Classifications

• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
  • C12M23/00—Constructional details, e.g. recesses, hinges
  • C12M23/02—Form or structure of the vessel
  • C12M23/18—Open ponds; Greenhouse type or underground installations
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
  • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
  • C12M21/02—Photobioreactors
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
  • C12M23/00—Constructional details, e.g. recesses, hinges
  • C12M23/26—Constructional details, e.g. recesses, hinges flexible
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
  • C12M31/00—Means for providing, directing, scattering or concentrating light
  • C12M31/10—Means for providing, directing, scattering or concentrating light by light emitting elements located inside the reactor, e.g. LED or OLED
• • C—CHEMISTRY; METALLURGY
  • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
  • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
  • C12M39/00—Means for cleaning the apparatus or avoiding unwanted deposits of microorganisms

Definitions

• the present invention relates to a method and a device for the cultivation of algae.
• algae designates, for convenience, any species of aquatic microscopic photosynthetic organism such as microalgae, cyanobacteria or microscopic angiosperms ("micro-crops” such as duckweeds).
• These algae may be from one of hundreds of thousands of naturally occurring species on the surface of the globe or have been genetically modified by techniques known to those skilled in the art.
• the algae can be grown as pure cultures (single species) or mixed cultures with several algae of different species, identified or not.
• the algae can be grown in fresh water, sea water or brackish water, fresh or used.
• the algae can be grown for themselves or to manufacture various chemical compounds (cellulose, sugars, alcohols, lipids, proteins) by recycling carbon dioxide in the form of organic matter via the photosynthesis reaction. These chemical compounds can be produced inside the algal or secreted cells.
• the cultivated algae are separated from the water which contains, continuously or by batch methods, by various methods known to those skilled in the art.
• some of the chemical compounds of interest produced or secreted can be incorporated into various products or supplements for the chemical industries (eg ethanol), food or feed (eg Omega 3), cosmetics and pharmaceuticals. .
• Some of these chemical compounds can be used to make biofuels such as bioethanol, biodiesel, and various "designer fuels" that can be directly substituted, partially or totally, for gasoline, diesel or jet fuel. in motor, rail, water and marine transport.
• the algae may also, by various methods known to those skilled in the art, be used for the production of biogas, biohydrogen or bioelectricity
• Biorefineries built around algae cultivation thus offer several important advantages in the fields of chemistry, energy, environment, food and health compared to current processes based on seaweed substrates. fossil origin.
• Algae cultivation in the absence of light is similar to fermentation and therefore relies on equipment and technologies adapted to well established fermentation industries.
• This approach has two major drawbacks. First, qualitatively, it is estimated that a small minority, of the order of 1 to 10% of natural algae can be adapted to such a type of heterotrophic culture.
• the substrate typically used for fermentation is sugar.
• world production of annual sugar, for all purposes including food and ethanol production is 170 million tons per year, or 17 kJ per g, a quantity of energy of the order of 2.9 10 At 18 Joules.
• World energy consumption worth 500 exajoules (5 10 TO 20 Joules) we see that this heterotrophic approach is not able to replace, on a large scale, fossil fuels used to produce the vast majority of energy currently used for human activities and production.
• heterotrophic algae involves a preliminary photosynthesis step allowing the fixation of organic matter (sugars or possibly other substrates) by terrestrial plants thanks to terrestrial plants. This stage involves significant amounts of fertilizer, water, energy, human labor and soils and its environmental record is very imperfect.
• Microalgae are grown in the laboratory in the presence of artificial light.
• the cost of this approach is prohibitive. It is limited here to consider the marginal cost of electricity needed for lighting, without taking into account neither the installation and operating costs of the culture device nor the price of the light sources and their replacement. It takes a low cost of electricity, 5 cts / kWh, a yield of the artificial light source of 20%, an overall efficiency of photosynthesis of 25% with this source of light (it takes eight photons to fix a CH2O molecule), an algal oil content of 50% (by mass). It is considered that the oil extracted from algae has an energy density close to that of diesel, ie 12 kWh / kg. Producing 1 kg of oil implies, under these quite optimistic conditions, a marginal cost in electricity of
• the average annual solar power received by the Earth's surface is about 90,000 TW.
• the human consumption of energy is equivalent to an average power of the order of 18 TW or 5000 times less. It is thus obvious that solar radiation is a renewable source of primary energy that is largely enough abundant to meet all the energy needs of man.
• Photosynthesis allows, with instantaneous yields measured between 0.02 and 10%, and more generally average yields observed between 0.1% and 2% for terrestrial plants, to directly convert solar energy into fuels and bioproducts, and therefore into a source of energy.
