of the dielectric. In this configuration, the discharge gap is not clearly defined so that the discharge propagates along the dielectric surface. There are also combinations of both volume and surface discharge configurations, such as the coplanar arrangement used in plasma display consoles. The coplanar discharge device is characterized by pairs of long parallel electrodes of opposite polarity embedded within a dielectric mass near a surface. In addition to these configurations, other variants of DBD are also used in various applications. The typical arrangements of DBD, as described by Nehra et al. have shown two major modes of discharge, either the filament mode, which is the usual form of discharge composed of many microdischarges randomly distributed across the electrode surface, or the homogeneous glow discharge mode, also because of similarity to DC glow discharges Atmoshsphärendruckglimmentladungsmodus is known.

In one embodiment, the focused energy flow is applied to the subject in a predetermined manner, in particular a pulsed or pulsed mode, which is determined according to the classification (ie taxonomic classification) of the subject being treated or according to the desirable effect. For example, a focused energy flow pulse that is designed to induce a regenerative effect or effect or biochemical effect on a plant may be different (ie, in terms of pulse duration, pulse intervals, or both) from the focused energy flow profile that is designed for it therapeutic or biological or biochemical or regenerative process or to induce a beneficial effect in a human and / or the focused energy flow profile designed to induce a biochemical effect on water or gas or crude oil or tissue or on a microorganism.

It is within the scope of the present invention to provide a method for affecting or effecting at least one biological pathway of a microorganism in a substrate. The aforesaid method comprises steps of: (a) providing a source of non-thermal plasma (NTP) for generating an NTP beam as defined in Table 1; (b) providing a power flow focusing device (EFFM) for transferring the NTP beam into a focused energy flow; (c) providing the Microorganism in the substrate; and (d) operating the EFFM having a predetermined pulsed profile as defined in Table 2 to emit the focused energy flow to the at least one microorganism.

It is further within the scope of the invention to provide the method as defined above, wherein the EFFM comprises an energy flux vector selected from the group consisting of an electric field vector, a magnetic field vector, an electromagnetic field vector, a piezoelectric field vector, a piezoelectric field vector, and any combination thereof ,

It is further within the scope of the invention to provide the method as defined above, wherein the EFFM comprises a focusing vector (or focusing vector).

It is further within the scope of the invention to provide the method as defined above, wherein the substrate is selected from the group consisting of a food, a liquid, a beverage, a suspension, a biological culture, a medium, a growth medium, an emulsion , a biological tissue, a biological organism, a human, an animal, a plant, a fluid, a soil, minerals, media, a gas, a mixture of gas and liquid, gas mixtures, cells, tissue cultures, organs, and any combination thereof.

It is further within the scope of the invention to provide the method as defined above, wherein the microorganism is selected from the group consisting of yeast, bacteria, archaea, algae, fungi, protozoa, viruses, spores, hype, prions and any combination thereof.

It is further within the scope of the invention to provide the method as defined above, wherein the microorganism is further selected from the group consisting of: Candida, corynebacteria, aerobic bacteria, anaerobic bacteria, Listeria monocytogenes, Escherichia coli, Salmonella, Enterobacteriaceae bacteria. Family, Listeriaceae family bacteria, Gram-positive bacteria, Gram-negative bacteria, Staphylococcus aureus, Pseudomonas aeruginosa, Staphylococcus epidermidis, Staphylococcus haemolyticus, Enterococcus faecalis, the Clostridiaceae family, Clostridium, Actinobacteria, and any combination thereof.

It is further within the scope of the invention to provide the method as defined above, additionally comprising steps of affecting or effecting the at least one biological pathway of the microorganism within up to 24 hours from the emission of the focused energy flow to the microorganism, particularly within to to 6 hours from the emission of focused energy flow to the microorganism.

It is further within the scope of the invention to provide the method as defined above, additionally comprising steps of comparing the effect on the at least one biological pathway of the microorganism having the effect in a control organism.

It is further within the scope of the invention to provide the method as defined above, wherein the effect on at least one biological pathway is selected from the group consisting of: activation effect, inactivation effect, metabolic effect, catabolic effect, amphibole effect, regeneration effect, regrowing effect, Effect on a biochemical pathway, effect on the generation of at least one biochemical composition, effect on the production of recombinant proteins, effect on protein mutation, effect on protein profile, effect on protein modification, sterilization, infertility, antimicrobial effect, antibacterial effect, deactivation effect, Survival effect, infertility effect, loss of viability effect, killing effect, destruction effect, induction effect, destruction of pathogenic effect, bacterial destruction effect, viral destruction effect, genetic effect, water or gas disinfection effect Effect on water or gas contamination parameters, effect on the chemical composition, effect on the water or any other fluid chemistry, bacterial concentration effect, phenol concentration, effect on the chemical composition of crude oil, effect on crude oil components and concentration, effect on production and / or the destruction of biofuel or biodiesel compositions and any combination thereof.

It is further within the scope of the invention to provide the method as defined above, wherein the effect on the at least one biological pathway is an effect on at least one biochemical pathway for the production of a composition selected from the group consisting of biofuel and / or products from them, recombinant proteins, mutant proteins, protein profile, biodiesel and / or products from them, organic compounds, metabolic compounds, metabolites, antibiotics, prodrugs, fermentation products, acid compositions, gas compositions, alcohol compositions, precursors of amino acids, ethanol, 1-propanol, 2-propanol, isopropanol, 2,3-butanedione, vicinal diketone, acetoin, acetone , 2,3-butanediol, methanol, isobutanol, butanol, 1-butanol, n-butanol, propanediol, diesel and any isomeric or racemic compound thereof, any intermediate pathway composition or any precursor thereof and any combination thereof.

It is further within the scope of the invention to provide the method as defined above, additionally comprising steps of effecting or affecting at least one of the biological pathway of the microorganism in the substrate while at least substantially the pH, the hydrogen peroxide concentration or a combination thereof Substrates is maintained.

It is further within the scope of the invention to provide a system operated in a method as defined in any of the above, the system comprising: (a) a source of non-thermal plasma (NTP) for generating an NTP beam as defined in Table 1; and (b) an energy flux focusing device (EFFM) for transitioning the NTP beam into a focused energy flow, the EFFM having at least one aperture for passage of the NTP beam, the EFFM comprising at least one energy flux vector selected from the group consisting of: ( i) at least one electric field vector, (ii) at least one magnetic field vector, (iii) at least one electromagnetic field vector, (iv) at least one piezoelectric field vector, and (v) at least one piezomagnetic field vector, wherein the system additionally comprises at least one focusing vector which is complementary thereto is configured to focus the plasma jet, and wherein the EFFM is operated in a predetermined pulsed profile as defined in Table 2 to emit the focused energy flow toward the at least one microorganism.

