text:
Patent application title: Natural Photodynamic Agents and their use
Merle E. Olson (Calgary, CA)
Merle E. Olson (Calgary, CA)
Nicholas D. Allan (Calgary, CA)
IPC8 Class: AA61K36906FI
Class name: Drug, bio-affecting and body treating compositions plant material or plant extract of undetermined constitution as active ingredient (e.g., herbal remedy, herbal extract, powder, oil, etc.) containing or obtained from zingiberaceae (e.g., afromonun, cardemon, ginger, turmeric, etc.)
Publication date: 2010-10-21
Patent application number: 20100266716
Patent application title: Natural Photodynamic Agents and their use
Merle E. Olson
Nicholas D. Allan
WILLIAM J BUNDREN;THE LAW OFFICE OF WILLIAM J BUNDREN
Origin: MILLERSVILLE, MD US
IPC8 Class: AA61K36906FI
Publication date: 10/21/2010
Patent application number: 20100266716
The present invention is safe photodynamic agents and their use in
treating microbial contamination.
1. A composition for the treatment infection or contamination comprising
at least one photodynamic agent comprising a natural or synthetic food
2. The composition of claim 1 wherein the food dye is selected from the group consisting of annatto extract, turmeric, porphyrin containing compositions, and combinations thereof.
3. The composition of claim 1 wherein the concentration of food dye is sufficient to inactivate one or more microorganisms when the food dye is activated by a source of light.
4. A method for treating infections and contaminants on surfaces comprising:a) Administering photodynamic agents selected from a group consisting of natural or synthetic food dyes;b) Activating the agent by exposing the agent to a source of light; andc) Inactivating organism and microorganisms.
5. The method of claim 4 wherein administering comprises administering to a surface comprising hard surfaces, food surfaces, and living human, animal and plant tissue.
6. The method of claim 4 wherein the treatment of infections and contaminants comprises treating one or more organisms or microorganisms.
7. The method of claim 4 wherein the organisms and microorganisms are viral, bacterial, fungal, algal or parasites.
This application is claims priority from U.S. Provisional
Application No. 60/982,419 filed Oct. 25, 2007.
I. FIELD OF THE INVENTION
This invention relates to compositions comprising natural photodynamic agents and methods for treating and preventing contamination.
This invention also relates to the use of photoactive natural products that can be safely applied to hard surfaces, food surfaces, animal and plant surfaces to prevent, treat, decontaminate or eliminate infections, contaminants, spoilage microorganisms and foodborne pathogens.
II. BACKGROUND OF THE INVENTION
The photoactive agent that is used for therapeutic applications is a photosensitizer. Photosensitizers have shown great promise in cancer therapy. See, for example, Peng et al, Ultrastructural Pathology, 20:109 (1996), Reddi, J. Photochem. Photobiol. B: Biol., 37:189 (1997), Jori, J. Photochem. Photobiol. B: Biol., 36:87 (1996), Calzavara-Pinton et al, J. Photochem. Photobiol. B: Biol., 36:225 (1996), Geze et al, J. Photochem. Photobiol. B: Biol., 20:23 (1993) and Spikes, J. Photochem. Photobiol. B: Biol., 6:259 (1990), the disclosures of each of which are hereby incorporated by reference herein in their entirety. Upon application of the appropriate light, photosensitizers can photochemically (e.g., through photooxidation, photoreduction and the like) change into a form that is toxic to the cell or surrounding tissue. For example, following excitation of a photosensitizer to a long-lived excited singlet and/or triplet state, a targeted tumor is destroyed either by free radical products (a Type I mechanism) generated by quantum energy transfer or by the highly reactive singlet oxygen species (a Type II mechanism). Major biological target molecules of the singlet oxygen species and/or free radical products include nucleic acids, enzymes and cell membranes. A secondary therapeutic effect of the present methods involves the release of pathophysiologic products such as prostaglandins, thromboxanes and leukotrienes by tissue exposed to the effects of activated photosensitizers.
To date, only Photofrin® has been approved by health boards in Canada, Japan, the Netherlands, and the United States. In spite of the favorable results obtained with Photofrin®, some important factors still limit the efficacy of PDT, including its complex composition, the low extinction coefficient in the red spectral region and the prolonged cutaneous phototoxicity. Therefore, there is a need for more ideally suited photosensitizers.
The present invention addresses the growing need for safe and effective methods of treatment of contaminants, spoilage microorganisms, and human, animal, plant, and food borne pathogens in a wide range of industries.
III. SUMMARY OF THE INVENTION
The present invention is the use of one or more food dyes or colors as a photodynamic agent (PDA), including in photodynamic therapy, to treat or remove microbial contamination. As used herein, food colors or dyes refers to natural, synthetic, Lake, and blended food colors.
It has been discovered that certain edible or ingestible food dyes are equal to or superior to synthetic chemical photodynamic agents. The advantages of such dyes are their non-toxic nature, their ability to be safely consumed, and their breakdown to safe and environmental friendly products. Such photoactive dyes are effective against pathogenic and spoilage microorganisms (bacteria, fungi, algae, viruses, protozoa).
This invention teaches a method to treat or decontaminate a surface, food product, infected animal or plant tissue through the use of a safe natural or synthetic food coloring agent that has photodynamic properties.
The present invention provides a composition for the treatment of infections and contaminants using one or more of the photodynamic agents of the present invention.
An advantage of the present invention is that it employs non-toxic agents, all of which are known to be safe for use and/or consumption.
IV. DETAILED DESCRIPTION OF THE INVENTION
The present invention comprises a composition comprising one or more natural or synthetic photoactive agents, typically one or more food dyes or colors. The present invention also involves using these photoactive agents to treat contamination or infection caused by microorganisms.
Some embodiments of the invention include contacting the microorganism with a composition comprising one or more natural or synthetic photoactive agents, and exposing the photoactive agent to a light source appropriate to activate the photoactive agent. In accordance with the present invention, contacting a microorganism with an activated photoactive agent results in microorganism apoptosis.
In preferred embodiments of the invention, the photoactive agents are selected from natural, synthetic, Lake, or blended food dyes or colors. In the most preferred embodiments of the invention, food dyes include one or more natural photoactive agents selected from the group consisting of annatto extract (contains the carotenoids bixin and norbixin), turmeric, heme, and porphyrin.
The present invention comprises administering at least one photodynamic agent(s) to one or more surfaces, activating said photodynamic agent by exposing the surface and/or microorganism to photoradiation, thereby inactivating one or more organisms or microorganisms.
Some embodiments of the invention include contacting or treating a smooth surface with a PDA of the present invention. In these embodiments of the invention, it has been found that treating smooth surfaces sometimes provides better results, due in part to ease with which light may contact the surface. That is, rough surfaces are sometimes more difficult to expose light sufficiently to decontaminate the surface.
In one embodiment of the present invention, the photodynamic agents are effective in killing microorganisms in the planktonic and biofilm state.
Upon irradiation with light, the agents of the present invention can generate active oxygen species, including singlet oxygen (1O2) superoxide anion radical (O2.sup.-), and hydroxyl radical (HO.sup.-), and possesses high photodynamic activity.
Photodynamic agents of the present invention may include compounds which preferentially adsorb to nucleic acids, thus focusing their photodynamic effect upon microorganisms and viruses with little or no effect upon accompanying cells or proteins.
Another embodiment of the present invention comprises photodynamic agents that provide treatment or elimination of planktonic and biofilm microorganism from surfaces including, but not limited to living and non-living hard surfaces, food surfaces and living tissue surfaces.
In a further embodiment of the present invention the organism and microorganisms are viral, bacterial, fungal, algal or parasitic. Microorganisms include viruses (both extracellular and intracellular), bacteria, bacteriophages, fungi, algae, parasites (nematode, cestode, trematode, arthropod), and insects.
Examples of microorganisms include but are not limited to Pseudomonas, E. coli, Campylobacter, Salmonella, Listeria, Staphylococcus, Aspergillus, and Fusarium.
The present invention also comprises a method of treating a disease, infection or contamination by administering a sufficient amount of at least one PDA agent of the present invention, and photoactivating the agent(s). Typically, the PDA agent may be activated, using known procedures, e.g., by exposing the derivative to a pre-determined wavelength of light, including natural sunlight.
The present invention also comprises a method of photodynamic treatment comprising administering a composition comprising one or more PDA agents of the present invention.
In the present invention photoradiation may comprise light in the visible spectrum, the ultraviolet spectrum, or light in both the visible and ultraviolet spectra. Any suitable wavelength or wavelengths of light may be used in any proportion and energy that produces the desired level of inactivation of microorganisms.
The surface that has been treated with the natural photodynamic agent is exposed to photoradiation of the appropriate wavelength to activate the photosensitizer, using an amount of photoradiation sufficient to activate the photosensitizer as described herein, but less than that which would cause non-specific damage to the hard surface, meat, vegetable produce or living biological tissue. The wavelength used will depend on the photosensitizer selected and composition of the surface. Nonspecific damage is damage that damages all components.
In the present invention, the photoradiation in both the ultraviolet and visible spectra may be supplied concurrently or sequentially, with the visible portion preferably being supplied first. The photoradiation source may be a simple lamp or may consist of multiple lamps radiating at differing wavelengths. The photoradiation source should be capable of delivering a sufficient amount of light to activate the photosensitizer.
In the present invention materials which may be treated include any surface where contamination may be found. Surface, as used herein, is intended to be broadly defined as including any place that can be exposed to light. It is intended to exclude in vivo applications, except where surgery might expose a surface. The following provides examples of such uses, a list that is not intended to limit the scope of the invention.
The photoactive agents of the present invention may be used on any surface where microbial contamination is an issue. Exemplary surfaces include, but are not limited to non-living materials such as hard surfaces (steel, plastic, glass, wood); a hard surface sanitizer; a hard surface disinfectant; a cold surface sanitizer (e.g., refrigerator, cold room, freezer); industrial biofouling inhibition and/or treatment (e.g., pulp and paper mills, mine tailings, oil/fuel/gas pipelines, milk transport lines, food transport lines, fuel lines, aviation fuel lines); medical devices; medical device coatings; food storage containers (including covers, treatment, or impregnated into the container material); Pule storage containers (including covers, treatment, or impregnated into the container material); contact lense compositions, including sanitizers and disinfectants; anti-corrosion agents, including but not limited to microbial induced corrosion;
