Superoxide dismutase soluble fiber compositions and methods of use

Abstract

Compositions are provided that include superoxide dismutase and soluble fiber. The compositions can additionally include other antioxidants, sweeteners, probiotics, vitamins and nutrients. The compositions can be used as food supplements and to improve health and wellness. Methods of using such compounds are also provided.

Description

[Technical field]

[0001] FIELD OF THEINVENTION The present disclosure generally relates to a composition comprising superoxide dismutase and soluble fiber. The composition can be used as a food supplement and to improve health and wellness. The present disclosure further relates to a method of using the composition comprising superoxide dismutase and soluble fiber. [Background technology]

[0002] 2. Background of the Invention Superoxide dismutase (SOD) is a group of metalloenzymes that protect cells from damage caused by reactive oxygen species (ROS). SOD converts the superoxide anion free radical (O2) into molecular oxygen and hydrogen peroxide (H2O2). - ) in cells. ROS can damage nucleic acids, proteins and lipids, leading to impaired cell function and possible apoptosis. Therefore, the ability to convert ROS into harmless molecules is crucial to protect cell function and overall health.

[0003] Almost all living organisms naturally produce some type of SOD, but the level of SOD produced in cells decreases with age or when the subject suffers from certain health disorders.Also, the presence of various pollutants and toxins in the environment can cause the elevated level of ROS in cells.Therefore, dietary supplementation of naturally produced SOD can be important for maintaining health.

[0004] Another important dietary component is soluble fiber. Soluble fiber absorbs water in the digestive tract and forms a gel. Soluble fiber has numerous benefits in the gut, including slowing the digestion of certain types of lipids and carbohydrates, helping to prevent the uptake of dietary cholesterol, and preventing spikes in blood sugar levels. Soluble fiber is also important for maintaining a healthy gut microbiome, as gut bacteria can ferment some types of soluble fiber. Modern diets are typically deficient in soluble fiber, and supplementation is often required for improved health.

[0005] Diabetes mellitus (diabetes) is a common carbohydrate metabolism disorder. In the United States, over 30 million people have diabetes. In the last 20 years, the number of adults diagnosed with diabetes has more than tripled as Americans age and become more overweight or obese. In people with diabetes, the body's normal ability to utilize glucose is impaired, resulting in elevated blood sugar levels. Diabetes is associated with an increased risk of cardiovascular or circulatory disease or disorders.

[0006] Dietary fiber, such as soluble fiber, has been shown to blunt the glycemic response to a meal. When fermented by gut bacteria, fiber also produces metabolic products of short-chain fatty acids [SCFAs] that are involved in glucose homeostasis. However, high-fiber foods can cause gastric discomfort, as can some fiber supplements.

[0007] Antioxidants have been shown to have beneficial effects in the human body in combating free radicals that are involved in many disease processes.However, the absorption of the effective metabolites of many antioxidants, such as polyphenols, depends on the microbial metabolism of antioxidants in the intestine.Therefore, for many nutritional supplements, these effective polyphenol metabolites are not produced, and the antioxidant activity of the supplement is limited. [Prior art documents] [Non-patent literature]

[0008] [Non-Patent Document 1] McCord, J.M. and Fridovich, I., J. Biol. Chem. 1969, 244:6049-6055. [Non-Patent Document 2] Weydert et al., Nature Protocols 2010, 5(1):51-66. [Non-Patent Document 3] https: / / www.sigmaaldrich.com / technical-documents / protocols / biology / enzymatic-assay-of-superoxide-dismutase.html Summary of the Invention

[0009] BRIEF SUMMARY OF THE PRESENTINVENTION One aspect of the present disclosure provides a liquid composition comprising: a) about 0.03 units / mL to about 0.5 units / mL of superoxide dismutase; b) about 1.3 mg / mL to about 23 mg / mL of soluble fiber; and c) water.

[0010] In an embodiment, the liquid composition comprises from about 0.05 units / mL to about 0.4 units / mL of superoxide dismutase, hi an embodiment, the liquid composition comprises from about 0.2 units / mL to about 0.3 units / mL of superoxide dismutase.

[0011] In embodiments, the liquid composition contains from about 2.7 mg / mL to about 12 mg / mL of soluble fiber. In embodiments, the liquid composition contains from about 5.55 mg / mL to about 11.11 mg / mL of soluble fiber.

[0012] In an embodiment, the superoxide dismutase is extracted from melon, beef liver, heterotrophic bacteria, or marine phytoplankton. In an embodiment, the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase, or nickel superoxide dismutase.

[0013] In an embodiment of the liquid composition, the ratio of superoxide dismutase to soluble fiber is about 1:100 to about 1:1000 by weight. In an embodiment, the ratio of superoxide dismutase to soluble fiber is about 1:500 to about 1:700 by weight.

[0014] In an embodiment of the liquid composition, the soluble fiber is a water-soluble polysaccharide. In an embodiment, the soluble fiber is selected from soluble corn fiber, inulin, dextrin, guar gum, oligopolysaccharides, galactopolysaccharides, fructooligosaccharides, lactulose, resistant starch, xylooligosaccharides and isomalto-oligossacharides. In an embodiment, the soluble fiber is soluble corn fiber. In an embodiment, the soluble corn fiber is resistant maltodextrin.

[0015] In an embodiment, the liquid composition further comprises from about 0.1 mg / mL to about 1.5 mg / mL of a simple sugar. In an embodiment, the liquid composition further comprises from about 0.1 mg / mL to about 1.5 mg / mL of d-ribose. In an embodiment, the liquid composition further comprises from about 0.40 mg / mL to about 0.85 mg / mL of d-ribose.

[0016] In an embodiment, the liquid composition further comprises from about 1.3 mg / mL to about 9.0 mg / mL of a sugar alcohol. In an embodiment, the liquid composition further comprises from about 1.3 mg / mL to about 9.0 mg / mL of erythritol. In an embodiment, the liquid composition further comprises from about 2.7 mg / mL to about 5.6 mg / mL of erythritol.

[0017] In an embodiment, the liquid composition further comprises from about 0.1 mg / mL to about 1.5 mg / mL of a pH adjuster. In an embodiment, the liquid composition further comprises from about 0.1 mg / mL to about 1.5 mg / mL of citric acid. In an embodiment, the liquid composition further comprises from about 0.4 mg / mL to about 0.7 mg / mL of citric acid.

[0018] In an embodiment, the liquid composition further comprises from about 0.05 mg / mL to about 0.75 mg / mL of a sweetener. In an embodiment, the liquid composition further comprises from about 0.05 mg / mL to about 0.75 mg / mL of a steviol glycoside. In an embodiment, the liquid composition further comprises from about 0.2 mg / mL to about 0.35 mg / mL of a steviol glycoside.

[0019] In an embodiment, the liquid composition further comprises a flavoring agent.

[0020] Another aspect of the present disclosure provides a composition comprising: a) about 10 units to about 200 units of superoxide dismutase; b) about 500 mg to about 8000 mg of soluble fiber; and c) a probiotic.

[0021] In an embodiment, the probiotic composition comprises from about 50 units to about 150 units of superoxide dismutase.In an embodiment, the probiotic composition comprises from about 70 units to about 100 units of superoxide dismutase.

[0022] In an embodiment, the probiotic composition contains from about 1000 mg to about 5000 mg of soluble fiber. In an embodiment, the probiotic composition contains from about 2000 mg to about 4000 mg of soluble fiber.

[0023] In an embodiment of the probiotic composition, the superoxide dismutase is extracted from melon, beef liver, heterotrophic bacteria, or marine phytoplankton. In an embodiment, the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase, or nickel superoxide dismutase.

[0024] In embodiments of the probiotic composition, the ratio of superoxide dismutase to soluble fiber is about 1:100 to about 1:1000 by weight. In embodiments, the ratio of superoxide dismutase to soluble fiber is about 1:500 to about 1:700 by weight.

[0025] In an embodiment of the probiotic composition, the soluble fiber is a water-soluble polysaccharide. In an embodiment, the soluble fiber is selected from soluble corn fiber, inulin, dextrin, guar gum, oligopolysaccharides, galactopolysaccharides, fructooligosaccharides, lactulose, resistant starch, xylooligosaccharides, and isomaltooligosaccharides. In an embodiment, the soluble fiber is soluble corn fiber. In an embodiment, the soluble corn fiber is resistant maltodextrin.

[0026] In an embodiment of the probiotic composition, the probiotic comprises bacteria of the genus Bifidobacterium. In an embodiment, the probiotic comprises bacteria of the genus Lactobacillus. In an embodiment, the probiotic comprises bacteria of the phylum Firmicutes·Lactobacillus, the phylum Actinobacteria·Bifidobacteriaceae, or a combination thereof.

[0027] In an embodiment of the probiotic composition, the composition is in the form of a gel. In an embodiment, the composition is in the form of a liquid. In an embodiment, the composition is in the form of a powder.

[0028] Another aspect of the present disclosure provides a method of increasing T cell activation in a subject, the method comprising orally administering to the subject a composition comprising: a) about 10 units to about 200 units of superoxide dismutase; and b) about 500 mg to about 8000 mg of soluble fiber; and wherein after administration of the composition, T cell activation is increased in the subject. In an embodiment of the method, the composition is administered in combination with an anti-cancer agent. In an embodiment of the method, the composition is administered in combination with an anti-viral agent.

[0029] Another aspect of the disclosure provides a method of increasing production of short chain fatty acids (SCFAs) in the digestive tract of a subject, comprising orally administering to the subject a composition comprising: a) about 10 units to about 200 units of superoxide dismutase; and b) about 500 mg to about 8000 mg of soluble fiber; and after administration of the composition, production of SCFAs is increased in the digestive tract of the subject. In an embodiment of the method, the SCFAs whose production is increased are acetate, propionate, butyrate, or lactate SCFAs, or combinations thereof. In an embodiment of the method, the SCFAs are increased to provide approximately the same ratio of acetate, propionate, butyrate, and lactate SCFAs compared to the ratio of acetate, propionate, butyrate, and lactate SCFAs before the increase.

[0030] Another aspect of the present disclosure provides a method for increasing the amount of bacteria of the genus Bifidobacterium or Lactobacillus in the digestive tract of a subject, comprising orally administering to the subject a composition comprising: a) about 10 units to about 200 units of superoxide dismutase; and b) about 500 mg to about 8000 mg of soluble fiber; and after administration of the composition, the amount of bacteria of the genus Bifidobacterium, Lactobacillus, or a combination thereof is increased in the digestive tract of the subject. In an embodiment of the method, the bacteria of the genus Bifidobacterium comprises a species of the family Bifidobacteria in the phylum Actinobacteria. In an embodiment of the method, the bacteria of the genus Lactobacillus comprises a species of the phylum Firmicutes Lactobacillus.

[0031] In embodiments of any of the methods herein, the composition comprises about 50 units to about 150 units of superoxide dismutase. In embodiments, the composition comprises about 70 units to about 100 units of superoxide dismutase.

[0032] In any of the embodiments of the methods herein, the composition comprises from about 1000 mg to about 5000 mg of soluble fiber. In embodiments, the composition comprises from about 2000 mg to about 4000 mg of soluble fiber.

[0033] In any of the embodiments of the methods herein, the superoxide dismutase is extracted from melon, bovine liver, heterotrophic bacteria, or marine phytoplankton. In embodiments, the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase, or nickel superoxide dismutase.

[0034] In any of the embodiments of the methods herein, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:100 to about 1:1000 by weight. In embodiments, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:500 to about 1:700 by weight.

[0035] In any of the embodiments of the methods herein, the soluble fiber is a water-soluble polysaccharide. In an embodiment, the soluble fiber is selected from soluble corn fiber, inulin, dextrin, guar gum, oligopolysaccharides, galactopolysaccharides, fructooligosaccharides, lactulose, resistant starch, xylooligosaccharides, and isomaltooligosaccharides. In an embodiment, the soluble fiber is soluble corn fiber. In an embodiment, the soluble corn fiber is resistant maltodextrin.

[0036] In embodiments of any of the methods herein, the composition is in the form of a gel. In embodiments, the composition is in the form of a liquid. In embodiments, the composition is in the form of a powder. BRIEF DESCRIPTION OF THE DRAWINGS [Brief description of the drawings]

[0037] [Figure 1] FIG. 1 is a plot of absorbance at 490 nm (representing cell viability as described in Example 1) for Jurkat cells treated with lipopolysaccharide (LPS), different components of the gel formulation of Example 1, gel combinations (finished formulation), and a negative control (no treatment). [Diagram 2]FIG. 1 is a plot showing T cell activation as measured by CD69 and CD3 expression as described in Example 1. Jurkat cells are abbreviated as JRK. [Diagram 3] 1 is a plot showing the effect of the gel of Example 1 and its components on CD4+ differentiation in Jurkat cells at the following concentrations: Fibersol prebiotic fiber 1.3 mg / mL; superoxide dismutase (SOD) 0.2 mg / mL; LPS 10 μg / mL; fruit juice 4.6 mg / mL and the combination (finished product) 5.6 mg / mL. [Figure 4] 1 is a plot showing the effect of the gel of Example 1 and its components on CD8+ differentiation in Jurkat cells at the following concentrations: Fibersol prebiotic fiber 1.3 mg / mL; superoxide dismutase (SOD) 0.2 mg / mL; LPS 10 μg / mL; fruit juice 4.6 mg / mL and the combination (finished product) 5.6 mg / mL. [Diagram 5] 1 is a plot showing the effect of the gel of Example 1 and its components on attenuating 8-isoprostane secretion from LPS-activated Jurkat cells as described in Example 1. [Figure 6] 1 is a plot showing the effect of the gel of Example 1 and its components on attenuating cyclooxygenase-2 (COX-2) secretion in LPS-activated Jurkat cells as described in Example 1. [Figure 7] 1 is a plot showing the effect of the gel of Example 1 and its components on attenuating interferon-gamma (IFN-γ) secretion in LPS-activated Jurkat cells as described in Example 1. [Figure 8] 1 is a plot showing the effect of the gel of Example 1 and its components on attenuating interleukin-6 (IL-6) secretion from LPS-activated Jurkat cells as described in Example 1. [Figure 9] 1 is a plot showing the effect of the gel of Example 1 and its components on attenuating transforming growth factor-beta (TGF-β) secretion from LPS-activated Jurkat cells as described in Example 1. [Figure 10]1 is a plot showing the effect of the gel of Example 1 and its components on attenuating tumor necrosis factor-alpha (TNF-α) secretion from LPS-activated Jurkat cells as described in Example 1. [Figure 11] 1 is a plot showing the effect of the gel of Example 1 and its components on attenuating C-X-C motif chemokine ligand 10 (CXCL10) secretion from LPS-activated Jurkat cells as described in Example 1. [Figure 12] 1 is a plot of bacterial density over time for cultures grown on goat intestines treated with the complete gel product (REVIVIFY® Finished Product) or components of the gel. [Figure 13] 1 is a plot of bacterial density over time for cultures grown on goat intestines treated with the complete gel product (REVIVIFY® Finished Product) or components of the gel. [Figure 14A] FIG. 1 is a plot of the concentrations of short chain fatty acids (SCFAs) acetate, propionate, butyrate, and lactate in in vitro gut microbial cultures when treated with the complete gel product (combination) or components of the gel as described in Example 3. [Figure 14B] FIG. 1 is a plot of the percentage of short chain fatty acids (SCFAs) acetate, propionate, butyrate, and lactate in in vitro gut microbial cultures when treated with the complete gel product (combination) or components of the gel as described in Example 3. [Figure 15] FIG. 1 is a plot showing the microbial profile of in vitro gut microbial cultures when treated with a complete gel product (combination) or components of the gel as described in Example 3. [Figure 16] 1 is a plot of the concentration of malondialdehyde (MDA) biomarker secreted by human brain microvascular endothelial cells (HBMEC) when treated with the complete gel product or components as described in Example 4. [Figure 17]1 is a plot of the concentration of 4-hydroxynonenal (HNE) biomarker secreted by HBMEC when treated with complete gel products or components as described in Example 4. [Figure 18] 1 is a plot of the concentration of protein carbonyl (PC) biomarker secreted by HBMEC when treated with complete gel products or components as described in Example 4. [Figure 19] 1 is a plot of the concentration of 3-nitrotyrosine (NT) biomarker secreted by HBMEC when treated with complete gel products or components as described in Example 4. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0038] Detailed Description of the Invention The present disclosure provides a composition comprising superoxide dismutase and soluble fiber.The composition disclosed herein can be used as a supplement to increase the level of superoxide dismutase and soluble fiber in a subject.As described herein, the composition can also include additional ingredients, such as antioxidants, vitamins or other nutrients, together with excipients and other formulating agents.