It is further within the scope of the invention to provide the method as defined in any of the above, the method providing an effect on improving the production of at least one biochemical compound in the at least one microorganism.

It is further within the scope of the invention to provide the method as defined in any of the above additionally comprising steps of harvesting at least one biochemical composition.

It is further within the scope of the invention to provide the method as defined in any of the above additionally comprising steps of selecting the biochemical composition from the group consisting of biofuel and / or products thereof, biodiesel and / or products thereof, organic compound, metabolic Compounds, metabolites, antibiotics, prodrugs, fermentation products, acid compositions, gas compositions, recombinant proteins, mutant proteins, proteins, alcohol compositions, precursors of amino acids and any combination thereof.

It is further within the scope of the invention to provide the method as defined in any of the above additionally comprising steps of selecting the biochemical composition from the group consisting of ethanol, 1-propanol, 2-propanol or isopropanol, 2,3-butanedione, vicinal diketone, acetoin, acetone, 2,3-butanediol, methanol, isobutanol, butanol, 1-butanol, n-butanol, propanediol, also referred to as propanediol or 1,3-propanediol, any isomeric or racemic compound thereof, any intermediate pathway composition or any precursor thereof, and any mixture thereof.

It is further within the scope of the invention to provide the method as defined in any of the above additionally comprising steps of inducing the production of at least one biochemical composition to a concentration of between about 5 mg / L to about 200 mg / L or more and more preferably between about 10 mg / L to about 100 mg / L or more.

It is further within the scope of the invention to provide the method as defined in any of the above, wherein the method provides an effect selected from the group consisting of sterilization, killing, deactivation, inactivation, infertility, and any combination thereof of the at least one microorganism.

It is further within the scope of the invention to provide the method as defined in any of the above additionally comprising steps of providing the at least one biological effect in at least one microorganism without a significant change in pH or pH Hydrogen peroxide concentration or a combination thereof of the substrate harboring the at least one microorganism.

It is further within the scope of the invention to provide a system for effecting at least one biological pathway of a microorganism in a substrate. The aforesaid system comprises: (a) a source of non-thermal plasma (NTP) for generating an NTP beam as defined in Table 1; and (b) a power flow focusing device (EFFM) for transitioning the NTP beam into a focused energy flow, the EFFM having at least one aperture for passage of the NTP beam, and wherein the EFFM comprises at least one energy flux vector selected from the group consisting of: ( i) at least one electric field vector, (ii) at least one magnetic field vector, (iii) at least one electromagnetic field vector, (iv) at least one piezoelectric field vector, and (v) at least one piezomagnetic field vector, wherein the system additionally comprises at least one focusing vector associated therewith is configured to focus the plasma jet and wherein the EFFM is operated in a predetermined pulsed profile as defined in Table 2 to emit the focused energy flow toward the at least one microorganism.

It is further within the scope of the invention to disclose the system as defined in any of the above, wherein the NTP emitting source is selected from the group consisting of a dielectric interface or barrier discharger, an atmospheric pressure glow discharge, a corona plasma discharger, a DC high voltage corona discharger High Voltage Corona Discharger with Negative DC Voltage, High Voltage Corona Discharger with Positive DC Voltage, Blanked Barrier or Barrier Discharger, Plasma Induced Arc (GD), and Plasma Jet.

It is further within the scope of the invention to disclose the system as defined in any of the above, wherein the at least one energy flow vector or the at least one coupling element at least partially comprises polyvinylidene fluoride, polyvinylidene difluoride (PVDF), PZT, lead zirconium titanate, ferroelectric oxides, Pb [Zr _{( x)} Ti _(1-x) ] O ₃ , PbZrO ₃ , PbTiO ₃ , barium titanate (BaTiO ₃ ), (Ba, Sr) TiO ₃ , Ba _(1-x) Sr _(x) TiO ₃ , a ferroelectric material characterized by at least one of piezoelectricity, pyroelectricity and storage properties, a permanent magnet, an electromagnet, a superconducting magnet, cobalt, magnetite (Fe ₃ O ₄ ), α-ferrite (α-Fe), iron, ferromagnetic alloys, piezomagnetic ferrite materials, magnetoelectric ceramic materials, and any combination thereof.

It is further within the scope of the invention to disclose the system as defined in any of the above, wherein the at least one focusing vector or the at least one focusing element comprises a material or element selected from the group consisting of a reflector coating, metals, iron, Metal alloys, stainless steel, aluminum, silver, gold and mixtures thereof, a dielectric coating, an ultraviolet coating, UV-reflective materials, 100-280 nm reflective materials, glass, amorphous materials, solid materials, insoluble materials, crystalline materials, polymers and any combination from that.

It is further within the scope of the invention to disclose the system as defined in any of the above, wherein the at least one energy flow vector or the at least one energy flow element is arranged in at least one group of pairs or triples or in at least one group of more than three elements ,

It is further in the scope of the invention to disclose the system as defined in any of the above, wherein in the focused energy flow is through at least one parameter selected from the group consisting of a dose range of between about 0.1 J / cm ² to about 4 J / cm ² , a frequency range of between about 100 Hz and about 20 MHz, a relative dielectric constant in a range of between about 500 and about 2500, a piezoelectric charge constant in a range of between about 100 (10 ^-12 C / N or 10 ^-12 m / V) to about 1000 (10 ^-12 C / N or 10 ^-12 m / V), a piezoelectric voltage constant in a range of between about 5 (10 ^-3 Vm / N or 10 ^-3 m ² / C) to about 50 (10 ^-3 Vm / N or 10 ^-3 m ² / C), a frequency constant in a range of between about 1000 (Hz · m or m / s) to about 5000 (Hz · m or m / s) and any combination thereof

It is further within the scope of the invention to disclose the system as defined in any of the above, wherein the focused energy flow is adapted to provide at least one biological or biochemical effect selected from the group consisting of at least one brain-related one Parameter, a protein fingerprint or profile, an enzymatic activity, a protein crystallization, at least one medicinal or therapeutic effect, at least one plant parameter, at least one water parameter, at least one air pollution parameter, at least one fluid or gas parameter, at least one oil or crude oil parameter, of the treatment gaseous emissions, ozone treatment, increased functional recovery after a disruptive effect, at least one immune system parameter, at least one skin related parameter, wound healing, recovery from bacterial infection, recovery from viral infection, tissue regeneration, pain relief, antioxidant activity , at least one rheological property, an in vivo effect, an in vitro effect and any combination thereof.