The photoactive agents of the present invention may be used on any food or plant, as an additive or as a sanitizer, disinfectant, or preservative. Exemplary uses include on meat (e.g., beef, pork, and poultry; processed meats; hot dogs, manufactured foods), vegetables (e.g., lettuce, sprouts, and potatoes), fruit (e.g., apples, pears, and peaches); living plants (vegetables, fruits, ornamentals, seeds, flowers, cereal crops, oil crops, trees, bushes, shrubs); as a foliar spray (e.g., anti-bacterial, anti-parasitic, anti-aphid, anti-locust, anti-fungal sanitizer, food sanitizer, disinfectant); as a seed treatment (e.g., anti-bacterial, anti-parasitic, anti-aphid, anti-locust, anti-fungal sanitizer, food sanitizer, disinfectant); as a food additive to extend the life of processed and non-processed foods (including extending the storage life).
The photoactive agents of the present invention may be used on human and animal tissues (e.g., skin, eyes, mucus membranes); human, animal, or plant wound treatment; human or animal mucosal infection treatment; nasal MSRA treatments; human or animal ocular infection treatments, alone or in combination with eye drops;
The photoactive agents of the present invention may be used as an additive to any other composition where it might be useful to include a disinfectant, sanitizer, or the like. Examples include but are not limited to cosmetics (e.g., as a preservative or additive), including for anti-microbial or anti-aging cosmetics; a paint or ink additive (e.g., a hard surface sanitizer); in fabrics or textiles (e.g., consumer products such as furniture, clothing, sprouting wear and equipment); as an odor preventative or treatment; for textile detergent additives; for pet litter (e.g., to prevent or treat odors and/or as an anti-toxiplasma agent); as a waste water additive; as a grey water additive; as a potable water additive; as a soap additive; in a shampoo composition; in any detergent composition; a paint or stain additive (e.g., to prevent staining).
The photoactive agents of the present invention may be used in human, animal, or plant wound treatment compositions and products, including but not limited to bandages, sutures, ointments or compositions.
The photoactive agents of the present invention may be used for peptic ulcer treatment systems (e.g., H. pylori induced). The photoactive agents of the present invention are particularly useful in any organic, green, environmentally friendly, natural, or natural based composition.
The photoactive agents of the present invention may also be used in combination with, or included in a kit for, a light system or source.
A kit in accordance with the present invention may include one or more food colors or dyes; one or media for dissolving the color; one or more media for applying, contacting, spraying, or distributing the color; one or more light systems or sources; and combinations of any of the above.
The photoactive agents of the present invention are preferably used as a hard surface cleaner, disinfectant, or sanitizer; as a food cleaner, disinfectant, or sanitizer; or as a seed or foliar cleaner, disinfectant, or sanitizer.
In a further embodiment of the present invention the treatment is particularly useful in greenhouses with natural or artificial lighting. The technology can be used to treat viral, fungal, bacterial and parasitic plant diseases which affect leaves or the seed/fruit itself. The advantage is the plant or seed can be safely consumed following application of the photodynamic agent.
The photoactive agents are used in the compositions of the present invention in any amount sufficient to treat microorganisms and/or to treat or prevent contamination. Typically, the appropriate amount or concentration is the amount needed to generate sufficient singlet oxygen to affect the microorganism. One skilled in the art will recognize that the appropriate concentration for a particular photodynamic agent will depend in part on the type of light being used to activate the agent, composition's ingredients, the particular microorganism being treated, the structure of the microorganism (e.g., biofilm or planktonic, among others), and/or the type of delivery (e.g., spray or immersion, among other). Further, the examples suggest that some of the photodynamic agents are effective only within a concentration range. One skilled in the art may determine the range for a particular photodynamic agent by using conventional assays to determine the concentration where the activated agent has little effect on the microorganism (i.e., the low end of the concentration range) and by using conventional assays to determine the concentration where the amount of photodynamic agents inhibits the light source from activating the agent.
Generally, a concentration range from about 0.01% to about 80%, by weight, is suitable for most photodynamic agents under most conditions. The preferred concentration range is from about 0.1% to about 10% by weight, more preferably about 0.01% to about 1% by weight. It is intended however that the invention should not be limited by the concentration of the photodynamic agent in the composition.
One skilled in the art can determine other effective combinations of light, photodynamic agent, and concentration by conducting experiments known in the art, including but not limited to high through put screening, MIC testing, MBEC testing, bioassays, dose titration experiments (manual or robotic), computer modeling. Exemplary assays are shown in the examples. The particular assays described in the examples and the patents cited therein are particularly useful because they are all high throughput assays. Therefore, a significant number of possible photodynamic agents may be determined for both effectiveness and concentration in a short amount of time using only simple experiments.
In accordance with the present invention, a composition comprising annatto, annatto extract, norbixin, annatto powder, or annatto derivatives may be used in a concentration range from about 0.01% to about 80%, by weight, preferably between about 0.01% and about 15%, most preferably between about 1% and about 10%. For example, the examples show that annatto extract is useful against Staph A at concentrations greater than about 1% using either LED light or fluorescent light; and against Salmonella at concentrations greater than about 0.01% using either LED light or fluorescent light.
In accordance with the present invention, a composition comprising heme (hemin and/or hemoglobin) may be used in a concentration range from about 0.01% to about 80%, by weight, preferably between about 0.01% and about 15%, most preferably between about 1% and about 10%. For example, the examples show that hemin and hemoglobin is useful against Salmonella at concentrations greater than about 0.01% using fluorescent light or greater than about 1% using LED light.
In accordance with the present invention, a composition comprising turmeric may be used in a concentration range from about 0.01% to about 80%, by weight, preferably between about 0.01% and about 15%, most preferably between about 1% and about 10%. For example, the examples show that turmeric against Salmonella at concentrations greater than about 0.01% using either fluorescent light and greater than about 1% using LED light; and against Staph A using greater than about 0.1% using fluorescent light.
In accordance with the present invention, a composition comprising porphyrin may be used in a concentration range from about 0.01% to about 80%, by weight, preferably between about 0.01% and about 15%, most preferably between about 1% and about 10%. For example, the examples show that porphyrin against Salmonella at concentrations greater than about 0.01% using either fluorescent light and greater than about 1% using LED light; and against Staph A using greater than about 0.1% using fluorescent light.
One skilled in the art can determine other effective combinations (e.g., of light, of food dye, and/or of food dye concentration) by conducting experiments known in the art. Exemplary tests include but are not limited to high throughput screening, MIC testing, MBEC testing, bioassays, dose titration experiments (manual or robotic), and computer modeling.
The compositions of the present invention may be prepared in various formulations depending on intended use and/or delivery mechanism, e.g., for topical or cutaneous administration. Topical or cutaneous delivery may also include aerosol formulations. The composition may be water based or oil based; and/or may be in the form of a liquid, gel, cream, ointment, lotion, or oil.
The compositions may also contain conventional pharmaceutical carriers and excipients appropriate for the type of administration contemplated. Other agents which may be included are stabilizing agents and skin penetration enhancing agents. Such solutions also may contain pharmaceutically acceptable buffers, emulsifiers, surfactants, and electrolytes such as sodium chloride. Acceptable carriers include aqueous buffer solutions, e.g., tris(hydroxymethyl)aminomethane and its salts, phosphate, citrate, bicarbonate, and the like, sterile water for injection, physiological saline, and balanced ionic solutions containing chloride and/or bicarbonate salts of normal blood plasma cations such as Ca2+, Na.sup.+, K.sup.+ and Mg2+. The carriers may contain a chelating agent, e.g., ethylenediaminetetraacetic acid, calcium disodium salt, or other pharmaceutically acceptable chelating agents. Compositions for oral administration may also contain flavoring agents and other ingredients for enhancing their organoleptic qualities. Formulations for topical delivery may also contain liquid or semisolid excipients to assist in the penetration or aerosol spray of the photoactive molecule.
The compositions and stabilizing materials of the present invention may also comprise or be used in combination with an additional bioactive agent. Suitable bioactive agents include, for example, antineoplastic agents, blood products, biological response modifiers, anti-fungal agents, hormones, steroids, vitamins, peptides, peptide analogs, enzymes, anti-allergenic agents, anti-coagulation agents, circulatory agents, anti-tubercular agents, anti-viral agents, anti-anginal agents, antibiotics, anti-inflammatory agents, analgesics, anti-protozoan agents, anti-rheumatic agents, narcotics, cardiac glycoside agents, chelates, neuromuscular blocking agents, sedatives (hypnotics), local anesthetic agents, general anesthetic agents, radioactive particles, radioactive ions, X-ray contrast agents, monoclonal antibodies, polyclonal antibodies and genetic material.
The compositions of the present invention may also include one or more food preservation agents, including but not limited to sodium benzoate and benzoic acid; calcium, sodium propionate, and propionic acid; calcium, potassium, sodium sorbate, sorbic acid; sodium and potassium sulfite. calcium, sodium ascorbate, and ascorbic acid; butylated hydroxyanisole (BHA) and butylated hydroxytoluene (BHT); lecithin; and sodium and potassium sulfite and sulfur dioxide. ethylenediamine-tetraacetic acid (EDTA), citric acid, sorbitol, and tartaric acid.
As one skilled in the art will recognize, any of the stabilizing materials and/or vesicle compositions may be lyophilized for storage, and reconstituted or rehydrated, for example, with an aqueous medium (such as sterile water, phosphate buffered solution, or aqueous saline solution), with the aid of vigorous agitation. Lyophilized preparations generally have the advantage of greater shelf life. To prevent agglutination or fusion of the lipids and/or vesicles as a result of lyophilization, it may be useful to include additives which prevent such fusion or agglutination from occurring. Additives which may be useful include sorbitol, mannitol, sodium chloride, glucose, dextrose, trehalose, polyvinyl-pyrrolidone and poly(ethylene glycol) (PEG), for example, PEG 400. These and other additives are described in the literature, such as in the U.S. Pharmacopeia, USP XXII, NF XVII, The United States Pharmacopeia, The National Formulary, United States Pharmacopeial Convention Inc., 12601 Twinbrook Parkway, Rockville, Md. 20852, the disclosure of which is hereby incorporated herein by reference in its entirety.
