Supeoxide dissoluble fiber compositions and methods of use

Abstract

The present invention provides a composition comprising a superoxide dismutase and a soluble fiber. The composition may additionally include other antioxidants, sweeteners, probiotics, vitamins, and nutrients. The compositions are useful as dietary supplements and for improving health and welfare. Methods of using such compounds are also provided.

Description

Technical Field

[0001] The present disclosure generally relates to compositions comprising superoxide dismutase and soluble fiber. The compositions can be used as dietary supplements and for improving health and well-being. The present disclosure further relates to methods of using the compositions comprising superoxide dismutase and soluble fiber. Background Art

[0002] Superoxide dismutase (SOD) is a group of metalloenzymes that prevent reactive oxygen species (ROS) from damaging cells. SOD catalyzes the superoxide anion radical (O2 - ) are dismutated into molecular oxygen and hydrogen peroxide (H2O2). ROS in cells can damage nucleic acids, proteins, and lipids, leading to decreased cell function and possible apoptosis. Therefore, the ability to convert ROS into harmless molecules is critical to protecting cell function and overall health.

[0003] While almost all organisms naturally produce some type of SOD, the level of SOD produced in cells decreases as the subject ages or when the subject suffers from certain health disorders. In addition, the presence of various pollutants and toxins in the environment can lead to increased levels of ROS in cells. Therefore, supplementing naturally produced SOD through the diet is important for maintaining health.

[0004] Another important dietary component is soluble fiber. Soluble fiber absorbs water and forms a gel in the digestive tract. Soluble fiber has many benefits for 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 certain types of soluble fiber. Modern diets are often low in soluble fiber, and supplementation is often necessary to improve health.

[0005] Diabetes is a common carbohydrate metabolism disorder. More than 30 million people in the United States have diabetes. Over the past 20 years, the number of adults diagnosed with diabetes has more than tripled as the U.S. population ages and becomes overweight or obese. In people with diabetes, the body's normal ability to use glucose is impaired, resulting in elevated blood sugar levels. Diabetes is associated with an increased risk of cardiovascular or circulatory disease or disorder.

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

[0007] Antioxidants have been shown to have beneficial effects in the body in combating free radicals, which are implicated in many disease processes. However, the absorption of the effective metabolites of many antioxidants, such as polyphenols, is dependent on microbial metabolism of the antioxidants in the gut. Therefore, for many nutritional supplements, these effective polyphenol metabolites are not produced and the antioxidant activity of the supplement is limited. Summary of the invention

[0008] One aspect of the present disclosure provides a liquid composition comprising: a) about 0.03 units / mL to about 0.5 units / mL superoxide dismutase; b) about 1.3 mg / mL to about 23 mg / mL soluble fiber; c) water.

[0009] In an embodiment, the liquid composition comprises about 0.05 units / mL to about 0.4 units / mL of superoxide dismutase. In an embodiment, the liquid composition comprises about 0.2 units / mL to about 0.3 units / mL of superoxide dismutase.

[0010] 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.

[0011] 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.

[0012] In embodiments of the liquid composition, the weight ratio of superoxide dismutase to soluble fiber is about 1:100 to about 1:1000. In embodiments, the weight ratio of superoxide dismutase to soluble fiber is about 1:500 to about 1:700.

[0013] 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, oligosaccharides, galactose polysaccharides, oligofructose, lactulose, digestion-resistant starch, oligoxylose and isomalto-oligosaccharides. In an embodiment, the soluble fiber is soluble corn fiber. In an embodiment, the soluble fiber is digestion-resistant maltodextrin.

[0014] In an embodiment, the liquid composition further comprises about 0.1 mg / mL to about 1.5 mg / mL of a simple sugar. In an embodiment, the liquid composition further comprises about 0.1 mg / mL to about 1.5 mg / mL of d-ribose. In an embodiment, the liquid composition further comprises about 0.40 mg / mL to about 0.85 mg / mL of d-ribose.

[0015] In an embodiment, the liquid composition further comprises about 1.3 mg / mL to about 9.0 mg / mL of sugar alcohol. In an embodiment, the liquid composition further comprises about 1.3 mg / mL to about 9.0 mg / mL of erythritol. In an embodiment, the liquid composition further comprises about 2.7 mg / mL to about 5.6 mg / mL of erythritol.

[0016] In an embodiment, the liquid composition further comprises a pH adjusting agent of about 0.1 mg / mL to about 1.5 mg / mL. In an embodiment, the liquid composition further comprises citric acid of about 0.1 mg / mL to about 1.5 mg / mL. In an embodiment, the liquid composition further comprises citric acid of about 0.4 mg / mL to about 0.7 mg / mL.

[0017] In an embodiment, the liquid composition further comprises about 0.05 mg / mL to about 0.75 mg / mL of a sweetener. In an embodiment, the liquid composition further comprises about 0.05 mg / mL to about 0.75 mg / mL of a steviol glycoside. In an embodiment, the liquid composition further comprises about 0.2 mg / mL to about 0.35 mg / mL of a steviol glycoside.

[0018] In an embodiment, the liquid composition further comprises a flavoring agent.

[0019] 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 8,000 mg of soluble fiber; and c) a probiotic.

[0020] In an embodiment, the probiotic composition comprises about 50 units to about 150 units of superoxide dismutase. In an embodiment, the probiotic composition comprises about 70 units to about 100 units of superoxide dismutase.

[0021] In embodiments, the probiotic composition comprises from about 1,000 mg to about 5,000 mg of soluble fiber. In embodiments, the probiotic composition comprises from about 2,000 mg to about 4,000 mg of soluble fiber.

[0022] 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.

[0023] In an embodiment of the probiotic composition, the weight ratio of superoxide dismutase to soluble fiber is about 1:100 to about 1:1000. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber is about 1:500 to about 1:700.

[0024] 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, oligosaccharides, galactose polysaccharides, oligofructose, lactulose, digestion-resistant starch, oligoxylose and isomalto-oligosaccharides. In an embodiment, the soluble fiber is soluble corn fiber. In an embodiment, the soluble fiber is digestion-resistant maltodextrin.

[0025] In an embodiment of the probiotic composition, the probiotic comprises a Bifidobacterium bacterium. In an embodiment, the probiotic comprises a Lactobacillus bacterium. In an embodiment, the probiotic comprises a Firmicutes Lactobacillus, an Actinobacteria Bifidobacterium, or a combination thereof.

[0026] 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.

[0027] Another aspect of the present disclosure provides a method of increasing T cell activation in 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; wherein, after administration of the composition, activation of T cells in the subject is increased. In an embodiment of the method, the composition is administered in combination with an anticancer agent. In an embodiment of the method, the composition is administered in combination with an antiviral agent.

[0028] Another aspect of the present disclosure provides a method for increasing the production of short-chain fatty acids (SCFA) in the digestive tract of a subject, comprising orally administering a composition to 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; wherein, after administering the composition, the production of SCFA in the digestive tract of the subject increases. In an embodiment of the method, the SCFA with increased production is acetic acid, propionic acid, butyrate or lactic acid SCFA or a combination thereof. In an embodiment of the method, SCFA is increased in a manner that provides approximately the same ratio of acetic acid, propionic acid, butyrate and lactic acid SCFA compared to the ratio of acetic acid, propionic acid, butyric acid and lactic acid SCFA before the increase.

[0029] Another aspect of the present disclosure provides a method for increasing the amount of Bifidobacterium or Lactobacillus bacteria 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; wherein, after administration of the composition, the amount of Bifidobacterium, Lactobacillus bacteria, or a combination thereof in the digestive tract of the subject increases. In an embodiment of the method, the Bifidobacterium bacteria comprises an Actinomyces species of Bifidobacterium. In an embodiment of the method, the Lactobacillus bacteria comprises a Firmicutes species of Lactobacillus.

[0030] In embodiments of any of the methods described herein, the probiotic composition comprises from about 50 units to about 150 units of superoxide dismutase. In embodiments, the composition comprises from about 70 units to about 100 units of superoxide dismutase.

[0031] In embodiments of any of the methods described herein, the composition comprises from about 1000 mg to about 5000 mg of soluble fiber. In embodiments, the composition comprises from about 2,000 mg to about 4,000 mg of soluble fiber.

[0032] In embodiments of any of the methods described herein, the superoxide dismutase is extracted from melon, beef liver, heterotrophic bacteria, or marine phytoplankton. In embodiments, the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase, or nickel superoxide dismutase.

[0033] In embodiments of any of the methods described herein, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:100 to about 1:1000. In embodiments, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:500 to about 1:700.

[0034] In embodiments of any method described herein, soluble fiber is a water-soluble polysaccharide. In embodiments, soluble fiber is selected from soluble corn fiber, inulin, dextrin, guar gum, oligosaccharides, galactose polysaccharides, oligofructose, lactulose, digestion-resistant starch, oligoxylose and isomalto-oligosaccharides. In embodiments, soluble fiber is soluble corn fiber. In embodiments, soluble fiber is digestion-resistant maltodextrin.

[0035] In an embodiment of any of the methods described herein, 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. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] Figure 1 is a graph of absorbance at 490 nm of Jurkat cells treated with lipopolysaccharide (LPS), different components of the gel formulation of Example 1, combinations of gels (finished formulations), and a negative control (no treatment) (indicating cell viability as described in Example 1).

[0037] Figure 2 is a graph showing T cell activation measured by CD69 and CD3 expression as described in Example 1. Jurkat cells are abbreviated as JRK.

[0038] Figure 3 is a graph showing the effect of the gel of Example 1 and its components at the following concentrations on CD4+ differentiation in Jurkat cells: Fibersol prebiotic fiber 1.3 mg / mL; superoxide dismutase (SOD) 0.2 mg / mL; LPS 10 μg / mL; juice 4.6 mg / mL, composition (finished product) 5.6 mg / mL.

[0039] Figure 4 is a chart showing the effect of the gel of Example 1 and its components at the following concentrations on CD8+ differentiation in Jurkat cells: Fibersol prebiotic fiber 1.3 mg / mL; superoxide dismutase (SOD) 0.2 mg / mL; LPS 10 μg / mL; juice 4.6 mg / mL, composition (finished product) 5.6 mg / mL.

[0040] Figure 5 is a graph showing the effect of the gel of Example 1 and its components on the attenuation of 8-Isoprostane secretion by LPS-activated Jurkat cells, as described in Example 1.

[0041] Figure 6 is a graph showing the effect of the gel of Example 1 and its components on the attenuation of cyclooxygenase 2 (COX-2) secretion from LPS-activated Jurkat cells, as described in Example 1.

[0042] Figure 7 is a graph showing the effect of the gel of Example 1 and its components on the attenuation of interferon-γ (IFN-γ) secretion by LPS-activated Jurkat cells, as described in Example 1.

[0043] Figure 8 is a graph showing the effect of the gel of Example 1 and its components on the attenuation of interleukin-6 (IL-6) secretion by LPS-activated Jurkat cells, as described in Example 1.

[0044] Fig. 9 is a graph showing the effect of the gel of Example 1 and its components on attenuation of transforming growth factor-β (TGF-β) secretion by LPS-activated Jurkat cells, as described in Example 1.

[0045] Fig.10 is a graph showing the effect of the gel of Example 1 and its components on attenuation of LPS-activated tumor necrosis factor alpha (TNF-α) secretion, as described in Example 1.

[0046] Fig.11 is a graph showing the effect of the gel of Example 1 and its components on the attenuation of CXC motif chemokine ligand 10 (CXCL10) secretion in LPS-activated Jurkat cells, as described in Example 1.

[0047] Fig.12 Is using a complete gel product ( Figure 2 is a graph showing the bacterial density over time of cultures grown on the intestines of goats treated with either the finished product (or the gel component).

[0048] Fig.13 Is using a complete gel product ( Figure 2 is a graph showing the bacterial density over time of cultures grown on the intestines of goats treated with either the finished product (or the gel component).

[0049] Fig.14A is a graph of the concentrations of the 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 gel components as described in Example 3.

[0050] Fig. 14B is a graph 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 gel components as described in Example 3.

[0051] Fig.15is a graph showing the microbial profile of in vitro intestinal microbial cultures when treated with the complete gel product (combination) or the gel component as described in Example 3.

[0052] Fig.16 is a graph of the concentration of malondialdehyde (MDA) biomarker secreted by human brain microvascular endothelial cells (HBMEC) when treated with the complete (finished) gel product or components as described in Example 4.

[0053] Fig.17 is a graph of the concentration of 4-hydroxynonenal (HNE) biomarker secreted by HBMECs when treated with the complete (finished) gel product or components as described in Example 4.

[0054] Fig.18 is a graph of the concentrations of protein carbonyl (PC) biomarkers secreted by HBMEC when treated with the complete (finished) gel product or components as described in Example 4.

[0055] Fig.19 is a graph of the concentration of 3-nitrotyrosine (NT) biomarker secreted by HBMEC when treated with the complete (finished) gel product or components as described in Example 4. DETAILED DESCRIPTION

[0056] The present invention provides compositions comprising superoxide dismutase and soluble fiber. The compositions disclosed herein can be used as supplements to increase the levels of superoxide dismutase and soluble fiber in a subject. As described herein, the compositions may also include additional components, such as antioxidants, vitamins or other nutrients, as well as excipients and other formulation ingredients.

[0057] It should be understood that the specific embodiments shown and described herein are examples, and are not intended to limit the scope of the present application in any way.

[0058] The published patents, patent applications, websites, company names, and scientific literature mentioned herein are hereby incorporated by reference in their entirety, to the extent that each is specifically and individually indicated to be incorporated by reference. Any conflict with any reference cited herein and the specific teachings of this specification should be resolved in favor of the latter. Similarly, any conflict between the definitions of the art-understood words or phrases specifically taught in this specification and the definitions of the words or phrases should be resolved in favor of the latter.

[0059] As used herein, "a" or "an" may refer to one or more than one. As used herein, when used in conjunction with the word "comprising", "a" or "an" may refer to one or more than one. As used herein, "another" or "another" may refer to at least a second or more.

