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WO2010019800A1 - Appareil de délivrance de lumière thérapeutique, procédé et système - Google Patents

Appareil de délivrance de lumière thérapeutique, procédé et système Download PDF

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Publication number
WO2010019800A1
WO2010019800A1 PCT/US2009/053752 US2009053752W WO2010019800A1 WO 2010019800 A1 WO2010019800 A1 WO 2010019800A1 US 2009053752 W US2009053752 W US 2009053752W WO 2010019800 A1 WO2010019800 A1 WO 2010019800A1
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WO
WIPO (PCT)
Prior art keywords
optical
optical element
light
optical fiber
fiber
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/US2009/053752
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English (en)
Inventor
Eric Bornstein
Eric Sinofsky
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
NOMIR MEDICAL TECHNOLOGIES Inc
Original Assignee
NOMIR MEDICAL TECHNOLOGIES Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by NOMIR MEDICAL TECHNOLOGIES Inc filed Critical NOMIR MEDICAL TECHNOLOGIES Inc
Publication of WO2010019800A1 publication Critical patent/WO2010019800A1/fr
Anticipated expiration legal-status Critical
Priority to US13/474,320 priority Critical patent/US8983257B2/en
Ceased legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B6/00—Light guides; Structural details of arrangements comprising light guides and other optical elements, e.g. couplings
    • G02B6/24—Coupling light guides
    • G02B6/26—Optical coupling means
    • G02B6/262—Optical details of coupling light into, or out of, or between fibre ends, e.g. special fibre end shapes or associated optical elements
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N5/0601—Apparatus for use inside the body
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N5/0613—Apparatus adapted for a specific treatment
    • A61N5/0624—Apparatus adapted for a specific treatment for eliminating microbes, germs, bacteria on or in the body
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
    • A61B18/00—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body
    • A61B18/18—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body by applying electromagnetic radiation, e.g. microwaves
    • A61B18/20—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body by applying electromagnetic radiation, e.g. microwaves using laser
    • A61B18/22—Surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body by applying electromagnetic radiation, e.g. microwaves using laser the beam being directed along or through a flexible conduit, e.g. an optical fibre; Couplings or hand-pieces therefor
    • A61B2018/2255—Optical elements at the distal end of probe tips
    • A61B2018/2261—Optical elements at the distal end of probe tips with scattering, diffusion or dispersion of light
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N2005/063—Radiation therapy using light comprising light transmitting means, e.g. optical fibres
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61N—ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY
    • A61N5/00—Radiation therapy
    • A61N5/06—Radiation therapy using light
    • A61N2005/0658—Radiation therapy using light characterised by the wavelength of light used
    • A61N2005/0659—Radiation therapy using light characterised by the wavelength of light used infrared
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B6/00—Light guides; Structural details of arrangements comprising light guides and other optical elements, e.g. couplings
    • G02B6/24—Coupling light guides
    • G02B6/26—Optical coupling means
    • G02B6/32—Optical coupling means having lens focusing means positioned between opposed fibre ends

Definitions

  • the present invention generally relates to methods and systems for generating infrared optical radiation in selected energies and dosimetries that will modify the bioenergetic steady-state trans-membrane and mitochondrial potentials of irradiated cells through a depolarization effect, and more particularly, relates to methods and systems for membrane depolarization to potentiate antibiotic compounds in bacterial cells, and particularly antibiotic resistant bacterial cells.
  • This invention also relates generally to phototherapy and, in particular, instruments employing optical fibers or other flexible waveguides to deliver radiation to a targeted biological site.
  • antibiotics not only prompts generation of resistant bacteria; such as, for example, methicillin-resistant staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE); but also creates favorable conditions for infection with the fungal organisms (mycosis), such as, Candida.
  • MRSA methicillin-resistant staphylococcus aureus
  • VRE vancomycin-resistant enterococci
  • mycosis fungal organisms
  • therapies for bacterial infections include administration of antibacterial therapeutics or, in some instances, application of surgical debridement of the infected area. Because antibacterial therapies alone are rarely curative, especially in view of newly emergent drug resistant pathogens and the extreme morbidity of highly disfiguring surgical therapies, it has been imperative to develop new strategies to treat or prevent microbial infections.
  • fiber optic phototherapy is an increasingly popular modality for the diagnosis and/or treatment of a wide variety of diseases.
  • infrared laser radiation will often be delivered to a surgical site via a hand-held instrument incorporating an optically transmissive fiber in order to coagulate blood or cauterize tissue.
  • Other uses for optical fiber-delivered radiation include treatment of atherosclerotic disease and prostatic disease.
  • U.S. Patent No. 4,878,492 issued to Sinofsky et al., incorporated herein by reference discloses the use of infrared radiation to heat blood vessel walls during balloon angioplasty in order to fuse the endothelial lining of the blood vessel and seal the surface.
  • Fiber optic delivery systems have been incorporated in endoscopic or catheter- based instruments to deliver radiation to a targeted biological site within a body lumen or cavity.
  • the fiber optic phototherapy device is inserted through an instrument lumen or catheter for delivery in-vivo.
  • Conventional optical fiber phototherapy devices can include an optical element, such as a focusing lens, that is coupled to the optical fiber by a cylindrical housing.
  • the housing is typically a metallic band or cuff, constructed from stainless steel or gold that is sized to hold both the lens and the optical fiber.
  • the housing can be glued to the optical fiber or can be threaded to facilitate connection to the fiber.
  • Flouropolymer housings in the prior art are thermally fused to the buffer of the fiber.
  • Baxter et al, in US 6, 102,905 discloses an optical system that is held together by thermoforming the cuff onto a fiber with the identical material as the fiber buffer.
  • This technique although effective, requires a complex thermoforming machine, and can damage the system's optical elements by exposing them to the 500 degrees C it takes to melt the fluoro-polymers together. This temperature exceeds the recommended temperature for both the optical fiber cladding and the grin lens. This technique can also not be used when the fiber and cuff are not similar materials.
  • optical fiber phototherapy devices have proven less than optimal, it is an object of the present invention to provide improved phototherapy devices provide a precise, stable, controlled illumination of multiple wavelengths.
  • a further object of the present invention is to provide phototherapy devices that inhibit the effects of heat cycling.
  • a further object of the present invention is to provide phototherapy devices that are simple and inexpensive to manufacture.
  • Another object of the present invention is to provide an improved method of making a phototherapy device.
  • the phototherapy devices of the present invention which include an integrating optical fiber, having an optical element which is optically coupled to the fiber disposed at its distal end, and a precisely defined, elastic housing physically coupling the optical fiber buffer to the optical element.
  • the phototherapy device of the present invention may further include two or more source optical fibers having a diameter less than that of the integrating fiber optically coupled to the proximal end of the integrating fiber.
  • the housing is made from a material having a coefficient of thermal expansion approximately equal to the coefficient of thermal expansion of the buffer. In this manner, both the housing and the buffer will thermally expand (and contract) approximately the same amount, thus minimizing the effects of heat cycling on the device.
  • the housing is made from a polymer material having a anisotropic, non-linear Young's modulus with the greater value co-axial with the optical fiber.
  • the housing is made with a low index of refraction material to act as a cladding to the encased optical component.
  • multiple source optical fibers having a combined packed cross-section less than that of the integrating optical fiber may be couple the integrating optical fiber.
  • An embodiment of a methed for making a phototherapy device includes the steps of providing an optical fiber with a perfluorinated polymer buffer coating and attaching an optical element to a distal end of the optical fiber.
  • the optical element can be attached to the distal end of the fiber by encasing the optical element in a housing and press fitting the housing to at least a portion of the buffer.
  • the housing is also press fitted to the buffer.
  • the step of attaching the optical element can further include the step of employing a precision, press-fit template to facilitate the stable assembly of the elements.
  • the present invention provides an apparatus, systems and methods for microbial reduction using optical energy.
  • Specific near infrared wavelength ranges photodamage cell membranes, causing oxidative stress and membrane depolarization.
  • Bacteria in the field of the optical beam are photodamaged in that ATP production is compromised, efflux pumps are inhibited, cell wall biosynthesis is disrupted, and the bacteria display increased sensitivity to antibiotics.
  • optical photodamage can reverse a drug resistance phenotype, permitting the (re)use of common antibiotics against even multiple drug resistance (MDR) strains.
  • MDR drug resistance
  • the invention includes a method of effectuating antimicrobial activity at a microbial colonization site in a subject, by applying a redox modifying and membrane depolarizing dosage of near infrared energy to the site, the near infrared energy having a first wavelength of about 870 nm and a second wavelength of 930 run, the dosage of near infrared energy being insufficient to cause thermolysis of subject tissues at the site; and applying one or more antimicrobial agents to the microbial colonization site, wherein at least a two-fold log reduction in microbial colonization is observed in the subject at the colonization site.
  • the invention provides a method of inhibiting bacterial viability at a microbial colonization site in a subject, by applying a peptidoglycan biosynthesis inhibiting dosage of near infrared energy to the site, the near infrared energy having a first wavelength of about 870 nm and a second wavelength of 930 nm, the dosage of near infrared energy being insufficient to cause thermolysis of subject tissues at the site; and applying one or more antimicrobial agents to the microbial colonization site wherin at least one of the antimicrobial agents binds the active site of a bacterial transpeptidase enzyme, wherein at least a two-fold log reduction in microbial colonization is observed in the subject at the colonization site.
  • the invention includes a method of inhibiting microbial viability at a microbial colonization site in a subject, comprising: a) applying a DNA replication and transcription inhibiting dosage of near infrared energy to the site, the near infrared energy having a first wavelength of about 870 nm and a second wavelength of 930 nm, the dosage of near infrared energy being insufficient to cause thermolysis of subject tissues at the site; and b) applying one or more antimicrobial agents to the microbial colonization site, wherein at least a two-fold log reduction in microbial colonization is observed in the subject at the colonization site.
  • the invention provides a method of reducing the number and viability of microbes at a microbial colonization site in a subject, comprising: a) applying a bacterial phospholipid biosynthesis inhibiting dosage of near infrared energy to the site, the near infrared energy having a first wavelength of about 870 ran and a second wavelength of 930 nm, the dosage of near infrared energy being insufficient to cause thermolysis of subject tissues at the site; and b) applying one or more antimicrobial agents to the microbial colonization site, wherein at least a two-fold log reduction in microbial colonization is observed in the subject at the colonization site.
  • the invention provides a method of decontaminating an area of a subject, comprising: a) identifying in or on a subject, a wound or infection site or a surgical location in need of a reduction in bacterial colonization; b) applying one or more photodamaging doses of optical radiation to the area without thermally damaging the area; c) applying an antimicrobial agent to the area.
  • diffuser tip assembly adapted to receive the distal end of optical fiber.
  • the assembly includes a reflective cavity including: a first reflector positioned proximal the distal end of the received fiber and including an aperture adapted to admit light emitted from the fiber into the cavity; and a second reflector positioned distal the first reflector.
  • the assembly also includes a diffuser tube positioned between the first and second reflectors about a cavity axis extending from the first reflector to the second reflector, the diffuser tube including an inner void surrounded by an outer portion including a diffusive scattering material.
  • the cavity and diffuser tube are arranged such that at least a portion of light admitted into the cavity is scattered by the diffusive scattering material out of the tip assembly through the outer portion in a direction transverse to the cavity axis.
  • the cavity and diffuser tube are configured such that light admitted into the cavity is directed from the aperture towards the second reflector; a portion of the light directed towards the second reflector impinges upon the diffusive scattering material and is scattered out of the tip assembly in a direction transverse to the axis; at least a portion of unscattered light impinges upon the second reflector and is reflected back towards the first reflector; and a portion of the light directed back towards the first reflector impinges upon the diffusive scattering material and is scattered out of the tip assembly in a direction transverse to the axis.
  • the cavity and diffuser tube are configured such that light admitted into the cavity travels multiple passes between the first and second reflectors; and on each pass, at least a portion of the light is scattered by the diffusive scattering material out of the tip assembly in a direction transverse to the axis.
  • the light scattered out of the tip assembly on each pass combine to produce a cumulative illumination pattern.
  • the cumulative illumination pattern is characterized by substantially uniform axial intensity profile along at least a portion of the diffuser tube.
  • the cumulative illumination pattern is characterized by a substantially uniform azimuthal illumination profile.
  • the cumulative illumination pattern is characterized by substantially proscribed illumination in the direction parallel to the axis.
  • the cumulative illumination pattern is a substantially uniform cylindrical illumination pattern emitted radially from the outer surface of the diffuser tube. In some embodiments, the cumulative illumination pattern is determined by at least on at least one chosen from the list consisting of: a length of the diffuser tube, the diameter of the inner void of the diffuser tube, a numerical aperture associate with the aperture in the first reflector. In some embodiments, the inner void is filled with a substantially transparent non-scattering material.
  • the at least one of the first and second reflectors includes a curved reflector.
  • the first reflector is a diffuse reflector and the second reflector is a specular reflector.
  • the ratio of the distance between the first reflector and the second reflector along the cavity axis to the outer diameter of the diffusion tube is about 10 or less, about 1 or less, or even about 0.1 or less.
  • diffusive scattering material includes a plastic, a glass, a polymer, or a fluid. In some embodiments, the diffusive scattering material includes PTFE. In some embodiments, the diffusive scattering material is adapted to scatter light in the near infrared.
  • the reflective cavity and the diffuser tube are autoclavable. Some embodiments include substantially transparent outer jacket adapted to contain the reflective cavity and the diffuser tube. In some embodiments, the outer jacket is detachable from the reflective cavity and the diffuser tube. 22. The diffuser tip assembly of any of the preceding claims, where the tip assembly is adapted to scatter about 80% or more of the light delivered from the fiber while absorbing about 20% or less of the light delivered from the fiber.
  • kits for treating an antimicrobial resistant biological contaminate at a treatment site which includes: a diffuser tip adapted to receive near infrared therapeutic light from a light delivery system and diffuse the light to illuminate at least a portion of the treatment site; a quantity of an antimicrobial agent; instructions to use the antimicrobial agent in conjunction with the therapeutic light to potentiate the antimicrobial agent to treat the biological contaminate; and suitable packaging.
  • the diffuser tip is a diffuser tip assembly of the any of the types described herein.
  • the therapeutic light includes optical radiation substantially in a first wavelength range from about 865 nm to about 875 ran or a second wavelength range having a wavelength from about 925 nm to about 935 nm, or both wavelength ranges.
  • the diffuser tip assembly is adapted to provide substantially uniform illumination of the illuminated portion of the treatment site. In some embodiments, the diffuser tip assembly is adapted to illuminate the illuminated portion of the treatment site at a power density and an energy density which potentiates the antimicrobial agent at the treatment site without causing substantial photothermal or photomechanical damage to the treatment site. In some embodiments, the diffuser tip assembly is adapted to illuminate the illuminated portion of the treatment site at a power density of about 0.2 W/cm 2 to about 1 W/cm 2 and an energy density from about 100 J/cm 2 to about 400 J/cm 2 at the illuminated target region. In some embodiments, the diffuser tip is adapted to operate for about 30 seconds or more at an operating temperature of HO 0 F or less.
  • the diffuser tip assembly is adapted for detachable connection to a distal end of an optical fiber which transmits the therapeutic light from a light source to the distal end of the fiber.
  • the quantity of antimicrobial agent includes a topical paste.
  • the antimicrobial agent includes an antibiotic or a pharmacologically acceptable salt thereof, selected from the group consisting of: /3-lactams, glycopeptides, cyclic polypeptides, macrolides, ketolides, anilinouracils, lincosamides, chloramphenicols, tetracyclines, aminoglycosides, bacitracins, cefazolins, cephalosporins, mupirocins, nitroimidazoles, quinolones and fluoroquinolones, novobiocins, polymixins, cationic detergent antibiotics, oxazolidinones or other heterocyclic organic compounds, glycylcyclines, lipopeptides, cyclic lipopeptides, pleuromutilins, and gramicidins, daptomycins, linezolids, ans
  • the biological contaminate includes an aberrant microbial colonization
  • the instructions include instruction to use the antimicrobial agent in conjunction with the therapeutic light to potentiate the antimicrobial agent to reduce the level of colonization at the treatment site.
  • the an aberrant microbial colonization prior to application of the treatment light, has a drug resistant phenotype with respect to the antimicrobial agent.
  • the diffuser tip is sterilized.
  • q therapeutic system for treatment of a biological contaminant at a treatment site including:an optical radiation generation device configured and arranged to generate near infrared therapeutic light; a controller operatively connected to the optical radiation generation device for controlling dosage of the therapeutic light transmitted to the treatment site at a dosimetry sufficient to produce photodamage in the biological contaminant without causing substantial photothermal or photomechanical damage to biological tissue at the treatment site; a delivery assembly including an optical fiber which directs the therapeutic light to be transmitted to the treatment site; and a diffuser tip adapted to receive the therapeutic light from the delivery assembly and diffuse the therapeutic light to illuminate at least a portion of the treatment site with a prescribed illumination pattern.
  • the diffuser tip is the diffuser tip of any of the types described herein.
  • the therapeutic light includes optical radiation substantially in a first wavelength range from about 865 nm to about 875 nm or a second wavelength range having a wavelength from about 925 nm to about 935 nm, or both wavelength ranges.
  • the diffuser tip assembly is adapted to provide substantially uniform illumination of at least a portion of the illuminated portion of the treatment site. In some embodiments, the diffuser tip assembly is adapted to illuminate the portion of the treatment site at a power density and an energy density which potentiates an antimicrobial application at the treatment site.
  • the diffuser tip assembly is adapted to illuminate the portion of the treatment site at a power density of about 0.3 W/cm 2 to about .7 W/cm 2 and an energy density from about 100 J/cm 2 to about 400 J/cm 2 at the illuminated portion of the treatment site.
  • the dif ⁇ iser tip is adapted to operate for about 30 seconds or more at an operating temperature of 110° F or less.
  • the diffuser tip assembly is adapted for detachable connection to a distal end of the optical fiber.
  • a method treatment of a biological contaminant at a treatment site includes: generating near infrared therapeutic light; controlling dosage of the therapeutic light transmitted to the treatment site at a dosimetry sufficient to produce photodamage in the biological contaminant without causing substantial photothermal or photomechanical damage to biological tissue at the treatment site; directing the therapeutic light to be transmitted to the treatment site; and using a diffuser tip, diffusing the therapeutic light to illuminate at least a portion of the treatment site with a prescribed illumination pattern.
  • the diffuser tip is a diffuser tip assembly of any of the types described herein.
  • the therapeutic light includes optical radiation substantially in a first wavelength range from about 865 nm to about 875 nm or a second wavelength range having a wavelength from about 925 nm to about 935 nm, or both wavelength ranges.