It is further within the scope of the invention that the EFFM include a controller for operating the EFFM in a predefined pulsed manner.

According to a main embodiment of the invention, the focused energy flow generated by the system of the present invention is applied in a predetermined pulsed manner.

It is within the scope of the invention that the NTP plasma emitting source of the system be mounted on the second opposite surface of the plasma jet plate as defined in any of the above.

It is within the scope of the invention that the system, as defined in any of the above, is further adapted to provide pulses of focused energy flow in a predetermined manner. In a specific embodiment, the system includes a control device for providing the focused energy flow pulses in a predetermined manner. Such a device may include a computer-aided algorithm or software.

It is within the scope of the invention that the at least one energy flux vector of the system, as defined in any of the above, comprises a ferroelectric material characterized by at least one of piezoelectricity, pyroelectricity and storage characteristics.

It is within the scope of the invention that the at least one focusing vector or element at least partially comprises a material or element selected from the group consisting of a reflector coating, metals such as iron and alloys thereof, stainless steel, aluminum, silver, gold and mixtures thereof, a dielectric coating, an extreme ultraviolet coating, high energy UV, glass, amorphous materials, solid materials, crystalline materials, polymers, and any combination thereof.

It is further within the scope of the invention that the at least one plasma jet opening be positioned in the center of the plasma jet plate.

It is further within the scope of the invention that the at least one focusing vector or the at least one focusing element is positioned in the center of the plasma jet plate.

It is further within the scope of the invention that the at least one focusing vector or the at least one focusing element is positioned radially around the plasma jet aperture.

It is further within the scope of the invention that the at least one energy flow vector or the at least one energy flow element is arranged in at least one group of pairs or triples or in at least one group of more than three elements.

It is further within the scope of the invention that the at least one energy flow vector or the at least one energy flow element provides a magnetic field in a range of 25 μT to 10 T and higher.

It is further within the scope of the invention that the at least one focusing vector or the at least one focusing element comprises one, two, three or more focusing vectors, or that the at least one focusing vector is configured in a ring-like shape surrounding the plasma jet aperture ,

It is further within the scope of the invention that the focused energy flow be applied in a sequence of pulses having predetermined frequencies and durations.

It is further within the scope of the invention that the focused energy flow be applied in a sequence of pulses characterized by a constant frequency value.

It is further within the scope of the invention that the focused energy flow be applied in a sequence of pulses characterized by increasing duration and / or growing pulse intervals.

It is further within the scope of the invention that the focused energy flow be applied in a sequence of pulses selected from the group consisting of nanosecond pulses, millisecond pulses, second pulses, and a combination thereof.

It is within the scope of the invention that the process as defined in any of the above is useful for treating objects, humans, animals, plants, foods and fluids such as water, oil, crude oil, milk, honey, emulsions, ketchup, blood, Suspensions, other media, gases and liquids and mixtures thereof.

It is further within the scope of the invention that the method is adapted to provide an effect on a biological pathway of cells, tissues, tissue cultures, organs and a combination thereof.

It is further within the scope of the invention that the method is adapted to provide an effect below the outer integument of the treated subject.

It is further within the scope of the invention that the method is adapted to provide a synergistic effect in terms of inducing a therapeutic or regenerative or biological or beneficial effect on the subject as compared to the effect derived from each of the energy flow vectors or inducing vectors when administered individually.

It is further within the scope of the invention that the outflow effect of the focused energy flow may be detected by detection means and methods including, among others, direct measurements such as gas discharge visualization (GDV). , Kirlian photography means, a digital visualization of the biofield (DVB), a scanning electron microscope (SEM), a transmission electron microscope (TEM) and by indirect measurements such as a SQUID ("Superconducting Quantum Interference Device"), a Biophotonmessung, a Biophotonbildgebung , a charge-coupled device (CCD), a photomultiplier tube (PT), magnetic resonance imaging (MRI), nuclear magnetic resonance spectroscopy (NMR), electrophotonic imaging or electro-photon imaging (EPI).

It is further within the scope of the invention that the improved water parameters are selected from the group consisting of an oxidizing effect, induction of degradation of organic compounds, water purification, destruction of pathogens such as bacteria and viruses, purification of radioactive isotopes and heavy metals sterilization, pH values, hydrogen peroxide values, water disinfection, water contamination parameters, action on mineral ions such as calcium and magnesium, oxidation of inorganic ions and any combination thereof.

It is further within the scope of the invention that the protein fingerprint or protein profile be associated with a cellular pathway or protein family selected from the group consisting of signal transduction, stress response, cell cycle, antioxidant, DNA repair, replication, the blood plasma proteins, the glycoproteins, the lypoproteins, the signaling proteins, the therapeutic proteins and any combination thereof.

It is further within the scope of the invention that the protein fingerprint or protein profile is associated with a protein member selected from the group consisting of transferin, amyloid serum A, XPA, PKB, IMP, MMR, XPA, hTLRs, NE, transthyretin, LRR, Ku, NLR, catalase, superoxide dismutase, peroxidases, PAT, LTP, Apm1, NLR, LPAF, beta-glucanases, ferredoxin and any combination.

In some aspects, the system of the present invention includes a gas plasma flow regulator or a gas plasma monitoring device configured to control and / or monitor gas plasma flow parameters, such as flow, flow pressure, mass flow, and gas plasma composition ratios. The system may include a control device to provide focused energy flow pulses in a predetermined manner. Such a device may include an implemented computer-aided algorithm or software.

In order to understand the invention and to see how it can be put into practice, a plurality of preferred embodiments will now be described with reference to the following examples, but only by way of non-limiting example.

Example 1 - Effect on water chemistry parameters:

The effect of the focused energy flow treatment is assessed herein with respect to the chemical analysis of water parameters. It is within the scope of the present invention to test the system and method of the present invention for its effects on fluids, and more particularly, on water.

An exemplary system demonstrates an effect on water chemistry parameters. In this embodiment, an apparatus comprising an EFFM for applying a focused energy flow (ie, generated by magnetic and / or electric fields) is used to determine the effect of the discharged focused energy flow on the water chemistry, ie, the pH of the water and the H ₂ O ₂ concentration, test.