[0060] Throughout the application, the term "about" is used to represent a value that includes the inherent differences in the error of the device / method for determining the value or the differences present in the research subjects. Typically, the term "about" means including about or less than 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or 20% variability as the case may be.

[0061] The term "or" is used in the claims to mean "and / or" unless specifically indicated to refer to only alternatives, or the alternatives are mutually exclusive, but the disclosure supports a definition referring to only alternatives and "and / or."

[0062] As used herein, the terms "comprising" (and any variations or forms of comprising, such as "including" and "containing"), "having" (and any variations or forms of having, such as "containing" and "having"), "containing" (and any variations or forms of including, such as "including" and "containing"), or "containing" (and any variations or forms of containing, such as "containing" and "including") are inclusive or open-ended, and do not exclude additional, unrecited elements or method steps.

[0063] The use of the term "for example" and its corresponding abbreviation "such as" (whether italicized or not) means that the particular term recited is representative examples and embodiments of the disclosure and is not intended to be limiting to the particular examples recited or cited unless expressly stated otherwise.

[0064] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. Reference is made herein to various methods and materials known to one of ordinary skill in the art.

[0065] As used herein, the term "superoxide dismutase" (sometimes abbreviated as SOD) refers to an enzyme that catalyzes the production of superoxide anion radicals (O - ) into molecular oxygen and hydrogen peroxide (H2O2). The enzyme class (EC) number of superoxide dismutase is 1.15.1.1. Superoxide dismutase is an important component in protecting cells from reactive oxygen species (ROS), such as O formed during the metabolism of oxygen in cells. 2- Therefore, superoxide dismutase, as an important antioxidant, is present in almost all types of cellular organisms. However, especially in complex organisms (such as mammals), the level of superoxide dismutase in cells decreases as the organism ages, making the organism more susceptible to cell damage by reactive oxygen species.

[0066] As used herein, the term "soluble fiber" refers to any type of water-soluble dietary fiber. After consumption, soluble fiber absorbs water and forms a gel in the intestinal tract of an organism, helping to slow down the metabolism of lipids and carbohydrates. Soluble fiber is also a prebiotic that can be fermented by intestinal bacteria and helps maintain a healthy intestinal microbiome.

[0067] As used herein, the term "antioxidant" refers to a substance that significantly reduces the adverse effects of reactive species such as reactive oxygen and nitrogen species by completely or partially neutralizing the reactive species. Antioxidants can be divided into "primary antioxidants" and "secondary antioxidants." Primary antioxidants delay or inhibit the initial steps of oxidation, while secondary antioxidants slow oxidation by removing substrates or by quenching oxygen free radicals.

[0068] 1) Superoxide dismutase

[0069] In embodiments, the amount of superoxide dismutase contained in the compositions disclosed herein is measured in units of activity per mg of protein. For enzyme activity, one unit (U) (in μmol / min) is defined as the amount of enzyme that catalyzes the conversion of 1 micromole of substrate per minute under specified conditions. Units of superoxide dismutase activity can be measured by any known method. For example, methods for determining superoxide dismutase activity in units can be found in McCord, JM and Fridovich, I., J. Biol. Chem. 1969, 244: 6049-6055; Weydert et al., Nature Protocols 2010, 5 (1): 51–66; and https: / / www.sigmaaldrich.com / technical-documents / protocols / biology / enzymatic-assay-of-superoxide-dismutase.html The technical solution of the present invention; the disclosure of each of which is incorporated herein by reference. In an embodiment, the unit of superoxide dismutase activity is defined as the amount of superoxide dismutase that inhibits the reduction rate of cytochrome c by 50% in a coupled system using xanthine and xanthine oxidase at pH 7.8, 25°C, and a reaction volume of 3.0 ml.

[0070] In embodiments, the concentration of the amount of superoxide dismutase in the composition disclosed herein is measured with the total amount of superoxide dismutase units in the composition. In embodiments, the composition comprises about 10 units to about 200 units of superoxide dismutase. In embodiments, 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. In embodiments, the composition comprises about 20 units to about 190 units of superoxide dismutase. In embodiments, the composition comprises about 30 units to about 180 units of superoxide dismutase. In embodiments, the composition comprises about 40 units to about 170 units of superoxide dismutase. In embodiments, the composition comprises about 50 units to about 160 units of superoxide dismutase. In embodiments, the composition comprises about 60 units to about 150 units of superoxide dismutase. In embodiments, the composition comprises about 70 units to about 140 units of superoxide dismutase. In embodiments, the composition comprises about 80 units to about 130 units of superoxide dismutase. In embodiments, the composition comprises about 90 units to about 130 units of superoxide dismutase. In embodiments, the composition comprises about 75 units to about 95 units of superoxide dismutase. In embodiments, the composition comprises about 80 units to about 90 units of superoxide dismutase. In embodiments, 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 embodiments, the composition comprises about 84 units of superoxide dismutase.

[0071] The superoxide dismutase used in the compositions disclosed herein can be obtained from any source of enzymes. In embodiments, 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 embodiments, the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase, or nickel superoxide dismutase.

[0072] In embodiments, superoxide dismutase is extracted from a plant. In embodiments, the plant is a fruit, a grain or a tuber. In embodiments, the plant is a fruit selected from melon, citrus fruit, peach, pear, apple or banana. In embodiments, the plant is a grain selected from wheat, barley, rye, millet, oats, spelt, bulger, sorghum and farro. In some embodiments, the plant is a tuber selected from horseradish, potato, yam, sweet potato, cassava or dahlia.

[0073] In an embodiment, the superoxide dismutase is extracted from an animal. In an embodiment, the animal is a cow, a pig, a sheep or a goat.

[0074] In an embodiment, the superoxide dismutase is extracted from a microorganism. In an embodiment, the microorganism is a phytoplankton or a bacterium. In an embodiment, the microorganism is a heterotrophic bacterium, for example, a bacterium that obtains sugars required for energy production from their environment.

[0075] 2) Soluble Fiber

[0076] The ratio of superoxide dismutase to soluble fiber can be adjusted as needed by changing the amount of either or both components in the composition. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the composition is from about 1:100 to about 1:1000. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the composition is from about 1:200 to about 1:800. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the composition is from about 1:300 to about 1:700. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the composition is from about 1:500 to about 1:700. In some embodiments, the weight ratio of superoxide dismutase to soluble fiber in the composition is from about 1:650 to about 1:675.

[0077] Without wishing to be bound by theory, the inventors have discovered that the presence of soluble fiber and superoxide dismutase in a specific ratio can synergistically improve stability and absorption in the digestive tract (intestines), as demonstrated in the examples below. One of the mechanisms for this improved stability and absorption is that soluble fiber forms a gel in the digestive tract, which captures and protects superoxide dismutase while also making it more easily absorbed through the intestinal wall. The soluble fiber and antioxidant preparations of the present disclosure provide a synergistic effect because the soluble prebiotic fiber acts as a fertilizer for bacteria in the colon, while the polyphenols are mostly absorbed in the colon and are extensively decomposed by the colonic microbiota. The presence of prebiotic fiber enhances the effects of the colonic microbiota, resulting in more efficient absorption of polyphenols in the colon. Most of the polyphenols from antioxidant supplements eventually enter the large intestine, where they are converted to active metabolites through microbial metabolism, where they can exert an antioxidant effect. Soluble fiber modulates intestinal microorganisms and maximizes polyphenol metabolism, resulting in a number of antioxidant, anti-inflammatory and anti-infective effects.

[0078] 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, providing a matrix for the proliferation of microorganisms in the intestine.

[0079] In embodiments of the compositions disclosed herein, various types of soluble fibers may be used, including mixtures of two, three, four, five, six or more different types of soluble fibers. In an embodiment, the soluble fiber is a water-soluble polysaccharide. In an embodiment, the soluble fiber is separated from corn, wheat, barley, rye, beans, apples, pears, peaches, citrus fruits, berries, peas, rice bran or oats. In an embodiment, the soluble fiber is selected from soluble corn fiber, inulin, dextrin, guar gum, oligosaccharides, galactose polysaccharides, oligofructose, lactulose, digestion-resistant starch, oligoxylose and isomaltooligosaccharides. In an embodiment, the soluble fiber is soluble corn fiber. In an embodiment, the soluble fiber is digestion-resistant maltodextrin. In an embodiment, the soluble fiber is sold by Archer Daniels Midland Company and Matsutani Chemical Industry Co., Ltd. In an embodiment, the soluble fiber is There are alternative types of Fibersol, such as Fibersol-2AG, Fibersol-LQ, Fibersol-2L, Fibersol-DLQ or non-GMO Fibersol.

[0080] In some embodiments, the soluble fiber is corn-based resistant maltodextrin (Fibersol-2) prebiotic fiber.

[0081] 3) Liquid composition

[0082] 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 superoxide dismutase; b) about 1.3 mg / mL to about 23 mg / mL soluble fiber; c) water.

[0083] In an embodiment, the liquid composition comprises a superoxide dismutase of about 0.05 units / mL to about 0.4 units / mL. In an embodiment, the liquid composition comprises a superoxide dismutase of about 0.2 units / mL to about 0.3 units / mL. In an embodiment, the liquid composition comprises a superoxide dismutase of about 0.1 units / mL to about 0.4 units / mL. In an embodiment, the liquid composition comprises a superoxide dismutase of about 0.15 units / mL to about 0.35 units / mL. In an embodiment, the liquid composition comprises a superoxide dismutase of about 0.05, 0.1, 0.15, 0.2, 0.25, 0.3, 0.35 or 0.4 units / mL. In an embodiment, the liquid composition comprises a superoxide dismutase of 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.

[0084] In an embodiment, the liquid composition comprises about 2.7 mg / mL to about 12 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises about 5.55 mg / mL to about 11.11 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises about 2 mg / mL to about 15 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises about 4 mg / mL to about 12 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises about 5 mg / mL to about 12 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises about 6 mg / mL to about 12 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises about 7 mg / mL to about 12 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises about 8 mg / mL to about 12 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises about 9 mg / mL to about 12 mg / mL of soluble fiber. In an embodiment, the liquid composition comprises about 10 mg / mL to about 12 mg / mL of soluble fiber. In embodiments, the liquid composition comprises about 2.78, 5.56, 8.33, 11.11, or 13.89 mg / mL of soluble fiber.

[0085] In embodiments, the liquid composition comprises any superoxide dismutase disclosed herein. In embodiments, the superoxide dismutase is extracted from melon, beef liver, heterotrophic bacteria, or marine phytoplankton. In embodiments, the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase, or nickel superoxide dismutase.

[0086] In an embodiment of the liquid composition, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:100 to about 1:1000. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:200 to about 1:800. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:300 to about 1:700. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:500 to about 1:700. In some embodiments, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:650 to about 1:675.

[0087] In an embodiment, the liquid composition comprises any soluble fiber 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, oligosaccharides, galactose polysaccharides, oligofructose, lactulose, digestion-resistant starch, oligoxylose and isomalto-oligosaccharides. In an embodiment, the soluble fiber is soluble corn fiber. In an embodiment, the soluble fiber is digestion-resistant maltodextrin.

[0088] In embodiments, the liquid composition may further comprise additional ingredients, including sweeteners, pH adjusters, flavoring agents, and other agents, including combinations of these agents.

[0089] 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.

[0090] In an embodiment, the sweetener is a simple sugar. In an embodiment, the simple sugar is ribose, glucose, fructose, sucrose, lactose, or a combination thereof. In an embodiment, the simple sugar is ribose. In an embodiment, the simple sugar is d-ribose.

[0091] In an embodiment, the liquid composition comprises a simple sugar of about 0.1mg / mL to about 1.5mg / mL. In an embodiment, the liquid composition comprises a simple sugar of about 0.2mg / mL to about 1.3mg / mL. In an embodiment, the liquid composition comprises a simple sugar of about 0.3mg / mL to about 1.0mg / mL. In an embodiment, the liquid composition comprises a simple sugar of about 0.4mg / mL to about 0.8mg / mL. In an embodiment, the liquid composition comprises a simple sugar of about 0.4mg / mL to about 0.85mg / mL. In an embodiment, the liquid composition comprises a simple sugar of about 0.4mg / mL to about 0.6mg / mL. In an embodiment, the liquid composition comprises a simple sugar of about 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8 or 0.85mg / mL.

[0092] In an embodiment, the liquid composition comprises about 0.1mg / mL to about 1.5mg / mL of d-ribose. In an embodiment, the liquid composition comprises about 0.2mg / mL to about 1.3mg / mL of d-ribose. In an embodiment, the liquid composition comprises about 0.3mg / mL to about 1.0mg / mL of d-ribose. In an embodiment, the liquid composition comprises about 0.4mg / mL to about 0.8mg / mL of d-ribose. In an embodiment, the liquid composition comprises about 0.4mg / mL to about 0.85mg / mL of d-ribose. In an embodiment, the liquid composition comprises about 0.4mg / mL to about 0.6mg / mL of d-ribose. In an embodiment, the liquid composition comprises about 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8 or 0.85mg / mL of d-ribose.

[0093] 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.

[0094] In an embodiment, the liquid composition comprises about 1.3 mg / mL to about 9.0 mg / mL of sugar alcohol. In an embodiment, the liquid composition comprises about 2.0 mg / mL to about 8.0 mg / mL of sugar alcohol. In an embodiment, the liquid composition comprises about 3.9 mg / mL to about 7.0 mg / mL of sugar alcohol. In an embodiment, the liquid composition comprises 2.7 mg / mL to about 5.6 mg / mL of sugar alcohol. In an embodiment, the liquid composition comprises 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.

[0095] In an embodiment, the liquid composition comprises erythritol from about 1.3 mg / mL to about 9.0 mg / mL. In an embodiment, the liquid composition comprises erythritol from about 2.0 mg / mL to about 8.0 mg / mL. In an embodiment, the liquid composition comprises erythritol from about 3.9 mg / mL to about 7.0 mg / mL. In an embodiment, the liquid composition comprises erythritol from 2.7 mg / mL to about 5.6 mg / mL. In an embodiment, the liquid composition comprises erythritol 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.