  • the diffusing the therapeutic light includes providing substantially uniform illumination of at least a portion of the illuminated portion of the treatment site.
  • Some embodiment further rinclude applying a quantity of antimicrobial agent the treatment site; and illuminating the portion of the treatment site at a power density and an energy density which potentiates the antimicrobial application.
  • the antimicrobial agent is ineffective for treating the antimicrobial resistant biological contaminate in the absence of the therapeutic light. In some embodiments, the antimicrobial agent is ineffective for treating the biological contaminate in the absence of the therapeutic light.
  • the biological contaminate includes an aberrant microbial colonization, and further including includes using the antimicrobial agent in conjunction with the therapeutic light to potentiate the antimicrobial agent to reduce the level of colonization at the treatment site. In some embodiments, the aberrant microbial colonization, prior to application of the treatment light, has a drug resistant phenotype with respect to the antimicrobial agent.
  • controlling the dosage of the therapeutic light includes controlling the illumination of the portion of the treatment site at a power density of about 0.2 W/cm 2 to about 1 W/cm 2 and an energy density from about 100 J/cm 2 to about 400 J/cm 2 at the illuminated region.
  • antimicrobial agents that are appropriate for use in conjunction with optical photodamage to reduce bacterial counts include common antibiotics and pharmacologically acceptable salt thereof, including /3-lactams, glycopeptides, cyclic polypeptides, macrolides, ketolides, anilinouracils, lincosamides, chloramphenicols, tetracyclines, aminoglycosides, bacitracins, cefazolins, cephalosporins, mupirocins, nitroimidazoles, quinolones and fluoroquinolones, novobiocins, polymixins, cationic detergent antibiotics, oxazolidinones or other heterocyclic organic compounds, glycylcyclines, lipopeptides, cyclic lipopeptides, pleuromutilins, and gramicidins, daptomycins, linezolids, ansamycins, carbacephems, carbapenem
  • an optical delivery apparatus including: an optical fiber extending between a distal end having a distal end face and a proximal end having a proximal end face, the fiber configured to receive light from at least one source at the proximal end face, transmit the light from the proximal end to the distal end, and emit the light from the distal end face; an optical element positioned to receive the light emitted from the distal end face and direct the light to an illumination region; and a non-metallic housing containing the optical fiber and the optical element and maintaining the relative position of the optical fiber and the optical element.
  • the non-metallic housing is an elastic housing.
  • the elastic housing is an elastic cuff stretched over the optical fiber and optical element. The compressive force from the elastic cuff maintains the relative position of the optical fiber and the optical element.
  • the elastic cuff is tubular elastic member having a resting inner diameter lass that that of the optical fiber and optical element.
  • the non-metallic housing includes non-metallic clamp member maintaining the relative position of the optical fiber and the optical element.
  • the index of refraction is less than 1.7, less than 1.6, less than 1.5, less than 1.4, less than 1.3, less than 1.2, less than 1.1, less than 1, or even less.
  • the optical fiber includes a outer buffer layer, a cladding and a core, the buffer layer is disposed about a cladding, and the cladding is disposed about the core; the coefficient of thermal expansion of the buffer layer is substantially matched to the coefficient of thermal expansion of the elastic housing.
  • the buffer layer includes a polymer, e.g., a perflourinated polymer.
  • the cladding and core extend beyond the buffer layer at the distal end of the optical fiber.
  • the optical element is proximal to the distal end of the fiber. In some embodiments, the optical element is spaced apart from the distal end of the fiber, and the housing maintains the spacing of the optical element from the distal end of the fiber. In some embodiments, the optical element abuts the distal end of the fiber.
  • the optical element includes at least one selected from the list consisting of: a lens, a GRIN lens, a diffractive element, a diffusive element, a hologram, a concentrating element, and a collimator.
  • the optical element directs the light to illuminate the illumination region with substantially uniform illumination.
  • the optical element forms a beam of light from the light from the distal end of the fiber, where the beam has a substantially non-gaussian beam profile. In some embodiments, the beam has a substantially uniform beam profile. In some embodiments, beam profile varies by less than 5% , less than 1%, or even less across the beam profile.
  • the at least one source includes at a first source and a second source, and where In some embodiments, the fiber is configured to receive light from the first and the second source at the proximal end face, transmit the light from the proximal end to the distal end, and emit the light from the distal end face. In some embodiments, the optical element directs the light from the first and second to illuminate the illumination region such that light from the first and second source overlaps at at least a portion of the illumination region. In some embodiments, the first and second light sources produce light having differing wavelengths.
  • Some embodiments include a first source fiber and a second source each having a proximal end located proximal a respective on of the first source and the second source, and each having a distal end located proximal to the proximal end of the optical fiber.
  • the first source fiber transmits light from the first source to the proximal end of the optical fiber and the second source fiber transmits light from the second source to the proximal end of the optical fiber.
  • the first and second source fibers have a combined diameter 9e.g. a combined packed cross sectional diameter) at their respective distal ends less than that of the optical fiber at its proximal end.
  • the first and second source fibers have a combined diameter at their respective distal ends less than that of the optical fiber at its proximal end.
  • the at least one source includes at least one selected from the list consisting of: a laser, a diode laser, a solid state laser, a dye laser, an LED, an OLED, and a lamp.
  • the optical element images the core of the optical fiber at the distal end face to the illumination region.
  • the optical element has a focal plane
  • the non-metallic housing maintains the relative position of the optical fiber and the optical element such that the core of the optical fiber at the distal end face is positioned near the focal plane
  • a therapeutic system for treatment of a biological contaminant at a treatment site including: an optical radiation generation device configured and arranged to generate near infrared therapeutic light; a controller operatively connected to the optical radiation generation device for controlling dosage of the therapeutic light transmitted to the treatment site at a dosimetry sufficient to produce photodamage in the biological contaminant without causing substantial photothermal or photomechanical damage to biological tissue at the treatment site; and a delivery assembly including any of the optical delivery devices described herein and configured to deliver the therapeutic light to the treatment site to illuminate at least a portion of the treatment site with a prescribed illumination pattern.
  • the optical radiation generation device includes the at least one source.
  • the therapeutic light includes optical radiation substantially in a first wavelength range from about 865 nm to about 875 nm or a second wavelength range having a wavelength from about 925 nm to about 935 nm, or both wavelength ranges.
  • the treatment light includes optical radiation at both wavelength ranges.
  • the delivery assembly is adapted to provide substantially uniform illumination of at least a portion of the illuminated portion of the treatment site. In some embodiments, the delivery assembly is adapted to illuminate the portion of the treatment site at a power density and an energy density which potentiates an antimicrobial application at the treatment site. In some embodiments, the delivery assembly is adapted to illuminate the portion of the treatment site at a power density of about 0.3 W/cm 2 to about .7 W/cm 2 and an energy density from about 100 J/cm 2 to about 400 J/cm 2 at the illuminated portion of the treatment site. In some embodiments, the delivery assembly is adapted to operate for about 30 seconds or more at an operating temperature of 110° F or less. In some embodiments,the delivery assembly is adapted for detachable coupling to the optical radiation generation device.
  • a method treatment of a biological contaminant at a treatment site including: generating near infrared therapeutic light; controlling dosage of the therapeutic light transmitted to the treatment site at a dosimetry sufficient to produce photodamage in the biological contaminant without causing substantial photothermal or photomechanical damage to biological tissue at the treatment site; and using a delivery assembly including the optical deliver apparatus of any of claims 1-28, directing the therapeutic light to be transmitted to the treatment site to illuminate at least a portion of the treatment site with a prescribed illumination pattern.
  • generating near infrared therapeutic light includes generating light from the at least one source.
  • the therapeutic light includes optical radiation substantially in a first wavelength range from about 865 nm to about 875 nm or a second wavelength range having a wavelength from about 925 nm to about 935 nm, or both wavelength ranges.
  • the treatment light includes optical radiation at both wavelength ranges.
  • directing the therapeutic light to be transmitted to the treatment site to illuminate at least a portion of the treatment site with a prescribed illumination pattern includes providing substantially uniform illumination of at least a portion of the illuminated portion of the treatment site.
  • Some embodiments include, applying a quantity of antimicrobial agent the treatment site; and illuminating the portion of the treatment site at a power density and an energy density which potentiates the antimicrobial application.
  • the antimicrobial agent is ineffective for treating the antimicrobial resistant biological contaminate in the absence of the therapeutic light.
  • the biological contaminate includes an aberrant microbial colonization, and further including includes using the antimicrobial agent in conjunction with the therapeutic light to potentiate the antimicrobial agent to reduce the level of colonization at the treatment site.
  • controlling the dosage of the therapeutic light includes controlling the illumination of the portion of the treatment site at a power density of about 0.2 W/cm 2 to about 1 W/cm 2 and an energy density from about 100 J/cm 2 to about 400 J/cm 2 at the illuminated region.
  • a system including a source of therapeutic light optically coupled to an optical delivery assembly including the optical delivery apparatus of any of the types described herein.
  • a method including using the optical delivery apparatus of any the types described herein to deliver therapeutic light to illuminate a treatment site.
  • a method of constructing an optical delivery device including: obtaining an optical fiber extending between a distal end having a distal end face and a proximal end having a proximal end face; obtaining an optical element; obtaining a non-metallic housing; and fitting the optical fiber and optical element within the housing such that the housing maintains the relative position of the optical fiber and the optical element.
  • the fitting the optical fiber and optical element within the housing includes press fitting an elastic cuff about the optical fiber and optical element.
  • the fitting the optical fiber and optical element within the housing is performed at temperatures less than 500 degrees C, less than 250 degrees C, less than 100 decrees C, less than 50 degrees C or even less.
  • the fitting the optical fiber and optical element within the housing includes using a removable template to position the optical fiber and optical element. In some embodiments, the fitting the optical fiber and optical element within the housing includes fitting the optical fiber and optical element within the housing without applying heat to shrink any portion of the housing.
  • the non-metallic housing is an elastic housing.
  • the elastic housing is an elastic cuff, and where the fitting the optical fiber and optical element within the housing includes stretching the cuff over the optical fiber and optical element such that compressive force from the elastic cuff maintains the relative position of the optical fiber and the optical element.
  • Figure 1 shows a typical phospholipid bilayer
  • Figure 2 shows the chemical structure of a phospholipid
  • Figure 3 shows dipole effects in phospholipid bilayer membranes ( ⁇ d);
  • Figure 4A shows a phospholipid bilayer in bacterial plasma membrane, mammalian mitochondrial membrane, or fugal mitochondrial membrane with a steady-state transmembrane potential prior to NIMELS irradiation.
  • Figure 4B shows a transient-state plasma membrane potential in bacterial plasma membrane, mammalian mitochondrial membrane, or fugal mitochondrial membrane after NIMELS irradiation;
  • Figure 5 shows a phospholipid bilayer with trans-membrane proteins embedded therein
  • Figure 6 shows a general depiction of electron transport and proton pump
  • Figure 7 shows the effects of NIMELS irradiation (at a single dosimetry) on MRSA trans-membrane potential which is measured by green fluorescence emission intensities in control and lased samples as a function of time in minutes post-lasing
  • Figure 8 shows the effects of NIMELS irradiation (at a single dosimetry) on mitochondrial membrane potential of human embryonic kidney cells, which is measured by red fluorescence emission intensities in control and lased samples
  • NMELS irradiation at a single dosimetry
  • Figure 9 shows the reduction in total glutathione concentration in MRSA as it correlates with reactive oxygen species (ROS) generation in these cells as the result of NIMELS irradiation (at several dosimetries); the decrease in glutathione concentration in lased samples is shown as percentage relative to the control;
  • Figure 10 shows the reduction in total glutathione concentration in human embryonic kidney cells as it correlates with reactive oxygen species (ROS) generation in these cells as the result of NIMELS irradiation (at two different dosimetries); the decrease in glutathione concentration in lased samples is shown as percentage relative to the control;
  • Figure 11 shows the synergistic effects of NIMELS and methicillin in growth inhibition of MRSA colonies; data show methicillin is being potentiated by sub-lethal NIMELS dosimetry; and
  • Figure 12 shows the synergistic effects of NIMELS and bacitracin in growth inhibition of MRSA colonies; arrows indicate the growth or a lack thereof of MRSA colonies in the two samples shown; images show that bacitracin is being potentiated by sublethal NIMELS dosimetry.
  • Figure 13 shows a bar chart depicting the synergistic effects, as indicated by experimental data, of NIMELS with methicillin, penicillin and erythromycin in growth inhibition of MRSA colonies.
  • Figure 14 illustrates the detection of decreased membrane potential in E. coli with sub-lethal NIMELS irradiation.
  • Figure 15 illustrates the detection of increased glutathione in E. coli with sub-lethal NIMELS irradiation.
  • Figure 16a illustrates five subjects initially culturing positive for erythromycin resistant MSSA, all showing positive responses to phototherapy.
  • Figure 16b illustrates three subjects initially culturing positive for erythromycin resistant MSRA, all showing positive responses to phototherapy.
  • Fig. 17 illustrates an exemplary NIMELS treatment system.
  • Figs. 18a-18d illustrate the delivery of treatment light from a NIMELS treatment system.
  • Fig. 19 shows the principal optical fiber, cladding, buffer, housing and optical element
  • Figs. 20a & b shows the core imaging principal and intensity distribution of the present invention
  • Fig. 21 shows the positive-locking, buffer-housing system
  • Fig. 22 shows a GRIN embodiment of the present invention
  • Fig. 23 shows a multiple-source optical fiber integration embodiment
  • Figs. 24a-24c shows assembly methodology including the principal optical fiber, buffer, housing and optical element and press-fit template.
  • Fig. 25 shows a compressible mechanical assembly
  • Fig. 26 is a perspective illustration of a diffusive fiber tip device.
  • Fig. 27 is a cross sectional representation of a diffusive fiber tip device that shows how the light emitted from a fiber optic initially interacts with a diffusion tube without the mirrors that form the reflective cavity.
  • Fig. 28 is a perspective illustration of the operation of the diffusing tube of Fig. 20 with the addition of the cavity mirrors.
  • Fig. 29 is a cross sectional illustration of a curved mirror embodiment of a diffusive fiber tip assembly.
  • Fig. 30 shows intensity profiles observed from the operation of the diffusive tip assembly.
  • Fig. 31 is a cross sectional illustration of an exemplary construction of a curved mirror embodiment of the diffusive tip assembly.
  • Fig. 32 is a cross sectional illustration of an exemplary construction of the diffusive reflector embodiment of the diffusive fiber tip assembly.
  • Fig. 33 is a drawing of a reusable diffusion tip encased in a disposable polypropylene outer jacket.
  • Fig 34 illustrates several exemplary embodiments of diffusion tips.
  • Fig 35 is an intensity CCD camera scan of a diffusion tip.
  • Fig. 36 is an illustration of a diffusion tip.
  • Fig. 37 is an illustration of a kit including a diffusion tip. DETAILED DESCRIPTION OF THE INVENTION
  • a NIMELS wavelength includes any wavelength within the ranges of the NIMELS wavelengths described, as well as combinations of such wavelengths.
  • the present invention is directed to methods and systems for enhancing bacterial succeptability to antimicrobial agents thereby reducing the minimum inhibitory concentration (MIC) of the antimicrobial agent necessary to attenuate or eliminate microbial related pathology and/or enabling therapeutic use of antimicrobial agents that would otherwise be ineffective due to bacterial resistance.
  • MIC minimum inhibitory concentration
  • NIMELS near infrared optical radiation in selected energies and dosimetries
  • ⁇ membrane potential
  • NIMELS effect can potentiate existing antimicrobial agents against microbes infecting and causing harm to human or animal hosts.
  • NIMELS effects will affect many cellular anabolic reactions (e.g., cell wall formation) and drug-resistance mechanisms (e.g., efflux pumps) that require chemiosmotic electrochemical energy to function.
  • any membrane bound cellular resistance mechanisms or anabolic reactions that makes use of the membrane potential ⁇ , proton motive force ⁇ p, or the phosphorylation potential ⁇ Gp for their functional energy needs, will be affected by the NIMELS effects, and accordingly provide therapeutic targets for the methods and systems of the present invention.
  • the methods and systems of the present invention utilize optical radiation to sensitize undesirable microbial cells (e.g., MRSA infection in skin) without substantial thermal or chemical damage to host tissues.
  • undesirable microbial cells e.g., MRSA infection in skin
  • the applied optical radiation used in accordance with methods and systems of the present invention includes one or more, and preferably two independent wavelengths ranging from about 850 nm to about 900 nm, at a NIMELS dosimetry, as described herein. In one aspect, wavelengths from about 865 nm to about 875 nm are utilized. In another aspect, such applied radiation has a wavelength from about 905 nm to about 945 nm at a NIMELS dosimetry. In one aspect, such applied optical radiation has a wavelength from about 925 nm to about 935 nm. In a particular aspect, a wavelength of (or narrow wavelength range including) 930 nm can be employed. In some aspects of the present invention, multiple wavelength ranges include 870 and 930 nm, respectively.
  • the methods and systems of the present invention are used in treating, reducing and/or eliminating the infectious entities known to cause cutaneous or wound infections such as staphyloccocci and enterococci.
  • Staphyloccoccal and enterococcal infections can involve almost any skin surface on the body, and is known to cause numerous skin conditions such as boils, carbuncles, bullous impetigo and scalded skin syndrome.
  • one objective of the invention is to prevent or treat staphyloccoccal and enterococcal infections of the host skin, thereby treating the aforementioned conditions. S.
  • aureus is also the cause of staphylococcal food poisoning, enteritis, osteomilitis, toxic shock syndrome, endocarditis, meningitis, pneumonia, cystitis, septicemia and postoperative wound infections. Accordingly, another objective of the invention is to prevent or treat such infections. S. aureus can be acquired while a patient is in a hospital or long- term care facility, and yet another object of the invention is to prevent or treat nosocomial infections in the host.
  • a further object of the invention is to prevent or treat drug resistant bacterial infections of the host, preferably but not limited to MRSA infections of the host.
  • ROS reactive oxygen species
  • the term also includes irradiating a cell to increase the sensitivity of the biological contaminant through the lowering of ⁇ with the concomitant generation of ROS of an antimicrobial or antineoplastic agent, wherein the contaminant is resistant to the agent otherwise.
  • potentiation of an antibacterial agent it is meant that the methods and systems of this invention counteract the resistance mechanisms in the microbe sufficiently for the agent to inhibit the growth and/or proliferation of said microbe at a lower concentration than in the absence of the present methods and systems.
  • potentiation means that the agent will inhibit the growth and/or proliferation of pathogenic cells at a therapeutically acceptable dosage, thereby treating the disease state.
