Method for insuring adequate intracellular glutathione in tissue

Abstract

A method is provided for increasing and/or maintaining the level of intracellular glutathione. Pursuant to the invention, compositions including denatured and at least partially hydrolyzed proteins are administered to a patient in an amount sufficient to increase glutathione levels in the patient.

Claims

We claim: 1. A method for increasing intracellular glutathione levels in a stressed patient comprising the steps of enterally administering to the patient a therapeutically effective amount of a denatured and at least partially hydrolyzed protein that includes at least 2.1% cysteine by caloric content. 2. The method of claim 1 wherein the protein includes whey. 3. The method of claim 1 wherein the protein includes egg white protein. 4. The method of claim 1 wherein the patient initially has a less than physiological normal level of glutathione. 5. The method of claim 1 wherein the patient is an intensive care patient. 6. The method of claim 1 wherein at least 1.2 grams of cysteine are administered to the patient daily. 7. The method of claim 1 wherein the patient has a compromised immune system. 8. A method for enhancing the immune system of a metabolically stressed patient comprising the steps of enterally administering to the patient a composition that includes at least 0.37% of its caloric content as cysteine. 9. The method of claim 8 wherein the protein includes whey. 10. The method of claim 8 wherein the protein includes egg white protein. 11. The method of claim 8 wherein the patient initially has a less than physiological normal level of glutathione. 12. The method of claim 8 wherein the patient is an intensive care patient. 13. The method of claim 8 wherein at least 1.2 grams of cysteine are administered to the patient daily. 14. A method for increasing intracellular glutathione levels in a metabolically stressed patient having reduced intracellular levels of glutathione comprising administering to the patient an enteral composition that includes a sufficient amount of a denatured and at least partially hydrolyzed whey protein to increase the intracellular glutathione level of the patient. 15. The method of claim 14 wherein the composition includes glutathione. 16. The method of claim 14 wherein at least 1.2 grams of cysteine are administered to the patient daily. 17. The method of claim 14 wherein the composition provides at least 0.37% of its caloric content as cysteine. 18. A method for increasing intracellular glutathione levels in a patient having a compromised immune system and reduced intracellular levels of glutathione comprising administering to the patient an enteral composition that includes at least 0.37% of its caloric content as cysteine and includes a denatured and partially hydrolyzed protein in an amount sufficient to increase the intracellular glutathione level of the patient. 19. The method of claim 18 wherein the composition includes glutathione. 20. The method of claim 18 wherein at least 1.2 grams of cysteine are administered to the patient daily.
This is a continuation of application Ser. No. 08/659,111, filed Jun. 3, 1996, now abandoned which is a continuation of application Ser. No. 08/306,133, filed Sep. 14, 1994, now abandoned which is a continuation-in-part of application Ser. No. 07/650,222, filed Feb. 4, 1991 now abandoned. BACKGROUND OF THE INVENTION The present invention relates to insuring adequate levels of intracellular glutathione. It has been suggested that by increasing intracellular glutathione levels certain benefits can be realized. Glutathione protects cells against free radicals, reactive oxygen intermediates, and toxic compounds that are both of endogenous and of exogenous origin. Meister, "New Aspects of Glutathione Biochemistry and Transport-Selective Alteration of Glutathione Metabolism", Nutrition Review, 42:397-410. The intracellular biosynthesis of glutathione is rate limited by cysteine. However, increasing cysteine per se is limited because of its instability, and the use of oxidized cystine is limited due to its low solubility. It has been proposed to infuse into a patient a precursor that will stimulate the intracellular synthesis of glutathione. To this end, N-acetylcysteine, a precursor of cysteine that is involved in the intracellular synthesis of glutathione has been infused, parenterally, in patients to raise the glutathione level. It is also known to infuse, parenterally, L-2-oxothiazolidine-4-carboxylate and glutathione esters to also stimulate intracellular glutathione production. See, U.S. Pat. Nos. 4,335,210; 4,434,158; 4,438,124; 4,647,571; 4,665,082; and 4,784,685. Although the parenteral infusion of cysteine precursors as well as glutathione esters is believed to be an effective way to increase or maintain a sufficient level of intracellular glutathione, it would of course be desirable if the intracellular glutathione level could be maintained or increased through an enteral diet. This is especially true in view of the fact that a number of chronic disease states exhibit reduced or below normal cellular glutathione levels. One of the difficulties in increasing through an enteral regimen intracellular glutathione levels is that it is not typically possible merely to provide an enteral amino acid solution rich in cysteine. Cysteine typically will crystallize out as cystine in solution, e.g., an amino acid solution. Cystine is not readily biologically available to cells. Therefore, cysteine is not biologically available as a pharmaceutical. Chung, et al., L-2-oxothiazolidine-4-carboxylate As A Cysteine Precursor: Efficacy for Growth and Hepatic Glutathione Synthesis in Chicks and Rats, American Institute of Nutrition (1990), sets forth experiments, by which they conclude, that orally administered L-2-oxothiazolidine-4-carboxylate is active as a cysteine precursor. In the experiments, they also administered, enterally, L-cysteine-HCl-H 2 O. The paper also noted that " i!t is well established that Cys cysteine! is toxic when it is provided in excess. Rats given Cys via intraperitoneal injection show signs of toxicity, whereas those given OTC (on an equimolar basis to Cys) do not." European published patent application 0 374 390 discusses a whey protein composition comprising a suitable concentrate that contains proteins in an essentially undenatured state. The application states that: "It was shown, in controlled experiments, for the first time, that whey protein feeding of mice specifically enhances the immune response to sheep red blood cells (SRBC) and their resistance to pneumococcal infection, inhibits the development of DMH-induced colon cancer and increases tissue glutathione (GSH) levels independently of its nutritional quality. The present invention shows the correlation between the undenatured conformation of whey protein concentrate (w.p.c.) and host immunoenhancement whereby chemical indices of denaturation are given and the demonstration that the same crucial role of molecular conformation (undenatured state) applies to GSH promotion, which is the other major biological activity of w.p.c. Equally important is the demonstration that another protein source such as egg white, with the same high cysteine content as w.p.c. does not enhance GSH synthesis, further demonstrating the specificity of w.p.c. with respect to the described biological activity. The GSH promoting activity of undenatured w.p.c. is sustained over time (3-4 months). Whey and whey protein have been utilized from time immemorial for nutritional purposes. In addition, whey was recommended in folk and ancient medicine for the treatment of various diseases.sup.