WO2008143880A2 | 2008-11-27 |
US20020045656A1 | 2002-04-18 | |||
US20100222421A1 | 2010-09-02 | |||
US20190350890A1 | 2019-11-21 |
CLAIMS 1. A method of treating a subject at risk of having a traumatic brain injury or ischemia- reperfusion injury, the method comprising administering to the subject an effective amount of 1- palmitoyl-2-linoleoyl-3-acetylrac-glycerol (PLAG) to mediate the effects of traumatic brain injury or ischemia-reperfusion injury. 2. The method of claim 1, wherein administering includes multiple doses before, after, or before and after the traumatic brain injury or ischemia-reperfusion injury. 3. The method of claim 1, further comprising administering a subsequent dose after the traumatic brain injury or ischemia-reperfusion injury. 4. The method of claim 1, wherein the effective among is administered within 1 hour of an activity that increases the risk of traumatic brain injury or ischemia-reperfusion injury. 5. A method of treating a human having suffered a traumatic brain injury or ischemia- reperfusion injury, the method comprising administering to the human a therapeutically effective amount of 1-palmitoyl-2-linoleoyl-3-acetylrac-glycerol (PLAG) and Vitamin E, alpha-tocopherol or alpha-tocopheryl acetate, to treat the traumatic brain injury or ischemia-reperfusion injury. 6. The method of claim 5, wherein administering occurs within 6 hour of the traumatic brain injury or ischemia-reperfusion injury. 7. The method of claim 5, wherein administering occurs at about 1 to 6 hours after the traumatic brain injury or ischemia-reperfusion injury. 8. The method of claim 5, wherein administering occurs at about 6 to 12 hours after the traumatic brain injury or ischemia-reperfusion injury. 9. A method of ameliorating the sequelae of a traumatic brain injury or ischemia- reperfusion injury, the method comprising administering to a subject an effective amount of 1- palmitoyl-2-linoleoyl-3-acetylrac-glycerol (PLAG) as a daily nutritional supplement. 10. A dietary supplement for mediating traumatic brain injury (TBI) or ischemia-reperfusion injury comprising an effective amount of 1-palmitoyl-2-linoleoyl-3-acetylrac-glycerol (PLAG) formulated for daily use. |
1-palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol Alpha-tocopherol [0046] In an embodiment, the treatment can include one or more doses of the agent(s) within about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 14, 16, 18, 20, 22, or 24 hour(s) before or after the traumatic brain injury. For example, the agent(s) at a constant or varying doses can be administered once or in intervals of about 30 min, 1, 2, 3, 4, 5, 6, 8, 12, or 24 hour(s), as needed for treatment. The dose amount, frequency, and the number of doses, can depend upon the nature of the traumatic brain injury (e.g., cause, severity, extent of injury, damage after the injury, and the like), the subject (e.g., age, health, sex, and the like), the time that the first dose is administered, as well as other medications taken before or after the traumatic brain injury. [0047] The phrase “traumatic brain injury” (also referred to as “an intracranial injury”) can be caused by an impact, strike, force, shock wave (e.g., from an explosion) and the like on the head and/or a sudden acceleration or deceleration so that brain impacts the skull. The traumatic brain injury includes not only the direct damage but also the damage caused by pressure within the skull, changes in the blood flow, and the like. The traumatic brain injury is an acquired brain injury or head injury. The injury can be focal or diffuse. The injury can be mild, moderate, or severe based on variables such as duration of loss of consciousness, Glassgow Coma Score, and/or post-traumatic stress amnesia, or the like. The injury can be chronic or acute. The traumatic brain injury can be the result from a traffic accident, crashing or falling, sports activities, result of explosions, military activities, combinations thereof, and the like. [0048] The phrase “ischemia-reperfusion injury” can be caused by the circulatory conditions associated with the sudden poor blood flow and subsequent restoration of the blood flow to the certain part of the body, such as brain, eye, lung, heart, intestine, liver, limbs or other organs. Initial hypoxic stage causes marked imbalance in metabolism and microvascular dysfunction. Subsequent reperfusion stage further enhances the cell death programs causing the tissue damage The example of this condition are ischemic heart disease, stroke, intestinal ischemia, limb ischemia, sickle cell disease, diabetic vasculopathy, trauma, and other status involving ischemia and reperfusion. This mechanism is also important for cadiac,thoracic, and transplant surgeries. 