  • Membrane Dipole Potential ⁇ d (in contrast to the Transmembrane Potential ⁇ ) refers to the potential formed between the highly hydrated lipid heads (hydrophilic) at the membrane surface and the low polar interior of the bilayer (hydrophobic). Lipid bilayers intrinsically possess a substantial Membrane Dipole Potential ⁇ d arising from the structural organization of dipolar groups and molecules, primarily the ester linkages of the phospholipids and water. ⁇ d does not depend upon the ions at the membrane surface and will be used herein to describe five different dipole potentials:
  • Trans-Membrane Potential refers to the electrical potential difference between the aqueous phases separated by a membrane (dimensions mV) and will be given by the symbol ( ⁇ ). ⁇ does depend upon the ions at the membrane surface and will be used herein to describe three different plasma trans-membrane potentials.
  • Mitochondrial Trans-Membrane Potential refers to the electrical potential difference between the compartments separated by the mitochondrial inner membrane (dimensions mV) and will be used herein to describe two different mitochondrial trans-membrane potentials.
  • bacterial plasma trans-membrane potential ( ⁇ -plas- bact) refers to the electrical potential difference in the bacterial cell plasma membrane.
  • the bacterial plasma membrane potential is generated by the steady-state flow (translocation) of electrons and protons (H + ) across the bacterial plasma membrane that occurs with normal electron transport and oxidative phosphorylation, within the bacterial plasma membrane.
  • a common feature of all electron transport chains is the presence of a proton pump to create a transmembrane proton gradient.
  • bacteria lack mitochondria, aerobic bacteria carry out oxidative phosphorylation (ATP production) by essentially the same process that occurs in eukaryotic mitochondria.
  • P-class ion pump refers to a trans-membrane active transport protein assembly which contains an ATP-binding site (i.e., it needs ATP to function). During the transport process, one of the protein subunits is phosphorylated, and the transported ions are thought to move through the phosphorylated subunit.
  • This class of ion pumps includes the Na + /K + -ATPase pump in the mammalian plasma membrane, which maintains the Na + and K + electrochemical potential ( ⁇ Na + /K + ) and the pH gradients typical of animal cells.
  • Another important member of the P-class ion pumps transports protons (H + ions) out of and K + ions in to the cell.
  • steady-state plasma trans-membrane potential refers to the quantitative Plasma Membrane Potential of a mammalian, fungal or bacterial cell before irradiation in accordance with the methods and systems of the present invention that would continue into the future in the absence of such irradiation.
  • the steady-state flow of electrons and protons across a bacterial cell membrane that occurs during normal electron transport and oxidative phosphorylation would be in a steady- state due to a constant flow of conventional redox reactions occurring across the membrane.
  • any modification of this redox state would cause a transient-state membrane potential.
  • ⁇ -steady will be used herein to describe three (3) different steady-state plasma trans-membrane potentials, based on species.
  • Transient-state plasma membrane potential refers to the Plasma Membrane Potential of a mammalian, fungal or bacterial cell after irradiation in accordance with the methods and systems of the present invention whereby the irradiation has changed the bioenergetics of the plasma membrane.
  • ⁇ -tran will also change the redox state of the cell, as the plasma membrane is where the ETS and cytochromes reside.
  • ⁇ -tran is a state that would not occur without irradiation using methods of the present invention.
  • ⁇ -tran will be used herein to describe three (3) different Transient-state plasma trans-membrane potentials based on species.
  • steady-state mitochondrial membrane potential refers to the quantitative Mitochondrial Membrane Potential of mammalian or fungal mitochondria before irradiation in accordance with the methods and systems of the present invention that would continue into the future, in the absence of such irradiation.
  • the steady-state flow of electrons and protons across mitochondrial inner membrane that occurs during normal electron transport and oxidative phosphorylation would be in a steady-state because of a constant flow of conventional redox reactions occurring across the membrane. Any modification of this redox state would cause a transient-state mitochondrial membrane potential.
  • ⁇ -steady-mito will be used herein to describe two (2) different steady-state mitochondrial membrane potentials based on species.
  • transient-state mitochondrial membrane potential ( ⁇ - tran-mito-mam or ⁇ -tran-mito-fungi) refers to the membrane potential of a mammalian or fungal cell after irradiation in accordance with the methods and systems of the present invention whereby the irradiation has changed the bioenergetics of the mitochondrial inner membrane.
  • ⁇ -tran-mito will also change the redox state of the cell, as the inner mitochondrial membrane is where the electron transport system (ETS) and cytochromes reside.
  • ETS electron transport system
  • ⁇ -tran-mito could also drastically affect (the Proton- motive force) ⁇ p-mito-mam and ⁇ p-mito-fungi, as these mitochondrial (H + ) gradients are generated in the mitochondria, to produce adequate ATP for a myriad of cellular functions.
  • ⁇ -tran-mito is a state that would not occur without irradiation in accordance with methods and systems of the present invention.
  • ⁇ -tran-mito will be used herein to describe two (2) different transient-state mitochondrial membrane potentials based on species.
  • proton electrochemical gradient (dimensions kJ mol-1) refers to the electrical and chemical properties across a membrane, particularly proton gradients, and represents a type of cellular potential energy available for work in a cell.
  • ⁇ H 4" is reduced by any means, it is a given that cellular anabolic pathways and resistance mechanisms in the affected cells are inhibited.
  • ⁇ n and Tn can be further enhanced with the simultaneous or sequential administration of a pharmacological agent configured and arranged for delivery to the target site (i.e., the co- targeting of an anabolic pathway with ( ⁇ n and Tn) + (pharmacological molecule or molecules)).
  • a pharmacological agent configured and arranged for delivery to the target site (i.e., the co- targeting of an anabolic pathway with ( ⁇ n and Tn) + (pharmacological molecule or molecules)).
  • ⁇ x+ refers to the electrical and chemical properties across a membrane caused by the concentration gradient of an ion (other than H + ) and represents a type of cellular potential energy available for work in a cell. In mammalian cells, the Na + ion electrochemical gradient is maintained across the plasma membrane by active transport OfNa + out of the cell.
  • ⁇ x + is reduced by any means, it is a given that cellular anabolic pathways and resistance mechanisms in the affected cells are inhibited.
  • This can be accomplished by combining ⁇ n and Tn to irradiate a target site alone, or can be further enhanced with the simultaneous or sequential administration of a pharmacological agent configured and arranged for delivery to the target site (i.e., the co- targeting of an anabolic pathway with ( ⁇ n and Tn) + (pharmacological molecule or molecules)).
  • co-targeting of a bacterial anabolic pathway refers to (the ⁇ n and Tn lowering Of (AuH + ) and/or ( ⁇ x + ) of cells at the target site to affect an anabolic pathway) + (a pharmacological molecule or molecules to affect the same bacterial anabolic pathway) and can refer to any of the following bacterial anabolic pathways that are capable of being inhibited with pharmacological molecules: wherein the targeted anabolic pathway is peptidoglycan biosynthesis that is co-targeted by a pharmacological agent that binds at the active site of the bacterial transpeptidase enzymes (penicillin binding proteins) which cross-links peptidoglycan in the bacterial cell wall.
  • the targeted bacterial anabolic pathway is peptidoglycan biosynthesis that is co-targeted by a pharmacological agent that binds to acyl-D-alanyl-D- alanine groups in cell wall intermediates and hence prevents incorporation of N- acetylmuramic acid (NAM)- and N-acetylglucosamine (NAG)-peptide subunits into the peptidoglycan matrix (effectively inhibiting peptidoglycan biosynthesis by acting on transglycosylation and/or transpeptidation) thereby preventing the proper formation of peptidoglycan, in gram-positive bacteria; wherein the targeted bacterial anabolic pathway is peptidoglycan biosynthesis that is co-targeted by a pharmacological agent that binds with C 55 -isoprenyl pyrophosphate and prevents pyrophosphatase from interacting with C 55 - isopren
  • the targeted anabolic pathway is bacterial protein biosynthesis that is co-targeted by a pharmacological agent that binds to a specific aminoacyl-tRNA synthetase to prevent the esterification of a specific amino acid or its precursor to one of its compatible tRNA's, thus preventing formation of an aminoacyl-tRNA and hence halting the incorporation of a necessary amino acid into bacterial proteins;
  • the term "proton-motive force (Ap)” refers to the storing of energy (acting like a kind of battery), as a combination of a proton and voltage gradient across a membrane.
  • the two components of Ap are ⁇ (the transmembrane potential) and ⁇ pH (the chemical gradient OfH + ).
  • Ap consists of the H + transmembrane potential ⁇ (negative (acidic) outside) and a transmembrane pH gradient ⁇ pH (alkaline inside).
  • This potential energy stored in the form of an electrochemical gradient is generated by the pumping of hydrogen ions across biological membranes (mitochondrial inner membranes or bacterial and fungal plasma membranes) during chemiosmosis.
  • the Ap can be used for chemical, osmotic, or mechanical work in the cells.
  • the proton gradient is generally used in oxidative phosphorylation to drive ATP synthesis and can be used to drive efflux pumps in bacteria, fungi, or mammalian cells including cancerous cells.
  • ⁇ p- plas-Bact Bacterial Plasma Membrane Proton-motive force
  • H + electrochemical gradient
  • ⁇ p-plas-Bact is used in oxidative phosphorylation to drive ATP synthesis in the bacterial plasma membrane and can be used to drive efflux pumps in bacterial cells.
  • phosphorylation potential refers to the ⁇ G for ATP synthesis at any given ATP, ADP and Pi concentrations (dimensions: kJ mol "1 ).
  • CCCP refers to carbonyl cyanide m- chlorophenylhydrazone, a highly toxic ionophore and uncoupler of the respiratory chain. CCCP increases the conductance of protons through membranes and acts as a classical uncoupler by uncoupling ATP synthesis from the ⁇ H* and dissipating both the ⁇ and ⁇ pH.
  • Reactive Oxygen Species includes one of the following categories: a) The Superoxide ion radical (O 2 " ) b) Hydrogen Peroxide (non-radical) (H 2 O 2 ) c) Hydroxyl radical (*OH) d) Hydroxy ion (OH " ) These ROS generally occur through the reaction chain:
  • the term "singlet oxygen” refers to (“1O 2 ”) and is formed via an interaction with triplet-excited molecules.
  • Singlet oxygen is a non-radical species with its electrons in anti -parallel spins. Because singlet oxygen 1O 2 does not have spin restriction of its electrons, it has a very high oxidizing power and is easily able to attack membranes (e.g., via polyunsaturated fatty acids, or PUFAs) amino acid residues, protein and DNA.
  • the term “NIMELS effect” refers to the modification of the bioenergetic "state” of irradiated cells at the level of the cell's plasma and mitochondrial membranes from ⁇ -steady to ⁇ -trans with the present invention. Specifically, the NIMELS effect can weaken cellular anabolic pathways or antimicrobial and/or cancer resistance mechanisms that make use of the proton motive force or the chemiosmotic potential for their energy needs.
  • periplasmic space or periplasm refers to the space between the plasma membrane and the outer membrane in gram-negative bacteria and the space between the plasma membrane and the cell wall in gram-positive bacteria and fungi such as the Candida and Trichophyton species.
  • This periplasmic space is involved in various biochemical pathways including nutrient acquisition, synthesis of peptidoglycan, electron transport, and alteration of substances toxic to the cell.
  • periplasmic space is of significant clinical importance as it is where ⁇ - lactamase enzymes inactivate penicillin based antibiotics.
  • efflux pump refers to an active transport protein assembly which exports molecules from the cytoplasm or periplasm of a cell (such as antibiotics, antifungals, or poisons) for their removal from the cells to the external environment in an energy dependent fashion.
  • efflux pump inhibitor refers to a compound or electromagnetic radiation delivery system and method which interferes with the ability of an efflux pump to export molecules from a cell.
  • the efflux pump inhibitor of this invention is a form of electromagnetic radiation that will interfere with a pump's ability to excrete therapeutic antibiotics, anti-fungal agents, antineoplastic agents and poisons from cells via a modification of the ⁇ -steady-mam , ⁇ -steady-fungi or, ⁇ -steady-bact.
  • a cell that "utilizes an efflux pump resistance mechanism” it is meant that the bacterial cell exports anti-bacterial agents from their cytoplasm or periplasm to the external environment of the cell and thereby reduce the concentration of these agents in the cell to a concentration below what is necessary to inhibit the growth and/or proliferation of the bacterial cells.
  • anti-bacterial molecule refers to a chemical or compound that is bacteriacidal or bacteriastatic. Another principal efficacy resides in the present invention's ability to potentiate anti-bacterial molecules by inhibiting efflux pump activity in resistant bacterial strains, or inhibiting anabolic reactions and/or resistance mechanisms that require the proton motive force or chemiosmotic potential for energy.
  • a "sub-inhibitory concentration" of an antibacterial agent refers to a concentration that is less than that required to inhibit a majority of the target cells in the population. Generally, a sub-inhibitory concentration refers to a concentration that is less than the Minimum Inhibitory Concentration (MIC).
  • MIC Minimum Inhibitory Concentration
  • Minimal Inhibitory Concentration is defined as the lowest effective or therapeutic concentration that results in inhibition of growth of the microorganism.
  • the minimum inhibitory concentration (MIC) of an antibacterial agent is therefore the maximum dilution of the agent that will still inhibit the growth of a test microorganism.
  • the minimum bactericidal concentration (MBCs) of an antibacterial agent is the lowest concentration of the antimicrobial agent that will prevent the growth of an organism after subculture on to antibiotic-free media.
  • the minimum lethal concentration (MLC) of an antibacterial agent is the maximum dilution of the product that will kill the test organism. MIC/MLC values can be determined by a number of standard test procedures.
  • the most commonly employed methods are the tube dilution method and agar dilution methods. Serial dilutions are made of the products in bacterial growth media. The test organisms are then added to the dilutions of the products, incubated, and scored for growth. This procedure is a standard assay for antimicrobials. The procedure incorporates the content and intent of the American Society for Microbiology (ASM) recommended methodology.
  • ASM American Society for Microbiology
  • the term "therapeutically effective amount" of an antibacterial agent refers to a concentration of an agent that will partially or completely relieve one or more of the symptoms caused by the target (pathogenic) cells.
  • a therapeutically effective amount refers to the amount of an agent that: (1) reduces, if not eliminates, the population of target microbial cells in the patient's body, (2) inhibits (i.e., slows, if not stops) proliferation of the target microbial cells in the patients body, (3) inhibits (i.e., slows, if not stops) spread of the infection (4) relieves (if not, eliminates) symptoms associated with the infection.
  • the NIMELS effect lowers the therapeutic threshold by sensitizing the microbial targets to the antibiotic agent.
  • the term "Interaction coefficient" is defined as a numerical representation of the magnitude of the bacteriastatic/bacteriacidal interaction between the NTMELS laser and/or the antimicrobial molecule, with the target cells.
  • Thermodynamics of Energy Transduction in Biological Membranes Thermodynamics of Energy Transduction in Biological Membranes
  • the present invention is directed to perturbing cell membrane biological thermodynamics (bioenergetics) and the consequent diminished capacity of the irradiated cells to adequately undergo normal energy transduction and energy transformation.
  • the methods and systems of the present invention optically alter and modify ⁇ - plas-mam, ⁇ d-mito-mam, ⁇ d-plas-fungi, ⁇ d-mito-fungi and ⁇ d-plas-bact to set in motion further alterations of ⁇ and ⁇ p in the same membranes. This is caused by the targeted near infrared irradiation of the C-H covalent bonds in the long chain fatty acids of lipid bilayers, causing a variation in the dipole potential ⁇ d.
  • membranes lipid bilayers, see, Figure 1
  • Figure 2 possess a significant dipole potential ⁇ d arising from the structural association of dipolar groups and molecules, primarily the ester linkages of the phospholipids (Figure 2) and water.
  • These dipolar groups are oriented such that the hydrocarbon phase is positive with respect to the outer membrane regions ( Figure 3).
  • the degree of the dipole potential is usually large, typically several hundreds of millivolts.
  • the second major potential a separation of charge across the membrane, gives rise to the transmembrane potential ⁇ .
  • the trans-membrane potential is defined as the electric potential difference between the bulk aqueous phases at the two sides of the membrane and results from the selective transport of charged molecules across the membrane.
  • the potential at the cytoplasm side of cell membranes is negative relative to the extracellular physiological solution ( Figure 4A).
  • the dipole potential ⁇ d constitutes a large and functionally important part of the electrostatic potential of all plasma and mitochondrial membranes.
  • ⁇ modifies the electric field inside the membrane, producing a virtual positive charge in the apolar bilayer center.
  • lipid membranes exhibit a substantial (e.g., up to six orders of magnitude) difference in the penetration rates between positively and negatively charged hydrophobic ions.
  • ⁇ d also plays an important role in the membrane permeability for lipophilic ions. Numerous cellular processes, such as binding and insertion of proteins (enzymes), lateral diffusion of proteins, ligand-receptor recognition, and certain steps in membrane fusion to endogenous and exogenous molecules, critically depend on the physical properties ⁇ d of the membrane bilayer.
  • the energy transduction in biological membranes generally involves three interrelated mechanisms: 1) The transduction of redox energy to "free energy" stored in a trans-membrane ionic electrochemical potential also called the membrane proton electrochemical gradient ⁇ FT. This proton electrochemical potential difference between the two sides of a membrane that engage in active transport involving proton pumps is at times also called a chemiosmotic potential or proton motive force. 2) In mammalian cells, the (Na + ) ion electrochemical gradient ⁇ x + is maintained across the plasma membrane by active transport Of (Na + ) out of the cell.
  • ⁇ Gp is the ⁇ G for ATP synthesis at any given set of ATP, ADP and P 1 concentrations.
  • thermodynamics a state function ⁇ state quantity
  • ⁇ a property or a system that depends only on the current state of the system. It does not depend on the way in which the system attained its particular state.
  • the present invention facilitates a transition of state in a trans-membrane and/or mitochondrial potential ⁇ , in a temporally dependent manner, to move the bioenergetics of a membrane from a thermodynamic steady- state condition ⁇ -steady to one of energy stress and/or redox stress in a transition state ⁇ -trans.
  • the individual photons of infrared radiation do not contain sufficient energy (e.g., as measured in electron-volts) to induce electronic transitions (in molecules) as is seen with photons of ultraviolet radiation. Because of this, absorption of infrared radiation is limited to compounds with small energy differences in the possible vibrational and rotational states of the molecular bonds.