• algae offer surface yields that are much higher than traditional terrestrial crops such as rapeseed or even palm oil (between 5 and 100 times).
• algae do not need agricultural soils, eliminating the problems, put forward for current biofuels, competition with food and negative environmental impact of land use changes and practices such as deforestation.
• Algae can finally be used to directly recycle concentrated carbon dioxide and wastewater.
• PAR photosynthetically active radiation
• the invention relates to a method and a device that improves the growth performance of algae. This yield improvement improves economic viability and environmental performance (life cycle analysis), particularly in the context of biorefineries.
• the invention relates to a device for the cultivation of algae in natural light, comprising an enclosure with a culture medium and the algae to be cultivated, characterized in that the device further comprises a substrate arranged to receive the solar radiation for photoconversion of said solar radiation, the substrate comprising at least one luminescent compound for reemitting a radiation whose spectrum is adapted to the optimization of a biological parameter of interest resulting from the photosynthesis of said algae.
• the substrate is interposed between the incident solar radiation and the enclosure.
• the enclosure is formed of a culture basin, at least partially covered by the substrate.
• the substrate constitutes a wall of the enclosure.
• the enclosure is formed by a tube circuit in which circulates the culture medium containing the algae in suspension.
• the enclosure is formed by a flexible bag forming the substrate, made of a material substantially transparent doped with at least one luminescent compound.
• the substrate comprises particles suspended in the culture medium, the luminescent compound (s) being incorporated into the particles.
• the substrate comprises at least 2 luminescent compounds.
• At least one of said luminescent compounds at least partially overlaps the emission spectrum of at least one of said other luminescent compounds.
• At least one of said luminescent compounds has an absorption spectrum covering the 300 - 360 nm band and an emission spectrum covering the 340 - 400 nm band.
• At least one of said luminescent compounds has an emission according to an anti-Stokes mechanism.
• the device integrates a source of CO2.
• the device further comprises an incident solar energy concentrator.
• the one or more luminescent compounds have wavelengths of absorption or emission that promote the photosynthesis of algae.
• the invention relates to a method of manufacturing an algae culture device according to the first aspect comprising: prior exposure of said algae to cultivation at different wavelengths;
• said biological parameter of interest is the growth rate of the algae.
• the biological parameter of interest is the production of oil by the algae.
• said biological parameter of interest is the production of a pigment given by the algae.
• the invention relates to a method for cultivating algae in natural light, comprising culturing the algae in an enclosure with a culture medium, characterized in that it further comprises the photoconversion of the solar light by means of a substrate comprising at least one luminescent compound for reemitting a radiation whose spectrum is adapted to the photosynthesis wavelengths of said algae.
• the method according to the invention leads to an increase of 20% to 100% or more in both theoretical and practical maxima, for a limited increase in capital and operational costs, and thus provides a major economic advantage.
• This technique is compatible with many and can be combined with other more conventional improvements proposed by various authors, including the use of selected or genetically modified algae strains.
• FIG. 2 a luminescent substrate integrated into the walls of a photobioreactor
• the invention relates to methods and devices for modifying sunlight to overcome the limit of PAR (photosynthetic active radiation) as presented above.
• the applicant has shown that it is precisely by increasing the PAR value beyond the 45.8% conventionally accepted by all authors, which is not suggested in the literature, that the method according to the invention makes it possible, with a very broad generality and adaptability and for a limited associated cost, to improve the yield of algae cultures beyond the maximum currently admitted by specialists.
• the invention is applicable to a large number of algae culture systems including photobioreactors and simpler basin type systems.
• the notion of PAR is a simplification because some wavelengths of PAR - such as green (range around 550 nm) for green algae - do not lead to efficient photosynthesis.
• the PAR is therefore an increase in the fraction of solar radiation that is actually converted.
• the method according to the invention makes it possible to modify the sunlight in order to limit these wavelengths of the PAR which are not very efficient in favor of more efficient wavelengths.
• the Applicant has shown that by spectrally modifying fine sunlight incidentally to adapt it to the photosynthetic needs of the algae or cultivated algae, the process according to the invention provides an original and quantitatively important improvement applicable to almost -total methods of cultivation of algae from sunlight described or envisaged in the literature.
• the invention relates, in its most general sense to a device for cultivating algae in natural light, comprising an enclosure with a culture medium and the algae to be cultivated, characterized in that the device further comprises a substrate arranged to receiving the solar radiation for photoconversion of said solar radiation, the substrate comprising at least one luminescent compound for reemitting a radiation whose spectrum is adapted to the optimization of a biological parameter of interest resulting from the photosynthesis of said algae .