For topical applications, the stabilizing materials and/or vesicles may be used alone, may be mixed with one or more solubilizing agents, such as dimethylsulfoxide (DMSO), or may be used with a delivery vehicle, and applied to the skin or mucosal membranes. Penetrating and/or solubilizing agents useful for the topical application of the stabilizing materials and/or vesicles are well known in the art. Stabilizing materials and/or vesicles formulated with penetration enhancing agents may be administered transdermally in a patch or reservoir with a permeable membrane applied to the skin. The use of rupturing ultrasound may increase transdermal delivery of photoactive agents. Further, a mechanism may be used to monitor and modulate delivery of the photoactive agents. For example, ultrasound may be used to visually monitor the bursting of the gas filled vesicles and modulate delivery of the photoactive agents and/or bioactive agents and/or a hydrophone may be used to detect the sound of the bursting of the gas filled vesicles and modulate delivery of the photoactive agents and/or bioactive agents.
Generally, the compositions of the invention are administered in the form of an aqueous suspension such as in water or a saline solution (e.g., phosphate buffered saline). The water may be sterile but is preferably not sterile. Also, preferably the saline solution is an isotonic saline solution, although, if desired, the saline solution may be hypotonic (e.g., about 0.3 to about 0.5% NaCl). The solution may be buffered, if desired, to provide a pH range of about 5 to about 7.4. Preferably, dextrose or glucose is included in the media. Other solutions that may be used for administration of the compositions of the present invention include oils, such as, for example, almond oil, corn oil, cottonseed oil, ethyl oleate, isopropyl myristate, isopropyl palmitate, mineral oil, myristyl alcohol, octyldodecanol, olive oil, peanut oil, persic oil, sesame oil, soybean oil, squalene and fluorinated oils. Suitable fluorinated oils are described in U.S. Pat. No. 5,344,930, the disclosure of which is hereby incorporated by reference herein in its entirety.
The PDAs and/or the compositions of the present invention may also include one or nutrients for growing the microorganism, including planktonic or biofilm microorganisms. Exemplary nutrients include but are not limited to a nutrient broth such as Mueller Hinton Broth and those described in PCT/CA2006/001226, and U.S. Pat. Nos. 7,041,470; 6,410,256; 6,326,190; 6,051,423; and 6,599,714, all incorporated herein by reference in their entirety.
In some embodiments, the photoactive agents may be covalently bonded or conjugated to the stabilizing materials described herein, including lipids, polymers, proteins and surfactants, preferably lipids or surfactants, such as lipid conjugated rhodamine, lipid conjugated fluorescein, lipid conjugated N-(lissamine-rhodamine-β-sulfonyl), and those described by MacDonald, J. Biol. Chem., 265(23): 13533-9 (1990), Leenhouts et al, Biochim. Biophys. Acta, 1237(2):121-6 (1995), Ahlers et al, Biophys. J., 63(3):823-38 (1992) and Ebato et al, Anal. Chem., 66(10):1683-9 (1994), the disclosures of which are hereby incorporated by reference herein in their entirety. Alternatively, where a --OH group or --COO.sup.-. group is at the terminus of the photoactive agent, the photoactive agent may esterify with a lipid.
The photodynamic therapy agents of the present invention may also be conjugated to binding agents that bind pre-determined cells or structures in vitro or in vivo. Conjugation with monoclonal antibodies (e.g., immunoconjugates) affords specificity with respect to the treatment of a variety of diseases and conditions.
As used herein, photodynamic agents or photoactive agents, or similar terms, are defined as any compound that absorbs radiation of one or more defined wavelengths and subsequently utilizes the absorbed energy to carry out a chemical process. These agents include but are not limited to any compound or material that is active in light or that responds to light, including, for example, chromophores (e.g., materials that absorb light at a given wavelength), fluorophores (e.g., materials that emit light at a given wavelength), photosensitizers (e.g., materials that can cause necrosis of tissue and/or cell death in vitro and/or in vivo), fluorescent materials, phosphorescent materials and the like, that may be used in diagnostic or therapeutic applications. "Light" refers to all sources of light including the ultraviolet (UV) region, the visible region and/or the infrared (IR) region of the spectrum.
In the present invention, the photodynamic agents are one or more food dyes or colors. The preferred food dyes or colors are well known to those skilled in the art, and include natural, synthetic, Lake, and blended food colors, and combinations thereof. In general these colors may be selected from the group consisting of chlorophylls, carotenoids, flavenoids, quinonoids, coumarins, indigoids, curcuminoids, befalins, acthocyanins, cyanines, indocyanines, phthalocyanines, rhodamines, phenoxazines, phenothiazines, phenoselenazines, fluoresceins, porphyrins, benzoporphyrins, squaraines, corrins, croconiums, azo compounds, methine dyes, and indolenium. Natural food dyes include but are not limited to annatto extract, anthocyanins, B-carotene, beta APO 8 Carotenal, black currant, burnt sugar, Canthaxanthin, caramel, carbo medicinalis, carmine, carmine blue, carminic acid, carrot, chlorophyll, chlorophyllin, cochineal extract, copper chlorophyll, copper chlorophyllin, curcumin, curcumin/CU-chloro, grape, hibiscus, lutein, mixed carotenoids, paprika, riboflavin, spinach, stinging nettle, titanium dioxide, turmeric, aronia/red fruit, beet juice, paprika extract, and paprika oleoresin. Synthetic food colors include but are not limited to allura red, amaranth, black PN, carmoisine, fast red E, erythrosine, green S, patent blue V, ponceau 4R, quinoline yellow, Red 2G, sunset yellow, and tartrazine. Examples of Lake food colors include but are not limited to Lake allura red, Lake amaranth, Lake brillian blue FCF, Lake carmosine, Lake erythrosine, Lake indigo carmine, Lake ponceau 4R, Lake quinoline yellow, Lake sunset yellow, and Lake tartrazine.
As noted above, the photodynamic agent may be natural or synthetic; "natural" as used herein refers to existing in or formed by nature. "Synthetic", as used herein, refers to artificial or formed through a chemical process by human agency. The PDS may also be any coloring agent derived from any of the above (e.g., annatto extract can be used to produce bixin (an oil-soluble apocarotenoid) or norbixin (a water-soluble derivative).
As used herein, the term "inactivation of a microorganism" means totally or partially preventing the microorganism from replicating, either by killing the microorganism or otherwise interfering with its ability to reproduce or function.
"Delivery vehicle" or "vehicle" refers to a composition, substance or material that can transport or carry in vivo or in vitro a photoactive agent, a bioactive agent and/or a targeting ligand. Suitable delivery vehicles include, for example, stabilizing materials, vesicles, liposomes, micelles, aerogels, clathrates, gas and/or gaseous precursor filled vesicles, emulsions, suspensions, dispersions, hexagonal H II phase structures, cochleates and the like.
"Stabilizing material" or "stabilizing compound" refers to any material which can improve the stability of compositions containing the photoactive agents, gases, gaseous precursors, liquids, bioactive agents and/or targeting ligands described herein, including, for example, mixtures, suspensions, emulsions, dispersions, vesicles, or the like. The improved stability involves, for example, the maintenance of a relatively balanced condition, and may be exemplified by increased resistance of the composition against destruction, decomposition, degradation, and the like. In the case of preferred embodiments involving vesicles filled with photoactive agents, gases, gaseous precursors, liquids, bioactive agents and/or targeting ligands, the stabilizing compounds may serve to either form the vesicles or stabilize the vesicles, in either way serving to minimize or substantially (including completely) prevent the escape of photoactive agents, gases, gaseous precursors, bioactive agents and/or targeting ligands from the vesicles until release is desired. The term "substantially" as used in the context of preventing escape of photoactive agents, gases, gaseous precursors, bioactive agents and/or targeting ligands from the vesicles means greater than about 50% is maintained entrapped in the vesicles until release is desired, preferably greater than about 60%, about 70%, about 80% or about 85% is maintained entrapped in the vesicles until release is desired, with about 90% being even more preferred. In particularly preferred embodiments, greater than about 95%, about 99% or about 100% of the photoactive agents, gases, gaseous precursors, bioactive agents and/or targeting ligands are maintained entrapped until release is desired. Exemplary stabilizing materials include lipids, proteins, polymers, carbohydrates and surfactants. The resulting mixture, suspension, emulsion or the like may comprise walls (e.g., films, membranes and the like) around the photoactive agent, targeting ligand, bioactive agent, gases and/or gaseous precursors, or may be substantially devoid of walls or membranes, if desired. The stabilizing may, if desired, form droplets. The stabilizing material may also comprise salts and/or sugars. In other embodiments, the stabilizing materials may be substantially (including completely) crosslinked. The stabilizing material may be neutral, positively or negatively charged.
As used herein, administering refers to any action that results in exposing or contacting one or more PDA agents of the present invention with a pre-determined cell, cells, or tissue, typically mammalian. As used herein, administering may be conducted in vivo, in vitro, or ex vivo. For example, a composition may be administered by spraying or washing. Administering also includes the direct application to cells of a composition according to the present invention.
As used herein, physiologically acceptable fluid refers to any fluid or additive suitable for combination with a composition containing a PDA agent. Typically these fluids are used as a diluent or carrier. Exemplary physiologically acceptable fluids include but are not limited to preservative solutions, saline solution, an isotonic (about 0.9%) saline solution, or about a 5% albumin solution or suspension. It is intended that the present invention is not to be limited by the type of physiologically acceptable fluid used. The composition may also include pharmaceutically acceptable carriers. Pharmaceutically accepted carriers include but are not limited to saline, sterile water, phosphate buffered saline, and the like. Other buffering agents, dispersing agents, and inert non-toxic substances suitable for delivery to an inert, dead, or living surface may be included in the compositions of the present invention. The compositions may be solutions, suspensions or any appropriate formulation suitable for administration, and are typically sterile and free of undesirable particulate matter. The compositions may be sterilized by conventional sterilization techniques.
The compounds of the present invention may also be used in conjunction with and conjugated to a number of other compounds, signaling agents, enhancers, and/or targeting agents.
Suitable enhancers include but are not limited to pKa modifiers, hypoxic cell radiosensitizers, and bioreductively activated anti-neoplastic agents, such as mitomycin C (preferably to effect or potentiate the toxicity of the compound in hypoxic cells or microorganisms). Suitable signaling agents include but are not limited to markers of apoptotic cell death or necrotic cell death, or regulatory molecules endogenous to cell cycle control or delay, preferably to potentiate the phototoxicity of the compound(s) by induction of apoptotic or necrotic cell death, or by inhibition of the repair of any form of lethal or potentially lethal damage (PLD).