  • the vibrations or rotations within the lipid bilayer' s molecular bonds that absorb the infrared photons must cause a net change in the dipole potential of the membrane. If the frequency (wavelength) of the infrared radiation matches the vibrational frequency of the absorbing molecule (i.e., C-H covalent bonds in long chain fatty acids) then radiation will be absorbed causing a change in ⁇ d. This can happen in ⁇ d-plas-mam, ⁇ d-mito-mam, ⁇ d-plas-fungi, ⁇ d-mito- fungi and ⁇ d-plas-bact. In other words, there can be a direct and targeted change in the enthalpy and entropy ( ⁇ H and ⁇ S) of all cellular lipid bilayers with the methods and systems described herein.
  • ⁇ H and ⁇ S enthalpy and entropy
  • the present invention is based upon a combination of insights that have been introduced above and are derived in part from empirical data, which include the following: It has been appreciated that unique, single infrared wavelengths (about 870 nm and about 930 nm) are each capable of killing bacterial cells (prokaryotes) such as E.coli and (eukaryotes) such as Chinese Hampster Ovary (CHO) cells, as a result of the generation and interaction of ROS and/or toxic singlet oxygen reactions.
  • prokaryotes such as E.coli
  • eukaryotes such as Chinese Hampster Ovary (CHO) cells
  • the present invention employs these infrared wavelengths, preferably in combination, but at 5 log less power density than is typically found in a confocal laser microscope such as that used in optical traps ( ⁇ to 500,000 w/cm 2 less power) to advantageously exploit the use of such wavelengths for therapeutic laser systems, to cause a bacteriostatic or bacteriocidal effect at an infection site, without causing thermal damage to the hosts tissues.
  • cytochrome chains With 870 nm, will additionally alter ⁇ - steady and the redox potential of the membranes that have cytochromes (i.e., bacterial plasma membranes, and fungal and mammalian mitochondria).
  • the NIMELS effect occurs in accordance with methods and systems described herein, importantly, without thermal or ablative mechanical damage to the cell membranes.
  • This combined and targeted low dose irradiation approach is a distinct variation and improvement from existing methods that would otherwise cause actual thermal or mechanical damage to all membranes within the path of a beam of energy.
  • Entropy in a membrane is a state function whose change in a reaction describes the direction of a reaction due to changes in (energy) heat input or output and the associated molecular rearrangements.
  • the NIMELS effect will modify the entropy "state" of irradiated cells at the level of the lipid bilayer in a temporally dependent manner.
  • This increase in entropy will alter the Yd of all irradiated membranes (mitochondrial and plasma) and hence, thermodynamically alter the "steady-state” flow of electrons and protons across a cell membrane ( Figures 6 and 7).
  • This will in turn change the steady-state trans-membrane potential ⁇ -steady to a transient-state membrane potential ( ⁇ -tran). This phenomenon will occur in:
  • Bacterial Plasma Membrane Proton-motive force ( ⁇ p-plas-Bact). Such phenomena can in turn decrease the Gibbs free energy value AG available for the phosphorylation and synthesis of ATP (AGp).
  • the present invention carries out these phenomena in order to inhibit the necessary energy dependent anabolic reactions, potentiating pharmacological therapies, and/or lowering cellular resistance mechanisms (to antimicrobial, antifungal and antineoplastic molecules) as many of these resistance mechanisms make use of the proton motive force or the chemiosmotic potential for their energy needs, to resist and/or efflux these molecules.
  • the action of chemical uncouplers for oxidative phosphorylation and other bioenergetic work is believed to depend on the energized state of the membrane (plasma or mitochondrial). Further, it is believed that the energized state of the bacterial membrane or eukaryotic mitochondrial inner membrane, is an electrochemical proton gradient ⁇ KT that is established by primary proton translocation events occurring during cellular respiration and electron transport. Agents that directly dissipate (depolarize) the ⁇ HT, (e.g., by permeabilizing the coupling membrane to the movement of protons or compensatory ions) short-circuits energy coupling, and inhibit bioenergetic work, by inducing a reduction in the membrane potential ⁇ -steady.
  • the present invention can act as an optical uncoupler by lowering the ⁇ H 1" and ⁇ p of the following irradiated membranes:
  • Lipid peroxidation is a prevalent cause of biological cell injury and death in both the microbial and mammalian world.
  • strong oxidents cause the breakdown of membrane phospholipids that contain polyunsaturated fatty acids (PUFA's).
  • PUFA's polyunsaturated fatty acids
  • Peroxidation of mitochondrial membranes will have detrimental consequences on the respiratory chains resulting in inadequate production of ATP and collapse of the cellular energy cycle.
  • Peroxidation of the plasma membrane can affect membrane permeability, disfunction of membrane proteins such as porins and efflux pumps, inhibition of signal transduction and improper cellular respiration and ATP formation (i.e., the respiratory chains in prokaryotes are housed in the plasma membranes as prokaryotes do not have mitochondria).
  • a free radical is defined as an atom or molecule that contains an unpaired electron.
  • An example of the damage that a free radical can do in a biological environment is the one- electron (via an existing or generated free radical) removal from bis-allylic C-H bonds of polyunsaturated fatty acids (PUFAs) that will yield a carbon centered free radical.
  • PUFAs polyunsaturated fatty acids
  • This reaction can initiate lipid peroxidation damage of biological membranes.
  • a free radical can also add to a nonradical molecule, producing a free radical product.
  • ROS Reactive Oxygen Species
  • Oxygen gas is actually a free radical species. However, because it contains two unpaired electrons in different ⁇ r-anti-bonding orbitals that have parallel spin in the ground state, the (spin restriction) rule generally prevents O 2 from receiving a pair of electrons with parallel spins without a catalyst. Consequently O 2 must receive one electron at a time.
  • the Reaction Chain is:
  • Superoxide for example, can either act as an oxidizing or a reducing agent.
  • the protonated form of superoxide hydroperoxyl radical has a lower reduction potential than (O 2 " ), yet is able to remove hydrogen atoms from PUFA's.
  • the pKa value of (HOO*) is 4.8 and the (acid) microenvironment near biologiocal membranes will favor the formation of hydroperoxyl radicals.
  • the reaction of superoxide (O 2 " ) with any free F 6 +3 will produce a "perferryl" intermediate which can also react with PUFA' s and induce lipid (membrane) peroxidation.
  • Hydrogen peroxide is not a good oxidizing agent (by itself) and cannot remove hydrogen from PUFA's. It can, however, cross biological membranes (rather easily) to exert dangerous and harmful effects in other areas of cells.
  • (H 2 O 2 ) is highly reactive with transition metals inside microcellular environments, (such as Fe +2 and Cu + ) that can then create hydroxyl radicals (*OH) (known as the Fenton Reaction).
  • An hydroxyl radical is one of the most reactive species known in biology. Hydroxyl Radical
  • Hydroxyl radicals will react with almost all kinds of biological molecules. It has a very fast reaction rate that is essentially controlled by the hydroxyl radical (*OH) diffusion rate and the presence (or absence) of a molecule to react near the site of (*OH) creation.
  • the standard reduction potential (EO') for hydroxyl radical (*OH) is (+2.31 V) a value that is 7 ⁇ greater than (H 2 O 2 ), and is categorized as the most reactive among the biologically relevant free radicals. Hydroxyl radicals will initiate lipid peroxidation in biological membranes, in addition to damaging proteins and DNA. Reactive Oxygen Species Created from the Peroxidation of PUFAs
  • alkyl hydroperoxides are not technically radical species but are unstable in the presence of transition metals such as such as Fe +2 and Cu + .
  • Alkyl peroxyl radicles and alkoxyl radicles are extremely reactive oxygen species and also contribute to the process of propagation of further lipid peroxidation.
  • the altered redox state of irradiated cells and generation of free radicals and ROS because of the ⁇ - steady + (NIMELS Treatment) ->-> ⁇ - trans phenomenon is another object of the present invention. This is an additive effect to further alter cellular bioenergetics and inhibit necessary energy dependent anabolic reactions, potentiate pharmacological therapies, and/or lower cellular resistance mechanisms to antimicrobial, antifungal and antineoplastic molecules.
  • ROS overproduction can damage cellular macromolecules, above all lipids.
  • Lipid oxidation has been shown to modify both the small-scale structural dynamics of biological membranes as well as their more macroscopic lateral organization and altered a packing density dependent reorientation of the component of the dipole moment ⁇ d. Oxidative damage of the acyl chains (in lipids) causes loss of double bonds, chain shortening, and the introduction of hydroperoxy groups. Hence, these changes are believed to affect the structural characteristics and dynamics of lipid bilayers and the dipole potential ⁇ d. Antimicrobial Resistance
  • Antimicrobial resistance is defined as the ability of a microorganism to survive the effects of an antimicrobial drug or molecule. Antimicrobial resistance can evolve naturally via natural selection, through a random mutation, or through genetic engineering. Also, microbes can transfer resistance genes between one another via mechanisms such as plasmid exchange. If a microorganism carries several resistance genes, it is called multi-drug resistant or, colloquially, a "superbug.” Multi-drug resistance in pathogenic bacteria and fungi are a serious problem in the treatment of patients infected with such organisms. At present, it is tremendously expensive and difficult to create or discover new antimicrobial drugs that are safe for human use. Also, there have been resistant mutant organisms that have evolved challenging all known antimicrobial classes and mechanisms. Hence, few antimicrobials have been able to maintain their long-term effectiveness. Most of the mechanisms of antimicrobial drug resistance are known.
  • micro-organisms exhibit resistance to antimicrobials are: a) Drug inactivation or modification; b) Alteration of target site; c) Alteration of metabolic pathway; and d) Reduced drug accumulation: by decreasing drug permeability and/or increasing active efflux on the cell surface.
  • Staphylococcus aureus is a good example of one of the major resistant bacterial pathogens currently plaguing civilization. This gram positive bacterium is primarily found on the mucous membranes and skin of close to half of the adult world-wide population. S. aureus is extremely adaptable to pressure from all known classes of antibiotics. S. aureus was the first bacterium in which resistance to penicillin was found in 1947. Since then, almost complete resistance has been found to methicillin and oxacillin. The "superbug" MRSA (methicillin resistant Staphylococcus aureus) was first detected in 1961, and is now ubiquitous in hospitals and communities worldwide. Today, more than half of all S. aureus infections in the United States are resistant to penicillin, methicillin, tetracycline and erythromycin. Recently, in what were the new classes of antibiotics
  • glycopeptides and oxazolidinones there have been reports of significant resistance (Vancomycin since 1996 and Zyvox since 2003).
  • CA-MRSA community-associated MRSA
  • Such potentiators and/or inhibitors if not toxic to humans, would be very valuable for the treatment of patients infected with pathogenic and drug-resistant microbes.
  • S. aureus In the United States, as many as 80% of individuals are colonized with S. aureus at some point. Most are colonized only intermittently; 20-30% are persistently colonized. Healthcare workers, persons with diabetes, and patients on dialysis all have higher rates of colonization.
  • the anterior nares are the predominant site of colonization in adults; other potential sites of colonization include the axilla, rectum, and perineum.
  • Daptomycin's mechanism of action involves a calcium-dependent incorporation of the lipopeptide compound into the cytoplasmic membrane of bacteria. On a molecular level, it is calcium binding between two aspartate residues (in the daptomycin molecule) that decreases its net negative charge and permits it to act better with the negatively charged phospholipids that are typically found in the cytoplasmic membrane of gram-positive bacteria. There is generally no interaction with fungi or mammalian cells at therapeutic levels, so it is a very selective molecule.
  • ⁇ p the main component of which is the transmembrane electrical potential gradient ⁇ tT
  • cells cannot make ATP or take up necessary nutrients needed for growth and reproduction.
  • the collapse of ⁇ tT explains the dissimilar (detrimental) effects produced by daptomycin (e.g., inhibition of protein, RNA, DNA, peptidoglycan, lipoteichoic acid, and lipid biosynthesis).
  • Multidrug resistance efflux pumps are now known to be present in gram-positive bacteria, gram-negative bacteria, and other eukaryotic cells. Efflux pumps generally have a poly-specificity of transporters that confers a broad-spectrum of resistance mechanisms. These can strengthen the effects of other mechanisms of antimicrobial resistance such as mutations of the antimicrobial targets or enzymatic modification of the antimicrobial molecules. Active efflux for antimicrobials can be clinically relevant for /3-lactam antimicrobials, macrolides, fluoroquinolones, tetracyclines and other important antibiotic familiess.
  • a microbe With efflux pump-based resistance, a microbe has the capacity to seize an antimicrobial agent or toxic compound and expel it to the exterior (environment) of the cell, thereby reducing the intracellular accumulation of the agent. It is generally considered that the over-expression of one or more of these efflux pumps prevents the intracellular accumulation of the agent to thresholds necessary for their biological activity. Universally in microbes, the efflux of drugs is coupled to the proton motive force that creates electrochemical potentials and/or the energy necessary (ATP) for the needs of these protein pumps. This includes:
  • ABC ATP-binding cassette
  • SMR small multidrug resistance subfamily of the DMT (drug/metabolite transporters) superfamily
  • MFS major facilitator superfamily
  • RND resistance/nodulation/division superfamily
  • the approach of the current invention to inhibit efflux pumps is a general modification (optical depolarization) of the membranes ⁇ within the irradiated area, leading to lower electrochemical gradients that will lower the phosphorylation potential ⁇ Gp and energy available for the pumps functional energy needs. It is also the object of the present invention to have the same photobiological mechanism inhibit the many different anabolic and energy driven mechanisms of the target cells, including absorption of nutrients for normal growth.
  • Reserpine inhibits the activity of Bmr and NorA, two gram-positive efflux pumps, by altering the generation of the membrane proton-motive force ⁇ p required for the function of MDR efflux pumps. Although these molecules are able to inhibit the ABC transporters involved in the extrusion of antibiotics (i.e., tetracycline), the concentrations necessary to block bacterial efflux are neurotoxic in humans.
  • ⁇ -plas-bact During normal cellular metabolism, protons are extruded through the cytoplasmic membrane to form ⁇ -plas-bact. This function also acidifies (lower pH) the narrow region near the bacterial plasma membrane. It has been shown in the gram positive bacterium Bacillus subtilis, that the activities of peptidoglycan autolysins are increased (i.e., no longer inhibited) when the electron transport system was blocked by adding proton conductors. This suggests that ⁇ -plas-bact and AuH + (independent of storing energy for cellular enzymatic functions) potentially has a profound and exploitable influence on cell wall anabolic functions and physiology.
  • ⁇ -plas-bact uncouplers inhibit peptidoglycan formation with the accumulation of the nucleotide precursors involved in peptidoglycan synthesis, and the inhibition of transport of N-acetylglucosamine (GIcNAc), one of the major biopolymers in peptidoglycan.
  • GIcNAc N-acetylglucosamine
  • tachyplesin that decreases ⁇ -plas-bact in gram positive and gram negative pathogens.
  • Antimicrobial compositions and pharmaceutical preparations thereof United States Patent 5,610,139, the entire teaching of which is incorporated herein by reference.
  • This compound was shown at sub-lethal concentrations to have the ability to potentiate the cell wall synthesis inhibitor /3-lactam antibiotic ampicillin in MRSA. It is desirable to couple the multiple influences of an optically lowered ⁇ -plas-bact (i.e., increased cell wall autolysis, inhibited cell wall synthesis, and cell wall antimicrobial potentiation) to any other relevant antimicrobial therapy that targets bacterial cell walls. This is especially relevant in gram positive bacteria such as MRSA that do not have efflux pumps as resistance mechanisms for cell wall inhibitory antimicrobial compounds.
  • the invention provides a method of modifying the dipole potential ⁇ d of all membranes within the path of a NIMELS beam ( ⁇ d-plas-mam, ⁇ d-mito-mam, ⁇ - plas-fungi, ⁇ d-mito-fungi, and ⁇ d-plas-bact) to set in motion the cascade of further alterations of ⁇ and ⁇ p in the same membranes.
  • ⁇ -steady-mam, ⁇ -steady-fungi, ⁇ -steady-Bact, ⁇ -steady-mito-mam and ⁇ -steady- mito-fungi are altered to ⁇ -trans values ( ⁇ -trans-mam, ⁇ -trans-fungi, ⁇ -trans-Bact, ⁇ -trans-mito-mam and ⁇ -trans-mito-fungi).
  • ⁇ -trans values ⁇ -trans-mam, ⁇ -trans-fungi, ⁇ -trans-Bact, ⁇ -trans-mito-mam and ⁇ -trans-mito-fungi.
  • such applied optical radiation may have a wavelength from about 850 nm to about 900 nm, at a NIMELS dosimetry, as described herein. In exemplary embodiments, wavelengths from about 865 nm to about 875 nm are utilized. In further embodiments, such applied radiation may have a wavelength from about 905 nm to about 945 nm at a NlMELS dosimetry. In certain embodiments, such applied optical radiation may have a wavelength from about 925 nm to about 935 nm. In representative non-limiting embodiments exemplified hereinafter, the wavelength employed is 930 nm.
  • Bioenergetic steady-state membrane potentials may be modified, in exemplary embodiments, as noted below, and may employ multiple wavelength ranges including ranges bracketing 870 and 930 nm, respectively.
  • NIMELS parameters include the average single or additive output power of the laser diodes and the wavelengths (870 nm and 930 nm) of the diodes. This information, combined with the area of the laser beam or beams (cm 2 ) at the target site, the power output of the laser system and the time of irradiation, provide the set of information which may be used to calculate effective and safe irradiation protocols according to the invention.
  • a new set of parameters are defined that will take into account the implementation of any different dosimetric value for the NIMELS laser and any MIC value for a given antimicrobial being examined. This can be simply tailored to the NIMELS laser system and methods by creating only a set of variables that quantify CFU' s of pathogenic organisms within any given experimental or treatment parameter with the NIMELS system.
  • NIMELS Potentiation Magnitude Scale NPMS
  • NIMELS lasers inherent phenomenon of reversing resistance and/or potentiating the MIC of antimicrobial drugs while also producing a measure of safety against burning and injuring adjacent tissues, with power, and/or treatment time.
  • the NPMS scale measures the NIMELS effect number (Ne) between 1 to 10, where the goal is to gain a Ne of >4 in reduction of CFU count of a pathogen, at any safe combination of antimicrobial concentration and NIMELS dosimetry.
  • Ne NIMELS effect number
  • CFU count is used here for quantifying pathogenic organism, other means of quantification such as, for example, dye detection methods or polymerase chain reaction (PCR) methods can also be used to obtain values for A, B, and Np parameters.
  • the NTMELS effect number Ne is an interaction coefficient indicating to what extent the combined inhibitory/bacteriostatic effect of an antimicrobial drug is synergistic with the NIMELS laser against a pathogen target without significant harm to healthy tissue at the site of pathogen infection.
  • the NIMELS potentiation number (Np) is a value indicating whether the antimicrobial at a given concentration is synergistic, or antagonistic, to the pathogen target without harm to healthy tissue.
  • N/7 CFU Count of pathogen with (NIMELS + Antimicrobial).