[0039] It should be understood that the specific implementations shown and described herein are examples and are not otherwise intended to limit the scope of the application in any way.

[0040] Published patents, patent applications, websites, company names, and scientific literature referenced herein are incorporated herein by reference in their entirety to the same extent as if each was specifically and individually indicated to be incorporated by reference. Any conflict from any reference cited herein and the specific teachings of this specification shall be resolved in favor of the latter. Similarly, any conflict from the art-understood definition of a word or phrase and the definition of the word or phrase when specifically taught herein shall be resolved in favor of the latter.

[0041] As used herein, "a" or "an" can mean one or more. As used herein, when used in conjunction with the word "comprising," the word "a" or "an" can mean one or more than one. As used herein, "another" or "further" can mean at least a second or more.

[0042] Throughout this specification, the term "about" is used to indicate that a value includes the inherent variation of error for the method / device being used to determine the value or the variation that exists between study subjects. Typically, the term "about" is meant to encompass a variability of approximately 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or 20% or less, depending on the context.

[0043] Use of the term "or" in the claims is used to mean "and / or," unless expressly indicated to refer to alternatives only, or the present disclosure supports a definition that refers to alternatives only and "and / or," but the alternatives are mutually exclusive.

[0044] As used herein, the terms "comprising" (and any variation or form of comprising, e.g., "comprise" and ("comprises"), "having" (and any variation or form of having, e.g., "have" and "has"), "including" (and any variation or form of including, e.g., "includes" and "include"), or "containing" (and any variation or form of containing, e.g., "contains" and "contain") are inclusive or open-ended and do not exclude additional, unspecified elements or method steps.

[0045] The use of the term "for example" and its corresponding abbreviation "eg" (whether italicized or not) means that the specific terms set forth are representative examples and embodiments of the present disclosure that are not intended to be limited to the specific examples referenced or cited, unless expressly stated otherwise.

[0046] Unless otherwise defined, technical and scientific terms used herein have the meaning commonly understood by one of ordinary skill in the art to which this application belongs. Reference is made herein to various methodologies and materials known to those skilled in the art.

[0047] The term "superoxide dismutase" (sometimes abbreviated as SOD), as used herein, refers to the conversion of the superoxide anion free radical (O2) to molecular oxygen and hydrogen peroxide (HO). - Superoxide dismutase refers to an enzyme that catalyzes the dismutation of reactive oxygen species (ROS), such as O2, that are formed during the cellular metabolism of oxygen. -Superoxide dismutase is an important component in protecting cells against damage from reactive oxygen species. Thus, superoxide dismutase functions as an important antioxidant and is present in almost all types of cellular organisms. However, especially in complex organisms, such as mammals, the cellular level of superoxide dismutase decreases as the organism ages, making the organism more susceptible to cellular damage from reactive oxygen species.

[0048] The term "soluble fiber" as used herein means any type of water-soluble dietary fiber. When ingested, soluble fiber absorbs water and forms a gel in the intestine of an organism, which helps slow down the metabolism of lipids and carbohydrates. Soluble fiber is also a prebiotic that can be fermented by intestinal bacteria, helping to maintain a healthy gut microbiome.

[0049] The term "antioxidant" as used herein means a substance that significantly reduces the harmful effects of reactive molecular species, e.g., reactive oxygen and nitrogen species, by completely or partially neutralizing the reactive molecular species. Antioxidants can be classified as "primary antioxidants" and "secondary antioxidants." Primary antioxidants delay or inhibit the initiation steps of oxidation, while secondary antioxidants slow down oxidation by scavenging substrates or by quenching free oxygen radicals.

[0050] 1) Superoxide dismutase The embodiments disclosed herein, the composition comprises an amount of superoxide dismutase as measured in units of activity per mg of protein. With respect to enzyme activity, one unit (U) (expressed in μmol / min) is defined as the amount of enzyme that catalyzes the conversion of one micromole of substrate per minute under specific conditions. The unit of superoxide dismutase activity can be measured by any known method. For example, methods for determining a unit of superoxide dismutase activity are described in McCord, JM and Fridovich, I., J. Biol. Chem. 1969, 244:6049-6055; Weydert et al., Nature Protocols 2010, 5(1):51-66; and in the technical protocols at https: / / www.sigmaaldrich.com / technical-documents / protocols / biology / enzymatic-assay-of-superoxide-dismutase.html; the disclosures of each of which are incorporated herein by reference. In embodiments, a unit of superoxide dismutase activity is defined as the amount of superoxide dismutase that inhibits the rate of reduction of cytochrome c by 50% in a coupled system with xanthine and xanthine oxidase at pH 7.8 at 25° C. in a 3.0 mL reaction volume.

[0051] In an embodiment, the concentration of the amount of superoxide dismutase in the compositions disclosed herein is measured in terms of the total amount of units of superoxide dismutase in the composition. In an embodiment, the composition comprises from about 10 units to about 200 units of superoxide dismutase. In an embodiment, the composition comprises from about 50 units to about 150 units of superoxide dismutase. In an embodiment, the composition comprises from about 70 units to about 100 units of superoxide dismutase. In an embodiment, the composition comprises from about 20 units to about 190 units of superoxide dismutase. In an embodiment, the composition comprises from about 30 units to about 180 units of superoxide dismutase. In an embodiment, the composition comprises from about 40 units to about 170 units of superoxide dismutase. In an embodiment, the composition comprises from about 50 units to about 160 units of superoxide dismutase. In an embodiment, the composition comprises about 60 units to about 150 units of superoxide dismutase. In an embodiment, the composition comprises about 70 units to about 140 units of superoxide dismutase. In an embodiment, the composition comprises about 80 units to about 130 units of superoxide dismutase. In an embodiment, the composition comprises about 90 units to about 130 units of superoxide dismutase. In an embodiment, the composition comprises about 75 units to about 95 units of superoxide dismutase. In an embodiment, the composition comprises about 80 units to about 90 units of superoxide dismutase. In an embodiment, the composition comprises about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, or 160 units of superoxide dismutase. In an embodiment, the composition comprises about 84 units of superoxide dismutase.

[0052] The superoxide dismutase used in the compositions disclosed herein can be obtained from any source of enzyme. In an embodiment, the superoxide dismutase is extracted from melon, beef liver, heterotrophic bacteria or marine phytoplankton. The superoxide dismutase can also be any type of enzyme. In an embodiment, the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase or nickel superoxide dismutase.

[0053] In an embodiment, the superoxide dismutase is extracted from a plant. In an embodiment, the plant is a fruit, grain or tuber. In an embodiment, the plant is a fruit selected from melon, citrus fruit, peach, pear, apple or banana. In an embodiment, the plant is a grain selected from wheat, barley, rye, millet, oats, spelt, bulgur, sorghum and farro. In an embodiment, the plant is a tuber selected from horseradish, potato, yam, sweet potato, cassava or dahlia.

[0054] In an embodiment, the superoxide dismutase is extracted from an animal, hi an embodiment, the animal is a cow, a pig, a sheep or a goat.

[0055] In embodiments, the superoxide dismutase is extracted from a microorganism. In embodiments, the microorganism is a phytoplankton or a bacterium. In embodiments, the microorganism is a heterotrophic bacterium, e.g., a bacterium that takes up sugars from their environment that they need for energy production.

[0056] 2) Soluble Fiber The ratio of superoxide dismutase to soluble fiber can be adjusted, if necessary, by varying the amount of either or both components of the composition. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:100 to about 1:1000 by weight. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:200 to about 1:800 by weight. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:300 to about 1:700 by weight. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:500 to about 1:700 by weight. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:650 to 1:675 by weight.

[0057] Without wishing to be bound by theory, the inventors have found that the presence of soluble fiber together with superoxide dismutase in a certain ratio synergistically allows improved stability and uptake in the digestive tract (intestine), as demonstrated in the following examples.One mechanism behind this improved stability and uptake is that soluble fiber forms a gel in the digestive tract, and this formed gel captures and protects superoxide dismutase, while also allowing it to be more easily absorbed through the intestinal wall.The soluble fiber and antioxidant formulation of the present disclosure provides a synergistic effect, where polyphenols are absorbed in a substantial part of the colon and undergo extensive catabolism by colonic microbiota, while soluble prebiotic fiber acts as fertilizer for bacteria in the colon.The presence of prebiotic fiber enhances the action of colonic microbiota, thereby resulting in more effective absorption of polyphenols in the colon. The majority of polyphenols from antioxidant supplements ultimately reach the large intestine where they can undergo microbial metabolism to their active metabolites and exert their antioxidant effects. Soluble fiber modulates gut microbes and maximizes polyphenol metabolism, resulting in multiple antioxidant, anti-inflammatory, and anti-infective effects.

[0058] In an embodiment, the soluble fiber is a prebiotic fiber. As used herein, "prebiotic fiber" is a soluble fiber that forms a matrix in the digestive tract that provides a substrate for the growth of microorganisms in the gut.

[0059] In embodiments of the compositions disclosed herein, different types of soluble fiber can be used, including mixtures of two, three, four, five, six or more types of soluble fiber. In embodiments, the soluble fiber is a water-soluble polysaccharide. In embodiments, the soluble fiber is isolated from corn, wheat, barley, rye, beans, apples, pears, peaches, citrus fruits, berries, peas, rice bran or oats. In embodiments, the soluble fiber is selected from soluble corn fiber, inulin, dextrin, guar gum, oligopolysaccharides, galactopolysaccharides, fructooligosaccharides, lactulose, resistant starch, xylooligosaccharides and isomaltooligosaccharides. In embodiments, the soluble fiber is soluble corn fiber. In embodiments, the soluble fiber is resistant maltodextrin. In an embodiment, the soluble fiber is Fibersol-2® as sold by Archer Daniels Midland and Matsutani Chemical Industry, Inc. In an embodiment, the soluble fiber is another type of Fibersol-2®, such as Fibersol-2AG, Fibersol-LQ, Fibersol-2L, Fibersol-DLQ, or non-GMO Fibersol.

[0060] In some embodiments, the soluble fiber is corn-based resistant maltodextrin (Fibersol-2) prebiotic fiber.

[0061] 3) Liquid composition In an embodiment, the present disclosure provides a liquid composition comprising superoxide dismutase, soluble fiber, and water. In an embodiment, the present disclosure provides a liquid composition comprising: a) about 0.03 units / mL to about 0.5 units / mL of superoxide dismutase; b) about 1.3 mg / mL to about 23 mg / mL of soluble fiber; and c) water.

[0062] In an embodiment, the liquid composition comprises from about 0.05 units / mL to about 0.4 units / mL of superoxide dismutase. In an embodiment, the liquid composition comprises from about 0.2 units / mL to about 0.3 units / mL of superoxide dismutase. In an embodiment, the liquid composition comprises from about 0.1 units / mL to about 0.4 units / mL of superoxide dismutase. In an embodiment, the liquid composition comprises from about 0.15 units / mL to about 0.35 units / mL of superoxide dismutase. In an embodiment, the liquid composition comprises from about 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35, or 0.4 units / mL of superoxide dismutase. In embodiments, the liquid composition comprises about 0.20, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29 or 0.30 Units / mL of superoxide dismutase.

[0063] In an embodiment, the liquid composition comprises from about 2.7 mg / mL to about 12 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises from about 5.55 mg / mL to about 11.11 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises from about 2 mg / mL to about 15 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises from about 4 mg / mL to about 12 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises from about 5 mg / mL to about 12 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises from about 6 mg / mL to about 12 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises from about 7 mg / mL to about 12 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises from about 8 mg / mL to about 12 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises from about 9 mg / mL to about 12 mg / mL of soluble fiber. In embodiments, the liquid composition comprises from about 10 mg / mL to about 12 mg / mL of soluble fiber, hi embodiments, the liquid composition comprises about 2.78, 5.56, 8.33, 11.11 or 13.89 mg / mL of soluble fiber.

[0064] In embodiments, the liquid composition comprises any of the superoxide dismutases disclosed herein. In embodiments, the superoxide dismutase is extracted from melon, bovine liver, heterotrophic bacteria, or marine phytoplankton. In embodiments, the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase, or nickel superoxide dismutase.

[0065] In an embodiment of the liquid composition, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:100 to about 1:1000 by mass. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:200 to about 1:800 by mass. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:300 to about 1:700 by mass. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:500 to about 1:700 by mass. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:650 to 1:675 by mass.

[0066] In an embodiment, the liquid composition comprises any of the soluble fibers disclosed herein. In an embodiment, the soluble fiber is a prebiotic fiber. In an embodiment, the soluble fiber is a water-soluble polysaccharide. In an embodiment, the soluble fiber is selected from soluble corn fiber, inulin, dextrin, guar gum, oligopolysaccharides, galactopolysaccharides, fructooligosaccharides, lactulose, resistant starch, xylooligosaccharides, and isomaltooligosaccharides. In an embodiment, the soluble fiber is soluble corn fiber. In an embodiment, the soluble corn fiber is resistant maltodextrin.

[0067] In embodiments, the liquid composition may further comprise additional ingredients including sweeteners, pH adjusters, flavorings and other agents including combinations of these agents.

[0068] In an embodiment, the liquid composition comprises a sweetener. In an embodiment, the liquid composition comprises a combination of sweeteners, examples of which are provided below.

[0069] In embodiments, the sweetener is a simple sugar. In embodiments, the simple sugar is ribose, glucose, fructose, sucrose, lactose, or a combination thereof. In embodiments, the simple sugar is ribose. In embodiments, the simple sugar is d-ribose.