[0096] In an embodiment, the sweetener is a steviol glycoside. In an embodiment, the steviol glycoside is rebaudioside A, stevioside, rebaudioside C, dulcoside A, rebaudioside B, rebaudioside D, rebaudioside E, steviolbioside or a combination thereof.

[0097] In an embodiment, the liquid composition comprises about 0.05 mg / mL to about 0.75 mg / mL of steviol glycosides. In an embodiment, the liquid composition comprises about 0.2 mg / mL to about 0.35 mg / mL of steviol glycosides. In an embodiment, the liquid composition comprises about 0.1 mg / mL to about 0.5 mg / mL of steviol glycosides. In an embodiment, the liquid composition comprises about 0.2 mg / mL to about 0.3 mg / mL of steviol glycosides. In an embodiment, 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 steviol glycosides.

[0098] In an embodiment, the liquid composition comprises about 0.05 mg / mL to about 0.75 mg / mL of steviol biglycoside A. In an embodiment, the liquid composition comprises about 0.2 mg / mL to about 0.35 mg / mL of steviol biglycoside A. In an embodiment, the liquid composition comprises about 0.1 mg / mL to about 0.5 mg / mL of steviol biglycoside A. In an embodiment, the liquid composition comprises about 0.2 mg / mL to about 0.3 mg / mL of steviol biglycoside A. In an embodiment, 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 steviol biglycoside A.

[0099] In an embodiment, the liquid composition further comprises a pH adjusting agent. In an embodiment, the pH adjusting agent is any food safety agent that can be used to change the pH of the liquid composition. In an embodiment, the pH adjusting agent is citric acid, acetic acid, hydrochloric acid, lactic acid, malic acid, phosphoric acid, tartaric acid or a combination thereof.

[0100] In an embodiment, the liquid composition comprises about 0.1 mg / mL to about 1.5 mg / mL of a pH adjuster. In an embodiment, the liquid composition comprises about 0.2 mg / mL to about 1.3 mg / mL of a pH adjuster. In an embodiment, the liquid composition comprises about 0.3 mg / mL to about 1.0 mg / mL of a pH adjuster. In an embodiment, the liquid composition comprises about 0.4 mg / mL to about 0.8 mg / mL of a pH adjuster. In an embodiment, the liquid composition comprises about 0.4 mg / mL to about 0.85 mg / mL of a pH adjuster. In an embodiment, the liquid composition comprises about 0.4 mg / mL to about 0.6 mg / mL of a pH adjuster. In an embodiment, the liquid composition comprises 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 pH adjuster

[0101] In an embodiment, the liquid composition comprises citric acid from about 0.1mg / mL to about 1.5mg / mL. In an embodiment, the liquid composition comprises citric acid from about 0.2mg / mL to about 1.3mg / mL. In an embodiment, the liquid composition comprises citric acid from about 0.3mg / mL to about 1.0mg / mL. In an embodiment, the liquid composition comprises citric acid from about 0.4mg / mL to about 0.8mg / mL. In an embodiment, the liquid composition comprises citric acid from about 0.4mg / mL to about 0.85mg / mL. In an embodiment, the liquid composition comprises citric acid from about 0.4mg / mL to about 0.6mg / mL. In an embodiment, the liquid composition comprises citric acid from about 0.4, 0.45, 0.5, 0.55, 0.6, 0.65, 0.7, 0.75, 0.8 or 0.85mg / mL.

[0102] In an embodiment, the liquid composition comprises a flavor. In an embodiment, the flavor is a natural flavor. In an embodiment, the flavor is an artificial flavor. In an embodiment, the flavor is a juice flavor. In an embodiment, the flavor is pomegranate, red grape, blueberry, black sweet cherry, sour cherry, wolfberry, acai berry, blackberry, raspberry, strawberry, gooseberry, cranberry, orange, grapefruit, watermelon, beet, apple, lemon, lime, lychee, pineapple, plum, mango or a combination thereof. In an embodiment, the liquid composition comprises a cola flavor.

[0103] In an embodiment, the liquid composition is formulated in a beverage. In an embodiment, the liquid composition is formulated as a hydration drink, a protein shake, juice, tea, coffee, milk, kefir, ice cream, yogurt, smoothie, broth or soup.

[0104] In an embodiment, the volume of the liquid composition is from about 15 mL to about 1500 mL. In an embodiment, the volume of the liquid composition is from about 30 mL to about 1200 mL. In an embodiment, the volume of the liquid composition is from about 50 mL to about 1000 mL. In an embodiment, the volume of the liquid composition is from about 100 mL to about 500 mL. In an embodiment, the volume of the liquid composition is from about 200 mL to about 400 mL. In an embodiment, the volume of the liquid composition is from about 200 mL to about 1000 mL. In an embodiment, the volume of the liquid composition is from about 300 mL to about 1000 mL. In an embodiment, the volume of the liquid composition is about 50, 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, or 1000 mL

[0105] 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.

[0106] In an embodiment, specific examples of liquid compositions are provided in Table 1:

[0107] Table 1

[0108] 1.SOD [melon extract powder, 14,000 units / g]--------------------6.0 mg [84 enzyme units]

[0109] 2.Fibersol-2---------------------------------4000.0mg[i can be reduced to 3 or 2g]

[0110] 3.D-Ribose-------------------------------------------------200.0mg

[0111] 4. Erythritol---------------------------------------------------1494.0mg

[0112] 5.Citric acid--------------------------------------------------200.0mg

[0113] 6. Stevioside A-------------------------------------------------100.0mg

[0114] 7. Flavoring agent--------------------------------------------------100.0mg

[0115] 8.Appropriate amount of water-------------------------------------------------------360ml.

[0116] 4) Powder composition

[0117] In some embodiments of the compositions disclosed herein, the composition is in powder form. In embodiments in which the composition is in powder form, it can be consumed in dry powder form or added to a beverage or food. In embodiments, the powder is mixed into water, a hydration beverage, a protein shake, juice, tea, coffee, milk, kefir, ice cream, yogurt, smoothie, broth, or soup before consumption.

[0118] In an embodiment, the powder composition comprises a high concentration of superoxide dismutase by weight, for example greater than 0.1ppm, greater than 0.5ppm, greater than 1ppm, greater than 2ppm, greater than 5ppm, greater than 10ppm, greater than 20ppm, greater than 50ppm, greater than 100ppm, greater than 200ppm, greater than 300ppm, greater than 400ppm, or greater than 500ppm. In an embodiment, the powder composition comprises a superoxide dismutase of about 0.1ppm to about 10ppm by weight. In an embodiment, the powder composition comprises a superoxide dismutase of about 0.5ppm to about 5ppm by weight. In an embodiment, the powder composition comprises a superoxide dismutase of about 0.7ppm to about 2ppm by weight. In an embodiment, the powder composition comprises a superoxide dismutase of about 0.8ppm to about 1.2ppm by weight. 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 superoxide dismutase.

[0119] In an embodiment, the powder composition comprises about 5ppm to about 15ppm by weight of superoxide dismutase. In an embodiment, the powder composition comprises about 7.5ppm to about 12.5ppm by weight of superoxide dismutase. In an embodiment, the powder composition comprises about 8ppm to about 11ppm by weight of superoxide dismutase. In an embodiment, the powder composition comprises about 9ppm to about 11ppm by weight of superoxide dismutase. In an embodiment, the powder composition comprises about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15ppm by weight of superoxide dismutase.

[0120] In an embodiment, the powder composition comprises about 1% to about 90% soluble fiber by weight. In an embodiment, the powder composition comprises about 1% to about 50% soluble fiber by weight. In an embodiment, the powder composition comprises about 5% to about 25% soluble fiber by weight. In an embodiment, the powder composition comprises about 10% to about 20% soluble fiber by weight. In an embodiment, the powder composition comprises about 12% to about 14% soluble fiber by weight. In an embodiment, the powder 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.

[0121] 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.

[0122] 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.

[0123] In an embodiment, the sweetener is a simple sugar. In an embodiment, the simple sugar is ribose, glucose, fructose, sucrose, lactose, or a combination thereof. In an embodiment, the simple sugar is ribose. In an embodiment, the simple sugar is d-ribose.

[0124] In an embodiment, the powder composition comprises about 1% to about 10% by weight of simple sugars. In an embodiment, the powder composition comprises about 2% to about 8% by weight of simple sugars. In an embodiment, the powder composition comprises about 2% to about 5% by weight of simple sugars. In an embodiment, the powder composition comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% by weight of simple sugars.

[0125] In an embodiment, the powder composition comprises about 1% to about 10% d-ribose by weight. In an embodiment, the powder composition comprises about 2% to about 8% d-ribose by weight. In an embodiment, the powder composition comprises about 2% to about 5% d-ribose by weight. In an embodiment, the powder composition comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% d-ribose by weight.

[0126] 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.

[0127] In an embodiment, the powder composition comprises about 10% to about 50% erythritol by weight. In an embodiment, the powder composition comprises about 15% to about 35% erythritol by weight. In an embodiment, the powder composition comprises about 20% to about 30% erythritol by weight. In an embodiment, the powder composition comprises about 15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% erythritol by weight.

[0128] In an embodiment, the sweetener is a steviol glycoside. In an embodiment, the steviol glycoside is steviol diglycoside A, steviol diglycoside C, dulcoside A, steviol diglycoside B, steviol diglycoside D, steviol diglycoside E, steviol diglycoside or a combination thereof.

[0129] In an embodiment, the powder composition comprises about 0.5% to about 5% steviol glycosides by weight. In an embodiment, the powder composition comprises about 1% to about 4% steviol glycosides by weight. In an embodiment, the powder composition comprises about 1% to about 2.5% steviol glycosides by weight. In an embodiment, the powder composition comprises about 1%, 1.5%, 1.6%, 2%, 2.5%, 3%, 3.5% or 4% steviol glycosides by weight.

[0130] In an embodiment, the powder composition comprises about 0.5% to about 5% by weight of steviol biglycoside A. In an embodiment, the powder composition comprises about 1% to about 4% by weight of steviol biglycoside A. In an embodiment, the powder composition comprises about 1% to about 2.5% by weight of steviol biglycoside A. 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 biglycoside A.

[0131] In an embodiment, the liquid composition further comprises a pH adjusting agent. In an embodiment, the pH adjusting agent is any food safety agent that can be used to change the pH of the liquid composition. In an embodiment, the pH adjusting agent is citric acid, acetic acid, hydrochloric acid, lactic acid, malic acid, phosphoric acid, tartaric acid or a combination thereof.

[0132] In an embodiment, the powder composition comprises from about 1% to about 10% by weight of a pH adjuster. In an embodiment, the powder composition comprises from about 2% to about 8% by weight of a pH adjuster. In an embodiment, the powder composition comprises from about 2% to about 5% by weight of a pH adjuster. In an embodiment, the powder composition comprises from about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% by weight of a pH adjuster.

[0133] In an embodiment, the powder composition comprises from about 1% to about 10% by weight of citric acid. In an embodiment, the powder composition comprises from about 2% to about 8% by weight of citric acid. In an embodiment, the powder composition comprises from about 2% to about 5% by weight of citric acid. In an embodiment, the powder composition comprises about 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10% by weight of citric acid.

[0134] In an embodiment, specific examples of powder compositions are provided in Table 2:

[0135] Table 2

[0136] Total amount: 6 grams.

[0137] 1.SOD [melon extract powder, 14,000 units / g]--------------------6.0 mg [84 enzyme units]

[0138] 2.Fibersol-2-------------------------------------------------4000.0mg

[0139] 3.D-Ribose-------------------------------------------------200.0mg

[0140] 4. Erythritol [excipient]-------------------------------------------1494.0 mg

[0141] 5.Citric acid [excipient]---------------------------------------------200.0 mg

[0142] 6. Sweetener [stevioside A], natural stevia------------------------------100.0mg

[0143] 5) Gel composition

[0144] In embodiments of the compositions disclosed herein, the composition is in the form of a gel. In embodiments in which the composition is in the form of a gel, it can be consumed directly in that form. In other embodiments, the gel can be added to a beverage or food. In embodiments, the gel is mixed into water, a hydration beverage, a protein shake, juice, tea, coffee, milk, kefir, ice cream, yogurt, smoothie, broth, or soup before consumption.

[0145] In an embodiment, the gel composition comprises about 0.005 mg / mL to about 5.0 mg / mL of superoxide dismutase. In an embodiment, the gel composition comprises about 0.01 mg / mL to about 2.5 mg / mL of superoxide dismutase. In an embodiment, the gel composition comprises about 0.05 mg / mL to about 1.0 mg / mL of superoxide dismutase. In an embodiment, the gel composition comprises 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 superoxide dismutase.

[0146] In an embodiment, the gel composition comprises about 50 mg / mL to about 1,000 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.

[0147] In an embodiment of the composition disclosed herein, the composition comprising superoxide dismutase and soluble fiber further comprises fruit juice. The fruit juice can provide additional antioxidants, soluble fiber, insoluble fiber, vitamins and nutrients to the composition. In an embodiment, the gel composition disclosed herein comprises fruit juice.

[0148] In an embodiment, the juice is pomegranate juice, red grape juice, blueberry juice, black sweet cherry juice, sour cherry juice, wolfberry juice, acai 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 types of juice selected from the group consisting of pomegranate juice, red grape juice, blueberry juice, black sweet cherry juice, sour cherry juice, wolfberry juice, acai 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.

[0149] In an embodiment, the juice may be concentrated, for example, by removing some of the water from the original juice. In an embodiment, the juice is concentrated pomegranate juice, red grape juice, blueberry juice, black sweet cherry juice, sour cherry juice, wolfberry juice, acai 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 concentrated juices selected from: concentrated pomegranate juice, concentrated red grape juice, concentrated blueberry juice, concentrated black sweet cherry juice, concentrated sour cherry juice, concentrated wolfberry juice, concentrated acai 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, concentrated prune juice. In embodiments, the concentrated juice has about 60% to about 97% of the water removed from the juice. In embodiments, the concentrated juice has about 85% to about 95% of the water removed from the 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. Any method known in the art for concentrating juice may be used to remove water from the juice.