Experimental procedure:

Partial samples of 10 ml of distilled water were subjected to focused energy flux treatments of 8, 15, 22, 29 cycles according to the pulse profiles defined in Table 2. The distance between the surface of the treated water and the plasma jet discharge was about 4.5 cm

pH measurements:

pH measurements were performed using a inoLab ^® Multi 9310 IDS multi-parameter system with IDS sensors (WTW - Germany Technological workshops GmbH) performed. The inoLab ^® Multi 9310 IDS was equipped with an ADA S7 / IDS adapter (WTW GmbH, Germany) and a SenTix ^® Mic pH electrode (Ø = 4.5 mm, WTW GmbH, Germany) to adjust the pH range from 0- 14 to examine.

hydrogen peroxide:

Hydrogen peroxide (H ₂ O ₂ ) was determined photometrically by reaction with titanyl (IV) sulfate in sulfuric acid solution. Absorption measurements of the corresponding end product peroxotitanyl sulfate were carried out with a UV-3100PC UV / VIS scan spectrophotometer (VWR International GmbH, Germany) at Ø = 405 nm.

In this embodiment, the energy flow focusing device (EFFM) is equipped with a ferroelectric plate as the energy flow vector. The ferroelectric plate is preferably characterized as follows: it has a diameter Ø of about 1 to 2.25 cm and is made of piezoelectric material such as APC 841 as described in Table 3 below.

Reference is now made to Table 3, which describes the physical and piezoelectric properties of APC materials disclosed in U.S. Pat http://www.americanpiezo.com/apc-materials/piezoelectric-properties.html Are available. Table 3: Selected Properties of Certain Embodiments of the EFFM

According to specific embodiments, this example relates to properties of the EEFM comprising a ferroelectric material APC841.

It is within the scope of the invention that the system of the present invention is configured to provide improved properties, ie, an improvement of at least 5% relative to the properties presented in Table 3.

It is recognized herein that the water quality characteristics are often used in relation to a group of standards against which performance can be assessed. The most common norms used to assess water quality relate to the health of the ecosystem, the safety of human contact and drinking water as well as the human medical process. Different uses against various concerns and therefore different standards are considered. The system and method of the present invention is provided in a non-limiting manner for water or any other fluid such as an emulsion, suspension (e.g., a microbial suspension), oil, crude oil, food, drink, milk, etc. or the like Gas cleaning, disinfection or the cleaning of heavy metals or pollution used. Other parameters which may be affected by exposure to the focused energy flux treatment disclosed herein include but are not limited to: pH, dissolved oxygen, oxygen peroxide, nitric oxide (NO), nitrate (NO ₃ ) and nitrite (NO ₂ ), biochemical compounds or metabolite production, conductivity, chemical composition, oxygen reduction potential (ORP), bacterial concentration, ie E. coli, and turbidity. In some aspects, the effects provided by the system and method of the present invention include any biological effect on a fluid or fluid containing any sample, subject or substrate whose major component is a fluid, such as biological tissue or a microbial suspension or fluid a food or oil.

In other embodiments, the system and method of the present invention may be used to cause cleaning effects on treated water or a treated fluid as compared to a control sample. The cleaning effect may be determined by parameters associated with water or any other fluid such as oil, crude oil, milk, etc. or gas quality or purification, disinfection or contamination. These parameters may be chemical, physical and biological characteristics of water or any other fluid such as oil, crude oil, milk, etc., or gas.

These effects can be achieved, for example, by the following method:

Plasma is provided by a plasma emitter, ie a device such as the 3A or 3B illustrated. A focused energy flow is generated and applied to the test subjects using the EFFM of the present invention, ie, as in FIG 1 illustrated.

In accordance with certain aspects of the invention, the focused energy flow is expelled in a sequence of pulses, i. H. as described in Table 2. According to one option, a group of pulses is provided which have an increasing pulse duration in the range of 1 and 5 seconds and increasing times between pulses in the range of 1 to 5 minutes.

Experimental Results: Experiment 1: cycles pH H ₂ O ₂ (mg / l) 0 6,052 0,000 8th 5,965 0,000 15 5,916 0,000 22 6,027 1,386 29 5,891 6,727 Experiment 2: cycles pH H ₂ O ₂ (mg / l) 0 6,047 0,000 8th 5,944 0,000 15 5,902 0.023 22 6.023 1,500 29 5,992 4,341 Experiment 3: cycles: pH H ₂ O ₂ (mg / l) 0 6,063 0,000 8th 5,953 0,000 15 5,911 0,000 22 6.017 1,159 29 6,009 3,545

It is shown herein that exposing water to the focused energy flow protocols of the present invention surprisingly results in no significant changes in the pH of the treated water compared to the control (0 pulses) for each of the used pulse cycles remains comparable to its neutral pH. Furthermore, the treatments of the present invention are unexpectedly provided in the case of treatments using less than 22 pulsed cycles (less than about 20 minutes exposure to the modified plasma) without the production of H ₂ O ₂ in the treated water. It is emphasized that these effects are achieved without introducing any chemical or biological material or object into the modified plasma-treated water or fluid. These results are unique and differ from currently available non-thermal plasma technologies which produce H ₂ O ₂ and / or show a pH drop of from about 6 or 7 to about 4 or less (e.g. Oehmigen, K., et al., 2010, "The Role of Acidification for Antimicrobial Activity of Atmospheric Pressure Plasma in Liquids", Plasma Processes and Polymers 7, 3-4, pp. 250-257 ; Satoshi Ikawa, Katsuhisa Kitano and Satoshi Hamaguchi, 2010, "Effects of pH on Bacterial Inactivation in Aqueous Solutions due to Low-Temperature Atmospheric Pressure Plasma Application", Plasma Processes and Polymers, 7, 1, pp. 33-42 ; Fuxiang Liu et al., 2010, "Inactivation of Bacteria in an Aqueous Environment by a Direct-Current, Cold-atmospheric Pressure Air Plasma Microjet", Plasma Process And Polymers, 7, pp. 231-236 ).

To summarize, it is submitted that the above-disclosed focused energy flow treatment protocols are configured to provide an effective biological or biochemical effect on water or other fluids, as well as humans, animals, and plants.

Example 2 - Effect on metabolic pathways in yeast

General purpose of the experiments:

The microbial production of higher alcohols through fermentation of sugars offers sustainable solutions as a potential source of gasoline substitutes and alternatives to the traditional petrochemical industry. Among the most outstanding examples of an industrial microbial cell factory is the yeast Saccharomyces cerevisiae (S. cerevisiae), which is widely used to produce ethanol as an alternative fuel. However, advanced biofuels such as n-butanol, isobutanol and isopropanol offer superior fuel characteristics over ethanol.