[0070] In an embodiment, the liquid composition comprises from about 0.1 mg / mL to about 1.5 mg / mL of a simple sugar. In an embodiment, the liquid composition comprises from about 0.2 mg / mL to about 1.3 mg / mL of a simple sugar. In an embodiment, the liquid composition comprises from about 0.3 mg / mL to about 1.0 mg / mL of a simple sugar. In an embodiment, the liquid composition comprises from about 0.4 mg / mL to about 0.8 mg / mL of a simple sugar. In an embodiment, the liquid composition comprises from about 0.4 mg / mL to about 0.85 mg / mL of a simple sugar. In an embodiment, the liquid composition comprises from about 0.4 mg / mL to about 0.6 mg / mL of a simple sugar. In an embodiment, the liquid composition comprises from about 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, or 0.85 mg / mL of a simple sugar.

[0071] In an embodiment, the liquid composition comprises from about 0.1 mg / mL to about 1.5 mg / mL of d-ribose. In an embodiment, the liquid composition comprises from about 0.2 mg / mL to about 1.3 mg / mL of d-ribose. In an embodiment, the liquid composition comprises from about 0.3 mg / mL to about 1.0 mg / mL of d-ribose. In an embodiment, the liquid composition comprises from about 0.4 mg / mL to about 0.8 mg / mL of d-ribose. In an embodiment, the liquid composition comprises from about 0.4 mg / mL to about 0.85 mg / mL of d-ribose. In an embodiment, the liquid composition comprises from about 0.4 mg / mL to about 0.6 mg / mL of d-ribose. In an embodiment, the liquid composition comprises from about 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, or 0.85 mg / mL of d-ribose.

[0072] In an embodiment of the liquid composition, the sweetener comprises a sugar alcohol. In an embodiment, the sugar alcohol is erythritol, mannitol, sorbitol, xylitol, lactitol, isomalt or a combination thereof. In an embodiment, the sugar alcohol is erythritol.

[0073] In an embodiment, the liquid composition comprises from about 1.3 mg / mL to about 9.0 mg / mL of sugar alcohol. In an embodiment, the liquid composition comprises from about 2.0 mg / mL to about 8.0 mg / mL of sugar alcohol. In an embodiment, the liquid composition comprises from about 3.9 mg / mL to about 7.0 mg / mL of sugar alcohol. In an embodiment, the liquid composition comprises from 2.7 mg / mL to about 5.6 mg / mL of sugar alcohol. In an embodiment, the liquid composition comprises from about 2.0, 2.5, 3.0, 3.5, 4.0, 4.15, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5 or 8.0 mg / mL of sugar alcohol.

[0074] In an embodiment, the liquid composition comprises from about 1.3 mg / mL to about 9.0 mg / mL of erythritol. In an embodiment, the liquid composition comprises from about 2.0 mg / mL to about 8.0 mg / mL of erythritol. In an embodiment, the liquid composition comprises from about 3.9 mg / mL to about 7.0 mg / mL of erythritol. In an embodiment, the liquid composition comprises from 2.7 mg / mL to about 5.6 mg / mL of erythritol. In an embodiment, the liquid composition comprises from about 2.0, 2.5, 3.0, 3.5, 4.0, 4.15, 4.5, 5.0, 5.5, 6.0, 6.5, 7.0, 7.5, or 8.0 mg / mL of erythritol.

[0075] In embodiments, the sweetener is a steviol glycoside. In embodiments, the steviol glycoside is rebaudioside A, stevioside, rebaudioside C, dulcoside A, rebaudioside B, rebaudioside D, rebaudioside E, steviolbioside, or a combination thereof.

[0076] In an embodiment, the liquid composition comprises from about 0.05 mg / mL to about 0.75 mg / mL of steviol glycoside. In an embodiment, the liquid composition comprises from about 0.2 mg / mL to about 0.35 mg / mL of steviol glycoside. In an embodiment, the liquid composition comprises from about 0.1 mg / mL to about 0.5 mg / mL of steviol glycoside. In an embodiment, the liquid composition comprises from about 0.2 mg / mL to about 0.3 mg / mL of steviol glycoside. In an embodiment, the liquid composition comprises from about 0.10, 0.15, 0.20, 0.25, 0.27, 0.30, 0.35, 0.40, 0.45, or 0.50 mg / mL of steviol glycoside.

[0077] In embodiments, the liquid composition comprises from about 0.05 mg / mL to about 0.75 mg / mL of rebaudioside A. In embodiments, the liquid composition comprises from about 0.2 mg / mL to about 0.35 mg / mL of rebaudioside A. In embodiments, the liquid composition comprises from about 0.1 mg / mL to about 0.5 mg / mL of rebaudioside A. In embodiments, the liquid composition comprises from about 0.2 mg / mL to about 0.3 mg / mL of rebaudioside A. In embodiments, the liquid composition comprises about 0.10, 0.15, 0.20, 0.25, 0.27, 0.30, 0.35, 0.40, 0.45, or 0.50 mg / mL of rebaudioside A.

[0078] In an embodiment, the liquid composition further comprises a pH adjuster. In an embodiment, the pH adjuster is any food-safe agent that can be used to modify the pH of the liquid composition. In an embodiment, the pH adjuster is citric acid, acetic acid, hydrochloric acid, lactic acid, malic acid, phosphoric acid, tartaric acid, or a combination thereof.

[0079] In an embodiment, the liquid composition comprises from about 0.1 mg / mL to about 1.5 mg / mL of the pH adjuster. In an embodiment, the liquid composition comprises from about 0.2 mg / mL to about 1.3 mg / mL of the pH adjuster. In an embodiment, the liquid composition comprises from about 0.3 mg / mL to about 1.0 mg / mL of the pH adjuster. In an embodiment, the liquid composition comprises from about 0.4 mg / mL to about 0.8 mg / mL of the pH adjuster. In an embodiment, the liquid composition comprises from about 0.4 mg / mL to about 0.85 mg / mL of the pH adjuster. In an embodiment, the liquid composition comprises from about 0.4 mg / mL to about 0.6 mg / mL of the pH adjuster. In an embodiment, the liquid composition comprises from about 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, or 0.85 mg / mL of the pH adjuster.

[0080] In an embodiment, the liquid composition comprises from about 0.1 mg / mL to about 1.5 mg / mL of citric acid. In an embodiment, the liquid composition comprises from about 0.2 mg / mL to about 1.3 mg / mL of citric acid. In an embodiment, the liquid composition comprises from about 0.3 mg / mL to about 1.0 mg / mL of citric acid. In an embodiment, the liquid composition comprises from about 0.4 mg / mL to about 0.8 mg / mL of citric acid. In an embodiment, the liquid composition comprises from about 0.4 mg / mL to about 0.85 mg / mL of citric acid. In an embodiment, the liquid composition comprises from about 0.4 mg / mL to about 0.6 mg / mL of citric acid. In an embodiment, the liquid composition comprises from about 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8, or 0.85 mg / mL of citric acid.

[0081] In an embodiment, the liquid composition comprises a flavoring. In an embodiment, the flavoring is a natural flavoring. In an embodiment, the flavoring is an artificial flavoring. In an embodiment, the flavoring is a fruit flavoring. In an embodiment, the flavoring is pomegranate, red grape, blueberry, dark sweet cherry, blackberry, goji berry, acai berry, blackberry, raspberry, strawberry, gooseberry, cranberry, orange, grapefruit, watermelon, beet, apple, lemon, lime, lychee, pineapple, prune, mango, or combinations thereof. In an embodiment, the liquid composition comprises a cola flavoring.

[0082] In an embodiment, the liquid composition is formulated into a beverage, hi an embodiment, the liquid composition is formulated as a rehydration drink, protein shake, juice, tea, coffee, milk, kefir, ice cream, yogurt, smoothie, broth or soup.

[0083] In an embodiment, the liquid composition has a volume of about 15 mL to about 1500 mL. In an embodiment, the liquid composition has a volume of about 30 mL to about 1200 mL. In an embodiment, the liquid composition has a volume of about 50 mL to about 1000 mL. In an embodiment, the liquid composition has a volume of about 100 mL to about 500 mL. In an embodiment, the liquid composition has a volume of about 200 mL to about 400 mL. In an embodiment, the liquid composition has a volume of about 200 mL to about 1000 mL. In an embodiment, the liquid composition has a volume of about 300 mL to about 1000 mL. In an embodiment, the liquid composition has a volume of about 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mL.

[0084] In an embodiment, the liquid composition is packaged in a bottle. In an embodiment, the bottle is a glass bottle. In an embodiment, the bottle is a plastic bottle. In an embodiment, the liquid composition is packaged in a can. In an embodiment, the liquid composition is packaged in a beverage box.

[0085] In embodiments, specific examples of liquid compositions are provided in Table 1.

[0086] [Table 1]

[0087] 4) Powder composition In some embodiments of the compositions disclosed herein, the compositions are in the form of a powder. In embodiments in which the compositions are in the form of a powder, the compositions can be consumed in dry powder or added to beverages or foods. In embodiments, the powder is mixed into water, hydration beverages, protein shakes, juices, tea, coffee, milk, kefir, ice cream, yogurt, smoothies, broths or soups prior to consumption.

[0088] In an embodiment, the powder composition comprises a high concentration of superoxide dismutase by mass, for example, greater than 0.1 ppm, greater than 0.5 ppm, greater than 1 ppm, greater than 2 ppm, greater than 5 ppm, greater than 10 ppm, greater than 20 ppm, greater than 50 ppm, greater than 100 ppm, greater than 200 ppm, greater than 300 ppm, greater than 400 ppm, or greater than 500 ppm. In an embodiment, the powder composition comprises from about 0.1 ppm to about 10 ppm of superoxide dismutase by mass. In an embodiment, the powder composition comprises from about 0.5 ppm to about 5 ppm of superoxide dismutase by mass. In an embodiment, the powder composition comprises from about 0.7 ppm to about 2 ppm of superoxide dismutase by mass. In an embodiment, the powder composition comprises from about 0.8 ppm to about 1.2 ppm of superoxide dismutase by mass. In embodiments, the powder composition comprises about 0.1, 0.25, 0.5, 0.75, 1.0, 1.1, 1.2, 1.25, 1.3, 1.4, 1.5, 1.6, 1.7, 1.75, 1.8, 1.9, 2.0, 2.25, 2.5, 2.75, 3.0, 3.25, 3.5, 3.75, 4.0, 4.25, 4.5, 4.75, 5.0, 5.25, 5.5, 5.75, 6.0, 6.25, 6.5, 6.75, 7.0, 7.25, 7.5, 7.75, 8.0, 8.25, 8.5, 8.75, 9.0, 9.25, 9.5, 9.75, or 10.0 ppm by weight of superoxide dismutase.

[0089] In an embodiment, the powder composition comprises about 5 ppm to about 15 ppm by weight of superoxide dismutase. In an embodiment, the powder composition comprises about 7.5 ppm to about 12.5 ppm by weight of superoxide dismutase. In an embodiment, the powder composition comprises about 8 ppm to about 11 ppm by weight of superoxide dismutase. In an embodiment, the powder composition comprises about 9 ppm to about 11 ppm by weight of superoxide dismutase. In an embodiment, the powder composition comprises about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 ppm by weight of superoxide dismutase.

[0090] In an embodiment, the powder composition comprises from about 1% to about 90% soluble fiber by weight. In an embodiment, the powder composition comprises from about 1% to about 50% soluble fiber by weight. In an embodiment, the powder composition comprises from about 5% to about 25% soluble fiber by weight. In an embodiment, the powder composition comprises from about 10% to about 20% soluble fiber by weight. In an embodiment, the powder composition comprises from about 12% to about 14% soluble fiber by weight. In embodiments, the power composition comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29% or 30% soluble fiber by weight.

[0091] In an embodiment, the powder composition comprises about 50% to about 75% soluble fiber by weight. In an embodiment, the powder composition comprises about 50% to about 70% soluble fiber by weight. In an embodiment, the powder composition comprises about 60% to about 70% soluble fiber by weight. In an embodiment, the powder composition comprises about 50% to about 80% soluble fiber by weight. In an embodiment, the powder composition comprises about 50%, 55%, 60%, 65%, 66%, 67%, 70%, 75%, 80%, 85% or 90% soluble fiber by weight.

[0092] In an embodiment, the powder composition comprises a sweetener. In an embodiment, the powder composition comprises a combination of sweeteners, examples of which are provided below.

[0093] In embodiments, the sweetener is a simple sugar. In embodiments, the simple sugar is ribose, glucose, fructose, sucrose, lactose, or a combination thereof. In embodiments, the simple sugar is ribose. In embodiments, the simple sugar is d-ribose.

[0094] In embodiments, the powder composition comprises from about 1% to about 10% by weight of a simple sugar. In embodiments, the powder composition comprises from about 2% to about 8% by weight of a simple sugar. In embodiments, the powder composition comprises from about 2% to about 5% by weight of a simple sugar. In embodiments, the powder composition comprises from about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% by weight of a simple sugar.

[0095] In embodiments, the powder composition comprises about 1% to about 10% by weight d-ribose. In embodiments, the powder composition comprises about 2% to about 8% by weight d-ribose. In embodiments, the powder composition comprises about 2% to about 5% by weight d-ribose. In embodiments, the powder composition comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% by weight d-ribose.

[0096] In an embodiment of the powder composition, the sweetener comprises a sugar alcohol. In an embodiment, the sugar alcohol is erythritol, mannitol, sorbitol, xylitol, lactitol, isomalt or a combination thereof. In an embodiment, the sugar alcohol is erythritol.

[0097] In an embodiment, the powder composition comprises about 10% to about 50% by weight of erythritol. In an embodiment, the powder composition comprises about 15% to about 35% by weight of erythritol. In an embodiment, the powder composition comprises about 20% to about 30% by weight of erythritol. In an embodiment, the powder composition comprises about 15%, 20%, 25, 30%, 35%, 40%, 45% or 50% by weight of erythritol.

[0098] In embodiments, the sweetener is a steviol glycoside. In embodiments, the steviol glycoside is rebaudioside A, stevioside, rebaudioside C, dulcoside A, rebaudioside B, rebaudioside D, rebaudioside E, steviolbioside, or a combination thereof.

[0099] In an embodiment, the powder composition comprises about 0.5% to about 5% by weight of steviol glycosides. In an embodiment, the powder composition comprises about 1% to about 4% by weight of steviol glycosides. In an embodiment, the powder composition comprises about 1% to about 2.5% by weight of steviol glycosides. In an embodiment, the powder composition comprises about 1%, 1.5%, 1.6%, 2%, 2.5%, 3%, 3.5%, or 4% by weight of steviol glycosides.

[0100] In embodiments, the powder composition comprises from about 0.5% to about 5% by weight rebaudioside A. In embodiments, the powder composition comprises from about 1% to about 4% by weight rebaudioside A. In embodiments, the powder composition comprises from about 1% to about 2.5% by weight rebaudioside A. In embodiments, the powder composition comprises about 1%, 1.5%, 1.6%, 2%, 2.5%, 3%, 3.5%, or 4% by weight rebaudioside A.