[0150] In an embodiment, the composition comprises about 5 mg / mL to about 200 mg / mL of concentrated fruit juice. In an embodiment, the composition comprises about 75 mg / mL to about 150 mg / mL of concentrated pomegranate juice. In an embodiment, the composition comprises about 75 mg / mL to about 150 mg / mL of concentrated red grape juice. In an embodiment, the composition comprises about 25 mg / mL to about 100 mg / mL of concentrated blueberry juice. In an embodiment, the composition comprises about 20 mg / mL to about 80 mg / mL of concentrated black sweet cherry juice. In an embodiment, the composition comprises about 20 mg / mL to about 80 mg / mL of concentrated sour cherry juice. In an embodiment, the composition comprises about 2 mg / mL to about 20 mg / mL of concentrated wolfberry juice. In an embodiment, the composition comprises about 2 mg / mL to about 20 mg / mL of concentrated acai juice.

[0151] In an embodiment of the gel composition disclosed herein, the composition comprising superoxide dismutase and soluble fiber further comprises aloe vera. Aloe vera can provide additional antioxidants, soluble fiber, insoluble fiber, vitamins and nutrients to the composition.

[0152] In some embodiments, the aloe is concentrated aloe. In embodiments, the aloe can be concentrated, for example, by removing some water from the aloe extracted from the aloe plant. In embodiments, the concentrated aloe removes about 60% to about 97% of the aloe water. In embodiments, the concentrated aloe removes about 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% of the aloe water. In embodiments, the composition comprises about 2 mg / mL to about 20 mg / mL of concentrated aloe.

[0153] In an embodiment of the gel composition disclosed herein, the composition comprising superoxide dismutase and soluble fiber further comprises green tea. Green tea can provide additional antioxidants, vitamins and nutrients to the composition.

[0154] 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.

[0155] In embodiments of the gel compositions disclosed herein, the composition comprising superoxide dismutase and soluble fiber further comprises resveratrol. Resveratrol can provide additional antioxidants to the composition. In some embodiments, the composition comprises about 0.5 mg / mL to about 6 mg / mL of resveratrol.

[0156] In an embodiment, the specific gel composition is provided in Table 3.

[0157] Table 3

[0158]

[0159] 6) Probiotic composition

[0160] In embodiments, disclosed herein are compositions comprising superoxide dismutase, soluble fiber, and probiotics. In embodiments, the probiotic composition comprises: a) about 10 units to about 200 units of superoxide dismutase; b) about 500 mg to about 8,000 mg of soluble fiber; and c) probiotics.

[0161] In an embodiment, the concentration of the amount of superoxide dismutase in the probiotic composition disclosed herein is measured by the total amount of superoxide dismutase units in the composition. In an embodiment, the composition 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 embodiments, the composition comprises about 90 units to about 130 units of superoxide dismutase. In embodiments, the composition comprises about 75 units to about 95 units of superoxide dismutase. In embodiments, the composition comprises about 80 units to about 90 units of superoxide dismutase. In embodiments, 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 embodiments, the composition comprises about 84 units of superoxide dismutase.

[0162] In an embodiment, the probiotic composition comprises about 1,000 mg to about 5,000 mg of soluble fiber. In an embodiment, the probiotic composition comprises about 2,000 mg to about 4,000 mg of soluble fiber. In an embodiment, the probiotic composition comprises about 1,000 mg to about 10,000 mg of soluble fiber. In an embodiment, the probiotic composition comprises about 2,000 mg to about 9,000 mg of soluble fiber. In an embodiment, the probiotic composition comprises about 3,000 mg to about 8,000 mg of soluble fiber. In an embodiment, the probiotic composition comprises about 2,000 mg to about 4,000 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.

[0163] In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the probiotic composition is about 1:100 to about 1:1000. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:200 to about 1:800. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:300 to about 1:700. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:500 to about 1:700. In some embodiments, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:650 to about 1:675.

[0164] The probiotic composition may comprise one or more superoxide dismutases disclosed herein. In embodiments, the superoxide dismutase is extracted from melon, beef liver, heterotrophic bacteria, or marine phytoplankton. In embodiments, the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase, or nickel superoxide dismutase.

[0165] The probiotic composition may include one or more soluble fibers 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, oligosaccharides, galactose polysaccharides, oligofructose, lactulose, digestion-resistant starch, oligoxylose and isomalto-oligosaccharides. In an embodiment, the soluble fiber is soluble corn fiber. In an embodiment, the soluble fiber is digestion-resistant maltodextrin.

[0166] In an embodiment, the probiotics of the probiotic composition comprise one or more beneficial microorganisms. In an embodiment, the probiotics comprise Bifidobacterium bacteria. In an embodiment, the probiotics comprise Lactobacillus bacteria. In an embodiment, the probiotics comprise Firmicutes Lactobacillus, Actinobacteria Bifidobacterium, or a combination thereof.

[0167] In an embodiment, the probiotic composition is in the form of a gel. Examples of gels suitable for supplementing with probiotics to prepare a probiotic composition are disclosed herein.

[0168] In an embodiment, the probiotic composition is in the form of a powder. Examples of powders suitable for supplementing with probiotics to prepare a probiotic composition are disclosed herein.

[0169] In an embodiment, the probiotic composition is in liquid form. Examples of liquids suitable for supplementing with probiotics to prepare a probiotic composition are disclosed herein.

[0170] 7) Excipients / Formulations

[0171] In an embodiment, the composition disclosed herein can be combined with one or more excipients. In an embodiment, the excipient is a gelling agent, a thickening agent, a carrier, a buffer or a filler. In an embodiment, the composition disclosed herein can be formulated into a beverage or food. In an embodiment, the composition is formulated into a juice, a hydration beverage (e.g., a sports drink), a protein shake, tea, coffee, milk, kefir, ice cream, yogurt, smoothie, broth or soup.

[0172] In an embodiment, the composition comprises a thickener / gelling agent carboxymethyl cellulose. In an embodiment, the composition is a gel comprising superoxide dismutase, soluble fiber and carboxymethyl cellulose. In an embodiment, the composition comprises about 0.5 mg / mL to about 5.0 mg / mL of carboxymethyl cellulose.

[0173] In an embodiment, the composition comprises a thickener / 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 mg / mL to about 5.0 mg / mL of xanthan gum.

[0174] 8) Methods

[0175] The present disclosure also relates to a method of increasing T cell activation in 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; wherein, after administration of the composition, activation of T cells in the subject is increased.

[0176] In an embodiment, the activation of T cells is measured by measuring biomarkers of T cell activation, as disclosed in the Examples below.

[0177] In an embodiment, the composition is administered in combination with an anticancer agent. In an embodiment, the composition is administered in combination with an antiviral agent.

[0178] In an embodiment of the method of increasing T cell activation, the method is used to prevent or treat viral infection, including infection by influenza A, influenza B, influenza C, influenza D, coronaviruses including SARS (severe acute respiratory syndrome), SARS-CoV-2 (causing COVID-19), MERS (Middle East respiratory syndrome), HIV, Ebola, rhinovirus, and respiratory syncytial virus.

[0179] The present disclosure also provides a method for increasing the production of short-chain fatty acids (SCFA) 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; wherein, after administration of the composition, the production of SCFA in the digestive tract of the subject is increased.

[0180] In an embodiment, the SCFA whose production is increased is acetic acid, propionic acid, butyric acid or lactic acid SCFA or a combination thereof. In an embodiment, the SCFA is increased in a manner that provides about the same ratio of acetic acid, propionic acid, butyric acid and lactic acid SCFA as compared to the ratio of acetic acid, propionic acid, butyric acid and lactic acid SCFA before the increase.

[0181] The present disclosure also provides a method for increasing the amount of Bifidobacterium or Lactobacillus bacteria 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; wherein after administration of the composition, the amount of Bifidobacterium, Lactobacillus bacteria, or a combination thereof in the digestive tract of the subject is increased.

[0182] In an embodiment of the method, the Bifidobacterium bacterium comprises a species Bifidobacterium actinomycetes. In an embodiment, the Lactobacillus bacterium comprises a species Lactobacillus Firmicutes.

[0183] 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 embodiments, the composition comprises about 75 units to about 95 units of superoxide dismutase. In embodiments, the composition comprises about 80 units to about 90 units of superoxide dismutase. In embodiments, 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 embodiments, the composition comprises about 84 units of superoxide dismutase.

[0184] 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 2,000 mg to about 4,000 mg of soluble fiber. In an embodiment, the composition comprises about 1,000 mg to about 10,000 mg of soluble fiber. In an embodiment, the composition comprises about 2,000 mg to about 9,000 mg of soluble fiber. In an embodiment, the composition comprises about 3,000 mg to about 8,000 mg of soluble fiber. In an embodiment, the composition comprises about 2,000 mg to about 4,000 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.

[0185] In an embodiment of the above method, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:100 to about 1:1000. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:200 to about 1:800. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:300 to about 1:700. In an embodiment, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:500 to about 1:700. In some embodiments, the weight ratio of superoxide dismutase to soluble fiber in the composition is about 1:650 to about 1:675.

[0186] In an embodiment of the above method, the composition may comprise one or more superoxide dismutases 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.

[0187] In an embodiment of the above method, the composition may include one or more soluble fibers 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, oligosaccharides, galactose polysaccharides, oligofructose, lactulose, digestion-resistant starch, oligoxylose and isomalto-oligosaccharides. In an embodiment, the soluble fiber is soluble corn fiber. In an embodiment, the soluble fiber is digestion-resistant maltodextrin.

[0188] In an embodiment of the above method, the composition is in the form of a gel. Examples of gels suitable for use in the above method are disclosed herein.

[0189] In an embodiment of the above method, the composition is in the form of a powder. Examples of powders suitable for use in the above method are disclosed herein.

[0190] In an embodiment of the above method, the composition is in liquid form. Examples of liquids suitable for use in the above method are disclosed herein.

[0191] In an embodiment of any method disclosed herein, the composition is administered to a mammal once a day. In an embodiment of any method disclosed herein, the composition is administered to a mammal twice a day. In an embodiment of any method disclosed herein, the composition is administered to a mammal three, four, five, six, seven, eight, nine, ten or more times a day.

[0192] In embodiments of any of the methods disclosed herein, the mammal is a human. In embodiments of any of the methods disclosed herein, the mammal is a primate (e.g., monkey, ape, gorilla, macaque), a domestic animal (e.g., dog, cat, rabbit, hamster, guinea pig, mouse, rat) or an agricultural animal (e.g., cattle, sheep, horse, goat, pig).

[0193] Example

[0194] Example 1: SOD and soluble fiber gel stimulate T cell activation, antioxidant and anti-inflammatory pathways in Jurkat cell in vitro experiments

[0195] Background and Purpose: Pro-vitality antioxidant gel mainly contains the antioxidant superoxide dismutase (SOD), prebiotic fiber, and various polyphenols from various fruit juices. The formula is shown in Table 4.

[0196] Table 4

[0197]

[0198]

[0199] SOD reduces superoxide anions produced by normal cellular activities. Polyphenols are phenolic compounds that have antioxidant, anti-inflammatory, and antiviral effects. They repair cells damaged by ROS / RNS reactive oxygen molecules. Dietary prebiotic fibers modulate beneficial gut ecological microbiota and provide many health benefits including enhanced immunity. The combination of these three components stimulates the immune system through T cell activation as well as antioxidant and anti-inflammatory pathways. The aim of this study was to evaluate Effects of the gel on in vitro T cell models.

[0200] method: Jurka Jurkat is an immortalized T lymphocyte cell line that is most commonly used as a prototype T cell line to study multiple events in T cell biology, including T cell signaling. Jurkat cells were seeded in 6-well plates. Cells were incubated in serum-free medium for 24 hours before treatment. Cells were treated with the following reagents for 48 hours: 1. Superoxide dismutase only; 2. Prebiotic fiber only; 3. Juice only; 4. Superoxide dismutase + prebiotic fiber + juice (combination); 5. Positive control: phorbol 12-myristate 13-acetate (PMA) combined with ionomycin; 6. Negative control: cell culture medium.

[0201] After treatment, the medium was removed from the cells and placed in a tube. The levels of CD-8+; CD-4+; interferon-γ (IFNγ); interleukin-6 (IL-6); interferon gamma-induced protein 10 (IP-10; also known as CXCL10); macrophage inflammatory protein 1α and 1β; monocyte chemoattractant protein 1 (MCP-1, also known as CCL2); and 8 isoprostanes were measured by commercially available ELISA kits.

[0202] Jurkat cell activation can be observed by upregulation of CD69 (measured using anti-CD69 antibody MCA2806A647, BioRad) expression on CD3 (measured using anti-CD3 antibody MCA463A488) positive cell populations. Lymphocytes were gated in the presence of Human Seroblock (BUF070A, BioRad). Jurkat cells were treated and stimulated for five days and stained with CytoTrack Red 628 / 643 using a cell proliferation assay kit (1351205, BioRad). Data were collected on a ZE5 cell analyzer. Data are expressed as mean ± SEM. The statistical significance of differences between treatment groups was assessed by ANOVA and Dunnett post hoc test, and treatments with negative control effects of p < 0.05 were considered significant. Results are expressed as mean ± SE. (n = 6, four replicates).

[0203] Results: As discussed in detail below, compared with culture medium, The gel and its components activated T cells by upregulating CD69 expression and activated CD4+ and CD8+ differentiation. The gel and its components attenuated the activation of 8-isoprostaglandin (8IP), COX-2, IFN-γ, IL-6, TGF-β, TNF-α and CXCL10 secreted by Jurkat cells induced by lipopolysaccharide.

[0204] Conclusion / Viewpoint: The gel contains superoxide dismutase, prebiotic fiber, and polyphenols and quercetin from the juice. This unique multidirectional approach is highly effective in preventing oxidative stress, maintaining the balance of pro- and anti-inflammatory, and stimulating immune response.