25 [0049] As mentioned above, the agent can include (i) monoacetyldiglyceride (e.g., 1- palmitoyl-2-linoleoyl-3-acetyl-rac-glycerol (PLAG)) (also known as EC-18); (ii) vitamin E, alpha-tocopherol or alpha-tocopheryl acetate; or (iii) 1-palmitoyl-2-linoleoyl-3-acetyl-rac- glycerol (PLAG) and vitamin E, alpha-tocopherol or alpha-tocopheryl acetate. [0050] The agent(s) can be contained in compositions, pharmaceutical compositions, liquid compositions, gel compositions, and the like, including without limitation controlled release and sustained release formulations. [0051] In certain embodiments, a simple and convenient dosing regimen for treating patients having a traumatic brain injury includes any regimen previously shown effective for use of the agent for its known uses. Dose forms of the agent can include 25 mg, 50 mg, 75 mg, 100 mg, 125 mg, 150 mg, and 200 mg unit dose forms. In one embodiment, the unit dose form is a tablet; in another, the unit dose form is a capsule. In one embodiment, sustained release formulations of agent are in the tablet and capsule form. In another embodiment, the unit dose form is a liquid. I. Nutritional Supplementation [0052] As used herein, the term “PLAG supplementation” refers to environmental intervention by delivering and/or administering PLAG to a subject in need thereof. In some embodiments, PLAG supplementation is a dietary component or dietary additive. PLAG supplementation may be delivered and/or administrated to a subject as part of a regular diet paradigm for a determined amount of time. For example, PLAG supplementation may be delivered and/or administered to a subject as part of a daily dietary paradigm including but not limited to once a day, twice a day, or three times a day. In some embodiments, PLAG supplementation is delivered and/or administered to a subject with food. In some embodiments, PLAG supplementation is delivered and/or administered to a subject without food. In some embodiments, PLAG supplementation is delivered and/or administered to a subject as part of a daily dietary routine over the course of including but not limited to, at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 8 weeks, at least 9 weeks, at least 10 weeks, at least 11 weeks, at least 12 weeks, at least 13 weeks, at least 14 weeks, at least 15 weeks, at least 16 weeks, at least 17 weeks, at least 18 weeks, at least 19 weeks, at least 20 weeks, at least 30 weeks, at least 40 weeks, or at least 50 weeks, at least 1 month, at least 2 months, at least 3 months, at least 4 months, at least 5 months, at least 6 months, at least 7 months at least 8 months, at least 9 months, at least 10 months, at least 11 months or at least 12 months. In certain aspects a PLAG supplementation can be administered or taken 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or more minutes, hours, or days before engaging in an activity that has an increased risk of TBI. Subjects at risk of TBI include, for example, those participating in a sport with an occurrence of concussions, such as football players, boxers and mixed martial arts fighters. Other subjects at risk of traumatic brain injury include combatants in armed conflicts, or undergoing dangerous or extreme sporting or adventure activities. [0053] PLAG supplementation, as used herein, is delivered and/or administered to a subject in an effective amount to ameliorate or treat TBI, including symptoms and sequelae of TBI. As used herein, the term “sequelae” refers to illness/disease and symptoms/signs that occur as a consequence of a condition and/or disease event. In some embodiments, sequelae occur long after the initial disease/illness has resolved. As used here, the term “effective amount” refers to the amount of each active agent required to confer therapeutic effect on the subject, either alone or in combination with one or more other active agents. Effective amounts vary, as recognized by those skilled in the art, depending on the particular condition being treated, the severity of the condition, the