[0101] In an embodiment, the liquid composition further comprises a pH adjuster. In an embodiment, the pH adjuster is any food-safe agent that can be used to modify the pH of the liquid composition. In an embodiment, the pH adjuster is citric acid, acetic acid, hydrochloric acid, lactic acid, malic acid, phosphoric acid, tartaric acid, or a combination thereof.

[0102] In an embodiment, the powder composition comprises about 1% to about 10% by weight of a pH adjuster. In an embodiment, the powder composition comprises about 2% to about 8% by weight of a pH adjuster. In an embodiment, the powder composition comprises about 2% to about 5% by weight of a pH adjuster. In an embodiment, the powder composition comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% by weight of a pH adjuster.

[0103] In an embodiment, the powder composition comprises about 1% to about 10% by weight citric acid. In an embodiment, the powder composition comprises about 2% to about 8% by weight citric acid. In an embodiment, the powder composition comprises about 2% to about 5% by weight citric acid. In an embodiment, the powder composition comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% by weight citric acid.

[0104] In an embodiment, specific examples of powder compositions are provided in Table 2.

[0105] [Table 2]

[0106] 5) Gel Composition In some embodiments of the compositions disclosed herein, the composition is in the form of a gel. In embodiments where the composition is in the form of a gel, it can be directly consumed in that form. In other embodiments, the gel can be added to a beverage or food. In embodiments, the gel is mixed into water, hydration beverages, protein shakes, juices, tea, coffee, milk, kefir, ice cream, yogurt, smoothies, broths, or soups prior to consumption.

[0107] In an embodiment, the gel composition comprises from about 0.005 mg / mL to about 5.0 mg / mL of superoxide dismutase. In an embodiment, the gel composition comprises from about 0.01 mg / mL to about 2.5 mg / mL of superoxide dismutase. In an embodiment, the gel composition comprises from about 0.05 mg / mL to about 1.0 mg / mL of superoxide dismutase. In an embodiment, the gel composition comprises from about 0.1 mg / mL to about 0.5 mg / mL of superoxide dismutase. In embodiments, the gel composition comprises about 0.005, 0.01, 0.05, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9 or 3.0 mg / mL of superoxide dismutase.

[0108] In an embodiment, the gel composition comprises about 50 mg / mL to about 1000 mg / mL of soluble fiber. In an embodiment, the gel composition comprises about 70 mg / mL to about 500 mg / mL of soluble fiber. In an embodiment, the gel composition comprises about 90 mg / mL to about 250 mg / mL of soluble fiber. In an embodiment, the gel composition comprises about 100 mg / mL to about 200 mg / mL of soluble fiber. In an embodiment, the gel composition comprises about 50, 75, 100, 110, 120, 125, 130, 133, 135, 140, 150, 160, 170, 175, 180, 190, 200, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, or 500 mg / mL of soluble fiber.

[0109] In an embodiment of the composition disclosed herein, the composition comprising superoxide dismutase and soluble fiber also comprises fruit juice. The fruit juice may provide additional antioxidants, soluble fiber, insoluble fiber, vitamins and nutrients to the composition. In an embodiment, the gel composition disclosed herein comprises fruit juice.

[0110] In an embodiment, the fruit juice is pomegranate juice, red grape juice, blueberry juice, dark sweet cherry juice, blackberry juice, raspberry juice, strawberry juice, gooseberry juice, cranberry juice, orange juice, grapefruit juice, watermelon juice, beet juice, apple juice, lemon juice, lime juice, lychee juice, pineapple juice, prune juice, or a combination thereof. In an embodiment, the composition comprises two, three, four, five or six of the juices selected from pomegranate juice, red grape juice, blueberry juice, dark sweet cherry juice, blackberry juice, raspberry juice, strawberry juice, gooseberry juice, cranberry juice, orange juice, grapefruit juice, watermelon juice, beet juice, apple juice, lemon juice, lime juice, lychee juice, pineapple juice, and prune juice.

[0111] In embodiments, the fruit juice may be concentrated, e.g., some of the water in the original fruit juice may have been removed. In embodiments, the fruit juice is concentrated pomegranate juice, red grape juice, blueberry juice, dark sweet cherry juice, black cherry juice, goji berry juice, acai berry juice, blackberry juice, raspberry juice, strawberry juice, gooseberry juice, cranberry juice, orange juice, grapefruit juice, watermelon juice, beet juice, apple juice, lemon juice, lime juice, lychee juice, pineapple juice, prune juice, or combinations thereof. In an embodiment, the composition includes two, three, four, five, or six types of concentrated fruit juice selected from concentrated pomegranate juice, concentrated red grape juice, concentrated blueberry juice, concentrated dark sweet cherry juice, concentrated black cherry juice, concentrated goji berry juice, concentrated acai berry juice, concentrated blackberry juice, concentrated raspberry juice, concentrated strawberry juice, concentrated gooseberry juice, concentrated cranberry juice, concentrated orange juice, concentrated grapefruit juice, concentrated watermelon juice, concentrated beet juice, concentrated apple juice, concentrated lemon juice, concentrated lime juice, concentrated lychee juice, concentrated pineapple juice, and concentrated prune juice. In an embodiment, the concentrated fruit juice has about 60% to about 97% of the water removed from the fruit juice. In an embodiment, the concentrated fruit juice has about 85% to about 95% of the water removed from the fruit juice. In embodiments, the concentrated juice has about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of the water removed from the juice. Water can be removed from the juice using any method known in the art for concentrating juice.

[0112] In an embodiment, the composition comprises from about 5 mg / mL to about 200 mg / mL of concentrated fruit juice. In an embodiment, the composition comprises from about 75 mg / mL to about 150 mg / mL of concentrated pomegranate juice. In an embodiment, the composition comprises from about 75 mg / mL to about 150 mg / mL of concentrated red grape juice. In an embodiment, the composition comprises from about 25 mg / mL to about 100 mg / mL of concentrated blueberry juice. In an embodiment, the composition comprises from about 20 mg / mL to about 80 mg / mL of concentrated dark sweet cherry juice. In an embodiment, the composition comprises from about 20 mg / mL to about 80 mg / mL of concentrated black cherry juice. In an embodiment, the composition comprises from about 2 mg / mL to about 20 mg / mL of concentrated goji berry juice. In an embodiment, the composition comprises from about 2 mg / mL to about 20 mg / mL of concentrated acai berry juice.

[0113] In embodiments of the gel compositions disclosed herein, the composition comprising superoxide dismutase and soluble fiber also comprises aloe vera, which may provide additional antioxidants, soluble fiber, insoluble fiber, vitamins and nutrients to the composition.

[0114] In some embodiments, the aloe vera is concentrated aloe vera. In embodiments, the aloe vera can be concentrated, e.g., extracted from the aloe vera plant, from which some of the water has been removed. In embodiments, the concentrated aloe vera has had about 60% to about 97% of the water removed from the aloe vera. In embodiments, the concentrated aloe vera has about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% of the water removed from the aloe vera. In embodiments, the composition comprises about 2 mg / mL to about 20 mg / mL of concentrated aloe vera.

[0115] In an embodiment of the gel composition disclosed herein, the composition comprising superoxide dismutase and soluble fiber also comprises green tea, which may provide additional antioxidants, vitamins and nutrients to the composition.

[0116] In an embodiment, the green tea is concentrated green tea. In an embodiment, the concentrated green tea is a green tea extract made from green tea leaves or green tea powder. In an embodiment, the green tea is added to the composition in powder form. In an embodiment, the composition comprises about 2 mg / mL to about 20 mg / mL of concentrated green tea.

[0117] In embodiments of the gel compositions disclosed herein, the composition comprising superoxide dismutase and soluble fiber also comprises resveratrol, which may provide additional antioxidants to the composition. In some embodiments, the composition comprises about 0.5 mg / mL to about 6 mg / mL of resveratrol.

[0118] In embodiments, specific gel compositions are provided in Table 3.

[0119] [Table 3]

[0120] 6) Probiotic Composition In an embodiment, disclosed herein is a composition comprising superoxide dismutase, soluble fiber, and probiotics. In an embodiment, the probiotic composition comprises: a) about 10 units to about 200 units of superoxide dismutase; b) about 500 mg to about 8000 mg of soluble fiber; and c) probiotics.

[0121] In an embodiment, the concentration of the amount of superoxide dismutase in the probiotic compositions disclosed herein is measured in terms of the total amount of units of superoxide dismutase in the composition. In an embodiment, the composition comprises from about 10 units to about 200 units of superoxide dismutase. In an embodiment, the composition comprises from about 50 units to about 150 units of superoxide dismutase. In an embodiment, the composition comprises from about 70 units to about 100 units of superoxide dismutase. In an embodiment, the composition comprises from about 20 units to about 190 units of superoxide dismutase. In an embodiment, the composition comprises from about 30 units to about 180 units of superoxide dismutase. In an embodiment, the composition comprises from about 40 units to about 170 units of superoxide dismutase. In an embodiment, the composition comprises from about 50 units to about 160 units of superoxide dismutase. In an embodiment, the composition comprises about 60 units to about 150 units of superoxide dismutase. In an embodiment, the composition comprises about 70 units to about 140 units of superoxide dismutase. In an embodiment, the composition comprises about 80 units to about 130 units of superoxide dismutase. In an embodiment, the composition comprises about 90 units to about 130 units of superoxide dismutase. In an embodiment, the composition comprises about 75 units to about 95 units of superoxide dismutase. In an embodiment, the composition comprises about 80 units to about 90 units of superoxide dismutase. In an embodiment, the composition comprises about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, or 160 units of superoxide dismutase. In an embodiment, the composition comprises about 84 units of superoxide dismutase.

[0122] In an embodiment, the probiotic composition comprises about 1000 mg to about 5000 mg of soluble fiber. In an embodiment, the probiotic composition comprises about 2000 mg to about 4000 mg of soluble fiber. In an embodiment, the probiotic composition comprises about 1000 mg to about 10000 mg of soluble fiber. In an embodiment, the probiotic composition comprises about 2000 mg to about 9000 mg of soluble fiber. In an embodiment, the probiotic composition comprises about 3000 mg to about 8000 mg of soluble fiber. In an embodiment, the probiotic composition comprises about 2000 mg to about 4000 mg of soluble fiber. In an embodiment, the probiotic composition comprises about 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, or 5000 mg of soluble fiber.

[0123] In an embodiment, the ratio of superoxide dismutase to soluble fiber in the probiotic composition is about 1:100 to about 1:1000 by weight. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:200 to about 1:800 by weight. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:300 to about 1:700 by weight. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:500 to about 1:700 by weight. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:650 to 1:675 by weight.

[0124] The probiotic composition may comprise one or more superoxide dismutases as disclosed herein. In an embodiment, the superoxide dismutase is extracted from melon, beef liver, heterotrophic bacteria or marine phytoplankton. In an embodiment, the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase or nickel superoxide dismutase.

[0125] The probiotic composition may comprise one or more soluble fibers as disclosed herein. In an embodiment, the soluble fiber is a water-soluble polysaccharide. In an embodiment, the soluble fiber is selected from soluble corn fiber, inulin, dextrin, guar gum, oligopolysaccharides, galactopolysaccharides, fructooligosaccharides, lactulose, resistant starch, xylooligosaccharides and isomaltooligosaccharides. In an embodiment, the soluble fiber is soluble corn fiber. In an embodiment, the soluble corn fiber is resistant maltodextrin.

[0126] In an embodiment, the probiotic of the probiotic composition comprises one or more beneficial microorganisms. In an embodiment, the probiotic comprises bacteria of the genus Bifidobacterium. In an embodiment, the probiotic comprises bacteria of the genus Lactobacillus. In an embodiment, the probiotic comprises Lactobacillus of the phylum Firmicutes, Bifidobacteriaceae of the phylum Actinobacteria, or a combination thereof.

[0127] In an embodiment, the probiotic composition is in the form of a gel. Examples of gels suitable for supplementing with probiotics to make the probiotic composition are disclosed herein.

[0128] In an embodiment, the probiotic composition is in the form of a powder. Examples of powders suitable for supplementing with probiotics to make the probiotic composition are disclosed herein.

[0129] In an embodiment, the probiotic composition is in the form of a liquid. Examples of liquids suitable for supplementing with probiotics to make the probiotic composition are disclosed herein.

[0130] 7) Excipients / Formulations In embodiments, the compositions disclosed herein can be combined with one or more excipients. In embodiments, the excipient is a gelling agent, a thickening agent, a carrier, a buffering agent or a bulking agent. In embodiments, the compositions disclosed herein can be formulated into beverages or foods. In embodiments, the compositions are formulated into juices, hydration drinks (e.g., sports drinks), protein shakes, teas, coffees, milks, kefirs, ice creams, yogurts, smoothies, broths or soups.

[0131] In an embodiment, the composition comprises a thickening / gelling agent carboxymethylcellulose. In an embodiment, the composition is a gel comprising superoxide dismutase, soluble fiber and carboxymethylcellulose. In an embodiment, the composition comprises about 0.5 to about 5.0 mg / mL carboxymethylcellulose.

[0132] In an embodiment, the composition comprises a thickening / gelling agent xanthan gum. In an embodiment, the composition is a gel comprising superoxide dismutase, soluble fiber and xanthan gum. In an embodiment, the composition comprises about 0.5 to about 5.0 mg / mL xanthan gum.

[0133] 8) Method The present disclosure is also directed to a method of increasing T cell activation in a subject, the method comprising orally administering to the subject a composition comprising: a) about 10 units to about 200 units of superoxide dismutase; and b) about 500 mg to about 8000 mg of soluble fiber; wherein after administration of the composition, T cell activation is increased in the subject.

[0134] In embodiments, T cell activation is measured by measuring biomarkers of T cell activation as disclosed in the Examples below.

[0135] In embodiments, the composition is administered in combination with an anti-cancer agent.In embodiments, the composition is administered in combination with an anti-viral agent.

[0136] In embodiments of the methods of increasing T cell activation, the methods are used to prevent or treat viral infections, including influenza A, influenza B, influenza C, influenza D, SARS (severe acute respiratory syndrome), SARS-CoV-2 (which causes COVID-19), coronaviruses including MERS (Middle East respiratory syndrome), HIV, Ebola, rhinovirus and respiratory syncytial virus infections.

[0137] The disclosure also provides a method for increasing production of short chain fatty acids (SCFAs) in the digestive tract of a subject, the method comprising orally administering to the subject a composition comprising: a) about 10 units to about 200 units of superoxide dismutase; and b) about 500 mg to about 8000 mg of soluble fiber; wherein after administration of the composition, production of SCFAs is increased in the digestive tract of the subject.

[0138] In an embodiment, the SCFAs whose production is increased are acetate, propionate, butyrate, or lactate SCFAs, or combinations thereof. In an embodiment, the SCFAs are increased to provide approximately the same ratio of acetate, propionate, butyrate, and lactate SCFAs compared to the ratio of acetate, propionate, butyrate, and lactate SCFAs before the increase.