(1,2) and, in one instance, lifetime feeding of hamsters with a whey protein diet has been shown to promote longevity with no explanation given.sup.(3,4). All these conditions appear to be somehow related to changes in glutathione which is a ubiquitous element exerting a protective effect against superoxide radicals and other toxic agents." (See page 2, lines 17-34.) The European application also states that the biological activity of the whey protein "is actually dependent on the undenatured conformation of the proteins" (see page 13, lines 21-23). Additionally, the European application states that "the administration of glutathione itself is of little consequence on tissue glutathione levels, because it apparently cannot be transported intact across the cell membrane" (see page 6, lines 23-24). The ability to give a composition orally that increases intracellular glutathione is highly desirable. However, all patient populations in need of same may not exhibit the same characteristics especially with respect to the bioavailability of the product to the patient. For patients having reduced or compromised gut function undenatured whey may not be absorbed in sufficient quantities so as to be readily bioavailable. Such patients with reduced gut function can include patients who suffer from: acquired immune deficiency syndrome (AIDS); Crohn's disease; chronic inflammatory bowel disease; short bowel syndrome; and inflammatory bowel reaction to radiation therapy. Not only do such patients typically have a reduced gut function, but, they may have reduced intracellular glutathione levels that should be elevated. SUMMARY OF THE INVENTION The present invention provides a method for insuring adequate intracellular glutathione levels in tissue. Pursuant to the present invention, a method for increasing intracellular glutathione levels in patients is provided comprising the steps of administering enterally to a patient having a compromised immune system a therapeutically effective amount of a solution including denatured proteins enriched in cysteine. In an embodiment of the invention, the solution includes hydrolyzed whey. In an embodiment of the invention, the solution includes hydrolyzed egg white protein. In an embodiment, the present invention includes a method for increasing intracellular glutathione levels in a patient comprising enterally administering to the patient a therapeutically effective amount of a solution including denatured whey or egg white protein. In an embodiment, a method for increasing glutathione levels in a patient is provided comprising the steps of administering enterally to the patient a sufficient amount of a denatured protein that includes at least 2.1% of its caloric content as cysteine. In an embodiment, a method for enhancing immune function in a patient is provided comprising administering enterally to the patient a solution having at least 0.37% of its caloric content as cysteine. An advantage of the present invention is that it provides a method for increasing glutathione levels in a patient. A further advantage of the present invention is that it provides a method for enhancing immune function of a patient. Additionally, an advantage of the present invention is that it provides a method for enhancing immune function of a patient who has a compromised immune function due to a disease state or trauma. Furthermore, an advantage of the present invention is that it provides a method for increasing glutathione levels that can increase glutathione levels to a greater extent than the administration of an intact protein. Still further, an advantage of the present invention is that it provides a method of treating stress in a patient. Furthermore, an advantage of the present invention is that it provides a method for treating intensive care patients. Additional features and advantages of the present invention are described in, and will be apparent from, the detailed description of the presently preferred embodiments. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1A illustrates graphically glutathione blood levels versus days for patients receiving a casein based formula pursuant to Example No. 1. FIG. 1B illustrates graphically glutathione blood levels versus days for patients receiving a denatured whey based formula pursuant to Example No. 1. FIG. 2A illustrates graphically glutathione blood levels versus days for patients receiving a casein based formula pursuant to Example No. 1. FIG. 2B illustrates graphically glutathione blood levels versus days for patients receiving a denatured whey based formula pursuant to Example No. 1. FIG. 3A illustrates graphically glutathione blood levels versus days for patients receiving a casein based formula pursuant to Example No. 1. FIG. 3B illustrates graphically glutathione blood levels versus days for patients receiving a denatured whey based formula pursuant to Example No. 1. FIG. 4 illustrates graphically glutathione blood levels versus days for patients receiving a casein or denatured whey product pursuant to Example No. 1. DETAILED DESCRIPTION OF THE PRESENTLY PREFERRED EMBODIMENTS The present invention provides a method for insuring adequate intracellular glutathione levels in tissue. Additionally, the present invention provides a method for increasing intracellular glutathione levels in a patient in need of same. The present invention can be used to treat stress patients, such as intensive care patients, patients having a compromised immune system, and/or patients having compromised gut function, for example. The method comprises, in an embodiment, enterally administering a therapeutically effective amount of a solution including completely denatured and at least partially hydrolyzed proteins enriched in cysteine. Because the proteins are denatured and at least partially hydrolyzed, they are readily absorbed, even in a patient having compromised gut function. In another embodiment, the method comprises administering to a patient a therapeutically effective amount of a solution including denatured protein and an amount of glutathione. The inventors have found, that by administering a hydrolyzed protein including a sufficient amount of cysteine, that it is possible not only to maintain sufficient levels of glutathione in patients in need of increased glutathione levels, indeed, it is possible to increase the intracellular glutathione levels in those patients who have a depressed level of glutathione. An advantage of present invention is that for a patient who requires increased levels of glutathione, it is possible to administer enterally the composition of the present invention and provide a sufficient level of intracellular glutathione. In fact, it is possible to increase