individual patient parameters including age, physical condition, size, gender and weight, the duration of the treatment, the nature of concurrent therapy (if any), the specific route of administration and like factors within the knowledge and expertise of the health practitioner. These factors are well known to those of ordinary skill in the art and can be addressed with no more than routine experimentation. It is generally preferred that a maximum dose of the individual components or combinations thereof be used, that is, the highest safe dose according to sound medical judgment. It will be understood by those of ordinary skill in the art, however, that a patient may insist upon a lower dose or tolerable dose for medical reasons, psychological reasons or for virtually any other reasons. [0054] Generally, for administration of the PLAG supplements an initial dosage can be greater than 500 mg/day. For the purpose of the present disclosure, a typical daily dosage might range from about any of 500 mg/day to 2,000 mg/day, 550 mg/day to 1,000 mg/day, 600 mg/day to 1,000 mg/day depending on the factors mentioned above. For repeated administrations over several days or longer, the treatment is sustained until a desired suppression of symptoms occurs or until sufficient therapeutic levels are achieved to alleviate a symptom or sequelae of TBI. An exemplary dosing regimen comprises administering dose of greater than about 500, 550, 600, 650, 700, 750, 800, 850, 900, 950, 1,000, 1,050, 1,100, 1,150, 1,200, 1,250, 1,300, 1,350, 1,400, 1,450, 1,500, 1, 550, 1,600, 1,650, 1,700, 1,750, 1,800, 1,850, 1,900, 1,950, or 2000 mg/day for 3 months, 6 months or a year. However, other dosage regimens may be useful, depending on the pattern of pharmacokinetic decay that the practitioner wishes to achieve. For example, dosing from one-four times a week is contemplated. The dosing regimen can vary over time. A subject suspected of having TBI or sequelae of TBI may be a subject having one or more clinical symptoms of TBI. A variety of clinical symptoms are known in the art. Symptoms of TBI include a loss of consciousness, headache, confusion, lightheadedness, dizziness, blurred vision or tired eyes, ringing in the ears, bad taste in the mouth, fatigue or lethargy, a change in sleep patterns, behavioral or mood changes, trouble with memory, concentration, attention, or thinking, vomiting or nausea, convulsions or seizures, an inability to awaken from sleep, dilation of one or both pupils of the eyes, slurred speech, weakness or numbness in the extremities, loss of coordination, and increased confusion, restlessness, and agitation. Symptoms of a TBI can be mild, moderate, or severe, depending on the extent of the damage to the brain. [0055] In some embodiments, the subject has a TBI. In some embodiments, the subject has a TBI and is undergoing a putative treatment for TBI. The methods described herein may be used to supplement the efficacy of a putative therapy for TBI, i.e., for increasing the responsiveness of the subject to a putative therapy for TBI. Based on this evaluation, the physician may continue the therapy, if there is a favorable response, or discontinue and change to another therapy if the response is unfavorable. [0056] As used herein, the term “treating” refers to the application or administration of a composition including one or more active agents to a subject, who has a TBI, a symptom of a TBI, or is at risk of TBI, with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve, or affect one or more symptom of TBI. [0057] Alleviating TBI or ischemia-reperfusion injury includes delaying the development or progression of the disease, or reducing disease severity. Alleviating the disease does not necessarily require curative results. As used therein, “delaying” the development of a disease means to defer, hinder, slow, retard, stabilize, and/or postpone progression of the disease. This delay can be of varying lengths of time, depending on the history of the disease and/or individuals being treated. A method that “delays” or alleviates the development of a disease, or delays the onset of the disease, is a method that reduces probability of developing one or more symptoms of the disease in a given time frame and/or reduces extent of the symptoms in a given time frame, when compared to not using the method. Such comparisons are typically based on