[0139] The disclosure also provides a method for increasing the amount of bacteria of the genus Bifidobacterium or Lactobacillus in the digestive tract of a subject, the method comprising orally administering to the subject a composition comprising: a) about 10 units to about 200 units of superoxide dismutase; and b) about 500 mg to about 8000 mg of soluble fiber; wherein after administration of the composition, the amount of bacteria of the genus Bifidobacterium, Lactobacillus, or a combination thereof is increased in the digestive tract of the subject.

[0140] In an embodiment of the method, the bacteria of the genus Bifidobacterium comprises a species of the family Bifidobacteria in the phylum Actinobacteria. In an embodiment, the bacteria of the genus Lactobacillus comprises a species of the phylum Firmicutes Lactobacillus.

[0141] In an embodiment of the above method, the composition used comprises about 10 units to about 200 units of superoxide dismutase. In an embodiment, the composition comprises about 50 units to about 150 units of superoxide dismutase. In an embodiment, the composition comprises about 70 units to about 100 units of superoxide dismutase. In an embodiment, the composition comprises about 20 units to about 190 units of superoxide dismutase. In an embodiment, the composition comprises about 30 units to about 180 units of superoxide dismutase. In an embodiment, the composition comprises about 40 units to about 170 units of superoxide dismutase. In an embodiment, the composition comprises about 50 units to about 160 units of superoxide dismutase. In an embodiment, the composition comprises about 60 units to about 150 units of superoxide dismutase. In an embodiment, the composition comprises about 70 units to about 140 units of superoxide dismutase. In an embodiment, the composition comprises about 80 units to about 130 units of superoxide dismutase. In an embodiment, the composition comprises about 90 units to about 130 units of superoxide dismutase. In an embodiment, the composition comprises about 75 units to about 95 units of superoxide dismutase. In an embodiment, the composition comprises about 80 units to about 90 units of superoxide dismutase. In an embodiment, the composition comprises about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, or 160 units of superoxide dismutase. In an embodiment, the composition comprises about 84 units of superoxide dismutase.

[0142] In an embodiment of the above method, the composition used comprises about 1000 mg to about 5000 mg of soluble fiber. In an embodiment, the composition comprises about 2000 mg to about 4000 mg of soluble fiber. In an embodiment, the composition comprises about 1000 mg to about 10000 mg of soluble fiber. In an embodiment, the composition comprises about 2000 mg to about 9000 mg of soluble fiber. In an embodiment, the composition comprises about 3000 mg to about 8000 mg of soluble fiber. In an embodiment, the composition comprises about 2000 mg to about 4000 mg of soluble fiber. In an embodiment, the composition comprises about 500, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, or 5000 mg of soluble fiber.

[0143] In an embodiment of the above method, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:100 to about 1:1000 by mass. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:200 to about 1:800 by mass. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:300 to about 1:700 by mass. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:500 to about 1:700 by mass. In an embodiment, the ratio of superoxide dismutase to soluble fiber in the composition is about 1:650 to 1:675 by mass.

[0144] In an embodiment of the above method, the composition can include one or more superoxide dismutases as disclosed herein. In an embodiment, the superoxide dismutase is extracted from melon, bovine liver, heterotrophic bacteria, or marine phytoplankton. In an embodiment, the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase, or nickel superoxide dismutase.

[0145] In an embodiment of the above method, the composition can include one or more soluble fibers as disclosed herein. In an embodiment, the soluble fiber is a water-soluble polysaccharide. In an embodiment, the soluble fiber is selected from soluble corn fiber, inulin, dextrin, guar gum, oligopolysaccharides, galactopolysaccharides, fructooligosaccharides, lactulose, resistant starch, xylooligosaccharides and isomaltooligosaccharides. In an embodiment, the soluble fiber is soluble corn fiber. In an embodiment, the soluble corn fiber is resistant maltodextrin.

[0146] In embodiments of the above methods, the composition is in the form of a gel. Examples of gels suitable for use in the above methods are disclosed herein.

[0147] In embodiments of the above methods, the composition is in the form of a powder. Examples of powders suitable for use in the above methods are disclosed herein.

[0148] In embodiments of the above methods, the composition is in the form of a liquid. Examples of liquids suitable for use in the above methods are disclosed herein.

[0149] In some embodiments of the methods disclosed herein, the composition is administered to the mammal once a day. In some embodiments of the methods disclosed herein, the composition is administered to the mammal twice a day. In some embodiments of the methods disclosed herein, the composition is administered to the mammal three, four, five, six, seven, eight, nine, ten or more times a day.

[0150] In any of the embodiments of the methods disclosed herein, the mammal is a human. In any of the embodiments of the methods disclosed herein, the mammal is a primate (e.g., monkey, ape, gorilla, macaque), domestic animal (e.g., dog, cat, rabbit, hamster, guinea pig, mouse, rat) or agricultural animal (e.g., cow, sheep, horse, goat, pig). EXAMPLES

[0151] Working Example Example 1 SOD and soluble fiber gel stimulate T cell activation, antioxidant and anti-inflammatory pathways as demonstrated in vitro in Jurkat cells Background and Objectives: REVIVIFY® Provitality Antioxidant Gel is composed of the primary antioxidant superoxide dismutase (SOD), prebiotic fiber, and a variety of polyphenols derived from various fruit juices. REVIVIFY® has the formulation as shown in Table 4.

[0152] [Table 4]

[0153] SOD reduces superoxide anions produced due to normal cellular activity. Polyphenols are phenolic compounds that act as antioxidants, anti-inflammatory and antiviral agents. They repair damaged cells caused by ROS / RNS reactive oxygen molecules. Dietary prebiotic fiber alters the beneficial gut ecology microbiome and provides numerous health benefits including increased immunity. The combination of these three ingredients stimulates the immune system via T cell activation as well as antioxidant and anti-inflammatory pathways. The purpose of this study was to evaluate the effect of REVIVIFY® Gel on an in vitro T cell model.

[0154] Methods: The Jurkat cell line is an immortalized T lymphocyte cell line that has been most frequently used as a prototypic T cell line to study multiple events in T cell biology, including T cell signaling. Jurkat cells were seeded in 6-well plates. Prior to treatment, cells were incubated in serum-free medium for 24 hours. Cells were treated for 48 hours with the following agents: 1. Superoxide dismutase only; 2. Prebiotic fiber only; 3. Fruit juice only; 4. Superoxide dismutase + Prebiotic fiber + Fruit juice (combination); 5. Positive control: Phorbol 12-myristate 13-acetate (PMA) in combination with ionomycin; 6. Negative control: Cell culture medium.

[0155] After treatment, the medium was removed from the cells and placed in test tubes. The levels of CD-8+, CD-4+, interferon-gamma (IFNγ), interleukin-6 (IL-6), interferon-gamma-inducible protein 10 (IP-10, also known as CXCL10), macrophage inflammatory protein 1α and 1β, monocyte chemotactic protein 1 (MCP-1, also known as CCL2), and 8-isoprostane were measured by commercially available ELISA kits.

[0156] Activated Jurkat cells were seen by upregulated CD69 (measured using anti-CD69 antibody MCA2806A647, BioRad) expression on the CD3 (measured using anti-CD3 antibody MCA463A488) positive population of cells. Cells were gated on lymphocytes in the presence of Human Seroblock (BUF070A, BioRad). Jurkat cells were stimulated with treatments for 5 days and stained with CytoTrack Red 628 / 643 with a Cell Proliferation Assay Kit (1351205, BioRad). Data were acquired on a ZE5 cell analyzer. Data are presented as mean ± SE. Statistical significance was assessed by ANOVA and Duncan post-hoc test for differences between treatment groups, with treatments with p<0.05 versus the negative control effect considered significant. Results are presented as mean ± SE (n=6, quadruplicate).

[0157] Results: As discussed in detail below, REVIVIFY® and its components activated T cells as seen by upregulated CD69 expression and stimulated CD4+ and CD8+ differentiation compared to culture medium. The gel and its components attenuated lipopolysaccharide-induced activation of 8-isoprostane (8IP), COX-2, IFN-γ, IL-6, TGF-β, TNF-α and CXCL10 secretion by Jurkat cells.

[0158] Conclusion / Perspective: REVIVIFY® Gel contains superoxide dismutase, prebiotic fiber, and polyphenols and quercetin derived from fruit juice. This unique multifaceted approach to prevent oxidative stress, maintain a balance between pro- and anti-inflammatory, and stimulate the immune response is very fast and effective.

[0159] introduction Superoxide dismutase (SOD): SOD constitutes a very important antioxidant defense against oxidative stress in the body. This enzyme serves as a good therapeutic agent against reactive oxygen species-mediated diseases. SOD may have therapeutic effects in various physiological conditions and pathologies, such as cancer, inflammatory diseases, cystic fibrosis, ischemia, aging, rheumatoid arthritis, neurodegenerative diseases, and diabetes. However, this enzyme has certain limitations in clinical applications due to problems associated with absorption. Therefore, SOD conjugates and mimetics have been developed to increase its therapeutic efficiency. 1、2、3、4、5、6、7 .

[0160] Polyphenols: Polyphenols are secondary metabolites of plants and are generally involved in defense against UV radiation or attack by pathogens. In the last decade, there has been great interest in the possible health benefits of dietary plant polyphenols as antioxidants. Epidemiological studies and associated meta-analyses strongly suggest that long-term intake of a diet rich in plant polyphenols confers protection against the development of cancer, cardiovascular disease, diabetes, osteoporosis and neurodegenerative diseases. 8、9、10、11、12 .

[0161] Dietary Prebiotic Fiber: The health benefits of dietary fiber have long been appreciated. Higher intakes of dietary fiber have been linked to less cardiovascular disease, and fiber plays a role in intestinal health, with many effective laxatives actually isolated from fiber sources. Higher intakes of fiber are associated with lower body weight. Originally, only polysaccharides were included in dietary fiber, but more recent definitions include oligosaccharides as dietary fiber based on their physiological effects, rather than based on their chemical measurement as dietary fiber by the commonly accepted total dietary fiber (TDF) method. Inulin, fructooligosaccharides, and other oligosaccharides are included as fiber on food labels in the United States. In addition, oligosaccharides are the best known "prebiotics," "selectively fermented ingredients that allow specific changes in both composition and / or activity in the gastrointestinal microflora that confer benefits to the well-bringing and health of the host." To date, all known and putative prebiotics are carbohydrate compounds, primarily oligosaccharides, that are known to survive digestion in the human small intestine and reach the colon where they are fermented by the gut microflora. Studies have provided evidence that inulin and oligofructose (OF), lactulose, and resistant starch (RS) meet all aspects of the definition, including the stimulation of the beneficial bacterial genus Bifidobacterium. Other isolated carbohydrates and carbohydrate-containing foods, including galactooligosaccharides (GOS), transgalactooligosaccharides (TOS), polydextrose, wheat dextrin, gum arabic, psyllium, banana, whole wheat, and whole corn, also have prebiotic effects. 13、14、15、16、17 .

[0162] T cell activation and immunity: T cells originate in the thymus gland and are programmed to be specific for one particular foreign particle (antigen). Upon leaving the thymus, T cells circulate through the body until they recognize their antigen on the surface of antigen-presenting cells (APCs). CD4 + On helper T cells and CD8 +Both T cell receptors (TCRs) on a cytotoxic T cell bind to the antigen when held in a structure called the MHC complex on the surface of an APC. This initiates the initial activation of the T cell. CD4 and CD8 molecules then also bind to the MHC molecule, stabilizing the whole structure. This initial binding between a T cell specific for an antigen and its matching antigen-MHC initiates the full response, which usually occurs in secondary lymphoid organs. 18 .

[0163] Role of T cells in COVID-19 infection: Like antibody-producing B cells, T cells are key players in the immune response to viral infections 19 When the SARS-CoV-2 virus that causes COVID-19 infects epithelial cells, such as those found in the respiratory tract, it replicates inside the cell, using the host cell's biochemical machinery. This causes the host cell to undergo programmed cell death and release molecules (e.g., nucleic acids and oligomers) called damage-associated molecular patterns. 20 These molecules are recognized by macrophages and adjacent endothelial and epithelial cells, causing them to produce inflammatory cytokines, including the chemokines: interleukin-6 (IL-6); interferon gamma-inducible protein 10 (IP-10; also known as CXCL10); macrophage inflammatory proteins 1α and 1β; and monocyte chemotactic protein 1 (MCP-1; also known as CCL2). Monocytes, macrophages, and T cells are then recruited to the site of infection by these chemokines and other cytokines, promoting further inflammation. As part of this inflammatory response, recruited T cells produce interferon-gamma (IFNγ).

[0164] Several types of T cells are involved in this response. CD4+ T helper (Th) cells interact with CD8+ T cells, which drive the cytotoxic response that kills virus-infected cells. CD8+ T cells directly recognize viral peptides displayed on the surface of infected cells, triggering apoptosis (a form of programmed cell death) and preventing the virus from spreading further. Follicular helper T (TFH ) cells are a specialized subset of CD4+ T cells that provide help to B cells through both cell-cell interactions and release of cytokines, leading to the production of antibodies by B cells. 19 These neutralizing antibodies are able to recognize the whole virus and function by blocking the virus from infecting cells. Alveolar macrophages recognize neutralized viruses and apoptotic cells (killed by CD8+ T cells) and remove them by phagocytosis. This subsequently results in recovery from viral infection. 20 .

[0165] Studies evaluating the clinical characteristics of patients infected with SARS-CoV-2 have reported an incubation period of 4 to 7 days for the onset of symptoms and an additional 7 to 10 days for progression to severe disease. 21 .

[0166] For many primary viral infections, it typically takes 7-10 days to prime and expand an adaptive T cell immune response to control the virus, which correlates with the typical time it takes for patients with COVID-19 to either recover or develop severe disease. 22 This raises the possibility that inadequate early T cell responses may contribute to SARS-CoV-2 persistence and severity, whereas early strong T cell responses may be protective.

[0167] CD4+ T cell responses in COVID-19: Some studies show that in patients with severe COVID-19, there is evidence of impaired CD4+ T cell function, including reduced IFNγ production, while other studies seem to suggest hyperactivation of these T cells. 23 .

[0168] Overall, the CD4+ T cell response in acute SARS-CoV-2 infection, whether impaired, overactivated, or inappropriate, and how this relates to disease outcome remain unclear and are important questions. In particular, CD4+ T cell responses specific to high frequencies of the viral spike protein were observed in patients who recovered from COVID-19, similar to what has been reported for influenza virus infections. 21 In one small study of 14 patients, circulating virus-specific CD4+ T cells were identified in all of those who recovered from SARS-CoV-2, also suggesting T cell memory. 24 and perhaps the potential to generate longer-term immunity.

[0169] CD8+ T cell responses in COVID-19: There appears to be heterogeneity in immune responses among patients. Some studies have reported that CD8+ T cells from patients with severe COVID-19 have reduced cytokine production after in vitro stimulation, and some have shown evidence of possible exhausted T cells; in contrast, other studies have reported overly aggressive CD8+ T cell responses or highly activated CD8+ T cells with increased cytotoxic responses in patients with COVID-19. 25 .