As used herein, anti-biofilm agent refers to any element, chemical, biochemical, or the like that is effective against a biofilm. Typical anti-biofilm agents are those that have anti-microbial, anti-bacterial, anti-fungal or anti-algal properties. While the invention has been described in some detail by way of illustration and example, it should be understood that the invention is susceptible to various modifications and alternative forms, and is not restricted to the specific embodiments set forth in the Examples. It should be understood that these specific embodiments are not intended to limit the invention but, on the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the invention.
Microbial growth and evaluation procedures are based on procedures described in PCT/CA2006/001226, incorporated herein by reference in its entirety.
Testing of Photocatalytic Activity of Food Dyes
Test microorganisms included Salmonella choleraesuis, Staphylococcus aureus, foodborne pathogens of humans. Test Compounds included Methylene blue, Hemoglobin (blood product), Hemin (blood product), Protoporphyrin (blood product), Annatto Extract (15% Norbixin), beet juice, Canthaxanthin (10%), Beta-carotene (1%), Purple carmine 50%, Carmine lake 60%, Black Carrot extract, Tomato lycopene (5%), sodium copper chlorophyllin, grape skin extract, paprika oleoresin, riboflavin, saffron, titanium dioxide, FD&C blue No1., turmeric oleoresin, FD&C blue No. 2, FD&C Green No. 3, FD&C Red No. 2 (Amaranth), FD&C Red No. 40, FD&C Yellow No. 5, and FD&C Yellow No. 6. Compounds were prepared as 1%, 0.1%, 0.01%, by weight, dissolved in water.
The light source was a full spectrum fluorescent light or LED light and the contact time was 1 and 16 hours. The study temperature was 37° C.
Approximately 106 organisms in 150 uL Cation Adjusted Mueller Hinton Broth (CAMHB) were placed in each of the wells of a 96 peg MBEC device. The lid of the 96 peg MBEC device was placed on the bottom plate containing organism. The device was placed on a shaker, set to 110 rpm, and incubated at 37° C. for 24 hours to produce biofilms.
The challenge plates were set up using a sterile 96-well microtitre plate using aseptic techniques in the following manner (one organism per plate, per light exposure time, per light source). Test solutions (200 uL) were placed into each well. The lid was transferred to the challenge plate and exposed to the various light sources for the prescribed exposure times (16 hours). Following the treatment, the pegs were transferred to the recovery media ("recovery plate") then wrapped in tin foil and incubated at 37° C. for 18 to 24 hours. The recovery plate(s) was wrapped in tin foil, incubated at 37° C. for 24 hours and read visually and by a plate reader, set to an optical density measurement of 630 nm, to determine Minimum Biofilm Eradication Concentration (MBEC) by looking at growth or no growth.
Both natural and artificial food dyes were effective as photodynamic agents in killing Salmonella choleraesuis and Staphylococcus aureus as biofilm organisms (Table 1). Effective food dyes included: Hemoglobin, Hemin, Protoporphyrin, Annatto extract, beet juice, purple carmine, carmine lake 6, black carrot extract, Grape skin, FD&C blue No1, and turmeric oleoresin. Most anti-microbial activity occurred after 16 hours of light exposure.
TABLE-US-00001 TABLE 1 Killing of Salmonella choleraesuis and Staphylococcus aureus by different photodynamic agents and broad spectrum LED or Fluorescent lights (16 Hours exposure) Salmonella cholerasuis Staph aureus 1% 0.1% 0.01% 1% 0.1% 0.01% Fluorescent Light Methylene blue - - - - - - Hemoglobin - - - + + + Hemin - - - + + + Protoporphyrin - - - + + + Annatto extract - - - - + + Beet Juice - - + + + + Canthaxanthin + + + + + + β-Carotene + + + + + + Purple Carmine - + + + + + Carmine Lake 6 - - - + + + Black Carrot - - - + + + Tomato + + + + + + chlorophyllin + + + + + + Grape Skin - + + + + + Paprika + + + + + + Riboflavin + + + + + + Saffron + + + + + + TiO2 + + + + + + Blue No. 1 - - - + + + Turmeric - - - - - + Blue No. 2 + + + + + + Green No. 3 + + + + + + Red No. 2 + + + + + + Red No. 40 + + + + + + Yellow No. 5 + + + + + + Yellow No. 6 + + + + + + LED Light Methylene blue - - - - - - Hemoglobin + + + + + + Hemin - + + + + + Protoporphyrin - + + + + + Annatto extract - - - - + + Beet Juice + + + + + + Canthaxanthin + + + + + + β-Carotene + + + + + + Purple Carmine + + + + + + Carmine Lake 6 - + + + + + Black Carrot + + + + + + Tomato + + + + + + chlorophyllin + + + + + + Grape Skin - + + + + + Paprika + + + + + + Riboflavin + + + + + + Saffron + + + + + + TiO2 + + + + + + Blue No. 1 + + + + + + Turmeric - + + + + + Blue No. 2 + + + + + + Green No. 3 + + + + + + Red No. 2 + + + + + + Red No. 40 + + + + + + Yellow No. 5 + + + + + + Yellow No. 6 + + + + + + "-" = biofilm eradication at the mentioned conditions (concentration, exposure time and Light source) "+" = biofilm growth
Some agents, e.g., beet juice and purple carmine, are effective under these conditions at higher concentrations, but are shown to be effective against gram negative microorganisms.
Testing of Photocatalytic Food Dyes Log Reduction
The test was conducted as shown in Example 1, but using Hemin (blood product), Protoporphyrin (blood product), Annatto Extract, and Turmeric. The group of tests organism also included E. Coli 0157H7.
All four photocatalytic dyes tested were effective anti-microbial agent, eliminating or reducing growth at all tested concentrations. Both broad spectrum light sources were effective but the fluorescent light provided superior antimicrobial activity when combined with the dyes.
TABLE-US-00002 TABLE 2 Average Log10 counts of Salmonella choleraesuis, E. coli 0157H7, Staphylococcus aureus in 1%, 0.1% and 0.01% concentrations of various photocatalytic agents when exposed to 16 hours of LED or Fluorescent light. Samples were run in triplicate. LED Light Fluorescent Light 1% 0.1% 0.01% 1% 0.1% 0.01% E coli Methylene blue (positive 0.00 0.00 1.77 0.35 0.79 3.41 control) Hemin 0.00 4.22 4.18 3.06 3.79 3.17 Protoporphyrin 3.60 3.48 4.04 0.00 0.57 0.44 Annatto Extract 0.00 4.04 4.45 0.00 0.00 1.34 Turmeric 4.61 3.26 2.91 0.00 0.73 2.05 Control 6.36 6.36 6.36 6.51 6.51 6.51 Staph. aureus Methylene blue (positive 4.84 5.40 5.71 0.00 0.00 3.20 control) Hemin 1.51 3.75 5.52 0.00 0.79 2.21 Protoporphyrin 3.10 0.50 1.29 0.00 0.00 0.00 Annatto Extract 0.00 4.28 4.56 0.00 0.00 3.90 Turmeric 4.33 5.35 5.63 0.00 0.00 4.50 Control 6.95 6.95 6.95 6.48 6.48 6.48 S. choleraesuis Methylene blue (positive 3.20 0.00 2.27 0.00 0.00 0.00 control) Hemin 4.74 4.48 4.02 4.55 3.85 2.79 Protoporphyrin 2.92 3.67 3.40 3.32 2.33 2.10 Annatto Extract 0.00 2.27 2.86 0.92 0.00 1.63 Turmeric 3.92 3.82 2.81 3.47 1.73 2.37 Control 7.28 7.28 7.28 7.28 7.28 7.28
Test microorganisms included Aspergillus niger and Fusarium solani. The test organisms were grown in potato dextrose broth. An organism suspension was transferred into potato dextrose broth and incubated at 28° C. for 72 hours. The test Compound was Annatto extract and Turmeric and was prepared as 1, 0.1, and 0.01%, by weight, in 30% Mueller Hinton Broth in phosphate buffered saline.
Using a sterile 96-well microtitre plate under the laminar flow hood, the following was done to set up the above challenge plate for 1 organism per plate/per light exposure per time point. To a 96 well nunc plate 150 μl of the potato dextrose broth containing the 72 hour culture was added to each well. The MBEC plate lid was placed into the inoculum. The devices were put on an orbital shaker set at 110 RPM and Incubated at 28° C. for 48 hours.
The compounds were added to distilled water in the challenge plate. The planktonic microorganisms were rinsed off by dipping the lid into distilled water for 1-2 minutes. The lid was transferred to the challenge plate and let sit for 20 minutes at room temperature with a tin foil cover.
The pegs with biofilm were exposed to light sources (full spectrum fluorescent light or LED light) by placing the devices on a rack. Both light sources were exposed for 16 hours. The fluorescent exposure was done in a cold room with 2 light fixtures with two fluorescent bulbs each. The entire assembly was covered in tinfoil. The LED exposure was done by placing the light source above the plate on the rack and then placing a tinfoil wrapped box over the rack. Following the 16 hour exposures, rinse plates were prepared by adding 0.9% sterile saline (200 μL per well) to each well of a sterile 96 well micro titre plate. The peg lids were rinsed twice in 0.9% sterile saline for 1 to 2 minutes.
The pegs were transferred to PDP recovery media, and then the recovery plate was sealed in tin foil (or other opaque covering) and incubated at 28° C. for 24 to 48 hours.
The MFC was determined by incubating the planktonic plate at 28° C. for 96 hours and read visually to determine MBC values. The MBEC was determined by incubating the recovery plate at 28° C. for 96 hours, and read visually to confirm MBEC values.
Both biofilm and planktonic forms of Aspergillus and Fusarium were killed by Turmeric or Annatto in combination with LED or fluorescent lights (Table 3). The data suggest a difference between the light source and the activity of the photodynamic agent.
TABLE-US-00003 TABLE 3 The number of test sites sterilized by the Photodynamic activity of Annatto extract or Turmeric with LED lights or Fluorescent Lights MBEC MBC 1% 0.1% 0.01% 1% 0.1% 0.01% LED Light Aspergillus Annatto 3/3 3/3 3/3 3/3 3/3 3/3 Extract Turmeric 3/3 3/3 3/3 3/3 3/3 3/3 Fusarium Annatto 0/3 0/3 0/3 1/3 2/3 2/3 Extract Turmeric 2/3 3/3 2/3 0/3 1/3 2/3 Fluorescent Light Aspergillus Annatto Extract 3/3 0/3 0/3 3/3 0/3 0/3 Turmeric 0/3 0/3 0/3 0/3 0/3 0/3 Fusarium Annatto Extract 1/3 0/3 0/3 2/3 0/3 1/3 Turmeric 2/3 2/3 0/3 0/3 0/3 0/3 MFC = Minimum Fungicidal Concentration MBEC = Minimum Biofilm Eradication Concentration MBC = Minimum Biocidal Concentration
Activity of Photodynamic Agent on Plant Bacterial Pathogens
Test microorganism included Pseudomonas syringae, a food spoilage microorganism and plant pathogen. Test Compounds included Hemin (blood product), Annatto Extract, and Turmeric. Compounds were prepared as 1%, 0.1%, 0.01%, by weight, in 30% Mueller Hinton Broth in phosphate buffered saline. The light source was a broad spectrum fluorescent light or an LED light and the contact time was 16 hours. The study temperature was 28° C.
Approximately 106 organisms in 150 uL Cationic
Adjusted Mueller Hinton Broth were placed in each of the wells of a 96 peg MBEC device. The lid of the 96 peg MBEC device was placed on the bottom plate containing organism. The device was placed on a shaker, set to 110 rpm, and incubated at 28° C. for 24 hours.
The challenge plates were set up using a sterile 96-well microtitre plate using aseptic techniques in the following manner (one organism per plate, per light exposure time, per light source). Test solutions (200 uL) were placed into each well. The lid was transferred to the challenge plate and exposed to the various light sources for the prescribed exposure times (16 hours). Following the treatment, the pegs were transferred to the recovery media ("recovery plate") then wrapped in tin foil and incubated at 28° C. for 18 to 24 hours.
The recovery plate(s) was wrapped in tin foil, incubated at 28° C. for 24 hours and read visually and by a plate reader, set to an optical density measurement of 630 nm, to determine MBEC growth.
Hemin, Annatto extract and Turmeric at 1%, 0.1% and 0.01% concentrations and 16 hours of LED or fluorescent exposure was an effective anti-microbial agent against P. syringae showing a reduction in growth (>90%) (Table 4).
TABLE-US-00004 TABLE 4 Average Log10 growth reductions (log10 CFU) of Pseudomonas syringae HB9 in 1%, 0.1% and 0.01% concentrations of various photocatalytic agents when exposed to 16 hours of LED light. Samples were run in triplicate. LED Light Fluorescent Light Test Compound 1% 0.1% 0.01% 1% 0.1% 0.01% Hemin 3.97 1.32 1.48 1.51 0.92 1.07 Annatto extract 1.51 1.16 1.64 3.75 1.50 1.04 Turmeric 1.39 1.63 1.20 0.94 1.32 1.85
Use of Photodynamic Agents for Sanitization of Hard Surfaces