[0205] introduction

[0206] Superoxide dismutase (SOD): SOD is a very important antioxidant defense against oxidative stress in the body. The enzyme is a good therapeutic agent against diseases mediated by reactive oxygen species. SOD has therapeutic effects on a variety of physiological and pathological conditions, such as cancer, inflammatory diseases, cystic fibrosis, ischemia, aging, rheumatoid arthritis, neurodegenerative diseases, and diabetes. However, due to absorption problems, the enzyme has certain limitations in clinical applications. Therefore, SOD conjugates and mimetics have been developed to improve its therapeutic efficiency. 1,2,3,4,5,6,7 .

[0207] Polyphenols: Polyphenols are secondary metabolites of plants that are often involved in protecting against ultraviolet radiation or pathogen invasion. In the past decade, there has been great interest in the potential health benefits of dietary plant polyphenols as antioxidants. Epidemiological studies and related meta-analyses strongly suggest that long-term consumption of a diet rich in plant polyphenols can protect against the development of cancer, cardiovascular disease, diabetes, osteoporosis, and neurodegenerative diseases. 8,9,10,11,12 .

[0208] Dietary Prebiotic Fiber: The health benefits of dietary fiber have long been widely recognized. Higher dietary fiber intakes have been associated with reduced cardiovascular disease, fiber plays an important role in gut health, and many effective laxatives are actually independent sources of fiber. Higher fiber intakes are associated with lower body weight. Dietary fiber originally included only polysaccharides, but more recent definitions have included oligosaccharides in the category of dietary fiber, based not on the chemical measurement of oligosaccharides as dietary fiber by the recognized total dietary fiber (TDF) method, but on the physiological effects of oligosaccharides. Inulin, oligofructose, and other oligosaccharides are included as fiber on food labels in the United States. In addition, oligosaccharides are the best known "prebiotics," "a selectively fermentable ingredient that produces specific changes in the composition and / or activity of the gastrointestinal microbiota that confer a benefit on the well-being and health of the host." To date, all known and suspected prebiotics are carbohydrate compounds, primarily oligosaccharides, which are known to be poorly digested in the human small intestine and fermented by the intestinal microbiota after reaching the colon. Studies have provided evidence that inulin, oligofructose (OF), lactulose, and resistant starch (RS) meet all aspects of the definition, including stimulation of the beneficial bacteria genus Bifidobacterium. Other isolated carbohydrates and carbohydrate-containing foods, including galacto-oligosaccharides (GOS), trans-galacto-oligosaccharides (TOS), polydextrose, wheat dextrin, acacia gum, psyllium, bananas, whole grain wheat, and whole grain corn also have prebiotic effects. 13,14,15,16,17 .

[0209] T Cell Activation and Immunity: T cells are generated in the thymus and are programmed to be specific for one particular foreign particle (antigen). Once they leave the thymus, they circulate throughout the body until they recognize the antigen on the surface of an antigen presenting cell (APC). The T cell receptors (TCRs) on CD4+ helper T cells and CD8+ cytotoxic T cells bind to the antigen as it is anchored in a structure of MHC complexes on the surface of the APC. This triggers the initial activation of the T cell. The CD4 and CD8 molecules then also bind to the MHC molecules, stabilizing the entire structure. This initial binding between a T cell that is specific for one antigen and its matching antigen-MHC initiates the entire response. This usually occurs in secondary lymphoid organs 18 .

[0210] The role of T cells in COVID-19 infection: Like antibody-producing B cells, T cells are central players in the immune response to viral infections 19 When SARS-CoV-2, the virus that causes COVID-19, infects epithelial cells (such as those found in the airways), it exploits the host cell's biochemical machinery to replicate inside the cell. This causes the host cell to undergo programmed cell death, releasing molecules called damage-associated molecular patterns (e.g., nucleic acids and oligomers) 20 These molecules are recognized by macrophages and neighboring endothelial and epithelial cells, causing them to produce proinflammatory cytokines, including the chemokines: interleukin-6 (IL-6); interferon gamma-induced protein 10 (IP-10; also known as CXCL10); macrophage inflammatory proteins 1α and 1β; and monocyte chemoattractant 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 and promote further inflammation. As part of the inflammatory response, recruited T cells produce interferon-gamma (IFNγ).

[0211] Multiple types of T cells participate in this response. CD4+ T helper (Th) cells interact with CD8+ T cells to drive a cytotoxic response that kills cells infected with the virus. CD8+ T cells directly recognize viral peptides on the surface of infected cells, leading to apoptosis (a form of programmed cell death) and preventing further spread of the virus. Follicular helper T (TFH) cells are a special subset of CD4+ T cells that provide help to B cells through cell-to-cell interactions and release of cytokines, leading to the production of antibodies by B cells. 19 These neutralizing antibodies can recognize the whole virus and work by blocking the virus from infecting cells. Alveolar macrophages recognize the neutralized virus and apoptotic cells (killed by CD8+ T cells) and clear them by phagocytosis. This leads to the resumption of viral infection 20 .

[0212] Studies evaluating the clinical characteristics of patients infected with SARS-CoV-2 have reported an incubation period of 4 to 7 days before symptom onset and 7 to 10 days before development of severe disease. 21 .

[0213] For many primary viral infections, it typically takes 7 to 10 days to initiate and expand an adaptive T cell immune response to control the virus, which correlates with the typical time it takes for COVID-19 patients to recover or develop severe disease. 22 This raises the possibility that a poor initial T cell response contributes to the persistence and severity of SARS-CoV-2, whereas an early strong T cell response may be protective.

[0214] CD4+ T cell responses in COVID-19: Some studies have shown evidence of impaired CD4+ T cell function, including reduced IFNγ production, in patients with severe COVID-19 22 , while other studies seem to indicate that these T cells are overactivated 23 .

[0215] Overall, the CD4+ T cell response in acute SARS-CoV-2 infection, whether impaired, overactivated, or inappropriate, and how it relates to disease outcome remains to be elucidated and is an important question. In recovered patients with COVID-19, a particularly high frequency of CD4+ T cell responses specific to the viral spike protein was observed, similar to what has been reported in influenza virus infection. 21 In a small study of 14 patients, circulating virus-specific CD4+ T cells were identified in all patients who recovered from SARS-CoV-2, also suggesting the potential for the formation of T cell memory 24 And may produce long-term immunity.

[0216] CD8+ T cell responses in COVID-19: There appears to be heterogeneity in immune responses between patients. Some studies have reported reduced cytokine production after in vitro stimulation of CD8+ T cells from patients with severe COVID-19, and some have shown evidence of possible T cell exhaustion; 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 .

[0217] Experimental procedures / research strategy

[0218] A model cell system was used to test this hypothesis. The Jurkat cell line is an immortalized T lymphocyte cell line originally derived from the peripheral blood of a boy with T cell leukemia. The Jurkat cell line is most commonly used as a prototype 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 The Jurkat cell line has been used to model and characterize signaling events in T cell activation (TCA), a key process for effective adaptive immune responses. 26 . As a model signaling axis, TCA involves surface signaling through the T cell receptor (TCR) and the cell surface accessory proteins CD3 and CD28, and initiates a series of molecular events that lead to the transcriptional activation of multiple genes, including the interleukin-2 (IL-2) gene, which is a classical T cell activation target gene. Steps in TCA include the activation of a series of kinases (e.g., LCK, JNK, PKC) and phosphatase proteins (calcineurin), as well as the activation of quiescent cytoplasmic transcription factors (e.g., NF-kB, NFAT), which upon activation undergo translocation into the nucleus to activate target genes. Each of these steps, as well as many other intermediate factors in the pathway, have been dissected using mutant subclones of the Jurkat T cell line, which has helped to map the signaling pathways and identify key players in T cell activation. 18,26 .

[0219] Jurkat cell line culture (Jurkat, clone E6-1 ( TIB-152 TM ; Human; Homo sapiens)

[0220] Complete Growth Medium: The basal medium for this cell line is RPMI-1640 medium prepared by ATCC, ATCC 30-2001. To prepare the complete growth medium, the following components were added to the basal medium: Fetal bovine serum (ATCC 30-2020) to a final concentration of 10%.

[0221] Subculture: To maintain a culture by adding fresh medium or replacing the medium. Alternatively, establish the culture by centrifugation and then culture at 1X 10 5 Resuspend at 10 viable cells / mL. The cell density should not exceed 3X 10 6 cells / mL. Recommended T-75 flasks are used to passage the product.

[0222] Interval: Cultures were maintained at 1X 10 5 Up to 1X 10 6 The cell concentration was 1 viable cell / mL.

[0223] Medium renewal: Add fresh medium every 2 to 3 days (depending on cell density).

[0224] Culture conditions: Atmosphere: air, 95%; carbon dioxide (CO2), 5%; temperature: 37°C

[0225] Treatment of Jurkat cells with a combination of a superoxide dismutase dietary supplement and prebiotic fiber containing polyphenols in juice

[0226] Effect of superoxide dismutase combined with prebiotic fiber containing juice polyphenols on Jurkat cells: Jurkat cells were seeded in 6-well plates. Before treatment, cells were incubated in serum-free medium for 24 hours. Cells were treated with the following reagents for 48 hours: 1. Superoxide dismutase alone; 2. Prebiotic fiber alone; 3. Juice alone; 4. Superoxide dismutase + prebiotic fiber + juice (combination); 5. Positive control: phorbol 12-myristate 13-acetate (PMA) combined with ionomycin; 6. Negative control: cell culture medium.

[0227] ELISA: After treatment, the culture medium was removed from the cells and placed in a tube. To assess T cell activation, the following factors were measured in the cell culture medium. CD-8+; CD-4+; interferon-γ (IFNγ); interleukin-6 (IL-6); interferon-γ-induced protein 10 (IP-10; also known as CXCL10); macrophage inflammatory proteins 1α and 1β; monocyte chemoattractant protein 1 (MCP-1, also known as CCL2); and levels of eight isoprostanes were determined by commercially available ELISA kits as described previously. 27,28,29,30,31,32,33,34 .

[0228] T cell stimulation for flow cytometry analysis: cells were treated for 48 hours with the following reagents: 1. Superoxide dismutase only; 2. Prebiotic fiber only; 3. Juice only; 4. Superoxide dismutase + prebiotic fiber + juice (combination); 5. Positive control: phorbol 12-myristate 13-acetate (PMA) combined with ionomycin; 6. Negative control: cell culture medium.

[0229] Activated Jurkat cells were detected as described above and statistical analysis of the results was performed as described above.

[0230] result

[0231] Cell viability assay: This assay measures the ability of viable cells to convert the redox dye resazurin into the fluorescent end product resorufin. Jurkat cells were seeded onto 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, juice only, superoxide dismutase + prebiotic fiber + juice (combination), or LPS (positive control). After 48 hours of incubation for each treatment, 20 μl of CellTiter-Blue reagent was added to each well. The absorbance at 520 nm was determined on a microtiter plate reader. The signal generated by the conversion of resazurin to resorufin is proportional to the number of viable cells. Figure 1 As shown, none of the gel components or combinations showed toxic effects in the cell viability assay.

[0232] By flow cytometry analysis, CD69 expression was upregulated on the CD3 positive cell population, indicating that the gel and its components activated T cells: T cell stimulation for flow cytometric analysis. The activation and proliferation protocol provides an effective method to determine immunocompetence and cell reactivity. Jurkat cells were then stimulated with vehicle (cell culture medium) or prebiotic fiber alone, superoxide dismutase alone, juice alone, superoxide dismutase + prebiotic fiber + juice (combination) or LPS (positive control). Lymphocytes were gated in the presence of Human Seroblock. Data are in ZE5 TM The cells were collected on a cell analyzer. Figure 2 As shown in Figure 2, CD69 expression is upregulated in the CD3-positive population, indicating activated T cells. Figure 2 As shown, this combination displayed surprisingly high levels of T cell activation as measured by CD69 expression.

[0233] Effect of the original components on CD4+ differentiation of Jurkat cells. Jurkat cells were then stimulated with vehicle (cell culture medium) or prebiotic fiber alone, superoxide dismutase alone, juice alone, superoxide dismutase + prebiotic fiber + juice (combination). CD4+ concentrations (ng / mL) were measured by ELISA according to the manufacturer's protocol. Figure 3 As shown, juice and superoxide dismutase + prebiotic fiber + juice (combination) significantly and surprisingly activated CD4+ differentiation compared to culture medium as determined by ELISA.

[0234] The original fractions acted on Jurkat cells CD8 +Jurkat cells were then stimulated with vehicle (cell culture medium) or prebiotic fiber alone, superoxide dismutase alone, juice alone, superoxide dismutase + prebiotic fiber + juice (combination). CD8 + The concentration of (pg / mL). Figure 4 As shown, juice and superoxide dismutase + prebiotic fiber + juice (combination) significantly and surprisingly activated CD4+ differentiation compared to culture medium as determined by ELISA.

[0235] CD4 + and CD8 + Concentration ratio:

[0236] CD4 + / CD8 + The concentration ratio is 48:1, which is higher than that in the peripheral blood of healthy adults and mice. 35 .CD4 + / CD8 + The ratio is the ratio of helper T cells (with the surface marker CD4) to cytotoxic T cells (with the surface marker CD8). + and CD8 + T cells consist of multiple subsets. 36 CD4 in peripheral blood of healthy adults and mice + / CD8 + The ratio is about 2:1, and changes in the ratio may indicate diseases related to immunodeficiency. 35 This large difference in ratios is attributed to the fact that the experiments were performed in a closed in vitro system. Stimulates T cells to differentiate into CD4 + These data suggest that follicular helper T (TFH) cells are a special subset of CD4+ T cells that provide help to B cells through cell-cell interactions and release of cytokines, leading to antibody production by B cells. 19 These neutralizing antibodies can recognize the whole virus and work by blocking the virus from infecting cells. Alveolar macrophages recognize the neutralized virus and apoptotic cells (killed by CD8+ T cells) and clear them by phagocytosis. This leads to the resumption of viral infection 20 In coronavirus disease 2019 (COVID-19), B cell, natural killer cell, and total lymphocyte counts decreased, but CD4+ and CD8+ cells decreased to a greater extent. 37Low CD4+ predicted a greater likelihood of intensive care unit admission, and CD4+ cell count was the only parameter that predicted the length of time to viral RNA clearance. 37

[0237] The gel has antioxidant activity: e.g. Figure 5 As shown in in vitro studies, And its components weaken the activation of lipopolysaccharide-induced Jurkat cells to secrete 8-isoprostane. Then use vehicle (cell culture medium) or only prebiotic fiber, only superoxide dismutase, only fruit juice, superoxide dismutase+prebiotic fiber+fruit juice (combination) to stimulate lipopolysaccharide-induced Jurkat cells. The level of 8IP in the culture medium is measured by ELISA. Its components attenuated lipopolysaccharide-induced activation of Jurkat cells to secrete 8-isoprostane (8IP). 8IP levels are considered a marker of antioxidant deficiency and oxidative stress. 38,39 .like Figure 5 As shown, this combination surprisingly reduced 8IP levels to below those observed in untreated cells.