[0170] Experimental Procedures / Research Strategy To test this hypothesis, a model cell line was used. The Jurkat cell line is an immortalized T lymphocyte cell line originally obtained from the peripheral blood of a boy with T cell leukemia. 26 The Jurkat cell line has been most frequently used as a prototypic T cell line to study multiple events in T cell biology, including a) T cell signaling and b) molecular events in the HIV infection life cycle. 18 In the literature, Jurkat cell lines have been used to model and characterize signaling events in T cell activation (TCA), a critical process in effective adaptive immune responses. 26As a model signaling axis, TCA involves surface signaling through the T cell receptor (TCR) on the surface of T cells and the accessory proteins CD3 and CD28, initiating a cascade of molecular events that result in the transcriptional activation of multiple genes, including the canonical T cell activation target gene, the interleukin-2 (IL-2) gene. Steps during TCA include the activation of a series of kinases (e.g., LCK, JNK, PKC) and phosphatase proteins (calcineurin), as well as the activation of dormant cytoplasmic transcription factors (e.g., NF-kB, NFAT) that translocate to the nucleus upon activation and activate target genes. Each of these steps, as well as numerous other intermediate factors in this pathway, have been investigated in detail using mutant subclones of the Jurkat T cell line, which have been instrumental in mapping the signaling pathway and identifying key players underlying T cell activation. 18、26 .

[0171] Jurkat cell line culture (Jurkat, clone E6-1 (ATCC® TIB-152™); human; Homo sapiens) Complete Growth Medium: The base medium for this cell line is ATCC preparation RPMI-1640 medium, ATCC 30-2001. To make the complete growth medium, the following components are added to the base medium: fetal bovine serum (ATCC 30-2020), final concentration 10%.

[0172] Subculture: Maintain the culture by adding fresh medium or replacing the medium. Alternatively, centrifugation and 1×10 5 Cultures are established by subsequent resuspension at 10 viable cells / mL. The cell density is 3 x 10 6 Do not allow to exceed 10 cells / mL. Corning® T-75 flasks are recommended for subculturing this product.

[0173] Interval: Culture, 1 x 10 5 ~1×10 6 The cells were maintained at a cell concentration of 1 viable cell / mL.

[0174] Medium renewal: Fresh medium was added every 2-3 days (depending on cell density).

[0175] Culture conditions: Atmosphere: air, 95%; carbon dioxide (CO2), 5%; Temperature: 37°C

[0176] Treatment of Jurkat cells with a dietary supplement of superoxide dismutase combined with prebiotic fibre containing fruit juice-derived polyphenols Effect of superoxide dismutase combined with prebiotic fiber containing fruit juice-derived polyphenols on Jurkat cells: Jurkat cells were seeded in 6-well plates. Prior to treatment, cells were incubated in serum-free medium for 24 hours. Cells were treated for 48 hours with the following agents: 1. superoxide dismutase only; 2. prebiotic fiber only; 3. fruit juice only; 4. superoxide dismutase + prebiotic fiber + fruit juice (combination); 5. Positive control: phorbol 12-myristate 13-acetate (PMA) combined with ionomycin; 6. Negative control: cell culture medium.

[0177] Enzyme-linked immunosorbent assay: After treatment, the medium was removed from the cells and placed in a test tube. To assess T cell activation, the following factors were measured in the cell medium: CD-8+; CD-4+; interferon-gamma (IFNγ); interleukin-6 (IL-6); interferon gamma-inducible protein 10 (IP-10; also known as CXCL10); macrophage inflammatory proteins 1α and 1β; monocyte chemotactic protein 1 (MCP-1; also known as CCL2); and 8-isoprostane levels were measured by commercial ELISA kits as previously described. 27、28、29、30、31、32、33、34 .

[0178] T cell stimulation for flow cytometry analysis: Cells were treated for 48 hours with the following agents: 1. Superoxide dismutase only; 2. Prebiotic fiber only; 3. Fruit juice only; 4. Superoxide dismutase + Prebiotic fiber + Fruit juice (combination); 5. Positive control: Phorbol 12-myristate 13-acetate (PMA) in combination with ionomycin; 6. Negative control: cell culture medium.

[0179] Activated Jurkat cells were detected as described above, and statistical analysis of the results was performed as described above.

[0180] result Cell viability assay: The assay determines the ability of viable cells to convert the redox dye resazurin to the fluorescent end product resorufin. Jurkat cells were seeded in 96-well plates in complete medium and allowed to adhere overnight at 37°C. The cells were then treated with vehicle (cell culture medium) or prebiotic fiber only, superoxide dismutase only, fruit juice only, superoxide dismutase + prebiotic fiber + fruit juice (combination) or LPS (positive control). After 48 hours of incubation with each treatment, 20 μL of Cell Titer-Blue reagent was added to each well. The absorbance at 520 nm was determined by a microtiter plate reader. The signal generated by the conversion of resazurin to resorufin is directly proportional to the number of viable cells. As shown in Figure 1, none of the gel components or combinations showed any toxic effects in the cell viability assay.

[0181] The gel and its components activate T cells as seen by upregulated CD69 expression on the CD3 positive cell population using flow cytometric analysis: T cell stimulation for flow cytometric analysis. The activation and proliferation protocol provides an effective method for determining immune competence and cell reactivity. Jurkat cells were stimulated with vehicle (cell culture medium) or prebiotic fiber only, superoxide dismutase only, fruit juice only, superoxide dismutase + prebiotic fiber + fruit juice (combination) or LPS (positive control). Cells were gated on lymphocytes in the presence of Human Seroblock. Data were acquired on a ZE5™ cell analyzer. As shown in Figure 2, activated T cells are seen by upregulated CD69 expression on the CD3 positive population. As shown in Figure 2, the combination showed a surprisingly high level of T cell activation as measured by CD69 expression.

[0182] Jurkat cell CD4 + Effect of the original components of REVIVIFY® on differentiation. Jurkat cells were stimulated with vehicle (cell culture medium) or prebiotic fiber alone, superoxide dismutase alone, fruit juice alone, superoxide dismutase + prebiotic fiber + fruit juice (combination). CD4 + The concentrations (ng / mL) were measured by ELISA according to the manufacturer's protocol. As shown in Figure 3, fruit juice and the combination of superoxide dismutase + prebiotic fiber + fruit juice significantly and surprisingly stimulated CD4+ differentiation compared to culture medium as determined by ELISA.

[0183] Jurkat cell CD8 + Effect of the original components of REVIVIFY® on differentiation. Jurkat cells were stimulated with vehicle (cell culture medium) or prebiotic fiber alone, superoxide dismutase alone, fruit juice alone, superoxide dismutase + prebiotic fiber + fruit juice (combination). CD8 +The concentrations of (pg / mL) were measured by ELISA according to the manufacturer's protocol. As shown in Figure 4, fruit juice and the combination of superoxide dismutase + prebiotic fiber + fruit juice significantly and surprisingly stimulated CD4+ differentiation compared to culture medium as determined by ELISA.

[0184] CD4 in REVIVIFY®-treated Jurkat cells + and CD8 + Concentration ratio of CD4 in REVIVIFY treated Jukart cells + / CD8 + The concentration ratio of β-lactamase in the peripheral blood of healthy adults and mice is 48:1. 35 CD4 + / CD8 + The ratio is the ratio of T helper cells (bearing the surface marker CD4) to cytotoxic T cells (bearing the surface marker CD8). + and CD8 + Both types of T cells contain several subsets. 36 CD4 in peripheral blood of healthy adults and mice + / CD8 + The ratio is approximately 2:1, and an altered ratio can indicate a disease associated with immune deficiency. 35 This large difference in ratio is due to the in vitro closed system experiments. + These data also demonstrate the high potency of REVIVIFY® to stimulate T cells for higher differentiation into CD4 T cells. These data suggest that follicular helper T (TFH) cells provide help to B cells through both cell-cell interactions and release of cytokines, leading to the production of antibodies by B cells. + Showing that it is a specialized subset of T cells 19 These neutralizing antibodies are able to recognize the whole virus and function by blocking the virus from infecting cells. Alveolar macrophages recognize neutralized viruses and apoptotic cells (killed by CD8+ T cells) and remove them by phagocytosis. This subsequently results in recovery from viral infection.20 In coronavirus disease 2019 (COVID-19), B cells, natural killer cells, and total lymphocyte counts are decreased, but both CD4+ and CD8+ cells are decreased to a much greater extent. 37 Low CD4+ predicted a greater likelihood of intensive care unit admission, and CD4+ cell count was the only parameter that predicted time to viral RNA clearance. 37 .

[0185] REVIVIFY® Gel has antioxidant activity: As shown in Figure 5, REVIVIFY® and its components attenuate lipopolysaccharide-induced activation of 8-isoprostane secretion by Jurkat cells in an in vitro study. Lipopolysaccharide (LPS)-stimulated Jurkat cells were treated with vehicle (cell culture medium) or prebiotic fiber only, superoxide dismutase only, fruit juice only, or superoxide dismutase + prebiotic fiber + fruit juice (combination). The levels of 8IP in the culture medium were measured by ELISA. REVIVIFY® and its components attenuated lipopolysaccharide-induced activation of 8-isoprostane (8IP) secretion by Jurkat cells. The levels of 8IP have been proposed as a marker of antioxidant deficiency and oxidative stress. 38、39 As can be seen in Figure 5, the combination surprisingly reduced 8IP levels below those seen in untreated cells.

[0186] REVIVIFY® Gel has anti-hypoxic activity: As shown in Figure 6, REVIVIFY® and its components attenuate lipopolysaccharide-induced activation of cyclooxygenase-2 (COX-2) secretion by Jurkat cells in an in vitro study. Lipopolysaccharide (LPS)-stimulated Jurkat cells were treated with vehicle (cell culture medium) or prebiotic fiber only, superoxide dismutase only, fruit juice only, superoxide dismutase + prebiotic fiber + fruit juice (combination). The levels of COX-2 in the culture medium were measured by ELISA. As can be seen in Figure 6, the combinations surprisingly reduced COX-2 levels below those seen in untreated cells.

[0187] REVIVIFY® and its components attenuated lipopolysaccharide-induced activation of COX-2 secretion by Jurkat cells (FIG. 6): Hypoxia increases COX-2 expression 40 Apigenin downregulates COX-2 expression in lupus T cells, B cells, and antigen-presenting cells, leading to their apoptosis. 41 Although no clear structure / function correlation has been established and we do not wish to be bound by theory, it appears that the C-2,3 double bond and hydroxyl substitutions on the A and B rings are important contributors to this inhibitory activity. 42 Animal data confirm downregulation of COX-2 expression in different inflammatory diseases 43 .

[0188] REVIVIFY® Gel has anti-inflammatory activity: REVIVIFY® and its components attenuate lipopolysaccharide-induced activation of inflammatory activity in an in vitro Jurkat cell study. Lipopolysaccharide (LPS)-stimulated Jurkat cells were treated with vehicle (cell culture medium) or prebiotic fiber only, superoxide dismutase only, fruit juice only, superoxide dismutase + prebiotic fiber + fruit juice (combination). Levels of interferon gamma (IFNγ, FIG. 7), interleukin 6 (IL-5, FIG. 8), transforming growth factor beta (TGF-β, FIG. 9), tumor necrosis factor (TNF, FIG. 10) and CXC motif chemokine ligand 10 (CXCL10, FIG. 11) in the culture medium were measured using commercially available ELISA kits.

[0189] REVIVIFY® and its components attenuate lipopolysaccharide-induced activation of IFN-γ secretion by Jurkat cells (FIG. 7): Interferon gamma (IFNγ) is a dimerizing soluble cytokine that is the only member of the type II class of interferons. 44 The existence of this interferon, also known as immune interferon early in its history, was described by E. F. Wheelock as a product of human leukocytes stimulated with phytohemagglutinin and by other authors as a product of antigen-stimulated lymphocytes. 45IFNγ, or type II interferon, is a cytokine that is critical for innate and adaptive immunity against viral, some bacterial and protozoan infections. IFNγ is an important activator of macrophages and an inducer of major histocompatibility complex class II molecules. Aberrant IFNγ expression is associated with several autoinflammatory and autoimmune diseases. The importance of IFNγ in the immune system is due in part to its ability to directly inhibit viral replication and, most importantly, its immunostimulatory and immunomodulatory effects. IFNγ is primarily activated by natural killer cells (NK) and natural killer T cells (NKT) as part of the innate immune response, and by CD4 Th1 and CD8 cytotoxic T lymphocyte (CTL) effector T cells during antigen-specific immunity as part of the adaptive immune response. 46、47 IFNγ is also produced by non-cytotoxic innate lymphoid cells (ILCs), a family of immune cells first discovered in the early 2010s. 48 As can be seen in FIG. 7, cells treated with SOD, fruit juice polyphenols and the combination surprisingly all had lower levels of IFNγ secretion than untreated cells, with the lowest levels of secretion seen in cells treated with the combination.

[0190] REVIVIFY® and its components attenuate lipopolysaccharide-induced activation of IL-6 secretion by Jurkat cells (FIG. 8): Interleukin 6 (IL-6) is an interleukin that acts as both a proinflammatory cytokine and an anti-inflammatory myokine. In humans, it is encoded by the IL6 gene. 49In addition, osteoblasts secrete IL-6 to stimulate osteoclast formation. Smooth muscle cells in the media of many blood vessels also produce IL-6 as a proinflammatory cytokine. The role of IL-6 as an anti-inflammatory myokine is mediated through its inhibitory effects on TNF-alpha and IL-1 and its activation of IL-1ra and IL-10. There is some early evidence that IL-6 can be used as an inflammatory marker for poor prognosis severe COVID-19 infection in the context of the broader coronavirus pandemic. 50 IL-6 is secreted by macrophages in response to specific microbial molecules called pathogen-associated molecular patterns (PAMPs). These PAMPs bind to an important group of innate immune system detector molecules called pattern recognition receptors (PRRs), including Toll-like receptors (TLRs). They are present on the cell surface and in intracellular compartments and induce intracellular signaling cascades that lead to inflammatory cytokine production. IL-6 is a key mediator of fever and acute phase response. IL-6 is responsible for stimulating acute phase protein synthesis and the generation of neutrophils in the bone marrow. IL-6 supports B cell growth and is antagonistic to regulatory T cells. As can be seen in Figure 8, the combination surprisingly reduced IL-6 activation levels below those seen in untreated cells.

[0191] REVIVIFY® and its components attenuate lipopolysaccharide-induced activation of TGF-β secretion by Jurkat cells (in FIG. 9): Transforming growth factor beta (TGF-β) is a multifunctional cytokine that belongs to the transforming growth factor superfamily, which includes three members 51 TGFB proteins are produced by all leukocyte lineages. Activated TGF-β complexes with other factors to form a serine / threonine kinase complex that binds to the TGF-β receptor. The TGF-β receptor is composed of both type 1 and type 2 receptor subunits. After binding of TGF-β, the type 2 receptor kinase phosphorylates and activates the type 1 receptor kinase, which activates the signaling cascade. 52This leads to the activation of different downstream substrates and regulatory proteins, which induce the transcription of different target genes that function in the differentiation, chemotaxis, proliferation, and activation of multiple immune cells. 52、53 .