Test microorganisms included Salmonella choleraesuis, E. coli 0157:H7, and Staphylococcus aureus. Test dye Compounds included Annatto Extract, and Turmeric. Compounds were prepared as 0.01%, by weight, in 30% Mueller Hinton Broth in phosphate buffered saline.
The light source was a broad spectrum Fluorescent light and the contact time was 16 hours. The study temperature was 4° C. The surfaces tested were wood, stainless steel, mild steel, rubber, silicone, laminated plastic, and ultra high molecular weight plastic. All materials are used in food processing operations. Equal sized samples of materials listed were prepared by mounting them onto the pegs in a 12-well BEST lid making sure that all samples formed extended approximately the same distance from the lid. The BEST lid and assay procedure is described in U.S. Pat. Nos. 7,041,470; 6,410,256; 6,326,190; 6,051,423; and 6,599,714, all incorporated herein by reference in their entirety.
An in-vitro assay system for evaluating adhesion, post adhesion killing and inhibition of biofilm formation on catheter surfaces using the BEST Assay. An inoculum in 3 mL sterile water matching a 0.5 McFarland Standard (1.5×108 cells per mL) was created in a glass test tube using a sterile cotton swab. The 0.5 McFarland solution was diluted 88 μl in 22 mL 30% Mueller Hinton Broth in PBS and 4 mL of the inoculums was placed into each designated well of a 12-well plate.
The BEST device with the attached samples was placed into the filled 12-well plate. The BEST device was resealed and placed on a rotary shaker (110 revolutions per minute) and incubated at 37±1° C. for 18 to 24 hours.
All photodynamic reagents were prepared by weighing the specific amount of dye using a sterile spatula and adding it to sterile 30% Mueller Hinton Broth in PBS. Challenge plates were prepared by filling a 12-well plate with 4.5 mL of photodynamic agent, which completely immersed the test surfaces. After 18 to 24 hours of biofilm growth, the BEST devices were rinsed once in sterile water, then transferred to the challenge plate. The test surfaces with the bacterial biofilms were allowed to contact the agent for 5 minutes in darkness. The test surfaces were then exposed to the test light source for the test exposure time.
After exposure, the samples on the peg lid were rinsed 3 times in sterile water by dipping the lid with samples attached into 3 consecutive bottom plate devices containing 5 ml of sterile water in each well. The BEST lid was then inserted into a 12 well plate containing 4.5 ml of universal neutralizer. The entire device was sonicated for 30 minutes. Following sonication, 100 μl from each well of the BEST plate was placed into the first 12 empty wells of the first row of a 96 well microtiter plate. 180 μl of 0.9% sterile saline was placed in the remaining rows. A serial dilution (10°-10-7) was prepared by moving 20_l down each of the 8 rows. 20 μl from each well was removed and spot plated on a prepared Trypticase Soy Agar plate. Plates were incubated at 37±1° C. and counted after approximately 24 hours of incubation.
The results in Table 5 correspond to Log10 reduction in number of viable bacterial biomass after exposure to photodynamic agents. This biomass was represented as cfu/mm2 on each surface.
Both turmeric and annatto extract were able to cause a significant reduction in biofilm on the surface of these materials. There was a difference in the preference of photodynamic agent depending on the challenge microorganism and surface. For example, Turmeric performed best for E. coli and Salmonella; Annatto extract was best for Staphylococcus aureus.
This data suggests that a combination of Turmeric and Annatto would be preferred. Wood surface is one of the most difficult surfaces to sanitise. Its porosity provides a good environment for biofilm to grow away from light.
TABLE-US-00005 TABLE 5 Log10 reduction values of various photocatalytic reagents applied to E. coli 0157:H7 growing on stainless steel, mild steel, UHMWP, laminated plastic, silicone, rubber, wood, and concrete after a 16 hour exposure time. E. coli 0157:H7 S. choleraesuis S. aureus Stainless Annatto 0.43 0.21 0.41 steel Turmeric 2.08 2.27 -0.03 Mild steel Annatto 0.64 -0.04 -2.90 Turmeric 0.79 2.09 -3.15 HMWP Annatto 1.79 0.66 1.71 Turmeric 4.33 3.79 4.80 Laminated Annatto 0.39 -3.65 1.88 Plastic Turmeric 1.20 -3.00 1.76 Silicone Annatto 1.24 0.50 1.25 Turmeric 3.88 2.66 1.84 Rubber Annatto 0.15 0.20 0.98 Turmeric 0.29 0.19 0.63 Wood Annatto -0.47 -0.58 1.61 Turmeric -0.44 -0.36 0.30 Concrete Annatto 0.56 0.29 1.65 Turmeric 1.58 0.55 1.41
Use of Photodynamic Agents for Sanitization of Foods
Test microorganisms included Salmonella choleraesuis, E. coli 0157:H7, Staphylococcus aureus, Campylobacter jejuni, and Listeria monocytogenes. The test Compound was Turmeric, prepared as 0.01%, by weight, in 30% Mueller Hinton Broth in phosphate buffered saline.
The light source was a broad spectrum Fluorescent light and the contact time was 16 hours. The study temperature was 4° C. The surfaces tested were beef and poultry. Using sterile shears the skin/surface of the meat products (Beef or poultry) was cut into pieces that fit into the bottom of a 9 cm Petri dish.
An 8 mm biopsy punch was used to take a sample of each piece of meat. The sample was used as the sterile control and to provide the background contamination count of the meat. 200 μl of a 0.5 McFarland standard, of an organism to be tested, was placed onto a single piece of meat in a Petri dish and spread using a sterile spreader. Inoculated samples were placed in an incubator for 1 hour incubation at Room temperature.
Following incubation, the piece of meat, were cut into 3 pieces (one for the positive control (no PDA applied) and two for the test compounds). One of the concentrations of one photodynamic dye was applied to one of the "Test compound section" of the meat using a spray bottle. A portion down the middle was left untreated and used as a growth control. This gave 2 dyes per piece with one common growth control.
The test device was exposed to the test light source for the test exposure time. Following the 16 hour light exposure an 8 mm biopsy punch was used to cut small sections out of the piece of meat. The piece of meat was homogenised in a 15 ml tube with 2 ml of sterile of CAMHB with 0.5% Tween 80. Following homogenisation the samples were serially diluted and spot plated on Trypticase Soy Agar plate or a Brain Heart Infusion Agar (For Listeria spp.). Plates were incubated at 37±1° C. and counted after approximately 24 hours of incubation. Data was evaluated as CFU/mm2. The results show that 0.1% Turmeric caused a reduced colonization of Salmonella choleraesuis, E. coli 0157:H7, Staphylococcus aureus, Campylobacter jejuni, and Listeria monocytogenes on the surface of chicken and beef (Table 6).
The results also appear to indicate that rough surfaces, e.g., the beef, may protect the bacteria from the light source. Slaughter house beef, beef covered with a layer of fat (smooth surface), allows light to better contact photodynamic agent, and therefore the PDA gives better results.
TABLE-US-00006 TABLE 6 Log10 Reduction of Bacterial Colonization on Food Surface using Turmeric as the Photodynamic agent and Fluorescent lights. Listeria Staph. Salmonella Campylobacter monocytogenes aureus E. coli cholerasuis jejuni Chicken Turmeric 0.62 1.09 0.02 1.29 1.32 0.1% Turmeric 0.44 0.87 0.53 0.64 -0.06 0.01% Beef Turmeric 0.06 0.22 0.70 -0.04 0.15 0.1% Turmeric 0.44 0.21 0.08 0.85 1.06 0.01%
Test microorganisms included Staphylococcus aureus, Listeria monocytogenes, E. coli ATCC 25922 and Salmonella choleraesuis ATCC 10708 foodborne pathogens of humans. Test Compounds are shown in Table 7. Compounds were prepared as described in Example 1.
TABLE-US-00007 TABLE 7 Dilution step to Stock get the start Chemical Solution conc. Ppm in Code V. Product Name State Content (ppm) 10% w/v CAMHB A. annatto powder, type 17.5G powder norbixin N/A 0.0125 g/5 mL B. annatto extract WSL liquid norbixin 69000 725 μL/5 mL C. natural annatto liquid "Y" liquid annatto 234000 215 μL/5 mL D. natural orange color liquid annatto 30000 1670 μL/5 mL E. natural egg shade liquid liquid Annatto/ 88000 570 μL/5 mL curcumin F. turmeric oleoresin WSL liquid curcumin 519000 95 μL/5 mL G. aquaresin turmeric paste curcumin 129000 390 μL/5 mL H. natural yellow shade liquid "IT4" liquid curcumin 20800 2400 μL/5 mL I. natural yellow shade liquid paste curcumin 43800 1140 μL/5 mL "CNW" J. natural yellow color I liquid curcumin 484000 100 μL/5 mL K. natural yellow color II liquid curcumin 48400 1030 μL/5 mL CAMBH = Cation Adjusted Mueller Hinton Broth
The dyes worked with both biofilm and planktonic bacteria, as shown in Tables 8 and 9.
TABLE-US-00008 TABLE 8 S. aureus Concentration that eliminated all Bacteria as a ATCC Biofilm (MBEC) or as a Planktonic organism (MBC) 29213 A B C D E F G H I J K MBC 2500 10000 9.8 10000 9.8 9.8 9.8 9.8 9.8 39.8 9.8 MBEC 2500 10000 10000 10000 10000 10000 19.5 1250 10000 156.2 10000
TABLE-US-00009 TABLE 9 Concentration that eliminated all Bacteria as a Biofilm L. monocytogenes (MBEC) or as a Planktonic organism (MBC) ATCC 29213 A B C D E F G H I J K MBC 2500 10000 10000 10000 10000 9.8 9.8 9.8 9.8 19.1 19.1 MBEC 2500 10000 10000 10000 10000 156 9.8 1250 156 19.1 19.1
TABLE-US-00010 TABLE 10 E. coli ATCC Compound cut-off point (ppm) Visual Cut-off 25922 A B C D E F G H I J K MBC >2500 10000 5000 >10000 10000 >10000 9.8 9.8 9.8 >10000 9.8 MBEC >2500 >10000 >10000 >10000 >10000 >10000 625 9.8 19.5 >10000 312.5
TABLE-US-00011 TABLE 11 Salmonella choleraesuis Compound cut-off point (ppm) Visual Cut-off ATCC 10708 A B C D E F G H I J K MBC 2500 10000 10000 >10000 10000 >10000 >10000 9.8 19.5 >10000 39.1 MBEC 2500 10000 10000 >10000 10000 >10000 10000 9.8 39.1 >10000 >10000
Chan Y, Lai C--H (2003) Bactericidal effects of different laser wavelengths on periodontopathic germs in photodynamic therapy. Laser Med Sci 18: 51-55. Abhay Sankar Chakraborti (2003) Interaction of porphyrins with heme proteins--a brief review. Molecular and Cellular Biochemistry 253: 49-54. Margaret E. Daub and Marilyn Ehrenshaft (2000) The Photoactivated Cercospora Toxin Cercosporin: Contributions to Plant Disease and Fundamental Biology Annual Reviews of Phytopatholology. 2000. 38:461-90. A. Egyed (1972) A Simple Method for Heme Isolation, Spcciaiia 15.11 1397-1398. J. R. Guimaraes and A. S. Barretto (2003) PHOTOCATALYTIC INACTIVATION OF Clostridium perfringens AND COLIPHAGES IN WATER Molecular and Cellular Biochemistry 253: 49-54. Michael R. Hamblin, Jennifer Viveiros, Changming Yang, Atosa Ahmadi, Robert A. Ganz and M. Joshua Tolkoff (2005) Helicobacter pylori Accumulates Photoactive Porphyrins and Is Killed by Visible Light Antimicrobial Agents and Chemotherapy, 49: 2822-2827. Giulio Joni and Stanley B. Brown (2004) Photosensitized inactivation of microorganisms Photochemistry and Photobiological Science 3: 403-405 A. Kubin, G. Alth, R. Jindra, G. Jessner, R. Ebermann, (1996) Wavelength-dependent photoresponse of biological and aqueous model systems using the photodynamic plant pigment hypedcin. Journal of Photochemistry and Photobiology B: Biology 36: 103-108. Yeshayahu Nitzan, Bracha Shainberg, and Zvi Malik (1997) Photodynamic Effects of Deuteroporphyrin on Gram-positive Bacteria Current Microbiology 251-258. Tony P. Paulino, Prislaine P. Magalhaes, Geraldo Thedei, Jr., Antonio C. Tedesco, and Pietro Ciancaglini (2005) Use of Visible Light-based Photodynamic Therapy to Bacterial Photoinactivation. Biochemistry and Molecular Biology Education 33: 46-49. C. R. Rovaldi, A. Pievsky, N. A. Sole, P. M. Friden, D. M. Rothstein, And P. Spacciapoli (2000) Photoactive Porphyrin Derivative with Broad-Spectrum Activity against Oral Pathogens In Vitro Antimicrobial Agents and Chemotherapy 44: 3364-3367. H. Wainwright, D. A. Phoenix, J. Marland, D. R. A. Wareing, F. J. Bolton (1997) A study of photobactericidal activity in the phenothiazinium series FEMS Immunology and Medical Microbiology 19: 7580
Patent applications by Merle E. Olson, Calgary CA
Patent applications in class Containing or obtained from Zingiberaceae (e.g., Afromonun, cardemon, ginger, turmeric, etc.)
Patent applications in all subclasses Containing or obtained from Zingiberaceae (e.g., Afromonun, cardemon, ginger, turmeric, etc.)