the intracellular glutathione level to normal levels. Typically, for example, an immune-compromised or intensive care unit patient has below normal cellular levels of glutathione. It is believed that a patient who has decreased glutathione levels is more susceptible to many disease states. It is therefore important to insure that cellular glutathione levels are maintained at near normal levels, or increased to meet those levels. A number of such patients having reduced glutathione levels also have impaired or compromised gut functions. Examples of such patients include those suffering from: acquired immune deficiency syndrome (AIDS); Crohn's disease; chronic inflammatory bowel disease; short bowel syndrome; and inflammatory bowel reaction to radiation therapy. Due to the compromised gut function providing an intact protein, such as casein, would not provide a sufficiently bioavailable source of glutathione to the patient. By providing, in an embodiment, hydrolyzed protein enriched in cysteine, it is possible to insure adequate glutathione levels in such patients. Intensive care patients typically are under physical stress from an event that led to their admission. Some of such stress is associated with oxidative damage. Glutathione plays a major role in protecting the body from oxidative damage. Additionally, cysteine which is rate limiting for glutathione synthesis, in vivo, may be conditionally essential in intensive care patients. Especially with respect to intensive care patients, such patients may require special solutions to be fed, such as enteral solutions. Accordingly, the protein in such enteral products may be the sole source of a patient's cysteine. Although protein may be available from a variety of sources, casein and whey, the inventors have found that they provide to the patient significantly different levels of glutathione and cysteine in vivo; this is demonstrated by Example No. 1 set forth below. Pursuant to the present invention, the metabolically stressed patient is fed a formulation that includes denatured, and at least partially hydrolyzed, protein. The protein includes at least 2.1% by caloric content cysteine. By way of example, hydrolyzed whey provides 2.3% of its calories as cysteine. Because the protein is denatured and includes cysteine at such a level, the glutathione levels and cysteine levels in the patient receiving same will be increased sufficiently to restore physiological glutathione levels and enhance immune function. Pursuant to the present invention, preferably, denatured whey protein is administered to the patient. However, other products can be administered, such as hydrolyzed egg white protein. A metabolically stressed patient should be provided with a minimum intake of cysteine of at least 0.3 grams (300 milligrams). This will maintain physiological levels of glutathione. However, if the patient's glutathione levels are depleted, pursuant to the present invention, the patient should be administered at least 1.2 grams (1,200 milligrams) of cysteine daily to restore depleted glutathione levels to normal physiological levels. For a patient having depleted glutathione levels, this requires providing a formulation that includes at least 0.37% of its calories as cysteine. In an embodiment of the present invention, whey protein hydrolysate is administered to a patient. An example of an enteral diet that can be administered to the patient is PEPTAMEN®, Clintec Nutrition Company, Deerfield, Ill. PEPTAMEN® includes whey protein hydrolysate that provides 0.37% of the total calories of the product as cysteine. The peptide distribution of PEPTAMEN® is as follows: ______________________________________Peptide Size:(number of amino Approxiinate Percentageacid residues) molecular weight by number______________________________________ 1 (free amino acid) 133 12%2-4 100-500 17%5-9 500-1000 38%10-40 1000-4600 28%>40 >4600 5%______________________________________ Average peptide size: 8 amino acid residues. The nutrient composition of PEPTAMEN® Liquid Elemental Diet is as follows: __________________________________________________________________________NUTRIENT INFORMATION Serving Sie One can (500 ml) PER 500 ml PER 2000 ml AMOUNT % U.S. RDA AMOUNT % U.S.RDA__________________________________________________________________________NUTRIENTCOMPOSITIONCALORIES kcal 500 ** 2000 **PROTEIN g 20.0 44 80 178CARBOHYDRATE g 63.5 ** 254 **FAT** g 19.5 ** 78 **Vitamin CompositionVITAMIN A IU 1875 37 7500 150VITAMIN D IU 100 25 400 100VITAMIN E IU 10 33 40 133VITAMIN K mcg 62.5 ** 250 **VITAMIN C mg 50 83 200 333THIAMINE (B.sup.1) mg 0.75 50 3 200RIBOFLAVIN (B.sub.2) mg 0.85 50 3.4 200NIACIN mg 10 50 40 200VITAMIN B.sub.6 mg 1.5 75 6 300FOLIC ACID mcg 200 50 800 200PANTOTHENIC ACID mg 5 50 20 200VITAMIN B.sub.12 mcg 3 50 12 200BIOTIN mcg 150 50 600 200CHOLINE mg 225 ** 900 **__________________________________________________________________________ Additionally, glutathione can be administered with the hydrolyzed protein to further assist in increasing intracellular glutathione levels. The glutathione can be added to the hydrolyzed protein that is enriched in cysteine. In an embodiment, approximately 1 to about 2.8 grams of glutathione is added per milliliter of PEPTAMEN®. By way of example, but not limitation, an experiment demonstrating the present invention will now be given. EXAMPLE NO. 1 Study Design This study was an open label, unblinded pilot study of the effects on whole blood glutathione and cysteine concentration of enteral feeding of critically ill patients. The study compared two enteral feeding formulas that differed only in the source of the protein component of the enteral feeding formula. Study Population Inclusion Criteria: 1. Greater than 18 years of age, 2. Expected to require at least eight days of enteral nutrition support, 3. Informed consent obtained from the patient or family. Exclusion Criteria: 1. Less than 18 years of age, 2. Unable to obtain informed consent, 3. Not expected to require enteral nutrition support for at least eight days. Patients enrolled in the study have the following general characteristics: 1. Closed head injury with GCS score of 4-8, and/or 2. Multiple trauma with multiple rib fractures and pulmonary contusions requiring mechanical ventilation, and/or 3. Twenty to fifty percent 2nd and 3rd degree burns, and/or 4. Multiple fractures requiring ventilatory support, and 5. Expected to require enteral nutrition support for a minimum of eight days. Experimental Procedures Nutrition Support. The calculated goal rate for enteral nutrition support was 1.5 gm protein/kg/day and 30-35 non protein kcal/kg/day. These goal rates are those currently employed for critically ill patients. Patients were advanced to the goal feeding rate as the feeding was tolerated according to routine clinical guidelines such as abdominal distention and gastric or esophageal reflux. Patients for evaluation had to have received a minimum of eight days of enteral nutrition support. Daily intakes and clinical signs of intolerance to the enteral feeding were recorded. Glutathione and Cysteine Measurements. Three (3) ml blood samples were drawn prior to initiation of enteral feeding