clinical studies, using a number of subjects sufficient to give a statistically significant result. [0058] Supplementation efficacy can be assessed by methods well-known in the art. II. Pharmaceutical Formulations and Routes of Administration [0059] Embodiments of the present disclosure include an agent formulated with one or more pharmaceutically acceptable excipients, diluents, carriers and/or adjuvants. In addition, embodiments of the present disclosure include the agent formulated with one or more pharmaceutically acceptable auxiliary substances. In particular, one or more agents can be formulated with one or more pharmaceutically acceptable excipients, diluents, carriers, and/or adjuvants to provide an embodiment of a composition of the present disclosure. [0060] A wide variety of pharmaceutically acceptable excipients are known in the art. Pharmaceutically acceptable excipients have been amply described in a variety of publications, including, for example, A. Gennaro (2000) “Remington: The Science and Practice of Pharmacy,” 20th edition, Lippincott, Williams, & Wilkins Pharmaceutical Dosage Forms and Drug Delivery Systems (1999) H. C. Ansel et al., eds., 7th ed., Lippincott, Williams, & Wilkins and Handbook of Pharmaceutical Excipients (2000) A. H. Kibbe et al., eds., 3rd ed. Amer. Pharmaceutical Assoc. [0061] The pharmaceutically acceptable excipients, such as vehicles, adjuvants, carriers or diluents, are readily available to the public. Moreover, pharmaceutically acceptable auxiliary substances, such as pH adjusting and buffering agents, tonicity adjusting agents, stabilizers, wetting agents and the like, are readily available to the public. [0062] In certain embodiments, the agent is administered to the subject using a convenient means capable of resulting in the desired effect. Thus, embodiments of the present disclosure provide an agent incorporated into a variety of formulations for therapeutic administration. For example, embodiments provide the agent formulated into pharmaceutical compositions by combination with appropriate, pharmaceutically acceptable carriers or diluents, and formulated into preparations in solid, semi-solid, liquid or gaseous forms, such as tablets, capsules, powders, granules, ointments, solutions, suppositories, injections, inhalants and aerosols. [0063] In pharmaceutical dosage forms, the agent may be present in the form of its pharmaceutically acceptable salts, or a subject active agent may be used alone or in appropriate association, as well as in combination, with other pharmaceutically active compounds. The following methods and excipients are merely an example and are in no way limiting. [0064] For oral preparations, the agent can be used alone or in combination with appropriate additives to make tablets, powders, granules or capsules, for example, with conventional additives, such as lactose, mannitol, corn starch or potato starch; with binders, such as crystalline cellulose, cellulose derivatives, acacia, corn starch or gelatins; with disintegrators, such as corn starch, potato starch or sodium carboxymethylcellulose; with lubricants, such as talc or magnesium stearate; and if desired, with diluents, buffering agents, moistening agents, preservatives and flavoring agents. [0065] In certain aspects the agent can be formulated into preparations for injection by dissolving, suspending or emulsifying them in an aqueous or nonaqueous solvent, such as vegetable or other similar oils, synthetic aliphatic acid glycerides, esters of higher aliphatic acids or propylene glycol; and if desired, with conventional additives such as solubilizers, isotonic agents, suspending agents, emulsifying agents, stabilizers and preservatives. [0066] Unit dosage forms for oral or rectal administration, such as syrups, elixirs, and suspensions, may be provided wherein each dosage unit, for example, teaspoonful, tablespoonful, tablet or suppository, contains a predetermined amount of the composition containing one or more agents. Similarly, unit dosage forms for injection or intravenous administration may comprise the agent in a composition as a solution in sterile water, normal saline or another pharmaceutically acceptable carrier. [0067] In certain aspects the agent can be formulated in an injectable composition in accordance with the present disclosure. Typically, injectable compositions are prepared as