The experiments described below aim to examine if and under what conditions different biofuel components or compounds or products can be produced by treating industrial microorganisms such as S. cerevisiae with the method and system and protocols as described, inter alia.

Experimental procedure:

The source of Saccharomyces cerevisiae used in this experiment is S. cerevisiae (DSM: 1333).

The freeze-dried yeast was inoculated into a culture in accordance with the instructions provided by the supplier. The yeast was grown in Caso Bouillon growth media at room temperature for 48 hours, spread on agar count plates and incubated for a further 48 hours at room temperature and kept refrigerated.

For these experiments, the yeast was grown for 48 hours at room temperature in a nutrient broth containing 20 g of glucose, 5 g of peptone and 3 g of yeast extract per 1 liter of "Gerolsteiner" mineral water without carbon dioxide.

• 1. Für insgesamt 5 Wiederholungen wurden zwei Proben von 5 ml Hefesuspension durch 3, 5 oder 7 Pulszyklen mit einer festen Menge an fokussiertem Energiefluss behandelt (d. h. erste fokussierte Energieflussbehandlung).
• 2. Das behandelte Volumen wurde zusammengefasst, um 10 ml Hefesuspension zu gewinnen, die einmal mit der gewählten Anzahl von Pulszyklen für jede der 5 Wiederholungen behandelt wurde.
• 3. 5 ml von jeder dieser 10 ml mit fokussiertem Energiefluss behandelten Hefesuspension wurde für jede Wiederholung genommen und über 6 Stunden bei Raumtemperatur bei 350 U/min konstantem Schütteln inkubiert.
• 4. Nach 6 Stunden Inkubation wurden 1,5 ml von jeder Probe für Gaschromatografieanalysen genommen.
• 5. Die verbleibende Menge Suspension wurde unter den gleichen Bedingungen über zusätzliche 18 und 42 Stunden inkubiert.
• 6. 24 und 48 Stunden nach der Plasmabehandlung wurde eine weitere 1,5 ml Probe für die Gaschromatografieanalyse genommen.
• 7. Die verbleibenden 5 ml von jeder Probe wurden 1 Stunde nach der ersten fokussierten Energieflussbehandlung über eine Sekunde behandelt (d. h. zweite fokussierte Energieflussbehandlung).
• 8. Nach der zweiten fokussierten Energieflussbehandlung (genau die gleiche Behandlung wie die erste fokussierte Energieflussbehandlung) wurden die Proben ebenfalls bei Raumtemperatur inkubiert, während sie mit 350 U/min geschüttelt wurden.
• 9. Proben für die Gaschromatografieanalyse wurden nach 6, 24 und 48 Stunden insgesamter Inkubation genommen.
• 10. Parallel zu den mit fokussiertem Energiefluss behandelten Proben wurden 5 ml unbehandelte Hefesuspension unter exakt den gleichen Bedingungen wie die behandelten Proben inkubiert und durch Gaschromatografie analysiert. Diese unbehandelten Proben wurden als die Kontrolle verwendet, der gegenüber die Messergebnisse verglichen werden.
• 11. Die unbehandelte Hefesuspension wurde verdünnt und auf Agar-Zählplatten ausgebreitet und bei Raumtemperatur über 48 Stunden inkubiert. Die Kolonien wurden gezählt und die Anfangskonzentration der Hefesuspension wurde berechnet.

The plasma treatment of the yeast suspension was carried out according to the following procedure or protocol:

• 1. For a total of 5 replicates, two samples of 5 ml of yeast suspension were treated by 3, 5 or 7 pulse cycles with a fixed amount of focused energy flow (ie first focused energy flux treatment).
• 2. The treated volume was pooled to obtain 10 ml of yeast suspension, which was treated once with the chosen number of pulse cycles for each of the 5 repetitions.
• 3. 5 ml of each of these 10 ml of focused energy flow treated yeast suspension was taken for each repetition and incubated for 6 hours at room temperature at 350 rpm constant shaking.
• 4. After 6 hours of incubation, 1.5 ml of each sample was taken for gas chromatographic analysis.
• 5. The remaining amount of suspension was incubated under the same conditions for an additional 18 and 42 hours.
• 6. At 24 and 48 hours after the plasma treatment, another 1.5 ml sample was taken for gas chromatographic analysis.
• 7. The remaining 5 ml of each sample was treated 1 hour after the first focused energy flux treatment for one second (ie second focused energy flow treatment).
• 8. After the second focused energy flux treatment (exactly the same treatment as the first focused energy flux treatment), the samples were also incubated at room temperature while being shaken at 350 rpm.
• 9. Samples for gas chromatographic analysis were taken after 6, 24 and 48 hours of total incubation.
• 10. In parallel with the focused energy flow treated samples, 5 ml of untreated yeast suspension were incubated under exactly the same conditions as the treated samples and analyzed by gas chromatography. These untreated samples were used as the control compared to the measurement results.
• 11. The untreated yeast suspension was diluted and spread on agar count plates and incubated at room temperature for 48 hours. The colonies were counted and the initial concentration of the yeast suspension was calculated.

It is noted that the protocol described above was repeated for each group of focused energy flow pulse cycles.

The concentrations of the yeast suspensions were as follows:
3 pulse cycles: 2.4 · 10 ^ 8 cfu · ml-1
5 pulse cycles: 2.3 · 10 ^ 8 cfu · ml-1
7 pulse cycles: 1.1 · 10 ^ 8 cfu · ml-1

The NTP source used in this experiment is preferably a plasma jet. The gas plasma discharge comprises a combination of argon, N ₂ and O ₂ , ie as defined in Table 1. The gas flow rate is between about 3-4 liters per minute. The plasma power is about 2 W.

It is further within the scope that the distance between the jet discharge and the treated sample be between about 1 cm and up to about 5 cm, more preferably between about 3 cm and about 5 cm.

The identification of the biofuel products was carried out by an analytical analysis method using a headspace sampler (i.e., 7697A, Agilent).

The separation was made with a 7890A GC system in combination with a 5975C mass detector (both Agilent).

The compounds identified with the NIST database included: ethanol, 2-propanol, 1-propanol, 2,3-butanedione, acetoin and 2,3-butanediol.

