CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to and any benefit of U.S. Provisional Application No. 61/925,407, filed January 9, 2014, the content of which is incorporated herein by reference in its entirety.

BACKGROUND

[0002] -hydroxy- -methylbutyrate (HMB) is a metabolite of the branched chain amino acid leucine. HMB has been shown to increase protein synthesis and decrease protein degradation. Traditionally, HMB has been used by athletes to enhance performance and build lean body mass. The presence of HMB signals muscle cells to preserve existing muscle, thereby promoting additional muscle growth and faster muscle development and recovery.

[0003] Recent studies have indicated that HMB may be effective in preserving muscle mass and strength in aged adults and in those subjected to immobility due to prolonged bed rest, and post- surgery. HMB may further be effective in preserving lean body mass and muscle strength in persons with chronic diseases, such as AIDS and cancer. It is postulated that the beneficial effects of HMB may be mediated by the phosphatidylinositol 3-kinase (P13K)/AKT serine/threonine protein kinase (AKT/mammalian target or rapamycin kinase (mTOR)) signaling pathway.

[0004] Traditionally, HMB has been administered through the delivery of leucine. As an essential amino acid, leucine cannot be synthesized by the human body and instead must be ingested.

[0005] The conversion of leucine to HMB is a 2-step enzymatic process. The first step involves the conversion of leucine to keto-isocaproate (KIC) in the muscle catalyzed by branched chain amino acid tranferase (BCAT). KIC is then converted to HMB in the cytosol of the liver catalyzed by KIC dioxygenase, more commonly known as 4-hydroxyphenylpyruvate dioxygenase.

[0006] However, the inventors have discovered an unexpected difference in the conversion efficiency of leucine to HMB in several different situations, which indicates that leucine may not be efficacious in all types of individuals. The inventors have further discovered that there are situations in which leucine may be converted to HMB efficiently, but the endogenously synthesized HMB is not able to be utilized by the body effectively.

SUMMARY

[0007] According to some exemplary embodiments, a method of increasing or maintaining lean body mass in a leucine conversion-compromised human is provided. The method includes administering an effective amount of P-hydroxy-P-methylbutyrate (HMB) to a human with compromised leucine conversion efficiency, whereby administration of the HMB composition to a leucine conversion-compromised human is effective to increase or maintain the human's lean body mass. In some exemplary embodiments, the composition is substantially free of leucine.

[0008] This embodiment also provides a nutritional composition for use in increasing or maintaining lean body mass in a subject having a reduced leucine-to-HMB conversion efficiency, the nutritional composition comprising an effective amount of a P-hydroxy-β- methylbutyrate (HMB). Administration of the composition is effective to increase or maintain the lean body mass of the subject.

[0009] This embodiment also provides use of a nutritional composition in the manufacture of a medicament for increasing or maintaining lean body mass in a subject having a reduced leucine- to-HMB conversion efficiency, the nutritional composition comprising an effective amount of a P-hydroxy-P-methylbutyrate (HMB). Administration of the composition is effective to increase or maintain the lean body mass of the subject.

[00010] In some exemplary embodiments, the leucine conversion-compromised human is a female needing to increase or maintain lean body mass. Administration of the HMB composition may be more effective than leucine for increasing protein synthesis in the female. Administration of the HMB composition may further be more effective than leucine for decreasing protein degradation in the female. The need for increasing or maintaining lean body mass in the human female subject may be attributed to sarcopenia, immobility, and chronic disease. Administration of the HMB composition may further be more effective than leucine for preventing age-related loss of lean body mass in the female.

[00011] In some exemplary embodiments, the leucine conversion-compromised human is a male or female experiencing conditions of muscle atrophy. A human experiencing conditions of muscle atrophy may include humans undergoing glucocorticoid treatment. Administration of the HMB composition may be more effective than leucine for preventing or decreasing the deleterious effects of muscle atrophy in the human subject. In some exemplary embodiments, the HMB composition and the glucocorticoid are administered simultaneously. In some exemplary embodiments, the HMB composition may be administered subsequent to treatment with a glucocorticoid.

[00012] In some exemplary embodiments, the HMB composition is administered orally.