[0192] TGF-β is secreted by many cell types, including macrophages, in a latent form complexed with two other polypeptides, latent TGF-beta binding protein (LTBP) and latency-associated peptide (LAP). Serum proteinases, such as plasmin, catalyze the release of active TGF-β from the complex. This often occurs on the surface of macrophages, where the latent TGF-β complex binds to CD36 via its ligand thrombospondin-1 (TSP-1). Inflammatory stimuli that activate macrophages enhance the release of active TGF-β by promoting the activation of plasmin. Macrophages can also endocytose IgG-bound latent TGF-β complexes secreted by plasma cells, subsequently releasing active TGF-β into the extracellular fluid. 54 Its key function is the regulation of inflammatory processes, especially in the intestine.[5] TGF-β also plays a crucial role in stem cell differentiation and T cell regulation and differentiation. 56、57 Due to its role in immune and stem cell regulation and differentiation, TGF-β is a highly studied cytokine in the areas of cancer, autoimmune disease, and infectious diseases.

[0193] The TGF-β superfamily contains endogenous growth inhibitory proteins; increased expression of TGF-β often correlates with aggressiveness of many cancers and with defects in the cell growth inhibitory response to TGF-β. Its immunosuppressive function subsequently becomes dominant, contributing to carcinogenesis. 58 Dysregulation of its immunosuppressive function is also involved in the pathogenesis of autoimmune diseases, but their effects are mediated by the milieu of other cytokines present. 55TGF-β induces apoptosis, or programmed cell death, in human lymphocytes and hepatocytes. The importance of this function is evident in TGF-β-deficient mice, which experience hyperproliferation and uncontrolled autoimmunity. 59 As can be seen in Figure 9, cells treated with SOD, fruit juice polyphenols and the combination surprisingly all had lower levels of TGF-β secretion than untreated cells, with the lowest levels of secretion seen in cells treated with the combination.

[0194] REVIVIFY® and its components attenuate lipopolysaccharide-induced activation of TNF-α secretion by Jurkat cells (in FIG. 10): Tumor necrosis factor (TNF, cachexin, or cachectin; often referred to as tumor necrosis factor alpha or TNF-α) is a cytokine, a small protein used by the immune system for cell signaling. When macrophages (a type of white blood cell) detect an infection, they release TNF to alert other immune system cells as part of the inflammatory response. TNF is a member of the TNF superfamily, which consists of a variety of transmembrane proteins with homologous TNF domains. TNF was thought to be produced primarily by macrophages, but 60 It is also produced by a wide range of cell types, including lymphoid cells, mast cells, endothelial cells, cardiac myocytes, adipose tissue, fibroblasts, and neurons. 61 Large amounts of TNF are released in response to lipopolysaccharide, other bacterial products, and interleukin-1 (IL-1). In the skin, mast cells appear to be the predominant source of preformed TNF that can be released upon inflammatory stimuli (e.g., LPS). 62 TNF, commonly together with IL-1 and interleukin-6 (IL-6), has several actions on various organ systems. A local increase in the concentration of TNF produces the cardinal signs of inflammation: heat, swelling, redness, pain and loss of function. High concentrations of TNF induce shock-like symptoms, while prolonged exposure to low concentrations of TNF can result in cachexia, a wasting syndrome. This can be seen, for example, in cancer patients. 63 .

[0195] Said et al. showed that TNF induces IL-10-dependent inhibition of CD4 T cell expansion and function by upregulating PD-1 levels on monocytes, which leads to IL-10 production by monocytes following engagement of PD-1 by PD-L. 64 Studies by Pedersen et al. show that TNF increases in response to sepsis are inhibited by exercise-induced production of myokines. To study whether acute exercise induces a true anti-inflammatory response, a model of "low-grade inflammation" was established in which a low dose of Escherichia coli (E. coli) endotoxin was administered to healthy volunteers who were randomized to either rest or exercise prior to endotoxin administration. In resting subjects, endotoxin induced a 2- to 3-fold increase in circulating levels of TNF. In contrast, when subjects performed 3 hours of ergometer bicycle exercise and received an endotoxin bolus at 2.5 hours, the TNF response was completely blunted. 65 This study provides some evidence that acute exercise can inhibit TNF production. 66 In the brain, TNF may protect against excitotoxicity 67 TNF strengthens synapses 68 TNF in neurons promotes their survival, whereas TNF in macrophages and microglia produces neurotoxins that induce apoptosis. 67 As can be seen in Figure 10, cells treated with fruit juice polyphenols and the combination surprisingly both had lower levels of TNF-alpha secretion than untreated cells, with the lowest levels of secretion seen in cells treated with the combination.

[0196] REVIVIFY® and its components attenuate lipopolysaccharide-induced activation of CXCL10 secretion by Jurkat cells (in FIG. 11): CXC motif chemokine ligand 10 (CXCL10), also known as interferon gamma-inducible protein 10 (IP-10) or small inducible cytokine B10, is an 8.7 kDa protein that in humans is encoded by the CXCL10 gene. 69、70 CXC motif chemokine 10 is a small cytokine that belongs to the CXC chemokine family. CXCL10 is secreted by several cell types in response to IFN-γ. These cell types include monocytes, endothelial cells, and fibroblasts. 69 CXCL10 has been ascribed several roles, including chemoattraction for monocytes / macrophages, T cells, NK cells, and dendritic cells, promotion of T cell adhesion to endothelial cells, antitumor activity, and inhibition of bone marrow colony formation and angiogenesis. 71、72 This chemokine exerts its effects through binding to the cell surface chemokine receptor CXCR3. 73 As can be seen in Figure 10, cells treated with SOD, fruit juice polyphenols and the combination surprisingly all had lower levels of CXCL10 secretion than untreated cells, with the lowest levels of secretion seen in cells treated with the combination.

[0197] A summary of all of the ELISA results discussed above is provided in Table 5.

[0198] [Table 5]

[0199] References for Example 1 [Table 6-1] [Table 6-2] [Table 6-3] [Table 6-4] [Table 6-5] [Table 6-6] [Table 6-7] [Table 6-8] [Table 6-9]

[0200] Example 2 Influence of superoxide dismutase absorption by altering gut microbiota background Superoxide dismutase (SOD) is a primary antioxidant enzyme with a very high molecular weight, which usually has problems associated with physical stability, gastric acid degradation, and most importantly, absorption. Although SOD is beneficial for reducing cellular oxidative stress and may be useful for many age-related dysfunctions, it has been difficult to produce an orally effective SOD formulation. REVIVIFY® contains a diverse group of functional molecules that affect each other's bioavailability, including SOD absorption. The acid stability and absorption of SOD are influenced by microbial changes provided by some components of REVIVIFY® gel - prebiotic fiber and polyphenols. To prove this hypothesis, studies were conducted to evaluate whether SOD alone and as part of the REVIVIFY® composition cause any microbial changes. The following studies show that changes in the gut microbial composition occur, which cause SOD absorption and availability.

[0201] Experimental Method The first objective of this study was to develop a scalable in vitro model for the maintenance of gut microbiome profiles. Bacterial cells were cultured in 96-deep well plates, with a silicone gel cover with holes on top of each well. This cover facilitates gas exchange with the outside environment in the chamber, thereby preserving the partial pressure of gases and volatile metabolites in each well, which in turn could preserve a constant level of dissolved gas molecules in the culture medium.

[0202] Treatment conditions: 1. SOD alone; 2. Fibersol (resistant maltodextrin) alone; 3. Polyphenols alone; 4. SOD + Fibersol; 5. SOD + Polyphenols; and 6. REVIVFY finished product. Using these treatments, it is possible to determine how Fibersol and polyphenols affect the absorption of SOD using an in vitro guy microbial system.

[0203] Fecal specimen collection and processing: Briefly, approximately 3 g of fresh fecal samples were collected from each individual using a 2.5 mL sterile sample collection spoon (Bel-Art, USA). Each spoon was dropped into a 50 ml Falcon tube containing 15 ml of sterile PBS previously reconstituted with 0.1% (w / v) L-cysteine ​​hydrochloride. Samples were immediately transferred to an anaerobic workstation (5% H2, 5% CO2, and 90% N2, 37°C). Optical density at 595 nm (OD ) was measured as a surrogate for microbial growth and biomass. 595 The microbiome was characterized at 0 h (immediately after inoculation), 3 h, 6 h, 9 h, 12 h, 24 h, 34 h, and 48 h by measurements of β-aminobutyric acid (β-aminobutyric acid), and by metaproteomic analysis. Results were determined using both the goat intestine and silicone mat microbiome systems.

[0204] Results: Bacterial growth gradually increases over time. Fibersol and SOD alone samples have no effect on bacterial growth compared to the control as shown by OD measurements (Figure 12, goat intestinal system; Figure 13, silicone mat system). In both Figures 12 and 13, the top line in the plot is for REVIVIFY® finished product, the middle two lines are for juice polyphenols only and SOD+Fibersol, and the bottom three lines are for the no treatment control, SOD only, and Fibersol only. However, the polyphenols in the juice significantly increased bacterial growth along with REVIVIFY® finished product, causing the largest increase in OD. These experiments demonstrated that Fibersol enhanced the absorption of SOD through both the silicone mat and fresh goat male experiments as demonstrated by increased bacterial growth.

[0205] Conclusion: This study shows the development of a scalable in vitro model for the maintenance of the gut microbiome profile. Furthermore, the absorption patterns of the different components of REVIVFY gel are shown, including the effect of Fibersol on the absorption of SOD. This in vitro model of gut microbes can be used for the evaluation of other drugs, prebiotics or nutraceuticals.

[0206] Example 3 Changes in the healthy gut microbiome and short-chain fatty acids assessed by an in vitro gut microbiome model Background: Gut health is very important for a healthy life and well-being. The microbial community in the gut plays a major role in the immune system, hormonal processes, neurological status, metabolism, mineral absorption, vitamin production, and several cellular processes. According to the American Diabetes Association, the gut microbiota is known to affect host physiology both inside and outside the gut. The gut microbiota is essential for immune system homeostasis in the gut, regulation of epithelial proliferation, and protection against opportunistic bacteria. The gut microbes present in the gastrointestinal tract have co-evolved within the human host and perform several functions without which the host would not be able to perform on its own. The predominant gut microbes are from the phyla Firmicutes and Bacteroidetes, followed by Actinobacteria and Protobacteria. The beneficial effects of gut microbes can be measured by the production of short chain fatty acids (SCFAs), mainly acetate, propionate, and butyrate, and lactate. This production depends on the type of fiber intake. In this example, the fiber is a soluble fiber known as Fibersol-2 with mixed fructooligosaccharides from various fruit concentrates. The hypothesis is that there should be a change in beneficial microorganisms with an increased ratio of short chain fatty acids that protects host health in multiple ways. This study evaluates the effect of REVIVIFY® finished product on gut microbiome changes and short chain fatty acids through an in vitro model of gut microbiome research.

[0207] Experimental Method: An in vitro gut microbiome culture model was established as described in Example 2 above. The first objective of this study is to develop a scalable in vitro model for the maintenance of gut microbiome profile. In vitro models that maintain the functional and compositional profile of the in vivo gut microbiome will be highly variable. In vitro model experiments were performed as described in Example 2. Gut microbes were cultured in 2ml 96-well plates and treated with control, SOD, prebiotic fiber, fruit juice, or finished REVIVIFY® product for 24 hours. Cultured microbiome samples were harvested at 24 hours for metaproteomic analysis. Culture aliquots were then collected for chemical analysis (SCFA content) and microbiome profiling.

[0208] Results: This study evaluated the changes in gut microbial composition and SCFAs when treated with four different compounds (SOD, prefibrotic fiber, fruit juice, and finished product). As can be seen in Figure 14, the results show that the amount of SCFAs increases significantly when treated with finished product (Figure 14A), but the ratio of SCFAs remains the same across all treatment groups, including the control (Figure 14B). In the control treatment, the concentrations of acetate, propionate, butyrate, and lactate were 30 μmol / ml, 9 μmol / ml, 15 μmol / ml, and 6 μmol / ml, respectively, which increased to 80 μmol / ml, 25 μmol / ml, 35 μmol / ml, and 12 μmol / lL when treated with finished product for 24 hours. This is an unexpectedly large 2.5-fold increase in SCFAs in response to finished product. When calculating the ratios of these SCFAs across the control and all four treatment groups, the ratios of acetate, propionate, butyrate, and lactate appeared to be consistently maintained around 53%, 15%, 24%, and 8%, respectively (see FIG. 14). Our study also demonstrated that pre-fibrotic fiber, fruit juice, and finished product promoted the growth of two beneficial gut microbes, Firmicutes Lactobacillus and Actinobacteria Bifidobacteriaceae, when compared to baseline and SOD (see FIG. 15; Firmicutes Lactobacillus is represented by vertical shading, and Actinobacteria Bifidobacteriaceae is represented by light gray shading). Both Lactobacillus and Bifidobacteria are beneficial microbes and are present in many food preparations, such as yogurt for digestive and immune benefits. Of these two types, the trend is towards Lactobacillus, which is reflected in the production of lactic acid in the SCFA graph. Lactobacillus also belongs to the species of the phylum Firmicutes. Normally, they are butyrate producers, which is reflected in the SCFA results.Microbiome profiles of cultures treated with the finished product showed a significant increase in Lactobacillus among the Firmicutes species in the gut microbial community, followed by Bifidobacterium among the Actinobacterium species.

[0209] Conclusion / Perspective: Dietary prebiotics are selectively fermented ingredients that result in specific changes in the composition of two beneficial microbiota: Lactobacillus in the Firmicutes phylum and Bifidobacterium in the Actinobacteria phylum. This is a remarkable finding where both beneficial microbes can positively impact the host by providing both homo- and hetero-fermentative results along with protein and carbohydrate digestion and metabolism, B vitamin and vitamin K synthesis, bile salt catabolism, enhancement of innate and adaptive immunity, inhibition of inflammatory mediators, and antibacterial activity against an array of pathogens such as Pseudomonas, Candida, E. coli, Aurous, Salmonella, as well as Shigella, C. difficile, and Helicobacter pylori. Lactobacillus may be a biomarker of vaginal health and is a major part of the vaginal microbiome. This study demonstrates that REVIVIFY® finished product increased the amount of SCFA in the intestine while maintaining the ratio of each SCFA (acetate, propionate, butyrate, and lactate) consistent with the control. This result shows a balanced increase in SCFA without in a consistent manner, which improves and maintains a healthy colonic environment. This study suggests that REVIVIFY® is a unique food supplement that produces relatively high amounts of butyrate, which contributes to multiple health benefits, intestinal epithelial cell integrity, immune cell integrity and response, enteric neuron bidirectional signaling of the gut-brain axis, and nutrient production and metabolism. Butyrate is the main energy source for colonocytes and is involved in maintaining colonic mucosal health.