  • Ne 1 then there is no potentiation effect. If Ne > 1 then there is a potentiation effect. If Ne >2 then there is at least a 50% potentiation effect on the antimicrobial. IfNe >4 then there is at least a 75% potentiation effect on the antimicrobial. IfNe >10 then there is at least a 90% potentiation effect on the antimicrobial.
  • the present invention provides systems and methods to reduce the MIC of antimicrobial molecules when the area being treated is concomitantly treated with the NIMELS laser system. If the MIC of an antimicrobial is reduced for a localized and resistant local infection (e.g., skin, diabetic foot, bedsore), the therapeutic efficacy of many of the older, cheaper and safer antimicrobials to treat these infections will be restored.
  • a localized and resistant local infection e.g., skin, diabetic foot, bedsore
  • this invention provides methods and systems that will reduced the MIC of antimicrobial molecules necessary to eradicate or at least attenuate microbial pathogens via a depolarization of membranes within the irradiated field which will decrease the membrane potential ⁇ of the irradiated cells.
  • This weakened ⁇ will cause an affiliated weakening of the proton motive force ⁇ p, and the associated bioenergetics of all affected membranes.
  • this "NIMELS effect" potentiate existing antimicrobial molecules against microbes infecting and causing harm to human hosts.
  • such applied optical radiation has a wavelength from about
  • wavelengths from about 865 nm to about 875 nm are utilized.
  • such applied radiation has a wavelength from about 905 nm to about 945 nm at a NIMELS dosimetry.
  • such applied optical radiation has a wavelength from about 925 nm to about 935 nm. In one aspect, the wavelength employed is 930 nm.
  • irradiation by the wavelength ranges contemplated are performed independently, in sequence, in a blended ratio, or essentially concurrently (all of which can utilize pulsed and/or continuous-wave, CW, operation).
  • NIMELS energy at NIMELS dosimetry to the biological contaminant is applied prior to, subsequent to, or concomitant with the administration of an antimicrobial agent.
  • said NIMELS energy at NTMELS dosimetry can be administered after antimicrobial agent has reached a "peak plasma level" in the infected individual or other mammal. It should be noted that the co-administered antimicrobial agent ought to have antimicrobial activity against any naturally sensitive variants of the resistant target contaminant.
  • the wavelengths irradiated according to the present methods and systems increase the sensitivity of a contaminant to the level of a similar non-resistant contaminant strain at a concentration of the antimicrobial agent of about 0.5 M or less, about 0.1 M or less, or about 0.01 M or less, about 0.005 M or less or about 0.005 M or less.
  • the methods of the invention slow or eliminate the progression of microbial contaminants in a target site, improve at least some symptoms or asymptomatic pathologic conditions associated with the contaminants, and/or increase the sensitivity of the contaminants to an antimicrobial agent.
  • the methods of the invention result in a reduction in the levels of microbial contaminants in a target site and/or potentiate the activity of antimicrobial compounds by increasing the sensitivity of a biological contaminant to an antimicrobial agent to which the biological contaminant has evolved or acquired resistance, without an adverse effect on a biological subject.
  • the reduction in the levels of microbial contaminants can be, for example, at least 10%, 20%, 30%, 50%, 70%, 100% or more as compared to pretreatment levels.
  • the invention provides a system to implement the methods according to other aspects of the invention.
  • a system includes a laser oscillator for generating the radiation, a controller for calculating and controlling the dosage of the radiation, and a delivery assembly (system) for transmitting the radiation to the treatment site through an application region.
  • Suitable delivery assemblies/systems include hollow waveguides, fiber optics, and/or free space/beam optical transmission components. Suitable free space/beam optical transmission components include collimating lenses and/or aperture stops.
  • the system utilizes two or more solid state diode lasers to function as a dual wavelength near-infrared optical source.
  • the two or more diode lasers may be located in a single housing with a unified control.
  • the two wavelengths can include emission in two ranges from about 850 nm to about 900 nm and from about 905 nm to about 945 nm.
  • the laser oscillator of the present invention is used to emit a single wavelength (or a peak value, e.g., central wavelength) in one of the ranges disclosed herein. In certain embodiments, such a laser is used to emit radiation substantially within the about 865-875 nm and the about 925-935 nm ranges.
  • Systems according to the present invention can include a suitable optical source for each individual wavelength range desired to be produced.
  • a suitable solid stated laser diode, a variable ultra-short pulse laser oscillator, or an ion-doped (e.g., with a suitable rare earth element) optical fiber or fiber laser is used.
  • a suitable near infrared laser includes titanium-doped sapphire.
  • Other suitable laser sources including those with other types of solid state, liquid, or gas gain (active) media may be used within the scope of the present invention.
  • a therapeutic system includes an optical radiation generation system adapted to generate optical radiation substantially in a first wavelength range from about 850 nm to about 900 nm, a delivery assembly for causing the optical radiation to be transmitted through an application region, and a controller operatively connected to the optical radiation generation device for controlling the dosage of the radiation transmitted through the application region, such that the time integral of the power density and energy density of the transmitted radiation per unit area is below a predetermined threshold. Also within this embodiment, are therapeutic systems especially adapted to generate optical radiation substantially in a first wavelength range from about 865 nm to about 875 nm.
  • a therapeutic system includes an optical radiation generation device that is configured to generate optical radiation substantially in a second wavelength range from about 905 nm to about 945 nm; in certain embodiments the noted first wavelength range is simultaneously or concurrently/sequentially produced by the optical radiation generation device. Also within the scope of this embodiment, are therapeutic systems especially adapted to generate optical radiation substantially in a first wavelength range from about 925 nm to about 935 nm.
  • the therapeutic system can further include a delivery assembly (system) for transmitting the optical radiation in the second wavelength range (and where applicable, the first wavelength range) through an application region, and a controller operatively for controlling the optical radiation generation device to selectively generate radiation substantially in the first wavelength range or substantially in the second wavelength range or any combinations thereof.
  • the delivery assembly comprises one or more optical fibers having an end configured and arranged for insertion in patient tissue at a location within an optical transmission range of the medical device, wherein the radiation is delivered at a NTMELS dosimetry to the tissue surrounding the medical device.
  • the delivery assembly may further comprise a free beam optical system.
  • the controller of the therapeutic system includes a power limiter to control the dosage of the radiation.
  • the controller may further include memory for storing a patient's profile and dosimetry calculator for calculating the dosage needed for a particular target site based on the information input by an operator.
  • the memory may also be used to store information about different types of diseases and the treatment profile, for example, the pattern of the radiation and the dosage of the radiation, associated with a particular application.
  • the optical radiation can be delivered from the therapeutic system to the application site in different patterns.
  • the radiation can be produced and delivered as a continuous wave (CW), or pulsed, or a combination of each.
  • CW continuous wave
  • pulsed pulsed
  • two wavelengths of radiation can be multiplexed (optically combined) or transmitted simultaneously to the same treatment site.
  • Suitable optical combination techniques can be used, including, but not limited to, the use of polarizing beam splitters (combiners), and/or overlapping of focused outputs from suitable mirrors and/or lenses, or other suitable multiplexing/combining techniques.
  • the radiation can be delivered in an alternating pattern, in which the radiation in two wavelengths are alternatively delivered to the same treatment site.
  • An interval between two or more pulses may be selected as desired according to NIMELS techniques of the present invention. Each treatment may combine any of these modes of transmission.
  • the intensity distributions of the delivered optical radiation can be selected as desired. Exemplary embodiments include top-hat or substantially top-hat (e.g., trapezoidal, etc.) intensity distributions. Other intensity distributions, such as Gaussian may be used.
  • biological contaminants include, but are not limited to, any bacteria, such as, for example, Escherichia, Enterobacter, Bacillus, Campylobacter, Cor ⁇ nebacterium, Klebsiella, Listeria, Mycobacterium, Neiseria, Pseudomonas, Salmonella, Streptococcus, Staphylococcus, Treponema, Vibrio and Yersinia.
  • bacteria such as, for example, Escherichia, Enterobacter, Bacillus, Campylobacter, Cor ⁇ nebacterium, Klebsiella, Listeria, Mycobacterium, Neiseria, Pseudomonas, Salmonella, Streptococcus, Staphylococcus, Treponema, Vibrio and Yersinia.
  • the target site to be irradiated need not be already infected with a biological contaminant. Indeed, the methods of the present invention may be used "prophylactically," prior to infection.
  • catheters e.g., IV catheter, central venous line, arterial catheter, peripheral catheter, dialysis catheter, peritoneal dialysis catheter, epidural catheter), artificial joints, stents, external fixator pins, chest tubes, gastronomy feeding tubes, etc.
  • irradiation may be palliative as well as prophylactic.
  • the methods of the invention are used to irradiate a tissue or tissues for a therapeutically effective amount of time for treating or alleviating the symptoms of an infection.
  • the expression "treating or alleviating” means reducing, preventing, and/or reversing the symptoms of the individual treated according to the invention, as compared to the symptoms of an individual receiving no such treatment.
  • the invention is useful in conjunction with a variety of diseases caused by or otherwise associated with any microbial, fungal, and viral infection (see, Harrison's, Principles of Internal Medicine, 13 th Ed., McGraw Hill, New York (1994), the entire teaching of which is incorporated herein by reference).
  • the methods and the systems according to the invention are used in concomitance with traditional therapeutic approaches available in the art (see, e.g., Goodman and Gilman's, The Pharmacological Basis of Therapeutics, 8th ed, 1990, Pergmon Press, the entire teaching of which is incorporated herein by reference.) to treat an infection by the administration of known antimicrobial agent compositions.
  • antibacterial composition refers to compounds and combinations thereof that are administered to an animal, including human, and which inhibit the proliferation of a microbial infection (e.g., antibacterial, antifungal, and antiviral).
  • a microbial infection e.g., antibacterial, antifungal, and antiviral.
  • the wide breath of applications contemplated include, for example, a variety of dermatological, podiatric, pediatric, and general medicine to mention but a few.
  • the interaction between a target site being treated and the energy imparted is defined by a number of parameters including: the wavelength(s); the chemical and physical properties of the target site; the power density or irradiance of beam; whether a continuous wave (CW) or pulsed irradiation is being used; the laser beam spot size; the exposure time, energy density, and any change in the physical properties of the target site as a result of laser irradiation with any of these parameters.
  • the physical properties e.g., absorption and scattering coefficients, scattering anisotropy, thermal conductivity, heat capacity, and mechanical strength
  • the target site may also affect the overall effects and outcomes.
  • a biological moiety e.g., a mammalian cell, tissue, or organ
  • NIMELS dosimetry parameters lie between known photochemical and photo-thermal parameters in an area traditionally used for photodynamic therapy in conjunction with exogenous drugs, dyes, and/or chromophores, yet can function in the realm of photodynamic therapy without the need of exogenous drugs, dyes, and/or chromophores.
  • the energy density also expressible as fluence, or the product (or integral) of particle or radiation flux and time — for medical laser applications in the art typically varies between about 1 J/cm 2 to about 10,000 J/cm 2 (five orders of magnitude), whereas the power density (irradiance) varies from about IxIO "3 W/cm 2 to over about 10 12 W/cm 2 (15 orders of magnitude).
  • laser exposure duration irradiation time is the primary parameter that determines the nature and safety of laser-tissue interactions.
  • This progression describes a suitable method or basic algorithm that can be used for a NIMELS interaction against a biological contaminant in a tissue.
  • this mathematical relation is a reciprocal correlation to achieve a laser-tissue interaction phenomena.
  • This ratioinale can be used as a basis for dosimetry calculations for the observed antimicrobial phenomenon imparted by NIMELS energies with insertion of NIMELS experimental data in the energy density and time and power parameters.
  • a practitioner is able to adjust the power density and time to obtain the desired energy density.
  • the optical energy is delivered through a uniform geometric distribution to the tissues (e.g., a flat-top, or top-hat progression).
  • a suitable NIMELS dosimetry sufficient to generate ROS can be calculated to reach the threshold energy densities required to reduce the level of a biological contaminant and/or to increase the sensitivity of the biological contaminant to an antimicrobial agent that said contaminant is resistant to, but below the level of "denaturization” and "tissue overheating”.
  • NIMELS dosimetries exemplified herein to target microbes in vivo were from about 125 J/cm 2 to about 700 J/cm 2 and preferably 150 J/cm 2 to about 400 J/cm 2 for approximately 100 to 700 seconds. These power values do not approach power values associated with photoablative or photothermal (laser/tissue) interactions.
  • the intensity distribution of a collimated laser beam is given by the power density of the beam, and is defined as the ratio of laser output power to the area of the circle in
  • the illumination pattern of a 1.5 cm irradiation spot with an incident Gaussian beam pattern of the area 1.77 cm 2 can produce at least six different power density values within the 1.77 cm 2 irradiation area.
  • Tn is from about 50 to about 300 seconds; in other embodiments, Tn is from about 75 to about 200 seconds; in yet other embodiments, Tn is from about 100 to about 150 seconds. In in vivo embodiments, Tn is from about 100 to about 1200 seconds.
  • NIMELS dosimetry encompasses ranges of power density and/or energy density from a first threshold point at which a subject wavelength according to the invention is capable of optically reducing ⁇ in a target site to a second end-point and/or to increase the sensitivity of the biological contaminant to an antimicrobial agent that said contaminant is resistant to via generation of ROS, immediately before those values at which an intolerable adverse risk or effect is detected (e.g., thermal damage such as poration) on a biological moiety.
  • a target site e.g., a mammalian cell, tissues, or organ
  • adverse effects and/or risks at a target site may be tolerated in view of the inherent benefits accruing from the methods of the invention.
  • the stopping point contemplated are those at which the adverse effects are considerable and, thus, undesired (e.g., cell death, protein denaturation, DNA damage, morbidity, or mortality).
  • the power density range contemplated herein is from about 0.25 to about 40 W/cm 2 . In other embodiments, the power density range is from about 0.5 W/cm 2 to about 25 W/cm 2 .
  • Currently preferred embodiments for decolonizing a microbial site on a subject utilize a power density range from about 0.3 W/cm 2 to about 0.7 W/cm 2 when antibacterial coumpounds are coadministered.
  • Currently preferred embodiments for decolonizing a microbial site on a subject utilize an energy density range from about 125 J/cm 2 to about 400 J/cm 2 when antibacterial coumpounds are coadministered.
  • power density ranges can encompass values from about 0.5 W/cm 2 to about 10 W/cm 2 .
  • Power densities exemplified herein are from about 0.5 W/cm 2 to about 5 W/cm 2 .
  • Power densities in vivo from about 1.5 to about 2.5 W/cm 2 have been shown to be effective for various microbes with or without coadministration of antibiotics.
  • Empirical data appears to indicate that higher power density values are generally used when targeting a biological contaminant in an in vitro setting (e.g., plates) rather than in vivo (e.g., toe nail).
  • the energy density range contemplated herein is greater than 50 J/cm 2 but less than about 25,000 J/cm 2 . In other embodiments, the energy density range is from about 750 J/cm 2 to about 7,000 J/cm 2 .
  • the energy density range is from about 1,500 J/cm 2 to about 6,000 J/cm 2 depending on whether the biological contaminant is to be targeted in an in vitro setting (e.g., plates) or in vivo (e.g., toe nail or surrounding a medical device). In certain embodiments (see, in vivo examples below), the energy density is from about 100 J/cm 2 to about 500 J/cm 2 . In yet other in vivo embodiments, the energy density is from about 175 J/cm to about 300 J/cm 2 . In yet other embodiments, the energy density is from about 200 J/cm 2 to about 250 J/cm 2 .
  • the energy density is from about 300 J/cm 2 to about 700 J/cm 2 . In some other embodiments, the energy density is from about 300 J/cm 2 to about 500 J/cm 2 . In yet others, the energy density is from about 300 J/cm 2 to about 450 J/cm 2 .
  • Power densities empirically tested for various in vitro treatment of microbial species were from about 1 W/cm 2 to about 10 W/cm 2 .
  • One of skill in the art will appreciate that the identification of particularly suitable NIMELS dosimetry values within the power density and energy density ranges contemplated herein for a given circumstance may be empirically done via routine experimentation. Practitioners (e.g., dentists) using near infrared energies in conjunction with periodontal treatment routinely adjust power density and energy density based on the exigencies associated with each given patient (e.g., adjust the parameters as a function of tissue color, tissue architecture, and depth of pathogen invasion).
  • a periodontal infection in a light-colored tissue e.g., a melanine deficient patient
  • a light-colored tissue e.g., a melanine deficient patient
  • the darker tissue will absorb near-infrared energy more efficiently, and hence transform these near- infrared energies to heat in the tissues faster.
  • antibiotic resistant bacteria may be effectively treated according to the methods of the present invention.
  • the methods of this invention may be used to augment traditional approaches, to be used in combination with, in lieu of tradition therapy, or even serially as an effective therapeutic approach. Accordingly, the invention may be combined with antibiotic treatment.
  • antibiotic includes, but is not limited to, 0-lactams, penicillins, and cephalosporins, vancomycins, bacitracins, macrolides (erythromycins), ketolides (telithromycin), lincosamides (clindamycin), chloramphenicols, tetracyclines, aminoglycosides (gentamicins), amphotericns, anilinouracils, cefazolins, clindamycins, mupirocins, sulfonamides and trimethoprim, rifampicins, metronidazoles, quinolones, novobiocins, polymixins, oxazolidinone class (e.g., linezolid), glycylcyclines (e.g., tigecycline), cyclic lipopeptides (e.g., daptomycin), pleuromutilins (e.g.,
  • tetracyclines include, but are not limited to, immunocycline, chlortetracycline, oxytetracycline, demeclocycline, methacycline, doxycycline and minocycline and the like.
  • aminoglycoside antibiotics include, but are not limited to, gentamicin, amikacin and neomycin, and the like.
  • a common tenet in the search for inhibitors of drug resistance systems in bacteria, or a potentiator of antimicrobial agents has always been that such agents are preferably nontoxic to the mammalian tissues that are infected, in order to have any intrinsic value.
  • most antimicrobials affect bacterial cellular processes that are not common to the mammalian host, and, hence, are less disruptive to host metabolic processes. If antimicrobials, potentiators, and/or resistance reversal entities were to also affect the mammalian cells in the same manner as they damage the pathogens, over similar concentrations, they could not be used safely as therapeutic agents.
  • the experimental data provided herein supports a universal alteration of ⁇ and ⁇ p among all cell types, and hence leads to the notion that not only the electro-mechanical, but also the electro-dynamical aspects of all cell membranes, have no differing properties that can adequately be separated. This indicates that all cells in the path of the beam are affected with depolarization, not only the pathogenic (non-desired) cells.