The term "1-propanol" as used herein in a nonlimiting manner refers to a primary alcohol having the formula CH ₃ CH ₂ CH ₂ OH. It is also known as propan-1-ol, 1-propyl alcohol, n-propyl alcohol and n-propanol.

The term "2-propanol" refers in a non-limiting manner herein to isopropyl alcohol and isopropanol.

The term "2,3-butanedione" herein refers, in a non-limiting manner, to diacetyl, butane-2,3-dione, vicinal diketone and 1,2-diketone.

The term 2,3-butanediol [(CH ₃ ) ₂ (CHOH) ₂ ] also refers to its three stereoisomers consisting of two enantiomers and one meso compound.

The term "acetoin" refers herein in a non-limiting manner to 3-hydroxybutanone or acetylmethylcarbinol or (R) -acetoin.

The present invention further includes isomers, stereoisomers and derivatives of the aforementioned compounds.

Experimental results:

It will be up now 4 Which presents a graphic illustration of the metabolic compounds produced by S. cerevisiae that undergo focused energy flux treatment ( 4A ) or two focused energy flux treatments of 5 pulse cycles each ( 4B ) was compared to an untreated (control) S. cerevisiae ( 4C ). It should be noted that the duration of each pulse cycle treatment is about 3.5 minutes. It is surprisingly shown that an enhanced production of advanced biofuel compounds is observed by exposure of the yeast S. cerevisiae to 5 cycles of focused energy flux treatment as compared to the untreated yeast. In particular, the signal level of 2,3-butanediol is increased after 24 hours recovery from one and / or two focused energy flux treatments ( 4A and 4B ). Furthermore, the levels of 1-propanol and 2-propanol are dramatically increased already after 6 hours after the one or two treatments ( 4A and 4B ). On the other hand, the observed levels of acetoin signaling produced by the treated yeast are as a result of 4A ) or two ( 4B ) focused energy flux treatments in comparison to the untreated control yeast ( 4C ) decreased.

It will be up now 5 , Which presents a graphic illustration of the metabolic compounds produced by S. cerevisiae which are a ( 5A ) or two ( 5B ) was subjected to focused energy flux treatments of 7 pulse cycles, compared to an untreated S. cerevisiae control ( 5C ). It can be seen that after 6 hours recovery from exposure to modified plasma treatment, the production signal levels of 1-propanol, 2-propanol and 2,3-butanedione in the treated yeast (one and two treatments, 5A respectively. 5B ) relative to the control yeast ( 5C ) have significantly increased. 24 hours after the focused energy flux treatment, the signal levels of 1-propanol and 2-propanol remained high ( 5A and 5B while the level of 2,3-butanediol is similar to the control ( 5C ) was.

It is therefore shown that the exposure of S. cerevisiae to about 3.5 to about 4.9 minutes of pulsed focused energy flux treatment according to the present invention, the production of 1-propanol, 2-propanol and 2,3-butanedione by the treated yeast induced relative to the untreated control yeast. The production of these biochemical compounds can be detected as early as about 6 hours or earlier after the treatment.

It will be up now 6 Which presents a graphical illustration of the concentration of metabolic compounds or biochemical compounds produced by S. cerevisiae that undergo focused energy flux treatment ( 6A ) or two focused energy flux treatments ( 6B ) was exposed to 5 pulse cycles compared to a S. cerevisiae control ( 6C ). These data clearly show the significantly improved production of 1-propanol and 2-propanol in the treated yeast. The concentration of 1-propanol and 2-propanol was undetectable in the control yeast during up to 24 Hours ( 6C ). On the other hand, in the treated yeast, the two focused energy flux treatments of 5 pulse cycles ( 6B ), the concentration of 1-propanol and 2-propanol after 6 hours recovery from the treatment was about 40 μl / l and about 30 μl / l, respectively, when the duration of each treatment was at most about 3.5 minutes.

It will be up now 7 Which presents a quantitative graphical illustration of the concentration of biochemical compounds produced by S. cerevisiae that undergo focused energy flux treatment ( 7A ) or two focused energy flux treatments ( 7B ) was exposed to 7 pulse cycles compared to an untreated S. cerevisiae control ( 7C ).). It can be seen that induction of 1-propanol and 2-propanol production is achieved in S. cerevisiae treated with the focused energy flow protocol of the present invention. The measured concentration of 1-propanol and 2-propanol detected 6 hours after treatment is approximately 40 μl / l and between approximately 20 to 25 μl / l, respectively.

The concentration of biofuel biofuel products produced in milligrams per liter is estimated in Table 4 as follows: TABLE 4 Determination of 1-propanol and 2-propanol concentration (mg / l) in the yeast suspension 1-propanol (mg / l) Number of pulse cycles A treatment Two treatments control 3 20.47 31.46 - 5 25.45 35.33 0 7 20.30 31.55 0 2-propanol (mg / l) Number of pulse cycles A treatment Two treatments control 3 15,30 22,50 - 5 18.78 24,99 0 7 16.03 20.60 0

The results described above clearly demonstrate that endogenous production of the compounds 2,3-butanediol, 1-propanol and 2-propanol is induced in S. cerevisiae exposed to the focused energy flux treatments of the present invention according to a predefined pulse protocol , It is further surprisingly shown that the improved production of this important and advanced generation of biofuel compounds is observed up to 6 to 24 hours incubation after yeast treatment. The duration of a treatment is between about 3.5 and up to about 4.9 minutes and about 7 to about 9.8 minutes for two treatments with 1 hour rest between treatments. It is also shown that 6 hours after the yeast treatment, the 1-propanol production reaches a concentration of between about 30 mg / L to about 40 mg / L and the 2-propanol production has a concentration of between about 20 mg / L to about 30 mg / l reached.

In summary, the present invention provides a method for producing advanced generation biofuel compounds, such as short chain alcohols, through the exposure of microorganisms, i. H. S. cerevisiae, ready for administration of a focused energy flow. It is noted that the production of the short chain alcohols (ie 2,3-butanediol, 1-propanol and 2-propanol) is achieved rapidly after up to about 6 to about 24 hours of a short treatment period of at most about 3 to about 6 minutes ( or about 7 to about 10 minutes for 2 treatments with 1 hour rest between treatments). It is further noted that the effect on the yeast's endogenous metabolic pathways is achieved without technical modification (i.e., DNA manipulation) of the yeast and without pretreatment with additional materials as reported so far. The generation of beneficial biofuels by use of S. cerevisiae treated by the process of the present invention provides a sustainable and / or renewable solution for the commercial production of alternative energy sources.