[0210] Example 4 Attenuation of oxidative damage in human brain microvascular endothelial cells (HBMEC) introduction Accumulating data suggests that oxidative stress and mitochondrial damage are involved in the pathogenesis of neurodegenerative disorders, including Parkinson's disease (PD), multiple sclerosis (MS), Alzheimer's disease (AD), and many other disorders. The brain uses about 20% of oxygen consumption and is therefore a high producer of reactive oxygen species (ROS). Brain cell membranes are also composed of relatively high amounts of unsaturated fatty acids (MUFA and PUFA), and are therefore relatively susceptible to lipid autoxidation by ROS. REVIVIFY® Gel may provide instant reduction of oxidative stress from multiple pathways and immediate effects induced by disease symptoms.

[0211] REVIVIFY® formulations neutralize the major oxidants: superoxide anion, hydroxyl radical, singlet oxygen, peroxynitrite, peroxy radical, and hypochlorite.

[0212] background Superoxide dismutase (SOD): SOD constitutes a very important antioxidant defense against oxidative stress in the body. This enzyme serves as a good therapeutic agent against reactive oxygen species-mediated diseases. The present review describes the therapeutic effects of SOD in various physiological conditions and pathologies, such as cancer, inflammatory diseases, cystic fibrosis, ischemia, aging, rheumatoid arthritis, neurodegenerative diseases, and diabetes. However, this enzyme has certain limitations in clinical applications. Therefore, SOD conjugates and mimetics have been developed to increase its therapeutic efficiency. 1、2、3、4、5、6、7 .

[0213] Polyphenols: Polyphenols are secondary metabolites of plants and are generally involved in the defense against UV radiation or attack by pathogens. In the last decade, there has been great interest in the possible health benefits of dietary plant polyphenols as antioxidants. Epidemiological studies and associated meta-analyses strongly suggest that long-term intake of a diet rich in plant polyphenols confers protection against the development of cancer, cardiovascular disease, diabetes, osteoporosis and neurodegenerative diseases. 8、9、10、11、12 .

[0214] Dietary Prebiotic Fiber: The health benefits of dietary fiber have long been appreciated. Higher intake of dietary fiber has been linked to less cardiovascular disease, and fiber plays a role in gut health, with many effective laxatives actually isolated from fiber sources. Higher intake of fiber is associated with lower body weight. Originally, only polysaccharides were included in dietary fiber, but more recent definitions include oligosaccharides as dietary fiber based on their physiological effects, rather than based on their chemical measurement as dietary fiber by the commonly accepted total dietary fiber (TDF) method. Inulin, fructooligosaccharides, and other oligosaccharides are included as fiber on food labels in the United States. In addition, oligosaccharides are the best known "prebiotics," "selectively fermented ingredients that allow specific changes in both composition and / or activity in the gastrointestinal microflora that confer benefits to the host's soundness and health." To date, all known and putative prebiotics are carbohydrate compounds, primarily oligosaccharides, that are known to survive digestion in the human small intestine and reach the colon where they are fermented by the gut microflora. Studies have provided evidence that inulin and oligofructose (OF), lactulose, and resistant starch (RS) meet all aspects of the definition, including the stimulation of the beneficial bacterial genus Bifidobacterium. Other isolated carbohydrates and carbohydrate-containing foods, including galactooligosaccharides (GOS), transgalactooligosaccharides (TOS), polydextrose, wheat dextrin, gum arabic, psyllium, banana, whole wheat, and whole corn, also have prebiotic effects. 13、14、15、16、17 .

[0215] The purpose of this study was to evaluate whether REVIVIFY® Gel attenuates human brain microvascular endothelial cells (HBMEC) from oxidative damage. The following biomarkers were evaluated in hypoxia-induced HBMEC culture medium: 1. Malondialdehyde (MDA) - a biomarker of lipid oxidative damage 2. 4-Hydroxynonenal, or 4-Hydroxy-2-nonenal, or 4-HNE, or HNE - a biomarker of lipid peroxidation 3. Protein carbonyls – biomarkers of protein oxidation 4. 3-Nitrotyrosine – A biomarker of cell damage associated with the degeneration of dopamine neurons.

[0216] Malondialdehyde (MDA) is an organic compound with the formula CH2(CHO)2. A colorless liquid, malondialdehyde is a highly reactive compound that exists as an enol [1]. It occurs naturally and is a marker for oxidative stress.

[0217] 4-Hydroxynonenal, or 4-hydroxy-2-nonenal, or 4-HNE, or HNE(CH 16 4-HNE (O2) is an α,β-unsaturated hydroxyalkenal generated by lipid peroxidation in cells. 4-HNE is the major α,β-unsaturated hydroxyalkenal formed in this process. 4-HNE has three reactive groups: an aldehyde, a double bond at carbon 2, and a hydroxy group at carbon 4.

[0218] Protein carbonyls: Protein carbonyl (PC) content in blood and tissues is a reliable indicator of protein oxidation. Traumatic brain injury (TBI) results from a head impact that disrupts normal brain function. Severe TBI can cause permanent brain damage or death. Diffuse axonal injury (DAI) is a typical pathological change after TBI and is closely associated with clinical prognosis. DAI has two distinct pathological features: large end bulbs due to swelling and excessive neurofilament aggregation. Secondary axonal injury resulting from skeletal muscle abnormalities is the most common cause of DAI.

[0219] Oxidative stress is a well-known factor involved in DAI, and mitochondrial phosphorylation capacity, concentration of nicotinic coenzyme pool, and oxidative / nitrosative stress are closely correlated with the severity of DAI. Carbonyl modification occurs as a direct result of oxidative damage to proteins, leading to protein dysfunction and the formation of protein aggregates. Protein carbonylation has been shown to contribute to the pathogenesis of several neurodegenerative diseases, such as multiple sclerosis, Parkinson's disease, and Alzheimer's disease. Under normal conditions, carbonylated proteins are thought to be degraded by the proteasome, whose main function is to recognize and degrade unused, damaged, or misfolded proteins. However, under pro-oxidative conditions, increased production of reactive oxygen species (ROS) or reactive carbonyl species may reduce the activity of the proteasome, leading to the accumulation of carbonylated proteins in diseased cells.

[0220] 3-Nitrotyrosine: Nitrotyrosine is the product of tyrosine nitration mediated by reactive nitrogen species such as peroxynitrite anion and nitrogen dioxide. Nitrotyrosine has been identified as an indicator or marker of cell damage, inflammation and NO (nitric oxide) production. Nitrotyrosine is formed in the presence of the active metabolite NO. In general, in many disease states, oxidative stress is caused by the production of superoxide (O2 - ) and NO production, and inhibits the production of the destructive free radical oxidant peroxynitrite (ONOO) - ) is formed. - The formation of ONOO is capable of oxidizing several lipoproteins and nitrating tyrosine residues in many proteins. - It is difficult to determine the formation of ONOO. -Nitrotyrosine is a detectable marker for indirectly detecting NO. It is detected in many pathologies and is considered to be a marker of NO-dependent reactive nitrogen species-induced nitrative stress. Nitrotyrosine is detected in biological fluids, such as plasma, lung aspirant-BALF (bronchoalveolar lavage fluid) and urine. Increased levels of nitrotyrosine are detected in rheumatoid arthritis, septic shock and celiac disease. In all these studies, nitrotyrosine was not detected in healthy subjects. Nitrotyrosine is also found in many other diseased tissues, such as the cornea in keratoconus. Peroxynitrite and / or nitrative stress may be involved in the pathogenesis of diabetes.

[0221] Nitrotyrosine, as a marker of reactive oxygen species, has also been linked to the degeneration of dopamine neurons. Tyrosine is the precursor to dopamine, a neurotransmitter important for motivation, attention, learning, circadian rhythms, and other biological processes.

[0222] Study Design: Human brain microvascular endothelial cells (HBMEC) were cultured in 6-well plates using hypoxic conditions (hypoxic conditions; 2% oxygen). Control HBMEC cells were cultured under normoxic conditions. Prior to treatment, cells were incubated in serum-free medium for 24 hours. Cells were treated for 48 hours with the following agents: 1. Superoxide dismutase only; 2. Prebiotic fiber only; 3. Fruit juice only; 4. Superoxide dismutase + prebiotic fiber + fruit juice (combination); and 5. Negative control.

[0223] Enzyme-Linked Immunosorbent Assay: After 48 hours of incubation, the medium was removed from the cells and placed in a test tube. To evaluate whether revivify gel attenuates human brain microvascular endothelial cells (HBMEC) from oxidative damage, the following biomarkers were evaluated in hypoxia-induced HBMEC culture medium: 1. malondialdehyde (MDA); 2. 4-hydroxynonenal, or 4-hydroxy-2-nonenal or 4-HNE or HNE; 3. protein carbonyl; and 4. 3-nitrotyrosine.

[0224] Results: As can be seen in Figures 16-19, some of the gel components showed improvement in the oxidative damage biomarkers, but the greatest improvement was seen with the finished product gel. As can be seen in Figure 16, each component of the gel provided some reduction in the MDA biomarker compared to the positive control, with the finished product gel surprisingly having lower MDA levels under hypoxic conditions than cells cultured under non-hypoxic control conditions. Similar results are seen in Figure 17 using HNE as the biomarker, and again, the finished product gel surprisingly has lower biomarker levels than cells cultured under non-hypoxic control conditions.

[0225] As can be seen in Figure 18, the levels of the protein carbonyl biomarker are reduced from the positive control for each individual gel component, with a surprising and significant reduction from the positive control for the finished gel product. Similar results are observed for the 3-nitrotyrosine biomarker as shown in Figure 19, with the finished product again showing a surprising and significant reduction compared to the positive control.

[0226] The results in this example demonstrate that the finished REVIVIFY® gel provides substantial and surprising protection of HBMEC from oxidative agents. These results suggest that absorbed SOD, when combined with soluble fiber and polyphenols, can provide significant protection of neural cells from oxidative damage.

[0227] References for Example 4 [Table 7-1] [Table 7-2]

[0228] Although certain embodiments have been illustrated and described herein, it is to be understood that the claims are not limited to the specific forms or arrangements of parts described and shown. Although exemplary embodiments have been disclosed and specific terms are used herein, they are used in a generic and descriptive sense only and not for purposes of limitation. Modifications and variations of the embodiments are possible in light of the above teachings. It is therefore to be understood that the embodiments may be practiced otherwise than as specifically described.

[0229] Although various embodiments have been described above, it should be understood that they are presented only as illustrations and examples of the present technology, and not for the purpose of limitation. It will be apparent to those skilled in the relevant art that various changes in form and detail can be made without departing from the spirit and scope of the present technology. Thus, the breadth and scope of the present technology should not be limited by any of the above-described embodiments, but should be defined only according to the appended claims and their equivalents. It will also be understood that each feature of the embodiments discussed in this specification and the references cited in this specification can be used in combination with the features of any other embodiment. All patents and publications discussed in this specification are incorporated herein by reference in their entirety.

Claims

1. a) about 0.03 units / mL to about 0.5 units / mL of superoxide dismutase; b) about 1.3 mg / mL to about 23 mg / mL of soluble fiber; and c) water A liquid composition comprising:

2. 10. The liquid composition of claim 1, comprising about 0.05 units / mL to about 0.4 units / mL of superoxide dismutase.

3. 3. The liquid composition of claim 1 or 2, comprising about 2.7 mg / mL to about 12 mg / mL of soluble fiber.

4. 3. The liquid composition of claim 1 or 2, wherein the superoxide dismutase is extracted from melon, beef liver, heterotrophic bacteria, or marine phytoplankton.

5. 3. The liquid composition according to claim 1, wherein the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase, or nickel superoxide dismutase.

6. 3. The liquid composition of claim 1, wherein the ratio of superoxide dismutase to soluble fiber is about 1:100 to about 1:1000 by weight.

7. 3. The liquid composition according to claim 1, wherein the soluble fiber is a water-soluble polysaccharide.

8. 3. The liquid composition of claim 1, wherein the soluble fiber is selected from soluble corn fiber, inulin, dextrin, guar gum, oligopolysaccharides, galactopolysaccharides, fructooligosaccharides, lactulose, resistant starch, xylooligosaccharides, and isomaltooligosaccharides.

9. 9. The liquid composition of claim 8, wherein the soluble fiber is soluble corn fiber.

10. 3. The liquid composition of claim 1 or 2, further comprising about 0.1 mg / mL to about 1.5 mg / mL of a simple sugar.

11. 3. The liquid composition of claim 1 or 2, further comprising about 0.1 mg / mL to about 1.5 mg / mL of d-ribose.

12. 3. The liquid composition of claim 1, further comprising about 1.3 mg / mL to about 9.0 mg / mL of a sugar alcohol.

13. 3. The liquid composition of claim 1, further comprising about 0.1 mg / mL to about 1.5 mg / mL of a pH adjusting agent.

14. 3. The liquid composition of claim 1 or 2, further comprising about 0.05 mg / mL to about 0.75 mg / mL of a sweetener.

15. 3. The liquid composition of claim 1 or 2, further comprising a flavoring agent.

16. a) about 10 units to about 200 units of superoxide dismutase; b) about 500 mg to about 8000 mg of soluble fiber; and c) Probiotics A composition comprising:

17. 17. The composition of claim 16, wherein the ratio of superoxide dismutase to soluble fiber is from about 1:100 to about 1:1000 by weight.

18. 18. The composition of claim 16 or 17, wherein the probiotic comprises bacteria of the genus Bifidobacterium.

19. 18. The composition of claim 16 or 17, wherein the probiotic comprises bacteria of the genus Lactobacillus.

20. 18. The composition of claim 16 or 17, wherein the composition is in the form of a gel.

21. 18. The composition of claim 16 or 17, wherein the composition is in liquid form.

22. 18. The composition of claim 16 or 17, wherein the composition is in the form of a powder.

23. 1. A composition for increasing T cell activation in a subject, the composition comprising: a) about 10 units to about 200 units of superoxide dismutase; and b) about 500 mg to about 8000 mg of soluble fiber Including, A composition, wherein T cell activation is increased in the subject after administration of the composition.

24. 24. The composition of claim 23, wherein the composition is administered in combination with an anti-cancer agent.

25. 24. The composition of claim 23, wherein the composition is administered in combination with an antiviral agent.

26. 1. A composition for increasing production of short chain fatty acids (SCFAs) in the gastrointestinal tract of a subject, the composition comprising: a) about 10 units to about 200 units of superoxide dismutase; and b) about 500 mg to about 8000 mg of soluble fiber Including, A composition, wherein after administration of the composition, production of SCFAs is increased in the digestive tract of the subject.

27. 1. A composition for increasing the amount of Bifidobacterium or Lactobacillus bacteria in the gastrointestinal tract of a subject, the composition comprising: a) about 10 units to about 200 units of superoxide dismutase; and b) about 500 mg to about 8000 mg of soluble fiber Including, A composition, wherein the amount of bacteria of the genus Bifidobacterium, Lactobacillus, or a combination thereof is increased in the digestive tract of the subject after administration of the composition.

28. 28. The composition of claim 27, wherein the composition is in the form of a gel.

29. 28. The composition of claim 27, wherein the composition is in liquid form.

30. 28. The composition of claim 27, wherein the composition is in the form of a powder.

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