liquid solutions or suspensions; solid forms suitable for solution in, or suspension in, liquid vehicles prior to injection may also be prepared. The preparation may also be emulsified or the active ingredient encapsulated in liposome vehicles. [0068] In certain aspects the agent is formulated for delivery by a continuous delivery system. The term “continuous delivery system” is used interchangeably herein with “controlled delivery system” and encompasses continuous (e.g., controlled) delivery devices (e.g., pumps) in combination with catheters, injection devices, and the like, a wide variety of which are known in the art. [0069] Drug release devices suitable for use in the disclosure may be based on any of a variety of modes of operation. For example, the drug release device can be based upon an erodible system (e.g., an erosion-based system). For example, the drug release device can be an osmotic bursting matrix, e.g., where the drug is incorporated into a polymer and the polymer provides for release of drug formulation concomitant with degradation of a drug-impregnated polymeric material (e.g., a biodegradable, drug-impregnated polymeric material). [0070] In some embodiments, the drug delivery device is an implantable device. The drug delivery device can be implanted at any suitable implantation site using methods and devices well known in the art. As noted herein, an implantation site is a site within the body of a subject at which a drug delivery device is introduced and positioned. Implantation sites include, but are not necessarily limited to a subdermal, subcutaneous, intramuscular, or other suitable site within a subject's body. [0071] Compositions of the present disclosure may be used with a sustained-release or controlled release matrix. In addition, embodiments of the present disclosure can be used in conjunction with other treatments that use sustained-release formulations. As used herein, a sustained-release matrix is a matrix made of materials, usually polymers, which are degradable by enzymatic or acid-based hydrolysis or by dissolution. Once inserted into the body, the matrix is acted upon by enzymes and body fluids. A sustained-release matrix desirably is chosen from biocompatible materials such as liposomes, polylactides (polylactic acid), polyglycolide (polymer of glycolic acid), polylactide co-glycolide (copolymers of lactic acid and glycolic acid), polyanhydrides, poly(ortho)esters, polypeptides, hyaluronic acid, collagen, chondroitin sulfate, carboxcylic acids, fatty acids, phospholipids, polysaccharides, nucleic acids, polyamino acids, amino acids such as phenylalanine, tyrosine, isoleucine, polynucleotides, polyvinyl propylene, polyvinylpyrrolidone and silicone. Exemplary biodegradable matrices include a polylactide matrix, a polyglycolide matrix, and a polylactide co-glycolide (co-polymers of lactic acid and glycolic acid) matrix. III. Dosages [0072] In certain embodiments the agent can be administered to a subject in one or more doses. In certain aspects the agent can be administered in an amount of about 10 mg to 1000 mg per dose, e.g., about 10 mg to 20 mg, about 20 mg to 25 mg, about 25 mg to 50 mg, about 50 mg to 75 mg, about 75 mg to 100 mg, about 100 mg to 125 mg, about 125 mg to 150 mg, about 150 mg to 175 mg, about 175 mg to 200 mg, about 200 mg to 225 mg, about 225 mg to 250 mg, about 250 mg to 300 mg, about 300 mg to 350 mg, about 350 mg to 400 mg, about 400 mg to 450 mg, about 450 mg to 500 mg, about 500 mg to 750 mg, or about 750 mg to 1000 mg per dose. [0073] In certain aspects the amount of the agent per dose is determined on a per body weight basis. For example, in an embodiment, the agent can be administered in an amount of about 0.5 mg/kg to 100 mg/kg, e.g., about 0.5 mg/kg to 1 mg/kg, about 1 mg/kg to 2 mg/kg, about 2 mg/kg to 3 mg/kg, about 3 mg/kg to 5 mg/kg, about 5 mg/kg to 7 mg/kg, about 7 mg/kg to about 10 mg/kg, about 10 mg/kg to 15 mg/kg, about 15 mg/kg to 20 mg/kg, about 20 mg/kg to 25 mg/kg, about 25 mg/kg to 30 mg/kg, about 30 mg/kg to 40 mg/kg, about 40 mg/kg to 50 mg/kg per dose, about 50 mg/kg to 60 mg/kg, about 60 mg/kg to 70 mg/kg, about 70 mg/kg to 80 mg/kg, about 80 mg/kg to 90 mg/kg, or about 90 mg/kg to 100 mg/kg, or more than about 100 mg/kg. [0074] Those of skill will readily appreciate that dose levels can vary as a function of the specific agent, the severity of