and every two days during the duration of enteral feeding support. Whole blood samples were prepared by a standard procedure developed by the Metabolic Assessment Laboratory at the University of Florida, Gainsville. Assays for whole blood glutathione and cysteine were performed using a HPLC procedure at the Metabolic Assessment Laboratory. Glutathione concentration as determined in whole blood was corrected for the patient's hematocrit and cysteine concentrations was corrected for the patient's "plasmacrit" (1-Hct). Results Baseline Analysis. Patient Characteristics. The patient characteristics at study entry thought to be of significance to glutathione status of critical care patients are presented below. ______________________________________ MEAN (range) YES NO______________________________________Age 48 (28-70)Injury severity score 23.8 (7-41)Days between injury and 19.5 (5-49)study entryOrgan dysfunction at study 0 11entryOn mechanical ventilation 8 3Septic at study entry 9 2Nutrition prior study entry 11 0Nutritionally deprived for 7 4greater than 3 daysGlutathione and cysteineconcentrations.______________________________________ Based on previous studies (unpublished) normal hematocrit corrected blood glutathione values are approximately 2300±300 micromoles/ml. As can be seen from Table 1 (below), eight of the fourteen patients analyzed in this study had glutathione concentrations within or above this range. Two patients (Patients 8 and 12) had greatly reduced glutathione concentrations in the range of those reported in severely stressed ARDS patients. Based on the same prior studies, the normal "plasmacrit" corrected blood cysteine concentrations are approximately 210±40 micromoles/ml. As seen in Table 1, six of the fourteen patients analyzed in this study had cysteine concentrations that were below this normal range. There was no clear correspondence between patients with low glutathione values and patients with low cysteine concentrations. TABLE 1______________________________________ GSH day 1 day 3 day 5 day 7 day 9 day 11______________________________________ 1 whey 2000.0 2060.2 2044.5 1659.1 1555.1 2179.5 1" whey 1653.3 2036.5 2415.1 2928.3 2400 2 whey 2037.7 3306.9 2000.0 2689.8 2540.1 2062.5 3 whey 2202.2 2368.8 2142.9 2222.9 2845.9 2530.6 4 whey 1240.3 1770.0 2281.3 2180.9 2619.6 1743.8 5 whey 1794.4 1715.3 2123.6 1930.1 1912.4 1395.312 whey 859.0 817.3 1381.5 1568.6 1356.0 1448.818 whey 2729.8 2639.7 2659.8 1839.3 2573.0 1596.8 14516.7 16714.8 17048.3 17018.9 17802.5 13743.8Mean whey 1814.6 2089.3 2131.0 2127.4 2225.3 1963.4 100.0 115.1 117.4 117.2 122.6 108.2 7 casein 1759.4 2186.7 1757.9 1677.8 2166.7 2568.2 8 casein 435.5 532.4 460.4 557.4 933.6 1026.710 casein 3611.9 2531.6 2080.8 2041.3 2273.6 3184.114 casein 2163.9 2126.2 226.6 2341.1 2547.4 1587.816 casein 2068.2 2090.6 1850.0 1948.6 1313.0 1541.120 casein 2274 2199 1824 1566 12313 11669 10202 10132 9234 9908Mean casein 2052 1945 1700 1689 1847 1982 100.0 94.8 82.9 90.0 96.6 77.3______________________________________ day 13 day 15 day 17 day 19 day 21______________________________________ 1 whey 1" whey 2 whey 2294.3 1894.3 2128.6 3080.0 3 whey 2429.9 2689.0 2290.6 2288.8 4 whey 1762.3 1827.7 5 whey12 whey 2154.2 1448.818 whey 8640.6 7859.7 4419.2 5368.8Mean whey 2160.2 1964.9 2209.6 2684.4 119.0 108.3 121.8 147.9 7 casein 2835.6 8 casein 766.7 910.0 1316.7 1393.3 1216.710 casein14 casein 955.1 1454.8 1327.8 1660.516 casein 1787.3 2025.420 casein 6345 4390 2644 3054 1217Mean casein 1586 1463 1322 1527 1217 77.3 71.3 64.4 74.4 59.3______________________________________ Protein and Cysteine Intake. At the protein intakes recorded in this study, the daily intake of cysteine was in the range of 1.45 to 1.85 grams per day in the patients receiving the hydrolyzed whey protein based diet (PEPTAMEN® elemental diet). Patients on the casein based diet (Nutren) received in the range of 0.13 to 0.23 grams of cysteine per day. Summary of Analyses Analysis of variance with repeated measures was used to analyze the data. The independent variables in the analysis were treatment (whey-vs. casein based diets), days on study, and the treatment by day interaction. Three analyses were performed; these analyses are illustrated graphically in FIGS. 1-3 specifically the figures illustrate the results graphically for: all patients in the study (FIGS. 1A and 1B); all patients who were in the study for at least 11 days (FIGS. 2A and 2B); and all patients who were in the study for at least 15 days (FIGS. 3A and 3B). Table 2 summarizes the p-values from these analyses. TABLE 2______________________________________Summary of p-values From Analysisof Variance with Repeated Measures Days in Treatment Treatment the by DayAnalysis Group study Interaction______________________________________All patients 0.35 0.95 0.46At least 11 days 0.43 0.94 0.78At least 15 days 0.10 0.55 0.57______________________________________ Table 3 (below) summarizes the whole blood GSH levels and Table 4 (below) summarizes the change from baseline of whole blood GSH for both groups. FIG. 4 graphically summarizes these same data comparing the results for a denatured and hydrolyzed whey diet versus an intact casein diet. TABLE 3______________________________________Summary of Whole Blood GSHTRT DAY N MEAN STD MIN MAX______________________________________casein 1 6 2052.00 1019.43 435 3612 3 6 1944.83 709.83 532 2532 5 6 1700.33 633.06 460 2229 7 6 1688.67 618.59 557 2341 9 5 1847.00 688.01 934 2547 11 5 1981.60 873.20 1027 3184 13 4 1586.25 943.72 767 2836 15 3 1463.33 557.55 910 2025 17 2 1322.50 7.78 1317 1328 19 2 1526.50 188.80 1393 1660 21 1 1217.00 1217 1217whey 1 8 1814.50 578.45 859 2730 3 8 2089.25 729.85 817 3307 5 8 2131.13 372.30 1381 2660 7 8 2127.38 481.49 1569 2928 9 8 2225.25 547.02 1356 2846 11 7 1963.43 399.54 1395 2531 13 4 2160.00 288.27 1762 2430 15 4 1965.00 520.97 1449 2689 17 2 2210.00 114.55 2129 2291 19 2 2684.50 559.32 2289 3080______________________________________ TABLE 4______________________________________Summary of GSH - Change From BaselineTRT DAY N MEAN STD MIN MAX______________________________________casein 1 6 0.000 0.00 0 0 3 6 -107.167 509.78 -1080 430 5 6 -351.667 609.40 -1531 65 7 6 -363.333 669.95 -1571 177 9 5 -160.600 835.68 -1338 499 11 5 -26.000 669.75 -576 809 13 4 -20.250 967.67 -1209 1077 15 3 -92.333 593.54 -709 475 17 2 23.000 1214.81 -836 882 19 2 227.000 1033.79 -504 958 21 1 782.000 782 782whey 1 8 0.000 0.00 0 0 3 8 274.750 460.07 -90 1269 5 8 316.625 423.51 -70 1041 7 8 312.875 715.77 -891 1275 9 8 410.750 568.59 -445 1380 11 7 125.857 776.78 -1133 1376 13 4 575.250 497.79 228 1295 15 4 380.250 352.79 -144 590 17 2 90.000 1.41 89 91 19 2 564.500 675.29 87 1042______________________________________ ______________________________________Listing of Whole Blood GSH DataTreatment PatientGroup ID Day GSH Day 11 Day 15______________________________________whey BM4 1 1240 * * 3 1770 * * 5 2281 * * 7 2181 * * 9 2620 * * 11 1744 * * 13 1762 * * 15 1828 * *whey JC1 1 2000 * 3 2060 * 5 2045 * 7 1659 * 9 1555 * 11 2179 *whey JC21 1 1653 3 2036 5 2415 7 2928 9 2400whey JG5 1 1794 * 3 1715 * 5 2124 * 7 1930 * 9 1912 * 11 1395 *whey JH2 1 2038 * * 3 3307 * * 5 2000 * * 7 2690 * * 9 1540 * * 11 2063 * * 13 2294 * * 15 1894 * * 17 2129 * * 19 3080 * *whey MS12 1 859 * * 3 817 * * 5 1381 * * 7 1569 * * 9 1356 * * 11 2235 * * 13 2154 * * 15 1449 * *______________________________________ *This patient's data included in analysis. ______________________________________List of Whole Blood GSH DataTreatmentGroup Patient ID Day GSH Day 11 Day 15______________________________________whey RH3 1 2202 * * 3 2369 * * 5 2143 * * 7 2223 * * 9 2846 * * 11 2531 * * 13 2430 * * 15 2689 * * 17 2291 * * 19 2289 * *whey WM18 1 2730 * 3 2640 * 5 2660 * 7 1839 * 9 2573 * 11 1597 *______________________________________ *This patient's data included in analysis. As illustrated graphically in FIG. 4, whole blood glutathione levels were increased and stayed increased using denatured and hydrolyzed whey protein as compared to intact protein (casein). Thus, the present invention provides a method for increasing glutathione levels, in vivo. As compared to the use of intact protein, the method of the present invention increases glutathione levels and maintains the increased levels. By way of example, but not limitation, contemplative examples of the present invention will now be given. Contemplative Example 1 A 32-year-old white male has been known to be HIV positive for seven years. He was essentially free of symptoms until two years ago when he had a complicated medical course following an upper respiratory tract infection. During the course of this illness he lost approximately 15% of his usual body weight. Following recovery from this illness, he was relatively free of symptoms except for occasional episodes of diarrhea of unknown etiology. Despite dietary and nutritional counseling he was unable to regain significant weight due, in part, to compromised gut function. The use of a hydrolyzed whey protein dietary supplement was recommended to aid in increasing nutrient intake and to aid in absorption of nutrients. He was able to drink approximately one liter of this supplement increasing his calorie intake by approximately 1000 kcal per day and to increase his protein intake by 40 grams per day. He gained six pounds in the first two weeks of dietary supplementation and continued to gain two to three pounds between biweekly clinic visits despite somewhat reduced intake of the supplement. He remained symptom-free and has had no further episodes of diarrhea. Plasma glutathione and cysteine levels were found to be at the levels seen in normal healthy individuals at 4 weeks following initiation of supplementation. Contemplative Example 2 A 54 -year-old white male has known to be HIV positive for five years. In the last six months he has had several opportunistic infectious episodes including two lung infections that required hospitalization. During the second hospitalization he was placed on total parenteral nutrition (TPN) for eight days to increase nutrient intake and restore a significant weight loss (18% of usual body weight). Before discharge from the hospital he was transitioned to oral intake using an enteral feeding formula based on hydrolyzed whey protein. He continued to use the preparation as an occasional dietary supplement after release from the hospital. During the period when he received TPN, a plasma amino acid profile was performed. Among several deviations from the normal amino acid profile was a 90% reduction in plasma cysteine concentration. This observation was followed by an analysis of plasma glutathione and white blood cell glutathione concentrations. Both plasma and white blood cell glutathione concentrations were less than 10% of normal values. These analyses were repeated one week after hospital discharge following two weeks of oral intake of the enteral supplement. At this time both plasma cysteine and glutathione were at the low end of normal values. White blood cell glutathione levels were at approximately 60% of normal values. It should be understood that various changes and modifications to the presently preferred embodiments described herein will be apparent to those skilled in the art. Such changes and modifications can be made without departing from the spirit and scope of the present invention and without diminishing its attendant advantages. It is therefore intended that such changes and modifications be covered by the appended claims.

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Patent Citations (51)

    Publication numberPublication dateAssigneeTitle
    US-4981704-AJanuary 01, 1991Union Des Cooperatives Laitieres D'isigny-Sur-Mer Et De Sainte-Mer-EglisePartial hydrolysate of whey proteins, enzymatic process for the preparation of this hydrolysate, and hypoallergenic dietetic milk food containing it
    US-4001437-AJanuary 04, 1977Givaudan CorporationProcess for making and flavorants from milk products and compositions containing same
    US-4016293-AApril 05, 1977Coughlin Robert W, Marvin CharlesMethod of carrying out enzyme catalyzed reactions
    US-5158883-AOctober 27, 1992Cornell Research Foundation, Inc.Method of using aminoarginine to block nitric oxide formation in vitro
    US-4752618-AJune 21, 1988New England Deaconess HospitalMethod of minimizing efects of infection through diet
    US-3755578-AAugust 28, 1973PfizerThiazoline and 5,6-dihydro-4h-1,3-thiazine anti-plant-viral agents
    US-5002953-AMarch 26, 1991Beecham Group P.L.C.Novel compounds
    US-2712202-AJuly 05, 1955Univ AlabamaMethod of treating tree diseases
    WO-9311104-A1June 10, 1993Ab AstraSels organiques de cystine de n, n'-diacetyle
    US-4868114-ASeptember 19, 1989Regents Of The University Of MinnesotaMethod for elevating glutathione levels
    US-5053387-AOctober 01, 1991Shriners Hospitals For Crippled ChildrenOmega-3 fatty acids in traumatic injury treatment
    US-4775675-AOctober 04, 1988Biogal GyogyszergyarThiazolidinecarboxylic acid derivatives and treatment of liver diseases therewith
    US-4175130-ANovember 20, 1979Yoshitomi Pharmaceutical Industries, Ltd.Oxazole- and thiazole-alkanoic acid compounds
    US-4798835-AJanuary 17, 1989Pfizer Inc.dl-5-[(2-benzyl-3,4-dihydro-2H-benzopyran-6-yl)methyl]thiazolidine-2,4-dione as an anti-atherosclerosis agent
    US-5039609-AAugust 13, 1991Research Technologies, Inc.Osmotic agents for peritoneal dialysis
    US-5208249-AMay 04, 1993Clintec Nutrition Co.Method for stimulating intracellular synthesis of glutathione using esters of L-2-oxothiazolidine-4-carboxylate
    US-4636388-AJanuary 13, 1987Stauffer Chemical CompanyPreparing protein for hydrolysis and product
    US-4482574-ANovember 13, 1984Stauffer Chemical CompanyProcess for the preparation of protein for hydrolysis
    US-4563471-AJanuary 07, 1986Warner-Lambert Company4-Oxothiazolidin-2-ylidene-acetamide derivatives as CNS agents
    US-5055446-AOctober 08, 1991University Of CincinnatiMethod to improve survival of patients during sepsis by diet composition
    US-3737536-AJune 05, 1973Bayer AgAnaesthetic compositions for and use in animals
    WO-9114424-A1October 03, 1991Yunis Adel A, Jimenez Joaquin J, Hussein Atif MMethods for protecting against chemotherapy-induced hair loss