• the luminescent compounds used are preferably fluorescent organic molecules and more preferentially "dye laser” type molecules. Molecules with high quantum yield are chosen
• the luminescent compounds used may include rare earths such as terbium or europium salts.
• the luminescent compounds used may include inorganic compounds of the quantum dots type.
• the luminescent compounds used can be selected from the following groups of compounds:
• Rhodamine 110 (Lambdachrome)
• a combination of several luminescent substances which comprises a substance of group A such as PPO or two substances of group A such as PPO and OB, and a substance of each of the following 1 to 3 groups (B, C, D, E or F).
• concentrations used preferably vary between 0.1 and 1000 ppm, preferably between 1 and 100 ppm.
• a serial emission-absorption system is used where the maximum emission wavelength of a substance corresponds to the maximum absorption wavelength of the next substance in the series.
• FRET fluorescence resonance energy transfer
• a group A substance, a group B substance and a group C substance are used.
• the concentrations employed decrease as the emission wavelength increases. This rule of nontrivial concentration is inspired by what is observed in phycobillisomes and it limits the phenomena of self-absorption.
• the substrate in which the phosphors are incorporated is a plastic, for example an acrylic plastic such as polymethylmethacrylate (PMMA) or else ethylene vinyl acetate (EVA), Apoliah (Arkema), polyvinylidene fluoride (PVDF) , polyethylene (PE), polycarbonate (PC).
• PMMA polymethylmethacrylate
• EVA ethylene vinyl acetate
• PVDF polyvinylidene fluoride
• PE polyethylene
• PC polycarbonate
• the luminescent substances are integrated in a resin or varnish that is spread on plates or glass tubes.
• particles of TiO 2 and / or aluminum oxide at 0.1-0.5% by weight are incorporated in the substrate, which act as light diffusers and UV reflectors.
• the luminescent substances are incorporated into millimetric beads of polystyrene, PMMA or another polymer, which are suspended in the culture medium.
• these beads also make it possible to clean the walls of the container containing the culture and to prevent their fouling.
• the beads thus constitute an internal light source in the culture medium and make it possible to remove a limitation of the devices of the prior art concerning the distribution of light within the culture volume.
• said millimeter balls are rendered phosphorescent by the use of so-called substances with a long remanence (> 10 ⁇ s).
• the phosphorescent beads which circulate in the culture medium, have been previously illuminated either by sunlight or by a UV flash or other source of high energy monochromatic artificial light. Such beads can be used day and night. They may comprise ZnS crystals with 10-1000 ppm of copper or silver dopants.
• one of the luminescent substances used is chosen by those skilled in the art so as to convert, by an anti-Stokes type mechanism, a portion of the infrared radiation (700-2000 nm) into visible radiation, preferably in red radiation (600-700 nm). Additional advantages provided by the process according to the invention are described in a nonlimiting manner below:
• luminescent substances according to the invention leads to a re-emission of solar light incident in all directions of space.
• a given phosphor re-emits the incident light anisotropically (with a "donut-like" distribution) but the orientation of the substance within the light-transforming material, which is itself random, leads to a re-emission.
• statistically isotropic at 4pi steradians This makes it possible to transform the incident direct sunlight into a diffuse light.
• the diffuse light thus obtained is favorable to the growth of algae because it limits photoinhibition phenomena.
• the method according to the invention promotes the growth of algae both when they are illuminated by direct sunlight and when the light that illuminates them is itself diffused or remanent, which is the case when the weather conditions include the presence of clouds and / or water vapor.
• the method according to the invention absorbs a part of the UV light of the sun (260-400 nm) which is re-emitted in visible wavelengths (400 nm and more). This makes it possible to limit the exposure of the algae to the UV which is known to those skilled in the art that it can limit the growth of said algae, or even, in certain cases, lead to mutations that can render genetically inhomogeneous and ultimately destabilize the species. cultivated. Thermal benefits
• the modification of the sunlight produced by the process according to the invention leads to an advantageous modification of the temperature profile to which the algal cultures are subjected.
• the effect depends on the chosen culture device (photobioreactor, greenhouse, bag or open basin) but it combines, with a greater or lesser intensity, on the one hand a decrease in the average value and the maximum of the temperature during the day and on the other hand an increase of the average value and the minimum of the temperature at night.