domain: chemistry


text:
Above: Tito Mboweni. File image © Government of South Africa, reproduced under CC licensing
- ZAR breaks 14.0 level amid hopes of Fed rate cuts, reforms.
- ING eyes further ZAR gains in short-term, risks further out.
- But RMB and Morgan Stanley says ZAR running out of steam.
- Morgan Stanley cuts forecasts for ZAR as rating risk mounts.
The Rand extended a stellar run of gains Friday and many tip it to rise even further in the weeks ahead but analysts at Rand Merchant Bank (RMB) and Morgan Stanley have warned that South Africa's currency risks running out of steam and head-on into the difficult demands of the economic reform process
South Africa's currency has been a significant beneficiary of an apparent decision by the Federal Reserve (Fed) to indulge market concerns about the U.S. economic outlook by cutting interest rates in order to head of an anticipated slowdown in growth.
The Fed's repeated signals that it intends to cut rates as soon as the end of July mean that U.S. assets are no longer quite so appealing to international investors, offering respite from the interest rate pressures that have sucked capital out of emerging markets and hurt the Rand in the last 18 months.
"The Rand, already strengthened by expectations of easier global financial conditions following the Fed statement on Wednesday, outperformed its EM peers, recording 0.23% in daily spot returns despite a better-than-anticipated US CPI print and a resurfacing of trade tensions expressed through yet another Trump tweet," says Nema Ramkhelawan-Bhana at Rand Merchant Bank.
Above: South African 10-year government bond yield, alongside U.S. equivalent (orange line, left axis).
It's not only expectations of respite from strong Dollar and high U.S. interest rates that have helped the Rand in recent days as domestic news-flow has also supported the South African unit. Thursday saw speculation of an imminent easing in funding pressures for state power utility Eskom lift the currency too.
Public Investment Corporation, South Africa's government pension fund and Eskom's largest bondholder, was said to be mulling over a debt-for-equity swap that would improve the condition of the ailing utility's balance sheet and reduce risks to the nation's top credit rating in the process.
Eskom is threatening South Africa's crucial 'investment grade' credit rating assigned by Moody's because the taxpayer is on the hook for R350bn of its debt that is guaranteed by the government. The cash-strapped utility is at risk of failing after years of mismanagement and underinvestment in its asset base.
"ZAR has been the best emerging market performer of the week, buoyed by: i) dovish Fed and soft dollar, ii) CNH stability and iii) news yesterday that Eskom’s biggest bondholder plans a debt-equity swap. Under 13.85, USD/ZAR could briefly trade to 13.50 (a SARB rate cut next week might help the ZAR via bond inflows), but ZAR gains could quickly evaporate if trade tensions re-escalate later this summer," says Chris Turner, head of FX strategy at ING.
Above: South African Rand performance Vs G10 this month. Source: Pound Sterling Live.
The USD/ZAR rate was quoted 0.10% lower at 13.94 Friday band is now down 2.8% for 2019, denoting weakness of the U.S. Dollar as well as recent strength in the Rand. The Pound-to-Rand rate was 0.06% lower Friday but has fallen 4.3% this year, with the Brexit saga continuing to haunt the British currency.
The prospect of the Fed cutting its interest rate at the end of this month and again in September, which is what the overnight-index-swap market was implying on Friday, could offer continued support to the Rand in the short term but RMB has warned clients the South African unit risks running out of steam.
"The market will be hard-pressed to keep the local unit trading below USD/ZAR14.00 and SAGB yields flatter, if risk sentiment peters out on the back of trade concerns and momentum is lost on Eskom’s turnaround plan," says Ramkhelawan-Bhana, in a research note posted Friday.
Ramkhelawan-Bhana is not alone in fearing the Rand could soon lose momentum because the analyst team at Morgan Stanley also cut their year-end forecasts for the currency this week, although even they say the South African unit can still outperform for a while longer in the short-term.
"With significant market attention focused on the path of US monetary policy,and whether the Fed will cut by 25bp or 50bp, we think there is a decent chance ZAR will rally in the short term as the USD declines," writes Hans Redeker, Morgan Stanley's head of FX strategy, in a recent missive to clients.
Above: USD/ZAR follows South African 10-year bond yield. Falling yield denotes selling of SA bonds.
Despite the upbeat outlook for the South African currency in the short-term, the challenges likely to be faced by the Rand in the Autumn months are significant and not least of all because November will bring the next Moody's decision on the nation's credit rating.
"The growth recovery in South Africa has been disappointing, while the scale of the reform challenges facing the South African government is considerable," Redeker says.
"Our economics team sees a significantly wider fiscal deficit relative to that forecast by the South African Treasury and also Moody's,and a very subdued growth forecast of 0.4% in 2019 and just 1.3% in 2020."
South Africa's government is under pressure from Moody's to reduce both its budget deficit as well as national debt pile, but the economy shrank in the first quarter and the outlook for it in the year ahead is highly uncertain. If the economy continues to shrink it will risk automatically lifting the debt-to-GDP ratio as well as necessitating higher government spending on things like welfare, which could prove too much for Moody's to tolerate.
Losing the Moody's rating would mean many international investors, particular those that benchmark to the Citi World Government Bond index, are automatically forced into selling their South African government bonds. This would force government funding costs higher by lifting bond yields and drive the Rand into the ground in the process as investors seek to swap capital back into their domestic currencies.
The above graph provides an indication of where the USD/ZAR rate might go if the 10-year yield is forced substantially higher because of foreigners selling their South African government bonds. The risk of a final quarter downgrade of the credit rating to 'junk' status is one of the reasons behind the Morgan Stanley forecast cuts.
"We think this justifies an adjustment to our USDZAR profile for the remainder of 2019. We make two adjustments.First, we shift our Q3 forecast to 13.6 (from 14.0) and second, we shift our Q4 forecast to 14.2 (from 13.2)," Redeker says.
Morgan Stanley forecasts the Pound-to-Rand rate will rise to 18.03 by year-end, from 17.50 on Friday.
Above: Pound-to-Rand rate at weekly intervals, alongside SA 10-year yield (purple line, left axis).
Time to move your money? Get 3-5% more currency than your bank would offer by using the services of foreign exchange specialists at RationalFX. A specialist broker can deliver you an exchange rate closer to the real market rate, thereby saving you substantial quantities of currency. Find out more here.

domain: news


A Lurgan man who apologised in court after being caught with cannabis has been handed a £450 fine.
The 22-year-old stated: “I am sorry for what I did, I have been treated with respect throughout.”
Jonathon Fee, of Silverwood Court, pleaded guilty to possession of a Class B drug at Craigavon Magistrates’ Court on Wednesday.
Court heard that police on patrol in the Oxford Island area of Lurgan spotted a Volkswagen Polo, with two males inside, in a car park at around 7.15pm on August 18.
Officers approached the vehicle to ask what the two men were up to.
When they got to the car the defendant in the passenger seat passed out a bag containing a green substance which was suspected to be cannabis.
Fee made no reply when cautioned.
During an interview, he stated the drugs were for personal use and that he had purchased them for £30.
The defendant, who represented himself in court, said: “I am sorry for what I did, I have been treated with respect throughout.”
Fee was ordered to pay a fine of £450, along with the offender’s levy of £15, within 12 weeks.
An Armagh teenager who assaulted his wife has avoided jail. The 19-year-old has been...
A former treasurer of Crossmaglen Rangers GAA club has had an additional ten charges,...
A Portadown man – whom a judge said had now added drugs to his...
/ 1 week ago
Kabosh presents The Box, a new play by Carlo Gébler, which brings to life...
/ 1 week ago
The Market Place Theatre, Armagh proudly hosts the opening of two exhibitions, ‘Designs for...


text:
Silent Buzz – Rumble Bees Fold After One Season
The logo is still in front of The Rink. Their website still remains active, but their Facebook page has been silent since late March. The gear has been sold. And now, it becomes official without a sound.
Just short of 11 months after they became members of the Federal Prospects Hockey League, the Battle Creek Rumble Bees are no more. There were many variables to the team's existence in the first place, but during a tumultuous season that resulted in just one win in 48 games, there may have been a slight chance to return after the FPHL ended their season amid the COVID-19 pandemic.
A recent story released on ESPN's website had several details that followed the season after a 1-45-2 inaugural season. General Manager/ Head Coach Adam Stio told ESPN he hadn’t heard a peep from team owners about the status of the team, but was surprised to receive a call recently from an official from another team asking for names of players on the Battle Creek roster worth selecting in the dispersal draft. That meant that the team was folding.
Hockey in some shape was a plan in Battle Creek in 2019. An upstart league of Michigan teams - the Interstate Hockey League - was in the works during the spring with plans to play at The Rink of Battle Creek. Initial plans were promising for the impending Battle Creek Calvary, but lack of funding from major contributors (in relation, the Alliance of American Football had the same issue) caused a quick cease of operations. The FPHL was able to make their way instead, needing a tenth team to fill their league in their tenth season of existence. Another expansion team, the Delaware Thunder, were pretty successful in year one. The Rumble Bees were owned by the ownership of the Danville (IL) Dashers.
The Rumble Bees took to the ice in their home opener October 25th, eight weeks after the team was formed. It was a 7-1 loss to Danville, but it was in front of what would be their largest crowd of the season at 672 (The Rink seats 1700). The crowds for the remaining 19 home games were not as large, peaking at 652 on February 21st. Their average was 372, ranked last and just 22% capacity. Delaware average 499 on the season, but the Delaware Fairgrounds Center Ice Arena seats just 700, filling the facility to 71% capacity.
There was also a shift quickly in the season, as head coach Clint Hagaman and the franchise parted ways eight games in. Stio would take over behind the bench for the remainder of the season.
The first win would have come at home against the Elmira (NY) Enforcers December 20th. A 5-1 lead was destroyed by a five-goal rally to pull the Enforcers ahead for the win 6-5. The Bees would get their revenge on January 3rd in a 2-1 win in Elmira - the lone win in the franchise. The win was epic, ending a 24-game slide and doing it on a promoted "Guaranteed Win Night" at First Arena.
Several home games started to be moved to road games in January due to logistics and scheduling. In most independent leagues, this can be a tell-tale sign of struggles financially. Bus League cited other examples in a story in mid-January. The Bees also made several trades throughout the season.
After the season was canceled in mid-March, all was quiet from the hockey landscape in the league. Out of the blue, a website that follows the independent hockey season, Bus League Hockey, made the announcement of a move to Berlin, New Hampshire, was in the works on April 1st. We reached out to Stio after the announcement to see if it was true and to get more details. Both Bus League and Stio stated that it was an April Fool's Day joke. Stio's return email also added some caution -
The team is in limbo.
Wednesday’s article by ESPN’s Chris Peters also had conversations with Stio, former Bees player and top scorer Ryan Alves (taken by the Columbus [GA] River Dragons in Monday's dispersal draft) and a fan that fully supported the Bees, even providing food for the players during the season. Stio worked tirelessly to keep the team for the upcoming season, but silence from ownership and the dreaded call to ask about current players in a dispersal draft led to the end.
Peters looked at some of the lowlights of the Rumble Bees only season, making a case for being the worst team in professional hockey history.
• One win in 48 tries, good for a .021 win percentage.
• Last in attendance with an average of 353 fans per game over 20 home dates.
• Outscored 304-94
• Broken-down buses, with players driving their cars to games.
• Practicing in soggy equipment.
• Getting chased off the ice for the home arena's popular public Sunday skates.
• 53 different skaters and seven different goalies


text:
Andice is a town in Williamson County, Texas. It is about 38 miles to the north of Austin. According to the 2005 census, it had a population of 25. Public education here is served by the Florence Independent School District.
Two New York teams are among the finalists (Long Island, NY) Alliance for Green Heat and the New York State Energy Research and Development Authority (NYSERDA) today announced that seven pellet stoves have been chosen as finalists in the Pellet Stove Design Challenge. This international competition, administered by the Alliance for Green Heat, identifies innovative low emissions and high efficiency..