    US-4963577-AOctober 16, 1990Behringwerke AktiengesellschaftUse of the thiazole derivative tiprotimod for the preparation of an agent for the therapy of virus infections
    US-4398026-AAugust 09, 1983Taisho Pharmaceutical Co., Ltd.4-Carboxy-2-acetamido thiazolidine derivatives
    US-4438124-AMarch 20, 1984Cornell Research Foundation, Inc.Cysteine delivery system
    US-4434158-AFebruary 28, 1984Cornell Research FoundationCysteine delivery system
    US-5089268-AFebruary 18, 1992Katz David PEgg phosphatide lipid emulsions altered for a specific therapeutic fatty acid composition
    EP-0002978-A2July 11, 1979SynthelaboDérivés de thiazolidinedione-2,4, leur préparation et leur application en thérapeutique
    US-4791125-ADecember 13, 1988Pfizer Inc.Thiazolidinediones as hypoglycemic and anti-atherosclerosis agents
    US-5028627-AJuly 02, 1991Cornell Research Foundation, Inc., Board Of Regents, The University Of Texas SystemMethod of using arginine derivatives to inhibit systemic hypotension associated with nitric oxide production or endothelial derived relaxing factor
    US-4780475-AOctober 25, 1988Cerra Frank B, Amen Ronald JPreparation for the prevention of catabolism
    US-4839387-AJune 13, 1989Poli Industria Chimica S.P.A.Derivative of thiazolidine-4-carboxylic acid, its preparation and pharmaceutical compositions containing it
    US-5095027-AMarch 10, 1992Clintec Nutrition Co.Method for treating reperfusion injury employing L-2-oxothiazolidine-4-carboxylic acid
    EP-0374390-A1June 27, 1990Immunotech Research Corporation Ltd.Composition de protéines de petit lait, procédé de production et application de la composition de protéines de petit-lait
    US-4665082-AMay 12, 1987Cornell Research FoundationCysteine delivery system
    EP-0318330-A2May 31, 1989Yamanouchi Pharmaceutical Co., Ltd.Glutathione esters for treating ventricular arrhythmia
    US-4710489-ADecember 01, 1987Cornell Research Foundation, Inc.Glutathione delivery system
    US-4427658-AJanuary 24, 1984Institut National De La Recherche AgronomiqueTotal enzymatic hydrolysate from whey proteins and process of obtaining the same
    EP-0373002-A2June 13, 1990Allergan, IncUse of 2-substituted-thiazolidine-4-carboxylic acids for treatment of cataract
    US-4784685-ANovember 15, 1988Cornell Research Foundation, Inc.Glutathione delivery system
    EP-0257992-A2March 02, 1988Yamanouchi Pharmaceutical Co., Ltd.Vorbeugende und therapeutische Mittel für zerebrale Ischämie
    US-4647453-AMarch 03, 1987Peritain, Ltd.Treatment for tissue degenerative inflammatory disease
    WO-8403625-A1September 27, 1984Baxter Travenol LabMonoesters nutritifs
    US-4338315-AJuly 06, 1982Eli Lilly And CompanyAntiviral method employing thiazinyl benzimidazole derivatives
    US-4335210-AJune 15, 1982Cornell Research FoundationMethod of producing L-cysteine
    US-4647571-AMarch 03, 1987Cornell Research FoundationCysteine delivery composition
    EP-0327263-B1September 07, 1994PROCTOR, Peter H.Hair growth stimulation with nitroxide and other radicals
    EP-0415598-A1March 06, 1991Unilever Plc, Unilever N.V.Cosmetic composition
    US-4420479-ADecember 13, 1983Eli Lilly And CompanyOlefinic benzimidazoles, formulations, and antiviral methods
    EP-0338459-A2October 25, 1989Trustees Of Boston University, Angio-Medical CorporationComposition for promoting hair growth in androgenetic alopecia and method thereof
    DE-2141765-A1March 01, 1973Henkel & Cie Gmbh4-chloro-4-thiazolin-2-ones - with microbicidal properties

NO-Patent Citations (88)

    Title
    A. Cantin et al., Glutathione Deficiency in the Epithelial Lining Fluid of the Lower Respiratory Tract in Idiopathic Pulmonary Fibrosis, Am. Rev. Respir. Dis. 1989; 139:370 372.
    A. Cantin et al., Normal Alveolar Epithelial Lining Fluid Contains High Levels of Glutathione, pp. 152 157.
    A. Cantin et al., Oxidants. Antioxidants and the Pathogenesis of Emphysema, Eur. J. Respi. Dist (l985) 66, Suppl. 139. pp.7 17.
    Abate, et al. Redox Regulation of Fos and jun DNA Binding Activity in Vitro, Science, Sep. 1990, vol. 249, pp. 1157 1161.
    American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference Committee, American College of Chest Physicians/Society of Critical Care Medicine Consensus Conference: Definitions for Sepsis and Organ Failure and Guidelines for the Use of Innovative Therapies in Sepsis, Critical Care Medicine, Jun. 1992, vol. 20, No. 6, pp. 864 874.
    Angier, From the Body Itself, Hope for a New Breed of Potent Antibiotics, New York Times, Feb. 26, 1991, pp. 26 27.
    Astor et al, Relationship, Between Intracellular GSH Levels and Hypoxic Cell Radiosensitivity, Pharmac. Ther., vol. 39, pp. 115 121 (1988).
    Bellin, et al., Purification of Glycosaminoglycens from Bovine Follicular Fluid, J. Dairy Sci., No. 9, 1987, vol. 70, pp. 1913 1919.
    Bjelton, et al. Availability of Cysteine and of L 2Oxo Thiazolidine4 Carboxylic Acid as a Source of Cysteine in Intravenous Nutrition, J. Parenter Enteral Nutr. Mar Apr. 1990, vol. 14, No. 2, pp. 177 182. (Abstract).
    Bone, et al. Definitions for Sepsis and Organ Failure, Critical Care Medicine, 1992, vol. 20, No. 6, pp. 724 725.
    Bounous et al. Immunoenhancing Property of Dietary Whey Protein in Mice: Role of Glutathione , Clin. Invest. Med., vol. 12, No. 3, pp. 154 161.
    Bounous et al., The influence of dietary where protein on tissue glutathione and the diseases of aging, Clinical Invest. Med., vol. 12, No. 6, pp. 343 349, 1989. (Abstract).
    Calvin, et al., Estimation and Manipulation of Glutathione Levels in Prepuberal Mouse Ovaries and Ova: Relevance to Sperm Nucleus Transformation in the Fertilized Egg, Gamete Research, 1986, vol. 14, pp. 265 275.
    Chang, et al., Cytoprotective Effect of Reduced Glutathione in Arsenical Iduced Endothelial Cell injury, Toxicology, 1991, vol. 69, No. 1, pp. 101 110. (Abstract).
    Chung et al, L 2 Oxothiazolidine 4 Carboxylate as a Cysteine Precursor: Efficacy for Growth and Hepatic Glutathione Synthesis in Chicks and Rats, American Institute of Nutrition, pp. 158 165 (1989).
    Clintec Nutrition Company, Carnation Liquid Elemental Diet Flavoring Ideas (1988).
    Clintec Nutrition Company, Evaluation of a Peptide Based Elemental Diet versus an Amino Acid Based Elemental Diet in Geriatric Tube Fed Patients (1987).
    Clintec Nutrition Company, Peptamen Brochure (1987).
    Clintec Nutrition Company, Peptamen Brochure (1990).
    Clintec Nutrition Company, The Case for a Balance Energy Substrate Formula (1988).
    Clintec Nutrition Company, The Case for Peptides in a Defined Formula (1988).
    Clintec Nutrition Company, Use of PEPTAMEN Liquid, Isotonic, Complete, Elemental Diet in a Patient with a High Output Ostomy (1987).
    Clintec Nutrition Company, Use of PEPTAMEN Liquid, Isotonic, Complete, Elemental Diet in The Transition from TPN and Long Term Management on Enteral Feeding (1987).
    Clintec Nutrition Company. Carnation Peptamen Liquid, Isotonic, Complete, Elemental Diet: Clinical Rationale (1987).
    Cretton, et al., Catabolism of 3 Axido 3 Deoxythymidine in Hepactocytes and Liver Microsomes, with Evidence of Formation of 3 Amino 3 Deoxythmidine, a Highly Toxic Catabolite for Human Bone Marrow Cells, Molecular Pharmacology, vol. 39, pp. 258 266, 1991.