[0238] The gel has anti-hypoxic activity: e.g. Figure 6 As shown in in vitro studies, and its components attenuated the activation of lipopolysaccharide-induced Jurkat cell secretion of cellular cyclooxygenase-2 (COX-2). LPS-induced Jurkat cells were then stimulated with vehicle (cell culture medium) or prebiotic fiber alone, superoxide dismutase alone, juice alone, superoxide dismutase + prebiotic fiber + juice (combination). The level of COX-2 in the culture medium was measured by ELISA. Figure 6 As shown, this combination surprisingly reduced COX-2 levels to below those observed in untreated cells.

[0239] and its components attenuated LPS-induced activation of COX-2 secreted by Jurkat cells. Hypoxia increases COX-2 expression 40 Apigenin downregulates COX-2 expression in lupus T cells, B cells, and antigen-presenting cells and induces their apoptosis 41 Although a clear structure / function relationship has not yet been established and without wishing to be bound by theory, it appears that the C-2,3-double bond and the hydroxyl substitutions on the A and B rings are important contributors to this inhibitory activity. 42 Animal data confirm that COX-2 expression is downregulated in different inflammatory diseases 43 .

[0240] The gel has anti-inflammatory activity: in vitro studies on Jurkat cells, and its components can attenuate the activation of lipopolysaccharide-induced inflammatory activity. Jurkat cells induced by lipopolysaccharide were then stimulated with vehicle (cell culture medium) or prebiotic fiber alone, superoxide dismutase alone, juice alone, superoxide dismutase + prebiotic fiber + juice (combination). Interferon gamma (IFNγ, Figure 7 ), interleukin-6 (IL-6, Figure 8 ), transforming growth factor β (TGF-β, Fig. 9 ), tumor necrosis factor (TNF, Fig.10 ) and CXC motif chemokine ligand 10 (CXCL10, Fig.11 ) levels were measured by commercially available ELISA kits.

[0241] and its components attenuated the activation of IFN-γ secreted by Jurkat cells induced by lipopolysaccharide ( Figure 7 Interferon gamma (IFNγ) is a dimeric soluble cytokine and the only member of the type II interferon. 44 The existence of this interferon, which was called immune interferon early in its history, was described by EF Wheelock as a product of human leukocytes stimulated by phytohemagglutinins, while others described it as a product of lymphocytes stimulated by antigens. IFNγ (or type II interferon) is a cytokine that is essential for innate and adaptive immunity against viral, some bacterial and protozoan infections. IFNγ is an important activator of macrophages and an inducer of the expression of major histocompatibility complex class II molecules. Aberrant IFNγ expression is associated with many autoinflammatory and autoimmune diseases. The importance of IFNγ in the immune system stems in part from its ability to directly inhibit viral replication and, most importantly, from its immunostimulatory and immunomodulatory effects. As part of the innate immune response, IFNγ is mainly produced by natural killer cells (NK) and natural killer T cells (NKT), and as part of the adaptive immune response, it is produced by CD4 Th1 and CD8 cytotoxic T lymphocytes (CTL) effector T cells after the development of antigen-specific immunity. 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 like Figure 7 As shown, surprisingly, cells treated with SOD, juice polyphenols and the combination all had lower levels of IFNγ secretion than untreated cells, with the lowest secretion levels observed in cells treated with the combination.

[0242] and its components attenuated the activation of IL-6 secretion by Jurkat cells induced by lipopolysaccharide ( Figure 8 ): Interleukin 6 (IL-6) is an interleukin that acts as both a pro-inflammatory cytokine and an anti-inflammatory myokine. In humans, it is encoded by the IL6 gene. 49 In addition, osteoblasts secrete IL-6 to stimulate osteoclast formation. Smooth muscle cells in the tunica media of many blood vessels also produce IL-6 as a pro-inflammatory cytokine. The effects of IL-6 as an anti-inflammatory myokine are mediated through its inhibitory effects on TNF-α and IL-1, and its activation of IL-1ra and IL-10. There is some early evidence that IL-6 could be used as an inflammatory marker for severe COVID-19 infection with a poor prognosis 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 detection molecules of the innate immune system 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 produce inflammatory cytokines. IL-6 is an important mediator of fever and acute phase responses. IL-6 is responsible for stimulating acute phase protein synthesis and the production of neutrophils in the bone marrow. It supports the growth of B cells and antagonizes regulatory T cells. Figure 8 As shown, this combination surprisingly reduced IL-6 activation levels below those observed in untreated cells.

[0243] and its components attenuated the activation of TGF-β secreted by Jurkat cells induced by lipopolysaccharide ( Fig. 9 ): Transforming growth factor β (TGF-β) is a multifunctional cytokine that belongs to the transforming growth factor superfamily, which includes three 51 . TGFB protein is 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 contains type 1 and type 2 receptor subunits. Upon binding to TGF-β, type 2 receptor kinase phosphorylates and activates type 1 receptor kinase, thereby activating the signaling cascade 52 This leads to the activation of different downstream substrates and regulatory proteins, inducing the transcription of different target genes that play a role in the differentiation, chemotaxis, proliferation and activation of many immune cells. 52,53

[0244] TGF-β is secreted by many cell types, including macrophages, in a latent form, in which it is complexed with two other polypeptides, latent TGF-β binding protein (LTBP) and latency associated peptide (LAP). Serum proteases, such as plasmin, catalyze the release of active TGF-β from the complex. This normally 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 latent TGF-β complexes bound to IgG secreted by plasma cells, and then release active TGF-β into the extracellular fluid. 54 One of its key functions is to regulate inflammatory processes, particularly in the intestine. [5] TGF-β also plays a crucial role in stem cell differentiation and T cell regulation and differentiation. 56,57 It is a much studied cytokine in the fields of cancer, autoimmune diseases, and infectious diseases due to its role in immune and stem cell regulation and differentiation.

[0245] The TGF-β superfamily includes endogenous growth inhibitory proteins; increased expression of TGF-β is often associated with the malignancy of many cancers and defects in the cellular growth inhibitory response to TGF-β. Its immunosuppressive function will dominate and lead to tumorigenesis. 58 Dysregulation of its immunosuppressive function has also been implicated in the pathogenesis of autoimmune diseases, although its effects are mediated by the presence of other cytokine milieu. 55 TGF-β can induce 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 unregulated autoimmunity. 59 like Fig. 9 As shown, surprisingly, cells treated with SOD, juice polyphenols, and the combination all had lower levels of TGF-β secretion than untreated cells, with the lowest secretion levels observed in cells treated with the combination.

[0246] and its components attenuated the activation of TNF-α secreted by Jurkat cells induced by lipopolysaccharide ( Fig.10): Tumor necrosis factor (TNF, cachexin, or cachectin; often referred to as tumor necrosis factor alpha or TNF-α) is a cytokine, which is a small protein used by the immune system for cell signaling. If macrophages (certain white blood cells) 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 multiple transmembrane proteins with homologous TNF domains. TNF is thought to be produced primarily by macrophages, 60 However, it is also produced by a variety of cell types, including lymphocytes, mast cells, endothelial cells, cardiomyocytes, adipose tissue, fibroblasts, and neurons. 61 Large amounts of TNF are released by lipopolysaccharide, other bacterial products, and interleukin-1 (IL-1). In the skin, mast cells appear to be the major source of preformed TNF, which can be released in response to inflammatory stimuli such as LPS. 62 It has multiple effects on various organ systems, often acting in conjunction with IL-1 and interleukin 6 (IL-6). Local increases in TNF concentrations result in the cardinal symptoms of inflammation: fever, swelling, redness, pain, and loss of function. High concentrations of TNF can cause shock-like symptoms, while prolonged exposure to low concentrations of TNF can lead to cachexia, a wasting syndrome. This can be seen, for example, in cancer patients. 63 .

[0247] Said et al. showed that TNF produces IL-10-dependent inhibition of CD4 T cell expansion and function by upregulating PD-1 levels on monocytes, leading to IL-10 production by monocytes following binding of PD-1 to PD-L. 64 Pedersen et al. showed that the sepsis-induced increase in TNF was suppressed by exercise-induced myokine production. To investigate whether acute exercise would induce a true anti-inflammatory response, a “low-grade inflammation” model was established in which a low dose of Escherichia coli endotoxin was administered to healthy volunteers who were randomized to rest or exercise prior to endotoxin administration. In resting subjects, endotoxin induced a 2- to 3-fold increase in circulating TNF levels. In contrast, when subjects underwent 3 h of ergometer cycling and received a bolus of endotoxin at 2.5 h, the TNF response was completely attenuated. 65 This study provides some evidence that intense exercise may inhibit TNF production. 66 In the brain, TNF protects 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 like Fig.10 As shown, surprisingly, cells treated with both juice polyphenols and the combination had lower levels of TNF-α secretion than untreated cells, with the lowest levels of secretion observed in cells treated with the combination.

[0248] and its components attenuated LPS-induced activation of CXCL10 secreted by Jurkat cells ( Fig.11 ): CXC motif chemokine ligand 10 (CXCL10), also known as interferon gamma-induced protein 10 (IP-10) or small inducible cytokine B10, is an 8.7 kDa protein encoded by the CXCL10 gene in humans. 69,70 CXC motif chemokine 10 is a small cytokine belonging to the CXC chemokine family. CXCL10 is secreted by a variety of cell types in response to IFN-γ. These cell types include monocytes, endothelial cells, and fibroblasts. 69 CXCL10 has multiple effects, such as chemoattraction of 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 by binding to the cell surface chemokine receptor CXCR3. 73 like Fig.10 As shown, surprisingly, cells treated with SOD, juice polyphenols, and the combination all had lower levels of CXCL10 secretion than untreated cells, with the lowest secretion levels observed in cells treated with the combination.

[0249] Table 5 summarizes all the above ELISA results.

[0250] Table 5. Effects of 6 replicates of each treatment on JRK cells (expressed as SE)

[0251]

[0252]

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[0389] Example 2: Effect of regulating intestinal microorganisms on superoxide dismutase absorption Background Art

[0391] Superoxide dismutase (SOD) is a major antioxidant enzyme with a very high molecular weight that often suffers from physical stability, gastric acid degradation, and most importantly, absorption issues. Although SOD helps reduce cellular oxidative stress and is helpful for many aging-related disorders, it has been difficult to produce an orally effective SOD dosage form. Contains a variety of functional molecules, which affect each other's bioavailability, including the absorption of SOD. The acid stability and absorption of SOD are affected by microbial regulation, while Certain components of the gel (prebiotic fibers and polyphenols) provide this regulation. To test this hypothesis, we conducted a study to evaluate the role of SOD itself and as a Whether SOD, which is part of the ingredients, has any regulatory effect on the microbiome. The study below showed that the composition of the intestinal microbiome was changed, which promoted the absorption of SOD and improved its efficacy.

[0392] Experimental methods

[0393] The first goal of this work was to develop a scalable in vitro model for maintaining the gut microbiome profile. Bacterial cells were cultured in 96-well plates and a perforated silicone gel cover was placed on top of each well. This cover facilitated gas exchange with the outside environment, thereby maintaining the partial pressure of gases and volatile metabolites in each well, thereby maintaining a certain level of dissolved gas molecules in the culture medium.

[0394] Treatment conditions: 1. SOD alone; 2. Fibersol (digestion-resistant maltodextrin) alone; 3. Polyphenols alone; 4. SOD + Fibersol; 5. SOD + polyphenols; 6. REVIVFY finished product. With these treatments, it was possible to determine how Fibersol and polyphenols affect the absorption of SOD using an in vitro microbial system.

[0395] Fecal sample collection and processing: Briefly, about 3 g of fresh fecal samples were collected from each individual using a 2.5 ml sterile sampling spoon (Bel-Art, USA). Each spoon was placed in a 50 ml Falcon tube containing 15 ml of sterile PBS pre-reduced with 0.1% (w / v) L-cysteine ​​hydrochloride. The sample was immediately transferred to an anaerobic workstation (5% H2, 5% CO2, and 90% N2 at 37°C). The optical density (OD595) at 595 nm (OD595) was measured as a representative of microbial growth and biomass, and the microbiome was characterized at 0 (immediately after inoculation), 3, 6, 9, 12, 24, 34 and 48 hours by macroproteome analysis. The results were determined using a goat intestinal and silica pad microbiome system.

[0396] Results: Bacterial growth increased gradually over time. As shown by OD measurements, samples treated with Fibersol and SOD alone did not affect bacterial growth compared to the control ( Fig.12 , goat intestinal system; Fig.13 , silicone pad system). Fig.12 and 13 The top line in the figure is Finished products, the middle two lines are juice polyphenols only and SOD+Fibersol, the bottom three lines are untreated control, SOD only, and Fibersol only. However, the polyphenols in the juice significantly increased bacterial growth, The finished product resulted in the largest increase in OD. These experiments showed that Fibersol promoted the absorption of SOD by silica mats and fresh goat intestine, as evidenced by the increase in bacterial growth.

[0397] Conclusion: This study demonstrates that we have developed a scalable in vitro model for maintaining the gut microbiome profile. In addition, the absorption patterns of different components in REVIVFY gel, including the effect of Fibersol on SOD absorption, are shown. This in vitro gut microbiome model can be used to evaluate other drugs, prebiotics, or nutraceuticals.