Example 3 - Effect on Deactivation of Bacteria

Listeria experiments:

Reference will now be made to an experiment showing the effect of focused energy flow treatment on Listeria monocytogenes.

Exemplary experimental protocol comprising the following steps:

1. Preparation of an overnight culture:

A colony of L. monocytogenes was suspended in a culture medium and incubated for 48 hours at 37 ° C.

2. Preparation of the stock solution:

The overnight culture was centrifuged at 4500 rpm. The supernatant was decanted and the cell pellet resuspended with a sterile NaCl solution.

3. dilution

The stock solution was diluted to give a concentration of ~ 10 ⁴ CFU / ml of the working solution.

4. Sampling

Partial samples (i.e., 100 μl) of the working solution were transferred to petri dishes together with 5 ml of sterile NaCl (diameter = 60 mm). Each sample was weighed prior to the focused energy flux treatment (since the treatment can cause evaporation)

5. Focused energy flow treatment

Gas mixture composition: argon, N ₂ and O ₂ , ie as defined in Table 1. Pulse cycles: 3, 5 and 11 (ie as defined in Table 2).

6. Adaptation for evaporation

After the plasma treatment, the samples were weighed again and fluid loss was balanced by sterile NaCl.

7. Spiral plate counting

50 μl of the plasma-treated samples were spiral plated onto XLD agar immediately after treatment (0), 1 hour after treatment (60) and 1.5 hours after focused energy flow treatment (90). The agar plates were incubated for 48 hours and at 37 ° C and the number of colonies counted (i.e., colony forming units = CFU or colony forming units).

Experimental results: Table 5: Mean and standard deviations of Listeria monocytogenes survival results from three experimental measurements (CFU measurements) Listeria monocytogenes, CFU mean (MV) and standard deviation (SD) cycles 3 5 11 control Time since treatment (min) MV SD MV SD MV SD MV SD 0 1510 682 27.4 31 0 0 35300 2610 60 47.8 65.1 0 0 0 0 13200 2940 90 0 0 0 0 0 0 9590 2560

The results described above show a significant effect of the focused energy flux treatments described in the present invention on bacterial sterilization. It is shown that the exposure of bacteria (ie Listeria monocytogenes) to the focused energy flux treatment (3 pulse cycles) for about 2 minutes results in deactivation of approximately 95% of the treated bacteria immediately after treatment compared to the untreated control bacteria (0). , It is further shown that exposure of bacteria (ie Listeria monocytogenes) over about 2 minutes to focused energy flux treatment (3 pulse cycles) results in destruction / inactivation of about 99% of the treated bacteria at 60 minutes post-treatment (0) and to inactivation of the treated bacteria 90 minutes after the 2 minute treatment (3 pulse cycles). A focused energy flow treatment over approximately 3.5 minutes (5 pulse cycles) results in a deactivation of approximately 99% of the treated bacteria immediately after treatment and a loss of viability of the treated bacteria 60 minutes after exposure to the focused energy flow treatment. It is further shown in Table 5 that the focused energy flux treatment for approximately 7.7 minutes results in deactivation of the treated Listeria bacteria known to be dangerous human pathogens immediately after exposure to the focused energy flux treatment.

E. coli experiments:

Reference will now be made to an experiment showing the effect of focused energy flow treatment on Escherichia coli (E. coli).

Exemplary experimental protocol comprising the following steps:

1. Preparation of an overnight culture:

A colony of E. coli was suspended in a culture medium and incubated at 37 ° C for 24 hours.

2. Preparation of the stock solution:

10 ml of the overnight culture was centrifuged at 4500 rpm. The cell pellet was resuspended with 10 ml of a sterile NaCl solution.

3. dilution

The stock solution was diluted with sterile NaCl, to obtain a working solution with a concentration of ~ 10 ⁴ CFU / ml.

4. Sampling

Partial samples (i.e., 100 μl) of the working solution were transferred to petri dishes together with 5 ml of sterile NaCl (diameter = 60 mm). Each sample was weighed prior to the focused energy flux treatment.

5. Focused energy flow treatment

Gas mixture composition: argon, N ₂ and O ₂ , ie as defined in Table 1. Pulse cycles: 8, 15, 22 and 29 (ie as defined in Table 2).

6. Adaptation for evaporation

After the plasma treatment, the samples were weighed again and fluid loss was balanced by sterile NaCl.

7. Spiral plate counting

50 μl of the focused energy flow-treated samples were spiral plated onto CaSo agar immediately after treatment (0), 1 hour after treatment (60) and 1.5 hours after focused energy flow treatment (90). The agar plates were incubated for 24 h and at 37 ° C. Table 6: Mean and standard deviations of E. coli survival results from three experimental measurements (CFU measurements) Escherichia coli in NaCl, CFU mean (MV) and standard deviation (SD) cycles 8th 15 22 29 control Time since treatment (min) MV SD MV SD MV SD MV SD MV SD 0 15100 10700 6100 4310 6460 9010 0 0 91700 871 60 4810 6710 0 0 0 0 0 0 94200 535 90 0 0 0 0 0 0 0 0 94100 899

The results described above show the significant effect of the focused energy flux treatments described in the present invention on bacterial sterilization. Exposure of bacteria (i.e., E. coli) to the focused energy flux treatment (8 pulse cycles) over about 5 minutes is shown to result in: deactivation of approximately 92% of the treated bacteria immediately after treatment compared to the control; and loss of approximately 95% of the treated bacteria (relative to the control) after a recovery of 60 minutes following treatment. Furthermore, deactivation of the treated bacteria is shown 90 minutes after the approximately five minute treatment. Focused energy flow treatment for approximately 10 minutes (15 pulse cycles) results in a deactivation of approximately 96% of the treated bacteria (relative to the control) immediately after the focused energy flow treatment and an approximately 99% loss of viability of the treated bacteria at 60 minutes after exposure towards the focused energy flow treatment. It is further shown in Table 6 that the focused energy flux treatment over about 20 minutes (29 pulse cycles) results in deactivation of the treated E. coli bacteria immediately after treatment.

In summary, the present invention unexpectedly demonstrates that by using the focused energy flow system and the focused energy flow method of the present invention, microbiological metabolic, anabolic, or amphibolic pathways can be effectively manipulated to alter metabolite production, for example, the microbiological production of highly important sustainable and renewable biofuels the next or advanced generation. It is further shown that bacterial sterilization or disinfection or antimicrobial activity can be achieved by administration of the relatively short and highly efficient focused energy flux treatments of the present invention.