    Darley Usmar, et al. Oxygen and Reperfusion Damage: an Overview, Free Radi. Res. Comms., vol. 7, No. 36, pp. 247 254, 1989.
    Duh, et al. Tumor Necrosis Factor Activates Human Immunodeficiency Virus Type 1 Through Induction of Nuclear Factor Binding to the NF K B Sites in the Long Terminal Repeat, Proc. Natl. Acad. Sci. USA, Aug. 1989, vol. 86, pp. 5974 5978.
    Feller et al, Effects of Three Liquid Diets on Nutrition Sensitive Plasma Proteins of Tube Fed Elderly Men, J. of Am. Geriatrics Soc y, No. 6, pp. 663 668 (1990).
    Flaherty, et al., Reperfusion Injury, Free Radical Biology & Medicine, 1988, Vol. 5, pp. 409 419.
    Frankova, The Effects of Amino Acids with Sulfhydryl Group on Herpest Viruses in Vitro, Acla Virol, Engl. Ed. (1967), vol. 11, No. 6, pp. 559 561.
    G. Bernard et al., Effect of N Acetylcysteine on the Pulmonary Response to Endotoxin in the Awake Sheep, and Upon In Vitro Granulocyte Function, J. Clin. Invest., vol. 73, Jun. 1984, pp. 1772 1784.
    Gordon et al., Applications of Micromanipulation to Human in Vitro Fertilization, Journal of In Vitro Fertilization and Embryo Transfer, 1988, vol. 5, No. 2, pp. 57 60.
    Guarding Against Cellular Glutathione Deficiency, Nutritional Reviews, vol. 48, No. 9, pp. 346 348 (1990).
    Gustafson, et al. Aids Antiviral Sulfolipids from Cyanobacteria (Blue Green Algae), J. Natl. Cancer Inst., 1989, vol. 81, No. 16, pp. 1254 1258. (Abstract).
    Handlon, et al. Thiol Reduction of 3 Aziodthymidine to 3 Aminothymidine: Kinetics and Biomedical Implications, Pharmaceutical Research, vol. 5, No. 5, pp. 297 299, 1988.
    Heinecke, et al. The Role of Sulfur containing Amino Acids in Superoxide Production and odification of Low Density Lipoprotein by Arterial Smooth Muscle Cells, The Journal of Biologicla Chemistry, 1987, Vool. 262, No. 21, pp. 10098 10103.
    I. Cotgreave et al. Lung and Systemic Thiol Homeostasis During an Acute Lung Inflammation in the Rat, Toxicology, 50 (1988), pp. 331 343.
    J. Strausz et al. Oxygen Radical Production bv Alveolar Inflammatory Cells in Idiopathic Pulmonary Fibrosis, Am. Rev. Respir. Dis. 1990; 141:124 128.
    J. Sun et al. Effects of Buthionine Sulfoximine on the Development of Ozone Induced Pulmonary Fibrosis, Experimental and Molecular Pathology 49 (1988), pp. 254 266.
    Jimenez, et al. Treatment with ImuVert/N Acetylcysteine Protects Rats from Cyclophosphamide/Cytarabine Induced Alopecia, Cancer Investigation, 1992, vol. 10, No. 4, pp. 271 276.
    Kalebic, eta l., Suppression of human immunodeficiency virus expression in chronically infected moncytic cells by glutathione, glutathione ester, and N acetycysteine, Proc. Natl. Acad. Sci, USA, 1991. (Abstract).
    Keller, et al. Decreased Hepatic Gluthatione Levels in Septic Shock, Predisposition of Hepatocytes to Oxidative Stress: an Experimental Approach, Arch. Surg. (Chicago), 1985, vol. 120, No. 8, pp. 941 945. (Abstract).
    Kermici, et al., Evidence for an Age Correlated Change in Glutathione Metabolism Enzyme Activities in Human Hair Follicle, Mechanisms of Aging and Development, 1990, vol. 53, pp. 73 84.
    Kilbourn et al., Inhibition of Interleukin 1 Induced nitric Oxide Synthase in Vascular Smooth Muscle and Full Reversal of Interleukin 1 induced Hypotension by N 107 Amino L Argine, Journal of the National Cancer Institute, Jul. 1, 1992, vol. 84, No. 13, pp. 1008 1016.
    Kuzuya, et al., Protective role of intracellular glutathione against oxidized low density lipoprotein in cultured endothelial cells, Biochem. Biophys. Res. Commun 163 (3) 1989 (Abstract).
    L. Smith et al. Effect of Fasting on Hyperoxic Lung Injury in Mice, Am. Rev. Respir. Dis. 1990; 141:141 149.
    Lachman, et al. The theory and Practice of Industrial Pharmacy, 1976, 2d Ed., pp. 513 524.
    Lamperth, et al. Abnormal Skeletal and Cardiac Muscle Mitochondria Induced by Zidovudine (AZT) in Human Muscle in vitro and in an Animal Model, Laboratory Investigation, vol. 65, No. 6, pp. 742 752, 1991.
    Lasalle, et al., Relationship Between Fertilizing Ability of Frozen Human Spermatozoa and Capacity for Heparin Binding and Nuclear Decondensation, J. Reprod. Fertil, 1992, vol. 95, No. 2, pp. 313 324.(Abstract).
    Levy, et al., Transport of Glutathione Diethyl Ester into Human Cells, Proc. Natl. Acad. Sci., USA, Oct. 1993, vol. 90, pp. 9171 9175.
    M.A. Passero et al., L 2 Oxothiazolidine 4 Carboxylic Acid Increases Glutathione in Mouse Lung, A. Rev. Respir. Dis. vol. 133, 1986, p. A395. (Abstract).
    M.F. Tsan et al. Enhancement of Intracellular Glutathione Protects Endothelial Cells Against Oxidant Damage, Biochemical and Biophysical Research Communications, vol. 127, No. 1, Feb. 28, 1985, pp. 270 276.
    M.F. Tsan et al. L 2 Oxothiazolidine 4 Carboxylate Protects Endothelial Cells Against Hperopia Induced Injury, Inflammation, vol. 12, No. 2, 1988, pp. 113 121.
    Mandel, et al. Intracellular Glutathione in the Protection from Anoxic injury in Renal Proximal Tubles, J. Clin. Invest., 1990, vol. 85, No. 2, pp. 316 324. (Abstract).
    Martensson et al, Glutathione Metabolism in the Lung: Inhibition of its Synthesis Leads to Lamellar Body an Mitochondrial Defects, Proc. Natl. Acad. Sci. USA, vol. 86, pp. 5296 5300 (1989).
    Martensson et al, Mitochondrial Damage in Muscle Occurs After Marked Depletion of Glutathione and is Prevented by Giving Glutathione Monoester, Proc. Natl. Acad. Sci. USA, vol. 86, pp. 471 475 (1989).
    Meister, Glutathione Esters Increase Cellular Glutathione Levels and are Thus Protective Against Oxidants and Other Compounds, Abstract, (undated).