domain: geography


The Hornady® Rapid Rack™ for 12ga shotguns is an empty chamber indicator that fits select shotguns.
*Works as an empty chamber indicator for most 12 gauge shotguns. Operates as a load assist device for the following semi-auto 12 gauge models: Benelli M1 Super 90 / Benelli M2 / Benelli Super Black Eagle II / Beretta 1201FP Remington 11-87 / Mossberg 930
Be the first to review this product! You'll be the talk of the town.
What started as a small dream has become a multi-million dollar international business selling ammunition, reloading supplies and accessories.Read Our Story


CommOptics has introduced a new, single mode fiber (SMF-28) collimating lens that delivers the precision focusing necessary to support telecommunications applications at OC-192 and above.
The lens enables precise focus to be achieved by positioning a fiber 250 µm from input surface of the lens, which is wedged to eliminate back reflections. Specifically, this 2.9 mm focal length, 1.8 mm diameter lens is produced with a focal length tolerance of ±0.1 percent. In contrast, commonly available fiber coupling lenses based on gradient index technology exhibit a focal length variation of as much as 3 percent, creating a coupling loss of in excess of 1 dB. (Consequently, gradient index lenses must often be individually repolished to move the point of focus to the correct position so as to achieve the fiber coupling required for efficient operation at 10 Gb/s or beyond.)
The lenses are supplied with an antireflection coating for C Band operation and deliver an insertion loss less than 0.5 dB; coatings for the S or L Bands are also available on request. An athermalized version, which maintains focus over a -20°C to 70°C temperature range, is also available.
CommOptics designs and manufactures high precision micro-optics, and precision housings and sub-assemblies for telecommunications applications. For more information, visit www.commoptics.com.


text:
Spanning the years 500 to 1000 A.D., this volume illustrates the conflict between brutality and civilization that seemed to characterize the period so often called—not improperly—the "Dark Ages." Islam and Byzantium, as much as Western Europe, figure in the twenty-two chapters of documents offered in this book, part of the ten-volume series, "Sources of Western Civilization."
Get our latest book recommendations, author news, and competitions right to your inbox.
Thank you for signing up, fellow book lover!
Tell us what you like and we'll recommend books you'll love.

domain: literature


text:
Request TOC of this Report- https://www.factmr.com/connectus/sample?flag=T&rep_id=3406
Urodynamic Catheter Market: Drivers and Restraints
Increasing prevalence of the older population which often requires urodynamic testing for possible cases of incontinence expected to drive the growth of the urodynamic catheter market. Increasing prevalence of the urinary tract infection further expected to drive the growth of the urodynamic catheter market. The infection affecting the urinary tract as well as bladder often requires evaluation on whether it has any effect on normal functioning of the urinary system even after recovery from the disease and to confirm that urodynamic catheter is often used. The advanced ability of the urodynamic catheter to measure the pressure and other required factors for evaluation of urinary tract functioning often makes the urodynamic catheter choice of urodynamic testing.
The ease of using urodynamic catheter without any major pain make it’s use feasible fuelling the growth of the urodynamic catheter market. Various conditions such as pregnancy, hepatic surgery are reported to cause temporary urinary continence and healthcare professional often examines the extent of urinary incontinence using urodynamic catheter which in turn expected to drive the growth of the urodynamic catheter market. Whereas, lack of skilled healthcare professionals to carry out the urodynamic testing using urodynamic catheter may hamper the growth of the market.
Urodynamic Catheter Market: Segmentation
The global urodynamic catheter market is segmented based on product type, end user and region
By product type, the global urodynamic catheter market is segmented as:
- Air Charged
- Water Charged
By end user, the global urodynamic catheter market is segmented as:
- Specialty Clinics
- Diagnostic Laboratories
Urodynamic Catheter Market: Regional Outlook
The global urodynamic catheter market is expected to be dominated by the North America due to higher number of the urodynamic tests in the region. Europe is expected to be the second most lucrative urodynamic catheter market due to comparatively higher prevalence of the urinary system related complications. Latin America urodynamic catheter market is expected to experience steady growth over due to increasing product availability in the region. Asia-Pacific is expected to witness significant growth due to increasing product adoption as well as rapidly increasing number of the skilled healthcare professionals in the region. Middle East & Africa is expected to be least lucrative urodynamic catheter market due to least product availability and low product adoption in the region.
To know more about the Urodynamic Catheter Market Trends, Visit the link- https://www.factmr.com/report/3406/urodynamic-catheter-market
Urodynamic Catheter Market: Overview
The urodynamic catheter market is expected to experience significant growth due increasing number of urodynamic tests. By product type, water charged urodynamic catheter expected to be the most lucrative segment in the market due to higher adoption as compared to air charged urodynamic catheter. By end user, hospitals expected to be the most dominant segment due to higher patient footfall. The manufacturers in the urodynamic catheter market are focusing on developing advanced as well as more sensitive urodynamic catheter which can precisely measure the pressure.
Urodynamic Catheter Market: Key Players
The key market players operating in the urodynamic catheter market are: LABORIE, Cook Medical, Teleflex Incorporated, Digitimer Ltd., Mediplus Ltd., MKS Medic, Stericom / P3 Medical, Medica S.p.A.,and others.
The research report presents a comprehensive assessment of the market and contains thoughtful insights, facts, historical data, and statistically supported and industry-validated market data. It also contains projections using a suitable set of assumptions and methodologies. The research report provides analysis and information according to market segments such as geographies, application, and industry.
The report covers exhaust analysis on:
- Market Segments
- Market Dynamics
- Market Size
- Supply & Demand
- Current Trends/Issues/Challenges
- Competition & Companies involved
- Value Chain
Regional analysis includes:
- North America (U.S., Canada)
- Latin America (Mexico. Brazil)
- Western Europe (Germany, Italy, France, U.K, Spain)
- Eastern Europe (Poland, Russia)
- Asia-Pacific excluding Japan (China, India, ASEAN, Australia & New Zealand)
- Middle East and Africa (GCC Countries, South Africa, Northern Africa)
The report is a compilation of first-hand information, qualitative and quantitative assessment by industry analysts, inputs from industry experts and industry participants across the value chain. The report provides in-depth analysis of parent market trends, macro-economic indicators and governing factors along with market attractiveness as per segments. The report also maps the qualitative impact of various market factors on market segments and geographies.
Request Brochure of this Report- https://www.factmr.com/connectus/sample?flag=B&rep_id=3406
FactMR is a fast-growing market research firm that offers the most comprehensive suite of syndicated and customized market insights reports. We believe transformative intelligence can educate and inspire businesses to make smarter decisions. We know the limitations of the one-size-fits-all approach; that's why we publish multi-industry global, regional, and country-specific research reports.
11140 Rockville Pike
Rockville, MD 20852
Read Industry News at - https://www.industrynewsanalysis.com/


text:
The Facebook exodus among young people is real, and disenchantment with the leading social media platform is extending to older users, too. According to new data from Pew Research Center that sampled US Facebook users aged 18 and up, 4 in 10 (42 percent) of those surveyed have taken a break from the social network for “several weeks or more” in the last year; a quarter of respondents said they’ve deleted the mobile app entirely from their smartphones.
Pew’s survey was conducted between May 29th and June 11th, so the burnout and frustrations stemming from Facebook’s Cambridge Analytica privacy scandal were still fresh in the minds of users. But it’s an alarming sign that shows people have a bad taste in their mouths from Facebook after…
from The Verge – All Posts https://ift.tt/2wJaOc5

domain: sociology


There are practical reasons for hats in summer: hair wet from a day at the beach, protection from the sun’s bright rays.
But those who make and love hats would ask that you look beyond the functionality.
“Hats are a wonderful opportunity for self-expression,” said Lore Emelio. “People who wear hats are more stylish, more self-expressed and a patron of a fine art.”
A Charlotte, N.C.-based designer, Emelio (www.LoreEmelio.com) made her first hat when she was 8. Now 35, she wishes everyone would wear them more often.
“People might have a bad past experience. The hat was too small or too big,” she says. “That painted their perspective. But it’s not that hats are not for you, it’s that that wasn’t the right hat.”
Choosing a hat is a personal thing, said Pablo Alvarez. Alvarez owns the Hat Lounge, a store that specializes in men’s hats, in Durham, N.C.
“It’s a feeling,” he said of finding the right topper. “You meet it and you greet it.”
A seasoned hat veteran can shop online, Alvarez says, but if you’re new to hats, go to a store and explore.
Emelio agreed, and added that the right hat for you depends on your hair, your face shape and, equally, your personality.
Her other tips: Hats should land about one-inch above the ear.
Make sure your vision is free and you feel comfortable.