the symptoms and the susceptibility of the subject to side effects. Preferred dosages for a given compound are readily determinable by those of skill in the art by a variety of means. [0075] In certain aspects multiple doses of the agent are administered. The frequency of administration of the agent can vary depending on any of a variety of factors, e.g., severity of the symptoms, and the like. [0076] The duration of administration of the agent, e.g., the period of time over which the agent is administered, can vary, depending on any of a variety of factors, e.g., patient response, etc. For example, the gent can be administered over a period of time of about 1 hour to a few hours, to 12 hours to one day to one week or more. IV. Routes of Administration [0077] Certain embodiments of the present disclosure provide methods for treating a traumatic brain injury comprising the administration of the agent to a subject (e.g., a human) using any available method and route suitable for drug delivery. Routes of administration include intranasal, intramuscular, intratracheal, subcutaneous, intradermal, topical application, intravenous, rectal, nasal, oral, and other enteral and parenteral routes of administration. Routes of administration may be combined, if desired, or adjusted depending upon the agent and/or the desired effect. An agent can be administered in a single dose or in multiple doses. The agent described herein is suitable for oral administration, which may be preferable for most patients. [0078] In certain aspects the agent can be administered to a subject using available conventional methods and routes suitable for delivery of conventional drugs, including systemic or localized routes. In general, routes of administration contemplated by the disclosure include, but are not limited to, enteral, parenteral, or inhalational routes. [0079] Parenteral routes of administration other than inhalation administration include, but are not limited to, topical, transdermal, subcutaneous, intramuscular, intraorbital, intracapsular, intraspinal, intrasternal, and intravenous routes, i.e., any route of administration other than through the alimentary canal. Parenteral administration can be carried to effect systemic or local delivery of the agent. Where systemic delivery is desired, administration typically involves invasive or systemically absorbed topical or mucosal administration of pharmaceutical preparations. [0080] In certain aspects the agent can also be delivered to the subject by enteral administration. Enteral routes of administration include, but are not limited to, oral and rectal (e.g., using a suppository) delivery. [0081] Methods of administration of the agent through the skin or mucosa include, but are not limited to, topical application of a suitable pharmaceutical preparation, transdermal transmission, injection and epidermal administration. For transdermal transmission, absorption promoters or iontophoresis are suitable methods. Iontophoretic transmission may be accomplished using commercially available “patches” that deliver their product continuously via electric pulses through unbroken skin for periods of several days or more. V. Examples [0082] The following examples as well as the figures are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the techniques disclosed in the examples or figures represent techniques discovered by the inventors to function well in the practice of the invention, and thus can be considered to constitute preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention. [0083] The studies described below assess prophylactic monoacetyldiglycerides and/or alpha- tocopherol treatment prior to TBI or ischemia-reperfusion injury. [0084] Drosophila melanogaster (the common fruit fly) is a known/characterized model organism with a lifespan of around two to three months with low maintenance requirements. Drosophila shares more than 50% of human genes and more than 75% of human disease genes. 21 Drosophila goes through the embryo, first instar larva, second instar larva, third instar larva, prepupa, pupa, and adult stages. 11 It is commonly found near unripe and rotted fruit. 