    Mihm et al. Inhibition of HIV 1 Replication and NF X B activity by Cysteine and Cysteine Derivatives, Aids 1991, vol. V, No. 5, pp. 497 503.
    Moslen, Protection by L 2 Oxothiazolidine 4 Carboxylate, a Cysteine Prodrug, Against 1.1 Dichlorethylene Hepatotoxicity in Rats is Associated with Decreased in Toxin Metabolism and Cytochrome P 450, J. Pharmacol. Exp. Ther., vol. 248, No. 1, pp. 157 63. (Abstract).
    Nappe, et al, Electrophoretic Analysis of Alkylated Proteins of Human Hair from Various Ethnic Groups, J. Soc. Cosmet, Chem., Mar./Apr. 1989, vol. 40, pp. 91 99.
    Oeriu, et al. 4 Thiazolidinecarboxylic Acids for Live Stock Raising, Ger. Offen, Oct. 22, 1970, p. 17. (Abstract).
    Osol, et al. Eds., Reminington s Pharmaceutical Sciences, 1980, Sixteenth Edition.
    P.H.S. Sporn et al. Complex Effects of In Vitro Hyperoxia on Alveolar Macrophage Arachidonic Acid Metabolism, American Journal of Respiratory Cell and Molecular Biology, vol. 2, No. 1, Jan. 1990, pp. 81 90.
    Pacht, et al. Deficiency of Alveolar Fluid Glutathione in Patients with Sepsis and the Adult Respiratory Distress Syndrome, Chest, 1991, vol. 100, No. 5, pp. 1397 1403 (Abstract).
    Parthasarathy, Oxidation of low density lipoprotein by thiol compounds leads to its recognition by the acetyl LDL receptor, Biochimica et Biophysica Acta, 917, 1987, pp. 337 340.
    Peristeris, et al. N Acetylcystein and Glutathione as Inhibitors of Tumor Necrosis Factor Production, Cell. Immunol., 1992, vol. 140, No. 2, pp. 390 399. (Abstract).
    Perreault, et al. Importance of Glutathione in the Acquisition and Maintenance of Sperm Nuclear Decondensing Activity in Maturing Hamster Oocytes, Developmental Biology, 1988, vol. 125, pp. 181 186.
    Perreault, et al. The Role of Disulfide Bond Reduction During Mammalian Sperm Nuclear Decondensation in Vivo, Developmental Biology, 1984, vol. 101, pp. 160 167.
    Perreault, et al. The Timing of Hamster Sperm Nuclear Decondensation and Male Pronucleus Formation is Related to Sperm Nuclear Disulfide Bond Content, Biology of Reproduction, 1987, vol. 36, pp. 239 244.
    Polk et al, Intermittent Administration of a Defined Formula Diet Induces Growth in Adolescents With Crohn s Disease and Growth Failure, Pediatric Res. Abstract No. 664, 113A (1990).
    Prendergast et al., Arachidonic Acid Binding Peptides, Antibodies Produced to these Peptides and unsaturated Fatty Acid Compounds Having Afinity for the Peptides for Therapy, Pharmaceuticals and Product Sterilization, Chemical Abstracts, 1992, vol. 116, p. 84, (Abstract).
    Pruche, et al., Changes in Glutathione Content in Human Hair Follicle Keratinocytes as a Function of Age of Donor: Relation with Glutathione Dependent Enzymes, International Journal of Cosmetic Science, 1991, vol. 13, pp. 117 124.
    Rao, et al., Synthesis and Characterization of Defensin NP 1, Int. J. Peptides Protein Res., 1992, vol. 40, pp. 507 514.
    Reyes, et al., Hesparin and Glutathione: Physiological Decondensing Agents of Human Sperm Nuclei, Gamete Research, 1989, vol. 23, pp. 39 47.
    Roberts et al, Prodrugs of L Cysteine as Protective Agents Against Acetaminophen Induced Hepatotoxicity. 2 (Polyhydroxyalkyl) and 2 (Polyacetoxvalkyl)Thiazolidine 4(R) Carboxylic Acids, J. Med. Chem., 30, pp. 1891 1896 (1987).
    Roderer et al, Cytokine Stimulated Human Immunodeficiency Virus Replication is Inhibited by N Acetvl L Cysteine, Proc. Natl. Acad. Sci. USA, vol. 87, pp. 4884 4888 (1990).
    Roseneld et al, Macrophage derived Foam Cells Freshly Isolated from Rabbit Athersclerotic Lesions Degrade Modified Lipoproteins. promote Oxidation of Low Density Lipoproteins, and Contain Oxidation specific Lip protein Adducts, The American Society for Clinical Investigation, Inc., 1991, vol. 87, pp. 90 99.
    S. Baldwin et al. Oxidant Activity in Expired Breath of Patients with Adult Respiratory Distress Syndrome, The Lancet, Jan. 4, 1986, pp. 11 14.
    Schnittman,et al., The Reservoir for HIV 1 in Human Peripheral Blood is a T Cell that Maintains Expression of CD4, Science, Jul. 21, 1989, vol. 245, pp. 305 308.
    Shapiro, The Control of Oxidant Stress at Fertilization, Science, Apr. 26, 1991, pp. 533 536.
    Slaweta, et al., The Effect of Glutathione on the Motility and Fertility of Frozen Bull Sperm, Amin. Reprod. Sci., 1987, vol. 13, No. 4., pp. 249 253. (Abstract).
    Staal, et al. Intracellular Thiols Regulate Activation of Nuclear Factor K B and Transcription of Human Immunodeficiency Virus, Proc. Natl. Acad. Sci. USA, Dec. 1990, vol. 87, pp. 9943 9947.
    Stevens, Human Herpesviruses, a Consideration of the Latent State, Microbiological Reviews, Sep. 1989, pp. 318 332.
    Suthanthiran et al, Glutathione Regulates Activation Dependent DNA Synthesis in Highly Purified Normal Human T Lymphocytes Stimulated Via the CD2 and CD3 Antigens, Proc. Natl. Acad. Sci. USA, vol. 87, pp. 3343 3347 (1990).
    Uhlig, et al. Glutathione Enhancement in Various Mouse Organs and Protection by Glutathione Isopropyl Ester Against Liver Injury, Jun. 15, 1990, pp. 1877 1880.
    W. Lucht et al., Prevention of Release of Granulocyte Aggregants into Sheep Lung Lymph Follownig Endotoxemia by N Acetylcysteine, The American Journal of the Medical Sciences, vol. 294, No. 3, Sep. 1987, pp. 161 167.
    Wellner et al, Radioprotection by Glutathione Ester: Transport of Glutathione Ester into Human Lymphoid Cells and Fibroblasts, Proc. Natl. Acad. Sci. USA, vol. 81, pp. 4732 4735 (1984).
    Zirkin, et al., In Vitro and In Vivo Studies of Mammalian Sperm Nuclear Decondensation, Gamete Research, 1985, vol. 11, pp. 349 365.

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