• These two thermal effects increase the average productivity of algae crops. They also make it possible to limit the occurrence of cases of unfavorable extreme temperatures which can lead to the extinction of algal cultures having been exposed to abnormally hot or abnormally cold temperatures.
• the method according to the invention makes it possible to modify the sunlight to adapt it to the needs of the various species of algae.
• green algae have a photosynthetic device that makes photosynthesis particularly effective in the presence of blue illumination (440 nm) and red (680 nm).
• the method according to the invention makes it possible, by using a combination of appropriate luminescent substances, to modify natural sunlight whose spectrum has a single maximum around 550 nm to obtain a light whose spectrum has two maxima, the first towards 440 nm and the second around 680 nm.
• a group A substance, a group B substance and a group D substance are used.
• red algae do not use blue light and red light well but reproduce with maximum efficiency when lit by green light, around 560 nm.
• the method according to the invention makes it possible, by using a combination of appropriate phosphors, to modify the solar spectrum to increase the intensity of its peak towards 550 nm and to decrease the intensity of one or more other regions of the spectrum.
• a group A substance, a group B substance and a group C substance are used.
• cyanobacteria do not have chloroplasts.
• These prokaryotic cells have a photosynthetic apparatus and a particular pigment equipment, which leads them to grow optimally when they are illuminated by a light enriched in wavelengths between 580 and 650 nm.
• the method according to the invention makes it possible to enrich the sunlight between 580 and 650 nm by converting wavelengths less than 580 nm (and / or wavelengths greater than 650 nm).
• a group A substance, a group B substance, a group E substance and a group F substance are used.
• Step 0 - We choose a species of alga given, isolated from a natural habitat or genetically modified, that we wish to cultivate.
• Step 1 - A broad-spectrum white light source and a monochromator or a light filtered by various interference filters and adjusted in power so that each color transmitted by the filter has the same intensity, or preferably, is used.
• colored light source for example blue, green, red LEDs, etc. of identical intensity.
• the algal culture is thus subjected to different wavelength ranges.
• Step 2 For each illumination condition, the photosynthetic activity of the algae is measured (for example by continuously measuring the oxygen produced) and / or the carbon dioxide consumed) and a spectrum of average productivity is deduced. (g / L / day) depending on the wavelength of incident light.
• Step 3 A combination of luminescent substances is selected in order to modify the sunlight, whose spectrum is otherwise easy to establish, to concentrate it in the wavelengths that have just been empirically observed to lead to a high productivity. maximum algal culture.
• Step 4 Luminescent substances whose composition has just been determined in step 3 are incorporated into a masterbatch.
• Step 5 Mixing said masterbatch, a monomer, and any additives known to those skilled in the art to make a plastic material.
• Step 6 Extruding said doped plastic material and thus producing plates and tubes or films from which a photobioreactor or cover element is constructed which accelerates the growth of the algae selected in step 0.
• Algae can allow the production of various compounds of interest such as pigments.
• One can empirically, by producing controlled algae cultures for various wavelength ranges (for example with twelve LEDs of the same power illuminating in ten wavelength ranges between 350 and 950 nm: 350-400 nm, 400 -450nm, 450-500nm etc.), identify an optimal wavelength interval that leads to obtaining a larger amount of the compound of interest.
• the phosphor compositions used in the process according to the invention are then adapted to modify the sunlight to concentrate it in the optimum range.
• Some species of algae may contain a significant amount of oil, up to 50% or more of their dry weight.
• the conditions that allow algae to grow at a maximum speed are different from the conditions that allow each cell to accumulate a large amount of lipids.
• the culture conditions of a culture of algae which grows rapidly are modified so as to favor, in a second time, the accumulation of lipids.
• Various types of stress are possible, including nitrogen deprivation stress.
• the method according to the invention offers the possibility of using a modification of the light to generate a stress favoring the production of lipids.
• the nature of the Soil spectrum adapted to the generation of said stress can be determined in the laboratory by an analysis of the response (lipid content per gram of dry matter) of a culture of the alga of interest exposed at different wavelengths. artificial light. Then, at the appropriate time of production, a large scale cultivation of the algae of interest can be subjected to sunlight modified by the process according to the invention in order to obtain a theoretical composition that is close to the Soil spectrum. previously determined ideal. This allows said culture to accumulate a large amount of lipids.
• the method according to the invention is applicable to hybrid culture systems combining the advantages of photobioreactors (controlled environment, high productivity) and ponds (lower cost).
• photobioreactors controlled environment, high productivity
• ponds low cost
• Different portions of the reactor may comprise differently doped materials.