[00013] In some exemplary embodiments, the HMB composition is administered as part of a nutritional composition. In some exemplary embodiments, the nutritional composition further comprises at least one of a source of protein, a source of fat, and a source of carbohydrate.

[00014] In some exemplary embodiments, the nutritional composition includes about 8 to about 100 grams of protein per serving. The source of protein may be intact, partially hydro lyzed or hydrolyzed forms of any of the following: one of a whey protein concentrate, a whey protein isolate, a whey protein hydrolysate, an acid casein, a sodium caseinate, a calcium caseinate, a potassium caseinate, a casein hydrolysate, a milk protein concentrate, a milk protein isolate, a milk protein hydrolysate, nonfat dry milk, condensed skim milk, skim milk powder, a soy protein concentrate, a soy protein isolate, a soy protein hydrolysate, a pea protein concentrate, a pea protein isolate, a pea protein hydrolysate, a collagen protein, collagen protein isolates, potato protein, insect protein isolates, earthworm protein, rice protein, quinoa protein, legume protein, grain protein, and combinations thereof.

[00015] In some exemplary embodiments, the nutritional composition includes about 15 to about 110 grams of carbohydrate per serving. The source of carbohydrate may be maltodextrin, hydrolyzed or modified starch or cornstarch, glucose polymer, corn syrup, corn syrup solid, rice- derived carbohydrate, sucrose, glucose, fructose, lactose, high fructose corn syrup, honey, sugar alcohol, isomaltulose, sucromalt, pullulan, potato starch, and other slowly-digested carbohydrates, dietary fibers including, but not limited to, fructooligosaccharides (FOS), galactooligosaccharides (GOS), oat fiber, soy fiber, gum arabic, sodium carboxymethylcellulose, methylcellulose, guar gum, gellan gum, locust bean gum, konjac flour, hydroxypropyl methylcellulose, tragacanth gum, karaya gum, gum acacia, chitosan, arabinoglactins, glucomannan, xanthan gum, alginate, pectin, low and high methoxy pectin, cereal beta-glucans (i.e., oat beta-glucan, barley beta-glucan), carrageenan and psyllium, Fibersol™, other resistant starches, and combinations thereof. [00016] In some exemplary embodiments, the nutritional composition includes about 0.5 grams to about 45 grams of fat per serving. The source of fat may include coconut oil, fractionated coconut oil, soy oil, corn oil, olive oil, safflower oil, high oleic safflower oil, high gamma-linolenic acid (GLA) safflower oil, MCT (medium chain triglycerides) oil, sunflower oil, high oleic sunflower oil, palm and palm kernel oils, palm olein, canola oil, marine oils, fish oil containing 40-70% combined eicosapentaenoic and docosahexaenoic acids, cottonseed oils, eicosapentaenoic acid, docosahexaenoic acid, gamma-linolenic acid, rice bran oil, wheat germ oil, algal oil, nut oil, conjugated linolenic acid from any source, and combinations thereof.

[00017] In some exemplary embodiments, the nutritional composition further includes one or more of fiber, fructooligosaccharide, milk oligosaccharide, a stabilizer, a sweetening agent, an anti-caking agent, a vitamin, a mineral, a masking agent, a preservative, an emulsifying agent, a buffer, creatine, a prebiotic, a probiotic, a pharmaceutical active, an anti-inflamatory agent, an additional nutrient, a colorant, a flavor, a thickening agent, and a lubricant.

[00018] In some exemplary embodiments, the nutritional composition is formulated as one of a beverage, bar, stick, cookie, frozen liquid, breakfast cereal, candy, snack chip, frozen entree, dietary supplement, meal replacement, chew, bite, capsule, gel, pill, tablet, lozenge, and powder.

[00019] In accordance with further exemplary embodiments, the leucine conversion to HMB may be accelerated, but the muscle mass is nonetheless reduced, suggesting that the body is resistant to the action of endogenous HMB. Administration of the exogenous HMB composition may be more effective than leucine for increasing or maintaining lean body mass in the diabetic human subject.