[0398] Example 3: Evaluation of healthy gut microbiome and regulation of short-chain fatty acids using an in vitro gut microbiome model

[0399] Background: Gut health is very important for healthy living and well-being. The microbial community in the gut plays an important role in the immune system, hormonal processes, nervous system status, metabolism, mineral absorption, vitamin production, and multiple cellular processes. According to the American Diabetes Association, the gut microbiota influences host physiology both inside and outside the gut. The gut microbiota is essential for the homeostasis of the intestinal immune system, the regulation of epithelial cell proliferation, and the protection against opportunistic bacteria. The gut microbes present in the gastrointestinal tract have co-evolved within the human host to perform many functions that the host cannot perform alone. The dominant gut microbes are the Firmicutes and Bacteroidetes, followed by the Actinobacteria and Protozoa. The beneficial effects of gut microbes can be measured by the production of short-chain fatty acids (SCFAs), mainly acetate, propionate, butyrate, and lactate. This production depends on the type of fiber consumed. In this case, the fiber was a soluble fiber called Fibersol-2 and mixed oligofructose from various fruit juice concentrates. It was hypothesized that the beneficial microbes should be regulated by increasing the proportion of short-chain fatty acids, thereby protecting host health in multiple ways. This study evaluated the beneficial effects of gut microbes by using an in vitro model for gut microbiome research. Effects of finished products on gut microbiome modulation and short-chain fatty acids.

[0400] Experimental methods : An in vitro gut microbiome culture model was established as described in Example 2 above. The first goal of this work was to develop a scalable in vitro model for maintaining the gut microbiome profile. An in vitro model that maintains the functional and compositional profile of the in vivo gut microbiome would be very valuable. In vitro model experiments were performed as described in Example 2. Gut microbes were cultured in 2 ml 96-well plates and treated with control, SOD, prebiotic fiber, juice or finished product. The products were treated for 24 hours. The cultured microbiome samples were harvested at 24 hours for metaproteome analysis. Then, culture aliquots were collected for chemical analysis (SCFA content) and microbiome profiling.

[0401] Results: This study evaluated the changes in gut microbial composition and SCFA when treated with four different compounds (SOD, pre-fiber, juice, and finished product). As shown in Figure 14, the results showed that the amount of SCFA increased significantly when treated with the finished product ( Fig.14A ), but the ratio of SCFA remained the same in all treatment groups including the control group ( Fig. 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 / ml after 24 hours of treatment with the finished product. Short-chain fatty acids SCFA unexpectedly increased by 2.5 times relative to the finished product. When the ratios of these SCFAs in the control group and all four treatment groups were calculated, the ratios of acetate, propionate, butyrate and lactate seemed to remain around 53%, 15%, 24% and 8%, respectively (see Figure 14). Our study also showed that pre-fiber fiber, juice and finished product promoted the growth of two beneficial gut microorganisms, Lactobacillus of the Firmicutes phylum and Bifidobacterium of the Actinobacteria phylum, compared with baseline and SOD (see Fig.15 ; where Lactobacillus from the Firmicutes phylum is represented by vertical hatching and Bifidobacterium from the Actinobacteria phylum is represented by light grey shading). Both Lactobacillus and Bifidobacterium are beneficial microorganisms that are present in many food preparations such as yogurt to improve digestibility and immunity. Of the two bacteria, the trend of changes in Lactobacillus mirrored the production of lactic acid in the SCFA graph. In addition, Lactobacillus belongs to the genus Firmicutes. Typically, these are producers of butyrate, which is reflected in the SCFA results. The microbiome profile of the culture treated with the finished product showed a significant increase in Lactobacillus from the Firmicutes phylum in the intestinal microbial community, followed by Bifidobacterium from the genus Actinobacillus.

[0402] Conclusion / Perspective: Dietary prebiotics are selectively fermented ingredients that induce specific changes in the composition of two beneficial microbiota, Lactobacillus from the Firmicutes phylum and Bifidobacterium from the Actinobacteria phylum. This important finding suggests that both beneficial microorganisms can have a positive impact on the host, they can produce homo- and heterofermentation, digest and metabolize proteins and carbohydrates, synthesize vitamins B and K, decompose bile salts, enhance innate and acquired immunity, inhibit pro-inflammatory mediators, and have antimicrobial activity against a range of pathogens (such as Pseudomonas, Candida, Escherichia coli, Bacillus, Salmonella, Shigella, Clostridium difficile, and Helicobacter pylori). Lactobacillus is a biomarker of vaginal health and is a major component of the vaginal flora. This study shows that The finished product increased the content of SCFA in the intestine while keeping the ratio of each SCFA (acetic acid, propionic acid, butyric acid and lactic acid) consistent with the control. This result shows that the balance of SCFA is increased, but it does not improve and maintain a healthy colon environment in a consistent manner. Studies have shown that It is a unique dietary supplement that produces relatively high levels of butyrate, which has many health benefits on intestinal epithelial cell integrity, immune cell integrity and response, bidirectional signaling of the enteric neuron gut-brain axis, and nutrient production and metabolism. Butyrate is the main energy source for colonocytes and is involved in maintaining colon mucosal health.

[0403] Example 4: Reducing oxidative damage in human brain microvascular endothelial cells (HBMEC)

[0404] introduction

[0405] Increasing data indicate that oxidative stress and mitochondrial damage are involved in the pathogenesis of neurodegenerative diseases, including Parkinson's disease (PD), multiple sclerosis (MS), Alzheimer's disease (AD), etc. The brain consumes about 20% of oxygen and is therefore a high producer of reactive oxygen species (ROS). In addition, brain cell membranes contain more unsaturated fatty acids (MUFA and PUFA), making them more susceptible to lipid auto-oxidation due to ROS. The gel can instantly reduce oxidative stress in multiple pathways and have an immediate effect on disease symptoms.

[0406] The preparation can neutralize major oxidants such as superoxide anion, hydroxyl radical, singlet oxygen, peroxynitrite, peroxyl radical and hypochlorite. Background Art

[0408] Superoxide dismutase (SOD): SOD is a very important antioxidant defense against oxidative stress in the body. The enzyme is a good therapeutic agent against diseases mediated by reactive oxygen species. This review describes the therapeutic effects of SOD on a variety of physiological and pathological conditions, such as cancer, inflammatory diseases, cystic fibrosis, ischemia, aging, rheumatoid arthritis, neurodegenerative diseases, and diabetes. However, the enzyme has certain limitations in clinical applications. Therefore, SOD conjugates and mimetics have been developed to improve its therapeutic efficiency. 1,2,3,4,5,6,7 .

[0409] Polyphenols: Polyphenols are secondary metabolites of plants that are often involved in protecting against UV radiation or pathogen invasion. In the past decade, there has been great interest in the potential health benefits of dietary plant polyphenols as antioxidants. Epidemiological studies and related meta-analyses strongly suggest that long-term consumption of a diet rich in plant polyphenols can protect against the development of cancer, cardiovascular disease, diabetes, osteoporosis, and neurodegenerative diseases. 8,9,10,11,12 .

[0410] Dietary Prebiotic Fiber: The health benefits of dietary fiber have long been appreciated. Higher dietary fiber intakes have been associated with reduced cardiovascular disease, fiber plays an important role in gut health, and many effective laxatives are actually independent sources of fiber. Higher fiber intakes are associated with lower body weight. Dietary fiber originally included only polysaccharides, but more recent definitions have included oligosaccharides in the category of dietary fiber, based not on their chemical measurement as dietary fiber by the accepted total dietary fiber (TDF) method, but on their physiological effects. Inulin, oligofructose, and other oligosaccharides are included as fiber on food labels in the United States. In addition, oligosaccharides are the best known "prebiotics," "a selectively fermentable ingredient that produces specific changes in the composition and / or activity of the gastrointestinal microbiota that confer a benefit on the well-being and health of the host." To date, all known and suspected prebiotics are carbohydrate compounds, primarily oligosaccharides, which are known to be poorly digested in the human small intestine and fermented by the intestinal microbiota once they reach the colon. Studies have provided evidence that inulin, oligofructose (OF), lactulose, and resistant starch (RS) meet all aspects of the definition, including stimulation of the beneficial bacteria genus Bifidobacterium. Other isolated carbohydrates and carbohydrate-containing foods, including galacto-oligosaccharides (GOS), trans-galacto-oligosaccharides (TOS), polydextrose, wheat dextrin, acacia gum, psyllium, bananas, whole grain wheat, and whole grain corn also have prebiotic effects. 13,14,15,16,17 .

[0411] The purpose of this study is to evaluate Whether the gel can alleviate oxidative damage in human brain microvascular endothelial cells (HBMECs). The following biomarkers were evaluated in hypoxia-induced HBMEC culture medium:

[0412] 1. Malondialdehyde (MDA) - a biomarker of lipid oxidative damage

[0413] 2.4-Hydroxynonenal, or 4-hydroxy-2-nonenal, 4-HNE or HNE - a biomarker of lipid peroxidation

[0414] 3. Protein carbonyls: a biomarker of protein oxidation

[0415] 4.3-Nitrotyrosine: a biomarker of cell damage associated with dopamine neuron degeneration

[0416] Malondialdehyde (MDA) is an organic compound with the chemical formula CH2(CHO)2. MDA is a colorless liquid that is a highly reactive compound in the enol form. [1] It occurs naturally and is a marker of oxidative stress.

[0417] 4-Hydroxynonenal, or 4-hydroxy-2-nonenal or 4-HNE or HNE, (C9H 16 O2), is an α,β-unsaturated hydroxyalkenal produced by lipid peroxidation in cells. 4-HNE is the main α,β-unsaturated hydroxyalkenal formed in this process. 4-HNE has three reactive groups: an aldehyde, a double bond at carbon 2, and a hydroxyl group at carbon 4.

[0418] Protein carbonyls: The protein carbonyl (PC) content in blood and tissues is a reliable indicator of protein oxidation. Traumatic brain injury (TBI) is caused by a blow to the head that disrupts normal brain function. Severe TBI may cause permanent brain damage or death. Diffuse axonal injury (DAI) is a typical pathological change after TBI and is closely related to clinical prognosis. DAI has two obvious pathological characteristics: large terminal enlargement and swelling due to excessive aggregation of neurofilaments. Secondary axonal injury caused by cytoskeletal abnormalities is the most common cause of DAI.

[0419] Oxidative stress is a well-known factor associated with DAI, and mitochondrial phosphorylation capacity, the concentration of nicotinic coenzyme pools, and oxidative / nitrosative stress are closely related to the severity of DAI. Carbonyl modification is a direct result of protein oxidative damage, leading to protein dysfunction and the formation of protein aggregates. Protein carbonylation has been shown to be involved in the pathogenesis of a variety of neurodegenerative diseases (such as multiple sclerosis, Parkinson's disease, and Alzheimer's disease). Under normal circumstances, carbonylated proteins are thought to be degraded by proteasomes, whose main function is to recognize and degrade unwanted, damaged, or misfolded proteins. However, under prooxidative conditions, increased production of reactive oxygen species (ROS) or reactive carbonyl species may reduce the activity of proteasomes, leading to the accumulation of carbonylated proteins in affected cells.

[0420] 3-Nitrotyrosine: Nitrotyrosine is a product of tyrosine nitration mediated by reactive nitrogen species such as peroxynitrite anions and nitrogen dioxide. Nitrotyrosine is considered an indicator or marker of cell damage, inflammation, and NO (nitric oxide) production. Nitrotyrosine is formed in the presence of the reactive metabolite NO. In general, oxidative stress increases superoxide (O2 - ) and NO, forming the destructive free radical oxidant, peroxynitrite (ONOO - ).ONOO - The production of ONOO can oxidize a variety of lipoproteins and nitrate tyrosine residues in many proteins. - Therefore, nitrotyrosine in protein is usually used as an indirect detection method for ONOO - It is detected under a large number of pathological conditions and is considered to be a marker of NO-dependent, reactive nitrogen species-induced nitrosative stress. Nitrotyrosine can be detected in biological fluids such as plasma, lung aspirate-BALF (bronchoalveolar fluid) and urine. Elevated levels of nitrotyrosine have been detected in rheumatoid arthritis, septic shock and celiac disease. In all of these studies, nitrotyrosine was not detected in healthy subjects. Nitrotyrosine is also present in many other disease-affected tissues, such as the cornea in keratoconus. Peroxynitrite and / or nitrosative stress may be involved in the pathogenesis of diabetes.

[0421] Nitrotyrosine, a marker of reactive oxygen species, has also been linked to the degeneration of dopamine neurons. Tyrosine is a precursor to dopamine, a neurotransmitter important for motivation, attention, learning, circadian rhythms, and other biological processes.

[0422] Study Design: Human brain microvascular endothelial cells (HBMEC) were cultured in six-well plates under hypoxic conditions (hypoxia; 2% oxygen). Control HBMEC cells were cultured under normoxic conditions. Cells were incubated in serum-free medium for 24 hours before treatment. Cells were treated for 48 hours with the following agents: 1. Superoxide dismutase only; 2. Prebiotic fiber only; 3. Juice only; 4. Superoxide dismutase + prebiotic fiber + juice (combination); 5. Negative control.

[0423] ELISA: After 48 h of incubation, the culture medium was removed from the cells and placed in a tube. To evaluate whether Revivify gel can attenuate oxidative damage in human brain microvascular endothelial cells (HBMECs), the following biomarkers were assessed in the culture medium of hypoxia-induced HBMECs: 1. malondialdehyde (MDA); 2. 4-hydroxynonenal, or 4-hydroxy-2-nonenal or 4-HNE or HNE; 3. protein carbonyls; and 4. 3-nitrotyrosine.

[0424] Results: Figure 16-19 As shown in Figure 2, while some components of the gel showed improvements in biomarkers of oxidative damage, the finished gel showed the greatest improvements. Fig.16 As shown, each component of the gel provided some degree of reduction in the MDA biomarker compared to the positive control, and surprisingly, the finished gels had lower MDA levels under hypoxic conditions than cells cultured under non-hypoxic control conditions. Fig.17 Similar results were seen using HNE as a biomarker in the 24h cells cultured under non-hypoxic control conditions, where the finished gels again unexpectedly had lower levels of the biomarker than cells cultured under non-hypoxic control conditions.