text:
A place where you can spend your vacation in complete comfort!
Bright and comfortable, this apartment is located right in the Centre of s. Agata Sui Due Golfi in a position that combines the tranquility of a large garden in a private park with the opportunity to walk to
Comfortable, spacious apt in center of Village. Grocer, Pastry, and homemade Pasta shop across the street. Restaurants within walking distance. Bus to Sorrento 20 mins. Although helpful to have a ca… More
“Perfect location to take in the Amazing Amalfi Coast and Sorrento”
Reviewed 2 Dec 2014
We had and unforgettable Thanksgiving holiday thanks to this place. It was the perfect location for seeing the entire peninsula and both gulfs- close to Sorrento, close to idyllic beaches, close to P… More
“Perfect solution to the madness of Amalfi”
Reviewed 13 Jun 2014
If you have a car and want to be close to Positanto, Sorrento, Amalfi , Pompeii and Heraculaneum but at the end of the day you want a comfortable spacious retreat with easy parking then this is the pl… More
Residence Sorrento and 'welcoming a tourist facility, located in the center of Sorrento, a short walk from the main square and close to both the Circumvesuviana to Porto. The apartment, very bright, is structured with two bedrooms, a living room with sofa bed, a kitchen
The apartment is situated on the Sorrentinian Peninsula, in Sant´Agata sui due Golfi, close to Sorrento and to Amalfi Coast. Embedded into the typical Mediterranean macchia vegetation,Li Galli is a part of the recently renovated former farm house, Antico Casale Ruoppo.
Li Galli is part of a lovely big converted farmhouse at the end of a windy road. The rooms are pretty spacious with high ceilings, and the balcony has an incredible view over the local bay.
The poo… More
“Sobald wie möglich zurück!”
Reviewed 16 Jun 2014
Wir haben in dem Apartment Li Galli eine sehr schöne Zeit verbracht. Alle Zimmer sind groß und haben einen wunderschönen Blick auf das offene Meer. Man kann urgemütlich auf der Terrasse frühstüc… More
Villa Il Buongiorno occupies a prime location in the peaceful hills surrounding Sorrento. A substantial and stylish two-storey villa of 450 square metres, Villa Il Buongiorno offers exceptional accommodation for a party of up to 11 people.
As you make the 15 minute driv
The apartment Libertà is located in a beautiful villa surrounded by olive threes and it is available garden and swimmingpool. The apartment has proper access from the street and from the garden, it is located on the first floor of the villa and it has 2 floors. On
Apartment in Villa '900 immersed in an elegant and stylish.
The house is accessed from the garden with - 30 steps .
The apartment consists of:
Living room with tv
Sofa - Bed,
3 bedrooms (2 double bedrooms and 1 bedroom with 2 single beds)
Maison Elvira is situated in one of the most picturesque spots in Italy, between the Sorrento coast and the Amalfi coast, exactly in the square of Torca (behind the church), a small village in Massa Lubrense. It is the ideal place for those who want to relax and visit t
“Lovely apartment, lovely people lovely area, love to go back and spend more time there.”
Reviewed 1 Oct 2017
Could not fault the apartment or owners. Only criticism was it was not particularly convenient for Sorrento as it required a 10 minute walk to the bus and then a half hour bus ride. Getting to a beach… More
Holiday Apartment Il Limoneto di Laura is a small independent house in a quiet position, in a lemon garden 600m from the center of Sorrento, station, 1 km from the sea. The house consists of a living room with kitchenette, two small armchairs and a table with chairs. Th
The profuse Mediterranean vegetation which encircles villa harmoniously melts with the characteristic elements of the property, creating a worthy atmosphere of a postcard. The villa, arranged on two levels, has a wide glass body that allows to live the thrilling
The apartment is situated on the upper floor, has three double bedrooms, 3 bathrooms of which 2 with Jacuzzi, a kitchen, a living room with a double sofa bed and spacious terraces with sea views of the bay of Marina del Cantone. The apartment is furnished in a be
Apartment situated in a comfortable private Residence at walking distance from the main Square of Sorrento (Piazza Tasso); although its closeness to the city centre, the Residence is actually located in a very quiet area surrounded by vegetation. The apartment is large
Sorrento Urban Beta
Explore the delights of Sorrento from this central apartment
Centre of Sorrento
2 double bedrooms, 1 twin bedroom; 2 bathrooms
Sorrento Urban Beta is a delightful apartment for up to six people in the centre of Sor
Superb penthouse with two terraces overlooking the Gulf of Naples and Sorrento. The apartment , 100 m from the city center , is equipped with all comfort, air-conditioned, park with swimming pool available from 15 June to 15 Settembre.There are two bedrooms with double
Villa Esposito is located in the proximities of the small town of Sant'Agata sui Due Golfi in an astonishing position facing the Gulf of Salerno, Li Galli and La Vetara. The Villa, on two levels, is surrounded by vineyards, lemon and olive trees. On the ground f
My friends and I loved every single bit of this beautiful Villa.
We had a BBQ by the pool and stayed by it most mornings.
The bedrooms and bathrooms are really clean and spacious.
Highly recommen… More
“Wonderful place, Wonderful people, Wonderful food at Fratoria Terranova”
Reviewed 13 Aug 2013
Beautiful, great views and very comfortable for 3 couples. We enjoy adventure when we travel and Villa Esposito provided that.… More
“Villa Esposito is idyllic!”
Reviewed 29 Jul 2013
Alex met us in Sant Agata and we followed him to the villa which is a little remote but with a wonderful view. There is a terrific restaurant just a 2 minute walk up the road - we had breakfast there … More
The apartment accommodates 6 people and has 3 double bedrooms, 2 bathrooms, one of which with Jacuzzi, kitchen, wide living room with exit to the terraces with sea view of the bay of Marina del Cantone. It also has a large solarium with lounge chairs and shower, where y
HOLIDAY APARTMENT - SAN FRANCESCO - MASSA LUBRENSE - SORRENTO COAST - 2 BEDROOMS - 2 BATHROOMS - SLEEPS 6 - SHARED POOL - SEA VIEW - TERRACE
In a lovely setting surrounded by lemon and olive groves, this new top floor 2-bedroom vacation apartment (friendly owner lives
This villa is in a very beautiful setting outside of Sorrento. We were a family with 2 teenagers and a 7 year old. The villa is quite isolated and remote from shops, cafes, restaurants etc. But it is … More
Thank you for being our guests and to have spent time in writing this to help orientate future guests
Villa Il Gioiello is a large villa, set within extensive gardens, in an exceptional location just 400m from the centre of Sorrento. The views from the balcony and terraces are simply magnificent, sweeping across the Gulf of Naples to Vesuvius. The villa is
Accommodation : Villa, 5 Double Bedrooms + 1 Triple Bedroom ( sleeps 13 people ), 6 bathrooms with shower, 1 kitchen, garden, sea view, jacuzzi outdoor pool. Villa Ivy has a wondeful view of the Gulfs of Naples and Massa Lubrense and lemon groves, olive
SORRENTO A 2 km DAL CENTRO IMMERSO NEL VERDE DELLA NATURA SULLE COLLINE UN OASI DI PACE CON VISTA MERAVIGLIOSA SUL GOLFO DI NAPOLI E SORRENTO ,ABILITA FACILE DA RAGGIUNGERE, POSTI FAMOSI DA VISITARE, COME COSTIERA AMALFITANA, POSITANO, RAVELLO, CAPRI
Villa Maia spreads over 350 square metres and is divided into 3 levels:
On the ground floor, with acces to garden and pool, there is a large kitchen, 1 dining room /2 living room areas with comfortable sofas, 1 double bedroom, 1 bathroom.
On the 1st floor there are 3 la
Villa Diamond is surrounded by terraces, balconies and a small garden with olive and citrus fruit trees. The wide terrace/solarium of approximately 200 square metres is also an ideal location for outdoor and dinner parties and is ideal for sunbathing in absolute
Villa Carlotta is located in the village of Torca (Sant'Agata sui due Golfi - Massa Lubrense); it is a charming private villa fully-equipped with all comforts and nice sea view (Marina del Cantone natural bay). The Villa, with wi-fi zone, is composed of two levels;
Beautiful and spacious villa well located in a small typical italian village with beautiful view of the ocean.Very clean and with all the accommodation that you need, it's the perfect place for a fami… More
i hope to see you again
“Beautiful views but far from everything”
Reviewed 18 Aug 2015
The home is comfortable and is excellent for large groups. There is pleanty of room to spread out. The views are beautiful. The location is probably the only negative about the property. It is far… More
Reviewed 20 Aug 2012
La Villa Carlotta est tout simplement splendide! La vue est superbe. La villa est parfaitement conçu, très grande, avec plusieurs salles de bain, personnes ne se pilent sur les pieds. Le seul inc… More
Villa Scarlet has been divided into two independent apartments and spreads over 180m2. The Villa has 3 Double Bedrooms + 1 Twin Bedroom + 2 double convertible beds ( Sleeps 8+4 ), 4 Bathrooms (1 Jacuzzi), 2 kitchens, 2 dining rooms, swimming pool. On the lower fl
The apartment accommodates 6 persons and has 3 double bedrooms, 2 bathrooms one of which with Jacuzzi, kitchen, wide hall and terraces with sea view on the bay of Marina del Cantone. Consisting of wide solarium with beach lounge chairs and shower, where taking t
Explore the delights of Sorrento from this central apartment Sorrento 1 double bedroom, 1 bedroom with a single bed and single folding bed, 1 bathroom Sleeps 3-4 Caruso 3 is a delightful apartment for up to four people in the centre of Sorrento. Part of
The house is an old farm house completely renovated beginning 2014, with kitchen completely equipped, laundry, TV and wifi. The view is wonderful and there're two big bedrooms each one with own bathroom. The living area is very spacious and you can have a dinner seeing

domain: travel


text:
Beaux Raymond Biography
Beaux Raymond is an England Legal Secretary, model, and reality television personality who is best known for being cast in the third season of Too Hot To Handle.
In February 2021, she became part of headlines after she had abused a British Airways staff. This happened while she was traveling to Miami on a flight with Harry Johnson and Matthew Mawhinney for filming and the three were spotted downing margaritas. They were also seen being intimate in their seats, the cabin attendants advised Johnson and Raymond to calm things down. None of them listened and when Mawhinney asked for more drinks, the crew members denied them thus they began abusing them and as a result, they were fined £3000 for their misbehavior.
Beaux Raymond Age
Raymond is a 25-year-old Kent, South East England native-born in 1997. However, she has neither disclosed the date nor the month she was born as she likes to keep her personal life private.
Beaux Raymond Height and Weight
Raymond stands at a height of 5 feet 4 inches (1.63 m| 163 cm) tall and weighs 51 kg (112 Pounds).
Beaux Raymond Family
Raymond’s mother is known as Vivien Raymond while her brother is known as Lewis Raymond. However, she has failed to disclose the name of her father.
Beaux Raymond Boyfriend
Raymond is currently single and did not disclose anything about her previous relationships as she likes to keep her personal life private.
Beaux Raymond Net Worth
Raymond has an estimated net worth of about $500,000 – $1 Million which she has earned through her career as a Legal Secretary and reality television cast.
Beaux Raymond Salary
Raymond earns a salary of about $25,000-$70,000.
Too Hot To Handle (season 3)
Too Hot To Handle is a British-American reality television dating game show which is 100% authentic. On the show, gorgeous singles meet and mingle on the shores of paradise. The show requires the cast who engage in meaningless flings and are unable to form long-lasting relationships to foster genuine connections between the participants while living under the same roof for four weeks. The casts have to follow a set of instructions given by their host Lana who is a cone-shaped virtual assistant and when rules are broken the $100,000 grand prize is reduced.
The third season of the show will premiere on January 19, 2022, filmed in the Turks and Caicos Islands. Raymond is a cast on the show among other nine casts who are;
- Georgia Hassarati a 26-year-old Brisbane, Australia resident
- Harry Johnson a 29-year-old Middlesbrough, England resident
- Holly Scarfone a 23-year-old Colorado, United States resident
- Izzy Fairthorne a 22-year-old Manchester, England resident
- Jaz Holloway a 25-year-old Virginia, United States resident
- Nathan Soan Mingomezulu is a 24-year-old Cape Town, South Africa resident
- Patrick Mullen a 29-year-old Hawaii, United States resident
- Stevan Ditter a 26 -year- old Los Angeles, California, United States resident
- Truth a 23 -year-old Texas, United States resident
Beaux Raymond Social Media Platform
Raymond is very active on her Tiktok and Instagram pages and she has 6k followers on Tiktok and 7k followers on Instagram.

domain: celebrity