  • MRSA EXPERIMENTS The following illustrates the general antibacterial methods according to the invention, using a MRSA model for the in vitro Experiments V and V1TI-XIL A.
  • Experiment Materials and Methods for MRSA :
  • HEK293 cells were seeded into appropriate wells of a 24-well plate at a density of 1 x 10 5 cells/ml (0.7ml total volume) in Freestyle medium (Invitrogen). Cells were incubated in a humidified incubator at 37 0 C in 8% CO 2 for approximately 48 hours prior to the experiment. Cells were approximately 90% confluent at the time of the experiment equating to roughly 3 x 10 5 total cells. Immediately prior to treatment, cells were washed in pre- warmed phosphate buffer saline (PBS) and overlaid with 2 ml of PBS during treatment. After laser treatment, cells were mechanically dislodged from the wells and transferred to 1.5 ml centrifuge tubes. Mitochondrial membrane potential and total glutathione was determined.
  • PBS phosphate buffer saline
  • EXAMPLE V NIMELS IN VITRO TESTS FOR ⁇ ALTERATION IN MRSA, AND E. COLI
  • fluorescent dyes that can be taken up by intact cells and accumulate within the intact cells within 15 to 30 minutes without appreciable staining of other protoplasmic constituents. These dye indicators of membrane potential have been available for many years and have been employed to study cell physiology. The fluorescence intensity of these dyes can be easily monitored, as their spectral fluorescent properties are responsive to changes in the value of the trans-membrane potentials ⁇ - steady.
  • ⁇ i fluorescence intensity in a control cell culture (no laser) subjected to carbocyanine dye
  • ⁇ 2 fluorescence intensity in the same cell culture pre-irradiated with sub-lethal dosimetry from the NIMELS laser
  • the data indicates that the fluorescence of cells is dissipated (less than control of unirradiated or "unlased” cells) by pre-treatment (of the cells) with the NlMELS laser system, indicating that the NIMELS laser interacted with respiratory processes and oxidative phosphorylation of the cells via the plasma membranes.
  • ⁇ i - ⁇ 2 0
  • BacLightTM Bacterial Membrane Potential Kit (B34950, Invitrogen U.S.).
  • the . ⁇ cLightTM Bacterial Membrane Potential Kit provides of carbocyanine dye DiOC2(3) (3,3'-diethyloxacarbocyanine iodide, Component A) and CCCP (carbonyl cyanide 3- chlorophenylhydrazone, Component B), both in DMSO, and a 1 x PBS solution
  • DiOC2(3) exhibits green fluorescence in all bacterial cells, but the fluorescence shifts toward red emission as the dye molecules self associate at the higher cytosolic concentrations caused by larger membrane potentials.
  • Proton ionophores such as CCCP destroy membrane potential by eliminating the proton gradient, hence causing higher green fluorescence.
  • Green fluorescence emission was calculated using population mean fluorescence intensities for control and lased samples at sub-lethal dosimetry: Table 7.
  • Red/green ratios were calculated using population mean fluorescence intensities for control and lased samples at sub-lethal dosimetry: 0
  • the data shows that ⁇ i - ⁇ 2 > 0 as the lased cells had less "Green fluorescence" as seen in Figure 19.
  • These E. coli samples showed clear alteration and lowering of ⁇ - steady-bact to one of ⁇ -trans-bact with sublethal NIMELS dosimetry.
  • ⁇ i fluorescence intensity in a mammalian control cell culture mitochondria (no laser) 0 subjected to a Mitochondrial Membrane Potential Detection Kit.
  • ⁇ 2 fluorescence intensity in the same mammalian cell culture pre-irradiated with sublethal dosimetry from the NTMELS laser and subjected to a Mitochondrial
  • APO LOGIX JC-I Mitochondrial Membrane Potential Detection Kit
  • JC-I mitochondrial membrane potential in cells.
  • JC-I (5,5',6,6'-tetrachloro-l,r,3,3'-tetraethylbenz- imidazolylcarbocyanine iodide) exists as a monomer in the cytosol (green) and also accumulates as aggregates in the mitochondria which stain red.
  • JC-I exists in monomelic form and stains the cytosol green.
  • HEK-293 Human Embryonic Kidney Cells
  • the (APO LOGK JC-I) kit measures membrane potential by conversion of green fluorescence to red fluorescence. The appearance of red color has been measured and plotted, which should only occur in cells with intact membranes, and the ratio of green to red is calculated for both control and lased samples.
  • ROS reactive oxygen species
  • Glutathione is the most abundant thiol (SH) compound in animal tissues, plant tissues, bacteria and yeast. GSH plays many different roles such as protection against reactive oxygen species and maintenance of protein SH groups. During these reactions, GSH is converted into glutathione disulfide (GSSG: oxidized form of GSH). Since GSSG is enzymatically reduced by glutathione reductase, GSH is the dominant form in organisms.
  • DTNB (5,5'-Dithiobis(2-nitrobenzoic acid)), known as Ellman's Reagent, was developed for the detection of thiol compounds.
  • glutathione recycling system by DTNB and glutathione reductase created a highly sensitive glutathione detection method.
  • DTNB and glutathione react to generate 2-nitro-5-thiobenzoic acid and glutathione disulfide (GSSG). Since 2-nitro-5- thiobenzoic acid is a yellow colored product, GSH concentration in a sample solution can be determined by the measurement at 412 nm absorbance.
  • GSH is generated from GSSG by glutathione reductase, and reacts with DTNB again to produce 2-nitro-5 -thiobenzoic acid. Therefore, this recycling reaction improves the sensitivity of total glutathione detection.
  • ROS glutathione antioxidant system
  • ⁇ -trans-bact is proof of generation of ROS with sub-lethal alteration of Trans-membrane ⁇ -steady-bact to one of ⁇ -trans-bact.
  • a reduction in total glutathione in C. albicans at sub-lethal NIMELS dosimetry that alters ⁇ -steady-mito-fungi to ⁇ -trans-mito-fungi and subsequently ⁇ -steady-fungi to one of ⁇ -trans-fungi, is proof of generation of ROS with sub-lethal alteration of Transmembrane ⁇ -steady-mito-fungi to ⁇ -trans-mito-fungi and subsequently ⁇ -steady-fungi to one of ⁇ -trans-fungi.
  • Erythromycin's mechanism of action is to prevent growth and replication of bacteria by obstructing bacterial protein synthesis. This is accomplished because erythromycin binds to the 23 S rRNA molecule in the 5OS of the bacterial ribosome, thereby blocking the exit of the growing peptide chain thus inhibiting the translocation of peptides. Erythromycin resistance (as with other marcolides) is rampant, wide spread, and is accomplished via two significant resistance systems:
  • Trimethoprim is an antibiotic that has historically been used in the treatment of urinary tract infections. It is a member of the class of antimicrobials known as dihydrofolate reductase inhibitors. Trimethoprim's mechanism of action is to interfere with the system of bacterial dihydrofolate reductase (DHFR), because it is an analog of dihydrofolic acid. This causes competitive inhibition of DHFR due to a 1000 fold higher affinity for the enzyme than the natural substrate.
  • DHFR bacterial dihydrofolate reductase
  • trimethoprim inhibits synthesis of the molecule tetrahydrofolic acid.
  • Tetrahydrofolic acid is an essential precursor in the de novo synthesis of the DNA nucleotide thymidylate.
  • Bacteria are incapable of taking up folic acid from the environment (i.e., the infection host) and are thus dependent on their own de novo synthesis of tetrahydrofolic acid. Inhibition of the enzyme ultimately prevents DNA replication.
  • Trimethoprim resistance generally results from the overproduction of the normal chromosomal DHFR, or drug resistant DHFR enzymes. Reports of trimethoprim resistance S. aureus have indicated that the resistance is chromosomally of the mediated type or is encoded on large plasmids. Some strains have been reported to exhibit both chromosomal and plasmid-mediated trimethoprim resistance.
  • trimethoprim In the gram positive pathogen S. aureus, resistance to trimethoprim is due to genetic mutation, and there have been no reports that trimethoprim is actively effluxed out of cells. Efflux Pumps in Bacteria
  • a major route of drug resistance in bacteria and fungi is the active export (efflux) of antibiotics out of the cells such that a therapeutic concentration in not obtained in the cytoplasm of the cell.
  • Active efflux of antibiotics (and other deleterious molecules) is mediated by a series of transmembrane proteins in the cytoplasmic membrane of gram positive bacteria and the outer membranes of gram negative bacteria.
  • antibiotic resistance that is mediated via efflux pumps, is most relevant in gram positive bacteria for marcolides, tetracyclines and fluoroquinolones.
  • ⁇ -lactam efflux mediated resistance is also of high clinical relevance.
  • ⁇ i sub-lethal dosimetry from the NIMEL laser system on MRSA as a control
  • ⁇ 2 is the same sub-lethal dosimetry from the NIMEL laser system on MRSA with the addition of trimethoprim at resistant MIC just below effectiveness level
  • ⁇ 3 is the same sub-lethal dosimetry from the NIMEL laser system on MRSA with the addition of erythromycin at resistant MIC just below effectiveness level.
  • Tetracycline is considered a bacteriostatic antibiotic, meaning that it hampers the growth of bacteria by inhibiting protein synthesis. Tetracycline accomplishes this by inhibiting action of the bacterial 30S ribosome through the binding of the enzyme aminoacyl-tRNA. Tetracycline resistance is often due to the acquisition of new genes, which code for energy-dependent efflux of tetracyclines, or for a protein that protects bacterial ribosomes from the action of tetracyclines. Rifampin:
  • Rifampin is a bacterial RNA polymerase inhibitor, and functions by directly blocking the elongation of RNA.
  • Rifampicin is typically used to treat mycobacterial infections, but also plays a role in the treatment of methicillin-resistant Staphylococcus aureus (MRSA) in combination with fusidic acid, a bacteriostatic protein synthesis inhibitor. There are no reports of rifampin resistance via efflux pumps in MRSA.
  • ⁇ i sub-lethal dosimetry from the NIMEL laser system on MRSA as a control
  • ⁇ 2 is the same sub-lethal dosimetry from the NIMEL laser system on MRSA with the addition of tetracycline at resistant MIC just below effectiveness level
  • ⁇ 3 is the same sub-lethal dosimetry from the NIMEL laser system on MRSA with the addition of rifampin at resistant MIC just below effectiveness level.
  • Methicillin is a /3-lactam that was previously used to treat infections caused by gram-positive bacteria, particularly /3-lactamase-producing organisms such as S. aureus that would otherwise be resistant to most penicillins, but is no longer clinically used.
  • MRSA methicillin-resistant S. aureus
  • methicillin acts by inhibiting the synthesis of peptidoglycan (bacterial cell walls). It has been shown in the gram positive bacterium Bacillus subtilis, that the activities of peptidoglycan autolysins are increased (i.e., no longer inhibited) when the ETS was blocked by adding proton conductors. This suggests that ⁇ -plas-bact and ⁇ tl* (independent of storing energy for cellular enzymatic functions) potentially has a profound and exploitable influence on cell wall anabolic functions and physiology.
  • ⁇ -plas-bact uncouplers inhibit peptidoglycan formation with the accumulation of the nucleotide precursors involved in peptidoglycan synthesis, and the inhibition of transport of N-acetylglucosamine (GIcNAc), one of the major biopolymers in peptidoglycan.
  • GIcNAc N-acetylglucosamine
  • Bacitracin will potentiate the multiple influences of an optically lowered ⁇ -plas- bact on a growing cell wall (i.e., increased cell wall autolysis, inhibited cell wall synthesis). This is especially relevant in gram positive bacteria such as MRSA, that do not have efflux pumps as resistance mechanisms for cell wall inhibitory antimicrobial compounds.
  • ⁇ i sub-lethal dosimetry from the NIMEL laser system on MRSA as a control
  • ⁇ 2 is the same sub-lethal dosimetry from the NIMEL laser system on MRSA with the addition of methicillin at resistant MIC just below effectiveness level and
  • ⁇ , - ⁇ 2 0
  • EXAMPLE XI Assessment of the impact of Sub-lethal doses of NIMELS Laser on MRSA with Bacitracin and ⁇ -plas-bact inhibition of cell wall synthesis
  • Bacitracin is a mixture of cyclic polypeptides produced by Bacillus subtilis. As a toxic and diff ⁇ cult-to-use antibiotic, bacitracin cannot generally be used orally, but is used topically.
  • Bacitracin interferes with the dephosphorylation of the C 55 -isoprenyl pyrophosphate, a molecule which carries the building blocks of the peptidoglycan bacterial cell wall outside of the inner membrane in gram negative organisms and the plasma membrane in gram positive organism.
  • ⁇ -plas-bact uncouplers inhibit peptidoglycan formation with the accumulation of the nucleotide precursors involved in peptidoglycan synthesis, and the inhibition of transport of N-acetylglucosamine (GIcNAc), one of the major biopolymers in peptidoglycan.
  • GIcNAc N-acetylglucosamine
  • Bacitracin potentiates the multiple influences of an optically lowered ⁇ -plas-bact on a growing cell wall (i.e., increased cell wall autolysis, inhibited cell wall synthesis). This is especially relevant in gram positive bacteria such as MRSA, that do not have efflux pumps as resistance mechanisms for cell wall inhibitory antimicrobial compounds.
  • ⁇ i sub-lethal dosimetry from the NIMEL laser system on MRSA as a control
  • ⁇ 2 is the same sub-lethal dosimetry from the NIMEL laser system on MRSA with the addition of bacitracin at resistant MIC just below effectiveness level and
  • ⁇ ! - ⁇ 2 0
  • the NIMELS laser and its concomitant optical ⁇ -plas-bact lowering phenomenon is synergistic with cell wall inhibitory antimicrobials in MRSA. Without wishing to be bound by theory, this most likely functions via the inhibition of anabolic (periplasmic) ATP coupled functions as MRSA does not have efflux pumps for bacitracin.
  • NIMELS approach to impact upon the viability of various commonly found microorganisms at the wavelengths described herein.
  • the microorganisms exemplified include E. coli K-12, multi-drug resistant E. coli, Staphylococcus aureus, methicillin-resistant S. aureus, Candida albicans, and Trichophyton rubrum.
  • NIMELS parameters include the average single or additive output power of the laser diodes, and the wavelengths (870 nm and 930 nm) of the diodes. This information, combined with the area of the laser beam or beams (cm 2 ) at the target site, provide the initial set of information which may be used to calculate effective and safe irradiation protocols according to the invention.
  • the power density of a given laser measures the potential effect of NIMELS at the target site. Power density is a function of any given laser output power and beam area, and may be calculated with the following equations: For a single wavelength:
  • Power Density (W/cm 2 ) Laser (1) Output Power + Laser (2) Output Power
  • Total energy distribution may be measured as energy density (Joules/cm 2 ). As discussed infra, for a given wavelength of light, energy density is the most important factor in determining the tissue reaction. Energy density for one NIMELS wavelength may be derived as follows:
  • the energy density may be derived as follows:
  • Energy Density (Joule/cm2) Power Density (1) (W/cm 2 ) * Time (Sees) + Power Density (2) (W/cm 2 ) * Time (Sees)
  • a practitioner may use either the energy density (J/cm 2 ) or energy (J), as well as the output power (W), and beam area (cm 2 ) using either one of the following equations:
  • Treatment Time (seconds) Energy (Joules)
  • the therapeutic system may also include a computer database storing all researched treatment possibilities and dosimetries.
  • the computer (a dosimetry and parameter calculator) in the controller is preprogrammed with algorithms based on the above-described formulas, so that any operator can easily retrieve the data and parameters on the screen, and input additional necessary data (such as: spot size, total energy desired, time and pulse width of each wavelength, tissue being irradiated, bacteria being irradiated) along with any other necessary information, so that any and all algorithms and calculations necessary for favorable treatment outcomes can be generated by the dosimetry and parameter calculator and hence run the laser.
  • the bacterial kill rate (as measured by counting Colony Forming Units or CFU on post-treatment culture plates) ranged from 93.7% (multi-drug resistant E. col ⁇ ) to 100% (all other bacteria and fungi).
  • E. coli Kl 2 liquid cultures were grown in Luria Bertani (LB) medium (25 g/L).
  • Plates contained 35 mL of LB plate medium (25 g/L LB, 15 g/L bacteriological agar). Culture dilutions were performed using PBS. All protocols and manipulations were performed using sterile techniques. B. Growth Kinetics
  • Liquid cultures of E. coli Kl 2 were set up as described previously. An aliquot of 100 ⁇ L was removed from the subculture and serially diluted to 1:1200 in PBS. This dilution was allowed to incubate at room temperature approximately 2 hours or until no further increase in O.D. 600 was observed in order to ensure that the cells in the PBS suspension would reach a static state (growth) with no significant doubling and a relatively consistent number of cells could be aliquoted further for testing.
  • C. albicans ATCC 14053 liquid cultures were grown in YM medium (21g/L, Difco) medium at 37°C.
  • a standardized suspension was aliquoted into selected wells in a 24-well tissue culture plate. Following laser treatments, 1 OO ⁇ L was removed from each well and serially diluted to 1 : 1000 resulting in a final dilution of 1 :5xlO 5 of initial culture. 3x100 ⁇ L of each final dilution were spread onto separate plates. The plates were then incubated at 37 0 C for approximately 16-20 hours. Manual colony counts were performed and recorded.
  • T. rubrum ATCC 52022 liquid cultures were grown in peptone-dextrose (PD) medium at 37 0 C.
  • PD peptone-dextrose
  • a standardized suspension was aliquoted into selected wells in a 24 -well tissue culture plate. Following laser treatments, aliquots were removed from each well and spread onto separate plates. The plates were then incubated at 37 0 C for approximately 91 hours. Manual colony counts were performed and recorded after 66 hours and 91 hours of incubation. While control wells all grew the organism, 100% of laser-treated wells as described herein had no growth.
  • a digital photograph of each plate was also taken.
  • Experimental data in vitro demonstrates that if the threshold of total energy into the system with 930 run alone of 5400 J and an energy density of 3056 J/cm 2 is met in 25% less time, 100% antibacterial efficacy is still achievable.
  • EXAMPLE XV DOSIMETRY VALUES FOR NMELS LASER WAVELENGTH 870 NM IN VITRO
  • This synergistic ability is significant to human tissue safety, as the 930 nm optical energy heats up tissues at a greater rate than the 870 nm optical energy, and it is beneficial to a mammalian system to produce the least amount of heat possible during treatment.
  • This simultaneous synergistic ability is significant to human tissue safety, as the 930 nm optical energy, heats up a system at a greater rate than the 870 nm optical energy, and it is beneficial to a mammalian system to produce the least amount of heat possible during treatment.