[0425] like Fig.18 As shown, the levels of protein carbonyl biomarkers were reduced relative to the positive control for each gel component, and unexpectedly significantly reduced compared to the positive control for the final gel product. Fig.19 As shown, similar results were observed for the 3-nitrotyrosine biomarker, with the finished product again showing an unexpectedly significant reduction compared to the positive control.

[0426] The results in this example show that the final The gel provided substantial and surprising protection to HBMECs from oxidants. These results suggest that absorbed SOD, when combined with soluble fiber and polyphenols, can significantly protect nervous system cells from oxidative damage.

[0427] References for Example 4

[0428] 1. Kangralkar VA, Patil SD, Bandivadekar RM. Oxidative stress and diabetes: A review. Intl J Pharm Appl. 2010;1:38–45.

[0429] 2. Yasui K, Baba A. Therapeutic potential of superoxide dismutase (SOD) for resolution of inflammation. Inflamm Res. 2006; 55: 359–63.

[0430] 3. Landis GN, Tower J. Superoxide dismutase evolution and life span regulation. Mech Ageing Dev. 2005;126:365–79.

[0431] 4. Noor R, Mittal S, Iqbal J. Superoxide dismutase–Applications and relevance to human diseases. Med Sci Monit. 2002;8:RA210–5.

[0432] 5. Inal ME, Kanbak G, Sunal E. Antioxidant enzyme activities and malondialdehyde levels related to aging. Clin Chim Acta. 2001; 305: 75–80.

[0433] 6. Riley DP. Functional mimics of superoxide dismutase enzymes as therapeutic agents. Chem Rev. 1999;99:2573–88.

[0434] 7. Salvemini D, Riley DP. Nonpeptidyl mimetics of superoxide dismutase in clinical therapies for diseases. Cell Mol Life Sci. 2000; 57: 1489–92.

[0435] 8. Scalbert A, Manach C, Morand C, Remesy C. Dietary polyphenols and the prevention of diseases. Crit Rev Food Sci Nutr. 2005; 45: 287–306.

[0436] 9. Beckman CH. Phenolic-storing cells: keys to programmed cell death and periderm formation in wilt disease resistance and in general defense responses in plants? Physiol. Mol. Plant Pathol. 2000;57:101–110.

[0437] 10. Arts ICW, Hollman PCH. Polyphenols and disease risk in epidemiologic studies. Am J Clin Nutr. 2005;81:317–325.

[0438] 11. Kondratyuk TP, Pezzuto JM. Natural Product Polyphenols of Relevance to Human Health. Pharm Biol. 2004; 42: 46–63.

[0439] 12. Shahidi F, Naczk M. Food phenolics, sources, chemistry, effects, applications. Lancaster, PA: Technonic Publishing Co., Inc.; 1995.

[0440] 13. Slavin JL Dietary fiber: Classification, chemical analyses, and food sources. J. Am. Diet. Assoc. 1987;87:1164–1171.

[0441] 14. Gibson GR, Roberfroid MB Dietary modulation of the human colonic microbiota: Introducing the concept of prebiotics. J. Nutr. 1995; 125: 1401–1412.

[0442] 15. Gibson GR, Probert HM, van Loo J., Rastall RA, Roberfroid MB. Dietary modulation of the human colonic microbiota: Updating the concept of prebiotics. Nutr. Res. Rev. 2004;17:259–275. doi:10.1079 / NRR200479.

[0443] 16. Roberfroid M., Gobson GR, Hoyles L., McCartney AL, Rastall R., Rowland I., Wolvers D., Watzl B., Szajewska H., Stahl B., et al. Prebiotic effects: Metabolic and health benefits. Br. J. Nutr. 2011;104:S1–S63.

[0444] 17. Leach JD, Sobolik KD High dietary intake of prebiotic inulin-type fructans in the prehistoric Chihauhuan desert. Br. J. Nutr. 2010;103:1158–1561.

[0445] It should be understood that although certain embodiments have been illustrated and described herein, the claims are not limited to the specific forms or arrangements of parts described and shown. In the specification, illustrative embodiments have been disclosed, and although specific terms are employed, they are used in a general and descriptive sense only and not for the purpose of limitation. Modifications and variations of the embodiments are possible in light of the above teachings. Therefore, it should be understood that the embodiments may be implemented in a manner other than that specifically described.

[0446] Although various embodiments are described above, it should be understood that they are only illustrations and examples of the present technology and are not intended to be limiting. It will be apparent to those skilled in the art that various modifications and changes may be made to the form and details of the present technology without departing from the spirit and scope of the present technology. Therefore, the breadth and scope of the present technology should not be limited to any of the above-described embodiments, but should only be defined in accordance with the appended claims and their equivalents. It will also be understood that each feature of each embodiment of each reference described and cited herein can be used in combination with the features of any other embodiment. All patents and disclosures cited herein are incorporated herein by reference in their entirety.

Claims

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

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

3. The liquid composition of claim 1, comprising about 0.2 units / mL to about 0.3 units / mL of superoxide dismutase.

4. The liquid composition of any one of claims 1-3, comprising from about 2.7 mg / mL to about 12 mg / mL of soluble fiber.

5. The liquid composition of any one of claims 1-3, comprising from about 5.55 mg / mL to about 11.11 mg / mL of soluble fiber.

6. The liquid composition according to any one of claims 1 to 5, wherein the superoxide dismutase is extracted from melon, beef liver, heterotrophic bacteria or marine phytoplankton.

7. The liquid composition according to any one of claims 1 to 6, wherein the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase, or nickel superoxide dismutase.

8. The liquid composition according to any one of claims 1 to 7, wherein the weight ratio of superoxide dismutase to soluble fiber is from about 1:100 to about 1:1000.

9. The liquid composition according to any one of claims 1 to 7, wherein the weight ratio of superoxide dismutase to soluble fiber is from about 1:500 to about 1:

700.

10. The liquid composition according to any one of claims 1 to 9, wherein the soluble fiber is a water-soluble polysaccharide.

11. The liquid composition according to any one of claims 1 to 10, wherein the soluble fiber is selected from the group consisting of soluble corn fiber, inulin, dextrin, guar gum, oligosaccharides, galactopolysaccharides, oligofructose, lactulose, digestion-resistant starch, xylo-oligosaccharides and isomalto-oligosaccharides.

12. The liquid composition of any one of claims 1-10, wherein the soluble fiber is soluble corn fiber.

13. The liquid composition of claim 12, wherein the soluble corn fiber is digestion-resistant maltodextrin.

14. The liquid composition of any one of claims 1-13, further comprising about 0.1 mg / mL to about 1.5 mg / mL of a simple sugar.

15. The liquid composition of any one of claims 1-13, further comprising about 0.1 mg / mL to about 1.5 mg / mL of d-ribose.

16. The liquid composition of any one of claims 1-13, further comprising about 0.40 mg / mL to about 0.85 mg / mL of d-ribose.

17. The liquid composition of any one of claims 1-16, further comprising about 1.3 mg / mL to about 9.0 mg / mL of a sugar alcohol.

18. The liquid composition of any one of claims 1-16, further comprising about 1.3 mg / mL to about 9.0 mg / mL of erythritol.

19. The liquid composition of any one of claims 1-16, further comprising about 2.7 mg / mL to about 5.6 mg / mL of erythritol.

20. The liquid composition of any one of claims 1-19, further comprising about 0.1 mg / mL to about 1.5 mg / mL of a pH adjuster.

21. The liquid composition of any one of claims 1-19, further comprising about 0.1 mg / mL to about 1.5 mg / mL of citric acid.

22. The liquid composition of any one of claims 1-19, further comprising about 0.4 mg / mL to about 0.7 mg / mL of citric acid.

23. The liquid composition of any one of claims 1-22, further comprising about 0.05 mg / mL to about 0.75 mg / mL of a sweetener.

24. The liquid composition of any one of claims 1-22, further comprising about 0.05 mg / mL to about 0.75 mg / mL of steviol glycosides.

25. The liquid composition of any one of claims 1-22, further comprising about 0.2 mg / mL to about 0.35 mg / mL of steviol glycosides.

26. The liquid composition of any one of claims 1-22, further comprising a flavoring agent.

27. A composition comprising: a) about 10 units to about 200 units of superoxide dismutase; b) from about 500 mg to about 8,000 mg of soluble fiber; and c) Probiotics.

28. The composition of claim 27 comprising about 50 units to about 150 units of superoxide dismutase.

29. The composition of claim 27 comprising about 70 units to about 100 units of superoxide dismutase.

30. The composition of any one of claims 27-29 comprising from about 1,000 mg to about 5,000 mg of soluble fiber.

31. The composition of any one of claims 27-29 comprising from about 2,000 mg to about 4,000 mg of soluble fiber.

32. The composition of any one of claims 27-31, wherein the superoxide dismutase is extracted from melon, beef liver, heterotrophic bacteria, or marine phytoplankton.

33. The composition of any one of claims 27-31, wherein the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase, or nickel superoxide dismutase.

34. The composition of any one of claims 27-33, wherein the weight ratio of superoxide dismutase to soluble fiber is from about 1:100 to about 1:1000.

35. The composition of any one of claims 27-33, wherein the weight ratio of superoxide dismutase to soluble fiber is from about 1:500 to about 1:

700.

36. The composition of any one of claims 27-35, wherein the soluble fiber is a water-soluble polysaccharide.

37. A composition according to any one of claims 27-35, wherein the soluble fiber is selected from the group consisting of soluble corn fiber, inulin, dextrin, guar gum, oligosaccharides, galactopolysaccharides, oligofructose, lactulose, digestion-resistant starch, xylo-oligosaccharides and isomalto-oligosaccharides.

38. The composition of any one of claims 27-35, wherein the soluble fiber is soluble corn fiber.

39. The composition of claim 38, wherein the soluble corn fiber is digestion-resistant maltodextrin.

40. The composition of any one of claims 27-39, wherein the probiotic comprises a Bifidobacterium bacterium.

41. The composition of any one of claims 27-39, wherein the probiotic comprises a Lactobacillus bacterium.

42. The composition of any one of claims 27-39, wherein the probiotic comprises Lactobacillus of the phylum Firmicutes, Bifidobacterium of the phylum Actinobacteria, or a combination thereof.

43. The composition of any one of claims 27-39, wherein the composition is in the form of a gel.

44. The composition of any one of claims 27-39, wherein the composition is in liquid form.

45. The composition of any one of claims 27-39, wherein the composition is in powder form.

46. ​​A method of increasing T cell activation in a subject, comprising orally administering to the subject a composition comprising: a) about 10 units to about 200 units of superoxide dismutase; and b) from about 500 mg to about 8,000 mg of soluble fiber; in, After administration of the composition, activation of T cells in the subject is increased.

47. The method of claim 46, wherein the composition is administered in combination with an anti-cancer agent.

48. The method of claim 46, wherein the composition is administered in combination with an antiviral agent.

49. A method of increasing short chain fatty acid (SCFA) production 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) from about 500 mg to about 8,000 mg of soluble fiber; in, After administration of the composition, the production of SCFAs in the digestive tract of the subject is increased.

50. The method of claim 49, wherein the SCFA whose production is increased is an acetic, propionic, butyric or lactic SCFA or a combination thereof.

51. The method of claim 50, wherein the SCFAs are increased in a manner that provides approximately the same ratio of acetate, propionate, butyrate and lactate SCFAs as compared to the ratio of acetate, propionate, butyrate and lactate SCFAs prior to the increase.

52. A method of increasing the amount of Bifidobacterium or Lactobacillus bacteria 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) from about 500 mg to about 8,000 mg of soluble fiber; in, After administration of the composition, the amount of Bifidobacterium, Lactobacillus, or a combination thereof in the digestive tract of the subject increases.

53. The method of claim 52, wherein the Bifidobacterium bacterium comprises a Bifidobacterium actinomycetemcomitans species.

54. The method of claim 52, wherein the Lactobacillus bacteria comprises a Lactobacillus species of the phylum Firmicutes.

55. The method of any one of claims 46-54, wherein the composition comprises about 50 units to about 150 units of superoxide dismutase.

56. The method of any one of claims 46-54, wherein the composition comprises about 70 units to about 100 units of superoxide dismutase.

57. The method of any one of claims 46-56, wherein the composition comprises from about 1000 mg to about 5000 mg of soluble fiber.

58. The method of any one of claims 46-56, wherein the composition comprises from about 2,000 mg to about 4,000 mg of soluble fiber.

59. The method of any one of claims 46-58, wherein the superoxide dismutase is extracted from melon, beef liver, heterotrophic bacteria, or marine phytoplankton.

60. The method of any one of claims 46-58, wherein the superoxide dismutase is copper / zinc superoxide dismutase, iron / manganese superoxide dismutase, or nickel superoxide dismutase.

61. The method of any one of claims 46-60, wherein the weight ratio of superoxide dismutase to soluble fiber in the composition is from about 1:100 to about 1:1000.

62. The method of any one of claims 46-60, wherein the weight ratio of superoxide dismutase to soluble fiber in the composition is from about 1:500 to about 1:

700.

63. The method of any one of claims 46-62, wherein the soluble fiber is a water-soluble polysaccharide.

64. The method according to any one of claims 46-62, wherein the soluble fiber is selected from the group consisting of soluble corn fiber, inulin, dextrin, guar gum, oligosaccharides, galacto-polysaccharides, oligofructose, lactulose, digestion-resistant starch, xylo-oligosaccharides and isomalto-oligosaccharides.

65. The method of any one of claims 46-62, wherein the soluble fiber is soluble corn fiber.

66. The method of claim 65, wherein the soluble corn fiber is digestion-resistant maltodextrin.

67. The method of any one of claims 46-66, wherein the composition is in the form of a gel.

68. The method of any one of claims 46-66, wherein the composition is in liquid form.

69. The method of any one of claims 46-66, wherein the composition is in powder form.

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