• Example 1 Tubular Photobioreactors
• the algae are grown in a device comprising a network of plastic tubes of diameter between 5 and 20 cm and a total length of up to several km.
• a sectional view of a portion of the device is shown schematically in Figure 2.
• Algae 20 are cultured in a tubular photobioreactor illuminated by natural light 30.
• the wall 15 of the photobioreactor receives sunlight and is composed of a material which comprises at least a luminescent compound therein for retransmitting radiation whose spectrum is adapted to algae.
• the device incorporates pumps and a concentrated carbon dioxide injection system.
• the plastic of the tubes is doped, before extrusion, by a combination of luminescent substances chosen so as to modify the sunlight according to the physiological requirements of the seaweed species considered, as previously determined experimentally.
• PMMA Polymethylmethacrylate
• acrylic plastic that offers excellent optical properties and allows for good integration of luminescent substances and can be used for the manufacture of tubes.
• a PMMA thickness of between 1 and 5 mm is used.
• green algae the following formula is used, for example, for 3 mm PMMA plates, for 1 kg of MMA:
• Example 2 - Bags A cheaper alternative for growing seaweed is using bags. These bags can be placed outdoors, in a closed area or float on the sea (semi-permeable bags are preferably used which can let out water and exchange nutrients with seawater but retain the seaweeds) .
• the plastic component of the bags may be a polyethylene-ethylene vinyl acetate polymer (PE-EVA), Apoliah (Arkema) or PMMA.
• the thickness of the bags is between 100 ⁇ m and 500 ⁇ m.
• the plastic is doped, before extrusion, by a combination of luminescent substances chosen so as to modify the sunlight according to the physiological needs of the seaweed species considered, as previously determined experimentally.
• the entire algae culture device which may include tubes, vats or panels, is integrated within an enclosed or semi-enclosed "shelter” type structure that plays a positive role in terms of thermal regulation. , bright, protection against parasites or predators and bad weather.
• the walls of the greenhouse are composed of glass whose inner face has been coated with a resin doped with a combination of luminescent substances chosen so as to modify the sunlight according to the physiological needs of the seaweed species considered, such as as previously determined experimentally.
• the coated glass can be replaced by doped PMMA plates.
• the algae are cultivated in an open-air device of the hypodrome basin type ("raceway") or trench type ("trough").
• a sectional view of a part of such a device is shown in FIG. 1.
• a trench 11 acting as a chamber illuminated by natural light 30 in which algae 20 are suspended in an aqueous medium of upper surface 12 is hollowed out.
• solar light is received and modified by a flexible film or a rigid plate 40 which comprises at least one luminescent compound for reemitting a radiation whose spectrum is suitable for algae.
• the basin elements or the trenches are covered by the flexible or slightly rigid film, for example plastic doped by a combination of luminescent substances chosen so as to modify the sunlight according to the physiological needs of the seaweed species considered, as previously determined experimentally.
• the film can be composed of PMMA, PE-EVA or PVDF.
• Figure 3 shows a sectional view of a portion of a device according to a variant.
• a closed system of tubular photobioreactor type 16 is illuminated by natural light to allow the cultivation of algae 20.
• particles 45 containing at least one luminescent compound which emits radiation whose spectrum is suitable for algae.
• the particles are for example formed of balls having a diameter of between 1 mm and 5 mm.
• These beads may be beads conventionally used to clean the wall of the tubes and prevent the formation of a film of algae adhering to the surface of the tubes and which would eventually prevent the penetration of light.
• a new function is given to said beads, which are doped with luminescent substances of short and / or long remanence (phosphorescent substances). allowing to work also in all darkness.
• the beads which are constantly agitated by the movement imposed on the fluid of the photobioreactor, realize, within the culture medium, a spectral adaptation and they constitute a source of diffuse internal light that promotes the growth of algae throughout the aqueous volume .
• the beads can be made from polymethylmethacrylate (PMMA), polypropylene, nylon, PVC or polyvinylacetate. Polystyrene beads or polymethylmethacrylate (PMMA) beads may be used. A porophore agent can be used to decrease the density of the beads while creating trapping vacuoles or photon concentration.
• PMMA polymethylmethacrylate
• PMMA polymethylmethacrylate
• a porophore agent can be used to decrease the density of the beads while creating trapping vacuoles or photon concentration.
• the detection device and the method according to the invention comprises various variants, modifications and improvements which will be obvious to those skilled in the art, it being understood that these various variants, modifications and improvements are within the scope of the invention, as defined by the following claims.

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• 2010
  • 2010-01-26 US US13/146,025 patent/US20110281295A1/en not_active Abandoned
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