  • Experimental in vitro data also demonstrates that when applied at safe thermal dosimetries, there is less additive effect with the 830 nm wavelength, and the NIMELS 930 nm wavelength when they are used simultaneously.
  • experimental in vitro data demonstrates that 17% less total energy, 17% less energy density, and 17% less power density is required to achieve 100 % E. coli antibacterial efficacy when 870 nm is combined simultaneously with 930 nm vs. the commercially available 830 nm. This, again, substantially reduces heat and harm to an in vivo system being treated with the NIMELS wavelengths.
  • microbial cells in 75% of the diabetic patients tested were all at least 100,000 CFU/gm, and in 37.5% of the patients, quantities of microbial cells were greater than 1,000,000 (lxlO 6 )CFU (see Brown et al., Ostomy Wound Management, 401:47, issue 10, (2001), the entire teaching of which is incorporated herein by reference).
  • the treated and a control untreated suspension were diluted and plated in triplicate on trypic soy agar with or without 30 ⁇ g/ml methicillm After 24hrs of growth at 37 °C colonies were counted. CFU (colony forming units) were compared between the plates with and without methicillin for both control (untreated) and treated MRSA.
  • the treated and a control untreated suspension were diluted and plated in tnplicate on trypic soy agar with or without 30 ⁇ g/ml methicillin. After 24hrs of growth at 37 0 C colonies were counted.
  • T +l 2 ml aliquots are dispensed into pre-designated wells in 24-well plates and transferred to NOMIR (8 24-well plates total)
  • the treated and a control untreated suspension were diluted and plated m pentuplicate on trypic soy agar with or without 30 ⁇ g/ml methicillin
  • the treated and a control untreated suspension were diluted and plated in pentuplicate on trypic soy agar with or without 30 ⁇ g/ml methicillin (Groups A4 and B4), 10 0 5 ⁇ g/ml penicillin G (Groups C4 and D4) or 4 ⁇ g/ml erythromycin (Groups E4 and F4)
  • Example XXI Laser Treatment for Microbial Reduction and Elimination of Nasal Colonization of MRSA
  • the Nomir Near Infrared Microbial Elimination Laser System (NOVEONTM Model 1120 dual-wavelength diode laser was employed for this study.
  • the laser operates in continuous wave format at two wavelengths, 870 nm (+/- 5 nm) and 930 nm (+/- 5 nm).
  • This device is a class II non-significant risk laser device.
  • the laser sources of this device are semiconductor laser arrays that are optically coupled to form a single fiber laser output.
  • the delivery system consists of a single flexible optical fiber. The device delivers continuous wave laser light only.
  • the device is designed specifically to effect microbial cell optical destruction, while preserving and without substantial damage optically or thermally to the human tissue at the infection site being irradiated.
  • the NOVEONTM system was designed to harness the known photo-lethal characteristics of these precise energies to kill pathogenic microorganism at far lower energy levels and heat deposition than is generally necessary to kill pathogens using laser-based thermal sterilization means.
  • topical intra-nasal antimicrobial agents are recognized as the preferred method for preventing (distal-site) infections because of their demonstrated effectiveness and widespread desire to minimize the use of systemic antimicrobials.
  • the design of this protocol includes a number of important factors have been considered. Foremost is the need to assure that the amount of energy used in the Nares is safe for the nasal and nares tissues. Furthermore, significant human and histological tests have been done with the Noveon laser in the areas that the study is treating
  • a cylindrical diffusing optical fiber tip for near infrared light delivery was fabricated specifically for uniform illumination of a length of 1.5 cm, to then be placed in a transparent catheter (of given width) to prevent placement too far anteriorly in the nostril, and guarantee a uniform power density at all tissues proximal to the catheter within the nostril.
  • the tip included an optically transmissive, light diffusing, fiber tip assembly having an entrance aperture through a proximal reflector, a radiation-scattering, transmissive material (e.g. a poly-tetrafluoroethylene tube) surrounding an enclosed cavity (e.g. a cylindrical void filled with air or another substantially non-scattering, transparent medium), and a distal reflective surface.
  • a radiation-scattering, transmissive material e.g. a poly-tetrafluoroethylene tube
  • enclosed cavity e.g. a cylindrical void filled with air or another substantially non-scattering, transparent medium
  • distal reflective surface e.g. a distal reflective surface
  • the scattering medium has a prescribed inner diameter. This inner diameter of the scattering material is designed such that the interaction with this material and the multiple reflections off of the cavity reflectors interact to provide a substantially proscribed axial distribution of laser radiation over the length of the tip apparatus. Suitable choices of tip dimensions provide control over the emitted axial and azimuthal energy distributions.
  • Diode lasers in the near infrared range have a very low absorption coefficient in water; hence, they achieve relatively deep optical penetration in tissues that contain 80% water (such as the dermis, the oral mucosa, bone and the gingiva.
  • the depth of penetration (before photon absorption) of the greatest amount of the incident energy is about 1.5 cm. This allows the near infrared laser energy to pass through water with minimal absorption, producing thermal effects deeper in the tissue and the photons are absorbed by the deeper tissue pigments.
  • This photobiology allows for controlled, deeper soft-tissue irradiation and decontamination, as the photons that emerge from the dispersion tip in a uniform dosimetry from the diffusing tip absorbed by blood and other tissue pigments.
  • This investigational protocol was designed to demonstrate that the Noveon Laser treatment is able to produce reduction in Nasal carriage of MRSA in patients with previously "culture positive" history.
  • This investigational protocol was an open-label study of subjects who are colonized with MRSA in the nares (nostril). The study was done in two parts.
  • H202 3% OTC hydrogen peroxide was applied to a cotton pledget for application to the subject prior to irradiation. This was inserted in the nose for 120 seconds and then removed. The subjects were then given doses of phototherapeutic near infrared radiation as described.
  • Application of generic topical Antimicrobial The subjects were first given doses of phototherapeutic near infrared radiation as described. Subsequently, 2% erythromycin paste was applied to a cotton tipped swab for application to the subject following irradiation. The swab was inserted approximately 1 cm in to the anterior nares and rotated 360 degrees several times and removed. Patients were instructed to perform the exact application procedure 3 times a day for the remaining 5 days.
  • the NOVEONTM laser was used for two (2) six-minute treatments in each nostril on day (1) and day (3) of the study.
  • the dosimetries used are shown in the Table TT, below.
  • the laser was calibrated before the first treatment of the day. Intermittent temperature testing of the treatment site was performed on each subject using a noncontact infrared thermometer (Raytek Minitemp), 30-60 second intervals. If a temperature of 110 F degrees was reached, or the patient complained of pain, the laser treatment was interrupted and only resumed when the patient was comfortable. Inturruption only occurred once in 40 treatments (20 nostrils x 2 treatments over three days), and was resumed 30 seconds later to completion.
  • Tables 42-44 represent the mean values of the triplicate CFU counts and plating of each swab from each nostril, pre and post laser therapy (for this data set the
  • a second human study was conducted, to further evaluate the therapeutic potential of the NOVEONTM laser system, including its ability to reverse drug resistance in bacteria.
  • the study was conducted in a similar manner as Part One, above. Outcome measures assessed included both laboratory study and clinical observations.
  • Positive anterior nares cultures were obtained in six patients (12 nostrils) having nasal colonization of MRSA or MSSA, before initiating bacterial photodamage through doses of phototherapeutic near infrared radiation.
  • One patient had MRSA only, 3 had MSSA only, and 2 had both MRSA and MSSA. All MRSA and MSSA were cultured and verified to be resistant to erythromycin.
  • Antimicrobial paste (generic 2% erythromycin) was placed on a cotton tipped swab for application after phototherapeutic near infrared radiation.
  • the swab was inserted approximately 1 cm in to the anterior nares of the subject, rotated 360 degrees several times and removed.
  • the application of erythromycin was maintained for 3 times a day for the remainder of the study.
  • the laser was calibrated before the first treatment of the day and between each patient.
  • the NOVEONTM laser was used for four six-minute treatments of the nares at the following sets of dosimetries (Tables 45), which were evaluated for safety in previous studies.
  • Tables 45 were evaluated for safety in previous studies.
  • each patient underwent exposure with the Noveon for 7 minutes (energy density - 207 J/cm2) to each anterior nostril on Day 1 and on Day 3.
  • the treatment was divided into two parts, an approximately 3-minute exposure using a combination of 870 run and 930 nm and an approximately 3-minute exposure of 930 nm
  • S. aureus was identified by colony morphology and StaphaurexTM latex agglutination test (Murex Biotech Limited, Dartford, Kent, UK). Samples were frozen and stored at - 2O 0 C. Results:
  • NOVEONTM laser exposure at a non-damaging energy density and approximately physiologic temperatures, re-sensitized erythromycin resistant MRSA and MSSA to 2% generic erythromycin paste. Photodamage to the organism results in sensitivity to antibiotics in otherwise drug resistant strains.
  • the NOVEONTM laser system provides for local reduction of drug resistant microbes and a concomitant reduction of bio-burden in: e.g., wounds, mucosal or cutaneous tissues, and other colonized or infected areas such as surgical sites and tissue/medical device interfaces, which are prone to contamination particularly by nosocomial strains of microbes frequently having multidrug resistance phenotypes.
  • FIG 17 illustrates a schematic diagram of a therapeutic radiation treatment device according one embodiment of the present disclosure.
  • the therapeutic system 110 includes an optical radiation generation device 112, a delivery assembly 114, an application region 116, and a controller 118.
  • the optical radiation generation device includes one or more suitable lasers, Ll and L2.
  • a suitable laser may be selected based on a degree of coherence.
  • a therapeutic system can include at least one diode laser configured and arranged to produce an output in the near infrared region.
  • Suitable diode lasers can include a semiconductor materials for producing radiation in desired wavelength ranges, e.g., 850nm-900nm and 905nm-945nm.
  • Suitable diode laser configurations can include cleave-coupled, distributed feedback, distributed Bragg reflector, vertical cavity surface emitting lasers (VCSELS), etc.
  • the delivery assembly 114 can generate a "flat-top" energy profile for uniform distribution of energy over large areas.
  • a diffuser tip 10 may be included which diffuses treatment light with a uniform cylindrical energy profile in an application region 116 (e.g. a nasal cavity as described in the example above).
  • the optical radiation generation device 112 can include one or more lasers, e.g., laser oscillators Ll and L2.
  • one laser oscillator can be configured to emit optical radiation in a first wavelength range of 850 nm to 900 nm
  • the other laser oscillator can be configured to emit radiation in a second wavelength range of 905 nm to 945 nm.
  • one laser oscillator is configured to emit radiation in a first wavelength range of 865 nm to 875 nm
  • the other laser oscillator 28 is configured to emit radiation in a second wavelength range of 925 nm to 935 nm.
  • the geometry or configuration of the individual laser oscillators may be selected as desired, and the selection may be based on the intensity distributions produced by a particular oscillator geometry or configuration.
  • the delivery assembly 114 includes an elongated flexible optical fiber 118 adapted for delivery of the dual wavelength radiation from the oscillators 26 and 28 to diffuser tip 10 to illuminate the application region 116.
  • the delivery assembly 14 may have different formats (e.g., including safety features to prevent thermal damage) based on the application requirements.
  • the delivery assembly 114 or a portion thereof e.g. tip 10) may be constructed with a size and with a shape for inserting into a patient's body.
  • the delivery assembly 114 may be constructed with a conical shape for emitting radiation in a diverging-conical manner to apply the radiation to a relatively large area.
  • Hollow waveguides may be used for the delivery assembly 114 in certain embodiments.
  • Other size and shapes of the delivery assembly 14 may also be employed based on the requirements of the application site, hi exemplary embodiments, the delivery assembly 114 can be configured for free space or free beam application of the optical radiation, e.g., making use of available transmission through tissue at NMELS wavelengths described herein. For example, at 930nm (and to a similar degree, 870nm), the applied optical radiation can penetrate patient tissue by up to 1 cm or more.
  • Such embodiments may be particularly well suited for use with in vivo medical devices as described herein.
  • the controller 118 includes a power limiter 124 connected to the laser oscillators Ll and L2 for controlling the dosage of the radiation transmitted through the application region 116, such that the time integral of the power density of the transmitted radiation per unit area is below a predetermined threshold, which is set up to prevent damages to the healthy tissue at the application site.
  • the controller 118 may further include a memory 126 for storing treatment information of patients.
  • the stored information of a particular patient may include, but not limited to, dosage of radiation, (for example, including which wavelength, power density, treatment time, skin pigmentation parameters, etc.) and application site information (for example, including type of treatment site (lesion, cancer, etc.), size, depth, etc.).
  • the memory 126 may also be used to store information of different types of diseases and the treatment profile, for example, the pattern of the radiation and the dosage of the radiation, associated with a particular type of disease.
  • the controller 118 may further include a dosimetry calculator 128 to calculate the dosage needed for a particular patient based on the application type and other application site information input into the controller by a physician.
  • the controller 118 further includes an imaging system for imaging the application site. The imaging system gathers application site information based on the images of the application site and transfers the gathered information to the dosimetry calculator 128 for dosage calculation. A physician also can manually calculate and input information gathered from the images to the controller 118.
  • the controller may further include a control panel 130 through which, a physician can control the therapeutic system manually.
  • the therapeutic system 10 also can be controlled by a computer, which has a control platform, for example, a WINDOWS TM based platform.
  • the parameters such as pulse intensity, pulse width, pulse repetition rate of the optical radiation can be controlled through both the computer and the control panel 30.
  • Figures 18a-18d show different temporal patterns of the optical radiation that can be delivered from the therapeutic system to the application site.
  • the optical radiation can be delivered in one wavelength range only, for example, in the first wavelength range of 850 nm to 900 ran, or in the range of 865 nm to 875 nm, or in the second wavelength range of 905 nm to 945 nm, or in the range of 925 nm to 935 nm, as shown in Figure 18a.
  • the radiation in the first wavelength range and the radiation in the second wavelength range also can be multiplexed by a multiplex system installed in the optical radiation generation device 112 and delivered to the application site in a multiplexed form, as shown in Figure 18b.
  • the radiation in the first wavelength range and the radiation in the second wavelength range can be applied to the application site simultaneously without passing through a multiplex system.
  • Figure 18c shows that the optical radiation can be delivered in an intermission-alternating manner, for example, a first pulse in the first wavelength range, a second pulse in the second wavelength range, a third pulse in the first wavelength range again, and a fourth pulse in the second wavelength range again, and so on.
  • the interval can be CW (Continuous Wave), one pulse as shown in Figure 18c, or two or more pulses (not shown).
  • Figure 18d shows another pattern in which the application site is first treated by radiation in one of the two wavelength ranges, for example, the first wavelength range, and then treated by radiation in the other wavelength range.
  • the treatment pattern can be determined by the physician based on the type, and other information of the application site. Delivery Apparatus
  • delivery devices that may be used in the delivery assembly 114 of the exemplary NIMELS system are described herein.
  • delivery devices of the types described herein may make up an end portion of the fiber 119 of the delivery assembly 114. These delivery devices may operate with or without the diffuser tip 10.
  • the improved delivery device 510 includes an optical fiber 512 having a fiber-optic core 514, a cladding layer 516 circumferentially disposed around the core 514, and an outer buffer coating 518 circumferentially disposed around the cladding layer 516.
  • the outer buffer coating 518 is removed from the emission end of the optical fiber 512, and the fiber-optic core 514 and cladding 516 extend to position close to but not in optical contact with the optical element 530.
  • the separation of these two elements determines the imaging properties of the emitted beam.
  • the optical element 530 may be a focusing lens having a focal plane located at or near the end face of the fiber optic core 514, thereby imaging the core to an image plane. In other embodiments, the optical element 530 may abut the end of the optical fiber 512.
  • housing 520 is adapted to accept the fiber optic 512 and the region having buffer coating 516 removed. At its distal end, the housing 520 firmly holds and aligns the optical element 530 with the fiber core 514.
  • housing 520 is an elastic cuff. The cuff is stretched perpendicular to the longitudinal axis of the optical fiber 512 providing a press fit that holds the optical components 512, 530 together.
  • the coupling and positioning between multiple source fibers, the integrating optical fiber, the housing, the buffer, and the optical element enables a substantially improved precise and stable uniform beam in a durable construction unaffected by the extreme thermal cycling of sterilization and other treatments.
  • the housing is made from a material having a coefficient of thermal expansion approximately equal to the coefficient of thermal expansion of the buffer. In this manner, both the housing and the buffer will thermally expand (and contract) approximately the same amount, thus minimizing the effects of heat cycling on the device.
  • the housing is made from a polymer material having an anisotropic, non-linear Young's modulus with the greater value co-axial with the optical fiber.
  • the housing is made with a low index (e.g. lower than that of the optical element) of refraction material to act as a cladding to the encased optical component.
  • the housing may have an index of refraction less than 1.7, less than 1.6, less than 1.5, less than 1.4, less than 1.3, or even less.
  • an image 514' of the core 514 of the optical fiber 512 is focused by optical element 530 at the design region of illumination 524. This results in a uniform spatial intensity or 'top hat' at the design region of illumination 524 as shown in graph of intensity verses axial spatial position in Fig. 20(b). Accordingly, in some embodiments, delivery device 510 produces a beam with a substantially non-gaussian beam profile.
  • the beam profile may vary in intensity by less than 10%, less than 5%, or even less than 1% across the substantially the entire profile of the beam). For example, as shown in Fig. 20(b) the beam is very uniform except at a small peripheral region where the intensity quickly drops to near zero.
  • Fig. 21 shows a construction of the delivery device 510 having a protrusion on the interior face of the 5 housing 520 locking at a recess on the buffer coating 518.
  • the recess 518 may be pre-fabricated or the result of the assembly of the components.
  • a GRTN lens 532 is used in place of the optical element 530.
  • the GRIN lens 532 normally abuts the coupling core 514.
  • any suitable optical element may be used to produce any desired illumination pattern at an illumination region.
  • the optical element may include a lens (e.g.
  • a spherical lens aspherical lens, compound lens, singlet, doublet, etc.
  • a GRIN lens a diffractive element, a diffusive element, a hologram, a concentrating element, and a collimator.
  • more than one optical element may be used.
  • Fig. 23 shows a preferred integrating embodiment of delivery device 510.
  • a multiplicity of sources 534, 534' may be of different wavelengths or wavelength ranges (e.g. either distinct ranges or partially overlapping ranges).
  • Each of the sources 534, 534' are optically coupled at an input end, respectively, to the source output optical fibers 536 and 538.
  • the source output fibers each transmit light to an output end optically couples to the core 514 of the principal integrating optical fiber 512.
  • This construction enables a transmission efficiency of up to about 92% or more and a uniform mixing of the individual sources during the transmission in the principal fiber 512.
  • sources 534, 534' may be lasers Ll and L2 of the NIMELS system shown in Fig. 17.