The present invention further demonstrates that the biological or biochemical effects provided by the focused energy flux treatments presented (ie, in particular, microbial sterilization or the production of biochemical compounds), inter alia, without significantly affecting or changing chemical parameters such as pH or the H ₂ O ₂ concentration of the treated sample can be achieved. The beneficial effects shown may result from synergistic characteristics of the focused energy flow system and the treatment regimen. It is within the scope that the system and method of the present invention is useful for providing biological effects below the outer integument of the treated or treated subject, e.g., human tissue. The effect can be achieved without leading to undesirable or detrimental effects such as lowering the pH or increasing the H ₂ O ₂ concentration.

QUOTES INCLUDE IN THE DESCRIPTION

This list of the documents listed by the applicant has been generated automatically and is included solely for the better information of the reader. The list is not part of the German patent or utility model application. The DPMA assumes no liability for any errors or omissions.

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Claims (11)

A method for affecting or effecting at least one biological pathway of a microorganism in a substrate, the method comprising steps of: a) providing a source of non-thermal plasma (NTP) for generating an NTP jet, said NTP jet containing argon, nitrogen and oxygen in a predetermined ratio; b) providing an energy flow focusing device (EFFM) for transferring the NTP beam into a focused energy flow; c) providing the microorganism in the substrate; and d) operating the EFFM with a predetermined pulsed profile to emit the focused energy flow to the at least one microorganism. The method of claim 1, wherein the predetermined ratio is selected from a table consisting of: No. Ar (%) N ₂ (%) O ₂ (%) R (%) 1 80-99 0-10 0-5 0 2 80-95 5-12 0-4 0 3 80-95 0-15 0-5 0 4 80-95 0-12 0-4 0-4 5 90 8th 2 0 6 85 9 6 0 7 90 6 4 0 8th 95 3 2 0 9 100a a = 0.7-1.3 100b b = 0.13-0.06 100c b = 0.01-0.03 0 10 100a a = 0.6-1.2 100b b = 0.075-0.22 100c b = 0.005-0.025 100r r = 0.005-0.075 and further wherein the predetermined pulsed profile is selected from a table consisting of:

 The method of claim 1, wherein the EFFM comprises a focusing vector and an energy flow vector selected from the group consisting of an electric field vector, a magnetic field vector, an electromagnetic field vector, a piezoelectric field vector, a piezoelectric field vector, and any combination thereof.  The method of claim 1, wherein the substrate is selected from the group consisting of a food, a liquid, a beverage, a suspension, a biological culture, a medium, a growth medium, an emulsion, a biological tissue, a biological organism, a Humans, an animal, a plant, a fluid, a soil or a solid, minerals, media, a gas, mixtures of gas and liquid, gas mixtures, cells, tissue cultures, organs and any combination thereof.  The method of claim 1, wherein the microorganism is selected from the group consisting of yeast, bacteria, archaea, algae, fungi, protozoa, viruses, spores, hype, prions, candida, corynebacteria, aerobic bacteria, anaerobic bacteria, Listeria monocytogenes, Escherichia coli, Salmonella, Enterobacteriaceae family bacteria, Listeriaceae family bacteria, Gram-positive bacteria, Gram-negative bacteria, Staphylococcus aureus, Pseudomonas aeruginosa, Staphylococcus epidermidis, Staphylococcus haemolyticus, Enterococcus faecalis, Clostridiaceae family, Clostridium, Actinobacteria and any combination thereof.  The method of claim 1, further comprising steps of affecting or effecting the at least one biological pathway of the microorganism within up to 24 hours from the emission of the focused energy flow to the microorganism, in particular within up to 6 hours from the emission of the focused energy flow the microorganism.  The method of claim 1, further comprising steps of comparing the effect on the at least one biological pathway of the microorganism having the effect in a control organism.  The method of claim 1, wherein the effect on at least one biological pathway is selected from the group consisting of: activation effect, inactivation effect, metabolic effect, effect on a biochemical pathway, effect on the generation of at least one biochemical composition, effect on the generation of recombinant pathways Proteins, effect on protein mutation, effect on protein expression, effect on protein modification, effect on protein profile, sterilization, infertility, antimicrobial effect, antibacterial effect, deactivation effect, survival effect, infertility effect, loss of viability effect, killing effect, destruction effect, induction effect, destruction of pathogenic effect, bacterial destructive effect, anti-viral effect, genetic effect, water or gas disinfection effect, effect on water or gas contamination parameters, effect on chemical composition, effect on water or any other fluid chemistry, bacterial concentration effect, phenol concentration, effect on the chemical composition of crude oil, effect on crude oil components and concentration, effect on the production and / or destruction of biofuel or biodiesel compositions and any combination thereof. The method of claim 8, wherein the effect on the at least one biological path is an effect on at least one biochemical path for the production of a composition selected from the group consisting of biofuel and / or products thereof, biodiesel and / or products thereof, organic compounds, metabolic compounds, metabolites, antibiotics, prodrugs, fermentation products, acid compositions, gas compositions, alcohol compositions, precursors of amino acids, ethanol, 1-propanol, 2- Propanol, isopropanol, 2,3-butanedione, vicinal diketone, acetoin, acetone, 2,3-butanediol, methanol, isobutanol, butanol, 1-butanol, n-butanol, propanediol, diesel and any isomeric or racemic compound thereof, any intermediates Pathway composition or any precursor thereof and any combination thereof.  The method of claim 1, further comprising steps of effecting or affecting the at least one biological pathway of the microorganism in the substrate while maintaining at least substantially the pH, hydrogen peroxide concentration, or a combination thereof of the substrate.  A method of affecting or effecting at least one biological pathway of a microorganism in a substrate in a system comprising: a) a source of non-thermal plasma (NTP) for generating an NTP jet, said NTP jet containing argon, nitrogen and oxygen in a predetermined ratio; and b) an energy flux focusing device (EFFM) for transferring the NTP beam into a focused energy flow, the EFFM having at least one aperture for passage of the NTP beam, and wherein the EFFM comprises at least one energy flux vector selected from the group consisting of: i) at least one electric field vector, ii) at least one magnetic field vector, iii) at least one electromagnetic field vector, iv) at least one piezoelectric field vector and v) at least one piezomagnetic field vector wherein the system additionally comprises at least one focusing vector configured to focus the plasma beam and wherein the EFFM is configured to provide focused energy flow pulses having a predetermined profile to at least one biological path of the microorganism in the substrate influence or effect.

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