21 A. Methods: [0085] There were 9 groups in this experiment, as shown in table 1. There were 3 supercategories: polysorbate 80 (0.01%, Velona Inc., Elk Grove Village, IL), lecithin (0.01%, Now Foods, Bloomingdale, IL), and no food additives. Within these, there were the subgroups vitamin E (200 µg/ml, Kirkland Signature, Seattle, Washington ), PLAG (278 µg/ml, Enzychem, South Korea), and no preventative agents. These agents were vigorously mixed and added to the Formula 4-24 Instant Drosophila culture medium (Carolina Biological Supply Company, Burlington, NC). Table 1. Experimental groups [0086] Traumatic brain injuries were induced through a High-Impact Trauma device. 7 For mortality, the number of deceased and living were measured. For climb up mobility, the number of Drosophila above the upper quartile (6 cm) was measured in 10 seconds and compared to the number below this region. For walking mobility, number of Drosophila reaching the peripheral wall of 35 mm Petri dish in 10 seconds was measured and compared to the number that could not reach, after applying Drosophila at the center of Petri dish. Relative risk was used for the statistical analysis of mortality and mobility by MedCalc Software Ltd. relative risk calculator. 12 [0087] Wild type D. melanogaster was divided into two groups: one group with PLAG 278 ug/ml for 5 days and the other group without PLAG. The vials of both groups were transferred to Eppendorf tubes covered with gauze and secured with a rubber band after removing their caps. The ischemia-reperfusion injury was induced through a hypoxic chamber infused with nitrogen gas at 1.5 l/min for 10 minutes. They remained in the hypoxia chambers for a total of 2.5 hours. Thereafter, they were transferred to the culture medium with room air. After 24 hours, mortality and negative geotaxis were measured. For the mild ischemia-reperfusion injury, they were only exposed to 1 hour of hypoxic chamber. B. Results [0088] Both PLAG and Vit E administered before TBI decreased mortality after a TBI (p<0.05). Moreover, the effects of PLAG appeared to be greater than Vit E (p<0.05). However, polysorbate 80 increased mortality after a TBI (p<0.05). PLAG with polysorbate 80 reduced mortality (p<0.05). Vit E with polysorbate 80 reduced mortality compared with the polysorbate 80-only group (p<0.05); however, it reduced to a degree comparable with the control group without food additives. Lecithin did not affect mortality after the TBI. Also, lecithin did not confer additional benefits from the combined use of PLAG or Vit E (p>0.05). Both PLAG and Vit E administered before TBI improved climb up mobility after the TBI (p<0.05). Polysorbate 80 and lecithin did not affect climb up mobility after the TBI (p>0.05). PLAG with polysorbate 80 reduced mortality (p<0.05), unlike Vit E with polysorbate 80 (p>0.05). However, with the presence of lecithin, Vit E improved climb up mobility after the TBI (p<0.05). PLAG and PLAG with Lecithin improved walking mobility after the TBI (p<0.05). PLAG with polysorbate did not improve walking mobility after TBI (p>0.05). Also, vitamin E did not affect walking mobility after the TBI (p>0.05). [0089] Only PLAG after TBI improved mortality with the treatment after a TBI (p<0.05). Vitamin E after TBI did not improve mortality (p>0.05). Table 2. Statistical Analysis of Mortality with Treatment Before a TBI Table 3. Statistical Analysis of Impaired Climb Up Mobility with Treatment Before a TBI
Table 4. Statistical Analysis of Impaired Walking Mobility with Treatment Before a TBI Table 5. Statistical Analysis of Mortality with treatment after A TBI M t lit R l ti 95% CI 95% CI Table 6. Statistical Analysis of Impaired Climb Up Mobility with the Treatment After a TBI [0090] PLAG has a protective effect on ischemia-reperfusion injury in Drosophila melanogaster. Mortality after an ischemia-reperfusion injury was 42% without PLAG, and 15% with PLAG (p<0.0001, Chi-square test). PLAG can reduce mortality by 64% in D. melanogaster. Negative geotaxis was slightly improved in PLAG group (49% vs 40%), but it was not statistically significant (p=0.3178, Chi-square test). However, in the mild ischemia-reperfusion injury without mortality, PLAG improved negative geotaxis significantly (79% vs 55%; p=0.0366, Chi-square test). Table 7. Mortality after ischemia-reperfusion injury (p<0.0001, Chi-square test) Table 8. Negative geotaxis after ischemia-reperfusion injury (p=0.3178, Chi-square test) Table 9. Negative geotaxis after the mild ischemia-reperfusion injury (p=0.0366, Chi-square test)
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