  • one or more optical elements may be used to couple light from the source output fibers 436 and 538 to integrating fiber 512.
  • each of the source output optical fibers may have a diameter less than that of the integrating fiber.
  • the total combined packed diameter of the source output optical fibers may be less than that of the integrating fiber.
  • the combined diameter of the output ends of the source output fibers 536 and 538 is less than the diameter of the core 514 of the integrating fiber 512.
  • Figs. 24a-24d show a method for the construction of the device described above.
  • Fig. 24a one end of the housing 520 is stretched over the end of the fiber 512.
  • an alignment template 540 is temporarily affixed to the optical fiber 512 enabling the accurate insertion and alignment of the optical element 530, as shown in Figs. 24b and 24c.
  • the alignment template 540 may be of a solid material and a split construction 540, 541'.
  • fig, 24d the alignment template 540 is removed.
  • the above described press-fit process provides the precise, controlled alignment and seating of the elements. It may be accomplished in a stable, temperature controlled environment, within the elastic modulus of the materials, further eliminating alignment errors and post manufacturing transitions. For example, the entire process may be accomplished ay room temperature, at temperatures below 500 C, below 400 C, below
  • the process has substantial advantages over a heat-shrink construction by eliminating the potential heat damage to the cladding, the heat-induced stresses in the optical element and the resulting misalignment from post- manufacturing cooling. Further, it permits the use of more stable optical, cladding, buffer and housing materials. These include but are not limited to new improved polymer optics whose operational temperature is less than 500 C.
  • fluoro-polymer materials such Teflon® materials and the like, are used as materials for the housing 520 to inhibit contact-adhesion between the tip assembly and biological tissue during procedures.
  • the Teflon® material is a Teflon® FEP material (a polyperfluoroethylene- 6 propylene copolymer).
  • Teflon® materials such as Teflon® PFA (a polytetra-fluoroethylene polymer with perfluoro-alkoxy side chains) and Teflon® PTFE (polytetrafluoroethylene) also can be useful in certain applications.
  • Fig. 25 shows an embodiment wherein two sides of a low index of refraction clamshell clamp 542, which when compressed holds and position the fiber and optical element.
  • the clamp may have an index of refraction less than the optical element 530, thereby serving as a cladding for the element.
  • delivery device 510 used with a NTMELS, but it is to be understood that the may be used with any phototherapeutic devices, e.g., any multi- wavelength device where it is desirable to integrate light output from multiple sources.
  • treatment system 110 employs a diffusion tip 10 to diffuse therapeutic treatment light delivered from a therapeutic light source by optical fiber 118.
  • the tip operates to provide a desired illumination profile (i.e. emitted intensity profile) at the application region 116.
  • a desired illumination profile i.e. emitted intensity profile
  • a substantially uniform cylindrical illumination profile is desirable.
  • Other embodiments of tip 10 may be used to direct treatment light to other areas such as tissue spaces (e.g. the periodontal pocket or within a joint e.g. in an orthopedic surgical procedure), interfaces between body tissue and other surfaces (e.g. the surface of an implantable medical device), over a wide area such as a dermal surface, etc.
  • tissue spaces e.g. the periodontal pocket or within a joint e.g. in an orthopedic surgical procedure
  • interfaces between body tissue and other surfaces e.g. the surface of an implantable medical device
  • diffuser tip 10 is an optically transmissive, light diffusing, fiber tip assembly having an entrance aperture through a proximal reflector, a radiation- scattering, transmissive material surrounding an enclosed void (e.g. a cylindrical cavity), and a distal reflective surface is disclosed.
  • a portion of the radiation is scattered in a cylindrical (or partly cylindrical) pattern along the distal portion of the fiber tip. Radiation, which is not scattered during this initial pass through the tip, is reflected by at least one surface of the assembly and returned through the tip. During this second pass, the remaining radiation, (or a portion of the returning radiation), is scattered and emitted from the proximal portion of the tube.
  • the scattering medium has a prescribed inner diameter.
  • This inner diameter of the scattering material is designed such that the interaction with this material and the multiple reflections off of the cavity reflectors interact to provide a substantially proscribed axial distribution of laser radiation over the length of the tip apparatus.
  • Various embodiments provide a diffusing tip with control over the emitted axial and azimuthal energy distributions.
  • the diffusion techniques and devices disclosed herein are generally applicable for diffusing radiation from an optical fiber to provide a larger exposure area for photo- illumination. Some embodiments are particularly useful as part of a fiber-optic based medical laser system in which a lower aspect ratio of length to diameter than typical diffuser designs is desirable. Suitable laser systems include those described herein along with those described in U.S. Patent Application No. 11/930,941 filed 31 October 2007 and U.S. Patent Application Serial No. 11/981,486 filed 31 October 2007; the entire contents of both of which applications are incorporated herein by reference.
  • Some embodiments provide substantially uniform energy distribution to a major portion of the exposure area. Some embodiments provide for constructing and implementing circumferential and/or sideways emitting diffusing tip assemblies for optical fibers to direct laser radiation in a radially outward pattern relative to the fiber's longitudinal axis.
  • optical fiber is intended to encompass optically transmissive waveguides of various shapes and sizes.
  • Some diffusing tip designs are intended for a higher aspect ratio of length to diameter. Typical aspect ratios for prior art diffusing tip technologies are usually from 20 to 1 and higher, (e.g. 1 mm diameter and 20 mm length). Some embodiments of the diffusion tips described herein allow for producing diffusing tip assemblies with aspect ratios of about 10 or less, about 1 or less, or about .1 or less. For example, one embodiment may be used to produce uniform emission from a diffusing tip with 10mm diameter and 10 mm length. The aspect ratio of this diffusing tip would be 1. In one embodiment, a diffusive tip assembly is disclosed for diffusing radiation from an optical fiber.
  • the tip assembly includes a light transmissive, tubular housing, alignable with, and adapted to receive, the distal end of the fiber and serve as a diffusive scattering medium for light that has been emitted by the optical fiber.
  • the assembly further includes a reflective cavity formed by reflectors on each side of the diffusive tube, such that the light is scattered by the tube on it's first pass through the tube, and is emitted outward to the illumination site. The un-scattered portion of the illumination is reflected back to further interact with the scattering tube. This second pass illumination is then scattered outward by the scattering tube to complement the light emitted on the first pass to produce the desired illumination profile. Additional 2nd, 3rd and 4th reflections with subsequent scattering from the diffusing tube can be added to produce additional homogeneity of the emitted axial energy profiles.
  • the reflective surfaces of the apparatus can also be modified to effect non-planar forms.
  • Reflective structures are disclosed which control the spatial distribution of the light emitted from the tip. These techniques and structures permit, for example, an evenly distributed orthogonal projection of the radiation.
  • the diameter of the tubular scattering material and/ or the length of the diffusing tip can be controlled such that the diffusion of the radiation during the initial and reflected paths are complementary.
  • the cumulative energy profile, or fluence, along at least a portion of the fiber tip can be rendered uniform.
  • substantially uniform is commonly used in the field of phototherapy to describe light diffusers that possess a uniformity of about +/-15% or less of the average intensity of light emitted from the diffusive tip assembly.
  • embodiments of tip 10 provide a mechanism for substantially uniform cylindrical illumination of biological structures and other illumination applications.
  • the diffuser tips described herein may be used to apply therapeutic light at NIMELS dosimetry and wavelengths (e.g. as described above) without exhibiting heating to temperatures which are unwanted or intolerable at the treatment site (i.e. temperatures that would cause substantial thermal damage at the site, or discomfort to a patient undergoing treatment).
  • the diffuser tip may absorb about 20% or less of the therapeutic light delivered from a therapeutic source at NIMELS dosimetry and wavelengths.
  • the diffuser tip may be operated to deliver therapeutic light at NIMELS dosimetry and wavelengths for treatment times on the order tens of seconds or on the order of minutes or more while remaining at an operating temperature of 110 F degrees or less, or 1 OOF degrees or less.
  • an optical fiber diffusive tip assembly 10 including and optical fiber 12 having a light transmissive core 14, a cladding 16, a proximal first mirror 18, a diffusing tube 20, and a distal second mirror 22.
  • the end face of fiber 12 is inserted through an aperture 24 in the first mirror 18.
  • the operation of diffuser tip 10 is shown where the radiation 28 from the fiber 12 expanding at angle defined by numerical aperture NA intersects the diffusing tube 20.
  • a distal portion 32 of diffusing tube 20 is illuminated by light which intersects the tub and is scattered radially outward.
  • the diffusing tube 28 shows the operation of diffuser tip 10 where the radiation 28 from the fiber 12 which does not interact with the diffusing tube 20 reflects back into the void 30 from the distal mirror 22.
  • the diffusing tube may be constructed from many materials with suitable optical and scattering properties, but preferably from those materials with low absorption (e.g. less than 20% of the incident intensity) and high scattering properties, of which expanded pory-tetrafluoroethylene (PTFE) is an example. Radiation which propagates into the diffusing material 20 is efficiently scattered in region 32 with a portion escaping the material 20 and emerging into external space. A portion of the radiation 28 is returned into the void 30 where it is propagated into another region of the diffusing tube 20 or continues to be reflected by first or second mirrors 18, 22.
  • PTFE expanded pory-tetrafluoroethylene
  • Fig. 29 shows the diffusive tip assembly 10. Shown are the optical fiber 12, a proximal first mirror 18, a diffusing tube 20, and a distal second mirror 22 having a curved reflective face.
  • Fig. 23 shows a graph of the tip's light intensity (ordinate) as a function of axial position (ordinate) for a planar distal mirror 22 (left graph) and for a curved distal second mirror 22 (right graph).
  • the void in the diffusion tube 20 was larger than preferred to provide a uniform axial intensity distribution. In this design regime a properly curved second mirror 22 can increase the uniformity of the tip's axial intensity profile.
  • Fig. 30 shows a method of construction of the curved second mirror 22 where a small form or sphere 34 is placed between the flexible reflective films 36 and the distal backing of the second mirror assembly 22.
  • a flexible reflective film is 3M Vikuity enhanced specular reflective film.
  • Fig. 32 shows a cross section of the fiber tip 10 having a proximal first mirror 18 with diffuse reflective surface 40.
  • An example of this kind of film is white Backlight reflector sheet made of a polyester film such as that produced by Kimoto.
  • This alternative construction enables an improved radiant uniformity when factors including, but not limited to, cost and simplicity of construction are considered.
  • the distal mirror 36 may be specular or diffuse, preferably specular.
  • the proximal mirror may be specular or diffuse, preferably diffuse.
  • Fig. 33 shows an embodiment of diffuser tip 10 where the diffusion tube is enclosed by a disposable, sterile, test tube sized appropriately for the diffusion tip assembly.
  • a preferred disposable tube is made from Polypropylene, due to its high transmission of visible and near infrared light, non shattering nature and ability to withstand high temperatures. Alternate materials could include polycarbonate or Pyrex glass.
  • diffuser tip 10 is autoclavable and reusable.
  • diffuser tip 20 is detachbly connected to fiber 12, and is disposable.
  • Fig. 34 shows how the size of void 30 may be adjusted relative to the length of tip 10 in order to maintain a substantially uniform axial intensity distribution.
  • the length of the tip that can be used to provide uniform illumination is increased up to the the point at which the wall thickness becomes extremely thin. Above that point, other properties of tip 10 (e.g. the curvature of one or more of the reflectors) must be adjusted to maintain uniform illumination.
  • Fig. 35 shows an intensity profile of a diffusion tip 10. Axial intensity distribution uniformity of +/- 8% is achieved using the techniques taught herein. Note also the lack of any light outside the cavity region. This indicates that the cumulative illumination pattern for this tip is a substantially uniform cylindrical pattern characterized by uniform axial intensity distribution directed outward in the radial direction along diffusion tube 20 and substantially proscribed illumination in the axial direction.
  • the preferred void size or preferred length for a given void size may be calculated as follows. Light from fiber 12 propagates radial outward in a cone from aperture 24 as defined by the numerical aperture NA and the length L of tube 20. The radius of this cone at the distal end of tube 20 is defined as R. The radius of void 30 is defined at R v0ld . In one embodiment, for uniform axial illumination, the area of the void is chosen to be chosen to be 40% of the area of the cone at the distal end of tube 20 such that
  • a kit 300 includes a diffuser tip 10, e.g.
  • Tip 10 may be may be detachably connected to the treatment system and may be configured to apply therapeutic light at NIMELS dosimetry and wavelengths without becoming subject to unwanted levels of heating at treatment site.
  • Kit 300 also includes antimicrobial (e.g. antibiotic or antifungal) application 301 which is potentiated by treatment light delivered from the diffuser tip .
  • antimicrobial application 301 may be potentiated by the treatment light to treat a resistant biological contaminate at a treatment site even though the antimicrobial application alone would be ineffective in treating the contaminate.
  • antimicrobial application 302 may be erythromycin potentiated by NIMELS treatment light to treat antibiotic resistant bacteria such as MRSA or MSSA.
  • antimicrobial application 301 may be a topical application (e.g. a paste) or may be administered in other ways known in the art (e.g. ingested orally, administered intravenously, etc.)
  • Kit 300 may also include instructions 302 instructing the use of antimicrobial application 301 in conjunction with potentiating treatment light diffused by tip 10.
  • the instructions may provide guidance as to the types of contaminates which may be treated using the kit, along with information regarding suitable dosimetry for the treatment light (e.g. NMELS dosimetry).
  • the instructions may be provided in any media including print or electronic formats.
  • Kit 300 may be contained in suitable packaging 303, e.g. a box or pouch.
  • packaging may include sterile packaging.
  • tip 10 may be sterilized and packed in a sterile container.
  • diffusion tip 10 may include any number of sensors, e.g., temperature sensors which may communicate (by wire or wirelessly) with therapeutic system 110. Information provided by these sensors may be used to control applied dosages of therapeutic light, e.g. for safety purposes. Tip 10 may also include one or more cooling devices (e.g. a thermoelectric cooler), or attachments suitable for engagement with external cooling devices (e.g. tubular plumbing for circulation of cooling fluids). It is to be understood that, as used herein, the phrases "light”, “optical”, etc. are not limited to the visible spectrum, but may refer to electromagnetic radiation at any wavelength including, e.g., the infrared.

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Abstract

L'invention concerne un appareil de délivrance optique, comprenant : une fibre optique s'étendant entre une extrémité distale ayant une face d'extrémité distale, et une extrémité proximale ayant une face d'extrémité proximale, un élément optique positionné pour recevoir la lumière émise depuis la face d'extrémité distale, et diriger la lumière vers une zone d'éclairage ; et un boîtier non métallique contenant la fibre optique et l'élément optique, et maintenant la position relative de la fibre optique et de l'élément optique.
PCT/US2009/053752 2002-08-28 2009-08-13 Appareil de délivrance de lumière thérapeutique, procédé et système Ceased WO2010019800A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US13/474,320 US8983257B2 (en) 2002-08-28 2012-05-17 Therapeutic light delivery apparatus, method, and system

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US8840108P 2008-08-13 2008-08-13
US61/088,401 2008-08-13

Related Parent Applications (2)

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US11/930,941 Continuation-In-Part US7713294B2 (en) 2002-08-28 2007-10-31 Near infrared microbial elimination laser systems (NIMEL)
US11/981,486 Continuation-In-Part US20090299263A1 (en) 2002-08-28 2007-10-31 Near-Infrared electromagnetic modification of cellular steady-state membrane potentials

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CN103038683A (zh) * 2010-05-19 2013-04-10 三菱铅笔株式会社 光学准直器和使用了光学准直器的光学连接器
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EP3532771A1 (fr) * 2016-10-25 2019-09-04 Rakuten Medical, Inc. Dispositifs de diffusion de lumière destinés à être utilisés en photo-immunothérapie
US10702706B2 (en) 2013-07-16 2020-07-07 Nomir Medical Technologies, Inc. Apparatus, system, and method for generating photo-biologic minimum biofilm inhibitory concentration of infrared light
CN114028733A (zh) * 2021-12-03 2022-02-11 固安翌光科技有限公司 Oled屏体及光疗装置
WO2022260650A1 (fr) * 2021-06-08 2022-12-15 Lumeda Inc. Applicateur de surface optique avec diffuseur intégré

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CN105629389A (zh) * 2010-04-16 2016-06-01 三菱铅笔株式会社 光学准直器以及光学准直器用保持构件
EP2560036A4 (fr) * 2010-04-16 2013-10-02 Mitsubishi Pencil Co Collimateur optique, connecteur optique le comprenant, et élément de support pour collimateur optique
CN102985861A (zh) * 2010-04-16 2013-03-20 三菱铅笔株式会社 光学准直器、使用了该光学准直器的光学连接器以及光学准直器用保持构件
KR101425298B1 (ko) * 2010-04-16 2014-08-01 미쓰비시 엔피쯔 가부시키가이샤 광콜리메이터 및 그를 이용한 광커넥터, 및, 광콜리메이터용 유지부재
CN103038683A (zh) * 2010-05-19 2013-04-10 三菱铅笔株式会社 光学准直器和使用了光学准直器的光学连接器
US9638860B2 (en) 2011-10-18 2017-05-02 Mitsubishi Pencil Company, Limited Optical coupling member and optical connector using the same, and optical coupling member holding member
CN103890626A (zh) * 2011-10-18 2014-06-25 三菱铅笔株式会社 光耦合构件和使用该光耦合构件的光连接器、以及光耦合构件用保持构件
US10702706B2 (en) 2013-07-16 2020-07-07 Nomir Medical Technologies, Inc. Apparatus, system, and method for generating photo-biologic minimum biofilm inhibitory concentration of infrared light
EP3532771A1 (fr) * 2016-10-25 2019-09-04 Rakuten Medical, Inc. Dispositifs de diffusion de lumière destinés à être utilisés en photo-immunothérapie
EP3663816A1 (fr) * 2016-10-25 2020-06-10 Rakuten Medical, Inc. Dispositifs de diffusion de lumière destiné à être utilisé en photoimmunothérapie
US10908341B2 (en) 2016-10-25 2021-02-02 Aspyrian Therapeutics Inc. Frontal light diffusing device for use in photoimmunotherapy
WO2022260650A1 (fr) * 2021-06-08 2022-12-15 Lumeda Inc. Applicateur de surface optique avec diffuseur intégré
US11666778B2 (en) 2021-06-08 2023-06-06 Lumeda Inc. Optical surface applicator with integrated diffuser
CN114028733A (zh) * 2021-12-03 2022-02-11 固安翌光科技有限公司 Oled屏体及光疗装置
CN114028733B (zh) * 2021-12-03 2024-02-13 固安翌光科技有限公司 Oled屏体及光疗装置

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