[00020] In particular, the method of administering a composition including an effective amount of HMB to a human undergoing treatment for diabetes may be effective to increase or maintain the human's lean body mass. This embodiment also provides a nutritional composition for use in increasing or maintaining lean body mass in a subject undergoing diabetic treatment for insulin resistance, the nutritional composition comprising an effective amount of β-hydroxy- β-methylbutyrate (HMB). This embodiment also provides use of a nutritional composition in the manufacture of a medicament for increasing or maintaining lean body mass in a subject undergoing diabetic treatment for insulin resistance, the nutritional composition comprising an effective amount of P-hydroxy-P-methylbutyrate. Administration of the composition to the composition to the subject is effective to increase or maintain lean body mass of the subject. In some exemplary embodiments, the treatment for diabetes includes any conventional diabetic treatment for controlling blood sugar in people with diabetes, such as metformin, thiazolidinediones, insulin releasing agents, etc. In some exemplary embodiments, the HMB composition and the diabetic treatment are administered simultaneously. In some exemplary embodiments, the HMB composition is administered subsequent to a diabetic treatment.

[00021] In some exemplary embodiments, administration of the HMB composition and diabetic treatment is effective to increase protein synthesis in the human undergoing treatment for diabetes. In some exemplary embodiments, administration of the HMB composition and diabetic treatment is effective to decrease protein degradation in the human with diabetes.

BRIEF DESCRIPTION OF THE FIGURES

[00022] Figure 1 illustrates a graph showing the HMB concentration over a 10 hour period in normal rats and rats undergoing glucocorticoid treatment.

[00023] Figure 2 illustrates a bar graph showing the Area Under the Curve ("AUC") of HMB concentration in normal rats compared to rats undergoing glucocorticoid treatment.

[00024] Figure 3 illustrates a bar graph showing the loss of lean mass in rats undergoing glucocorticoid treatment compared to normal rats.

[00025] Figure 4 illustrates a bar graph showing the HMB concentration in male rats vs. female rats when administered an equal dose of leucine.

[00026] Figure 5 illustrates a bar graph showing the Area Under the Curve ("AUC") of HMB concentration in male rats compared to female rats when administered an equal dose of leucine.

[00027] Figure 6 illustrates a bar graph showing the plasma concentration of HMB in Zucker Diabetic Fatty (ZDF) lean rats compared to ZDF obese rats.

[00028] Figure 7 illustrates a bar graph showing the Area Under the Curve ("AUC") of HMB concentration in ZDF lean rats compared to ZDF obese rats when administered an equal dose of leucine.

DETAILED DESCRIPTION

[00029] Unless otherwise defined, all 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 pertains. Although other methods and materials similar or equivalent to those described herein may be used in the practice or testing of the exemplary embodiments, exemplary suitable methods and materials are described below. In case of conflict, the present specification, including definitions, will control. In addition, the materials, methods, and examples are illustrative only and not intended to be limiting of the general inventive concepts.

[00030] The terminology as set forth herein is for description of the exemplary embodiments only and should not be construed as limiting the application as a whole. Unless otherwise specified, "a," "an," "the," and "at least one" are used interchangeably. Furthermore, as used in the description of the application and the appended claims, the singular forms "a," "an," and "the" are inclusive of their plural forms, unless contradicted by the context surrounding such.

[00031] The recitations of numerical ranges by endpoints include all numbers subsumed within that range (e.g. , 1 to 5 includes 1 , 1.5, 2, 2.75, 3, 3.80, 4, 5, etc.).

[00032] The expression "effective amount" as used herein, refers to a sufficient amount of HMB to increase or maintain lean body mass in a subject as defined herein. The exact amount required will vary from subject to subject, depending on the species, age, and general condition of the subject, the particular therapeutic agent, its mode and/or route of administration, and the like.

[00033] It will be understood, however, that the total daily usage of the compounds and compositions disclosed herein can be decided by an attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend upon a variety of factors including the disorder being treated and the severity of the disorder; the activity of the specific compound employed; the specific composition employed; the age, body weight, general health, sex and diet of the subject; the time of administration, route of administration, and rate of excretion of the specific composition employed; the duration of the treatment; drugs used in combination or coincidental with the specific composition employed; and like factors well known in the medical arts.

[00034] The subject is preferably a mammal, such as a human, a domesticated farm animal (e.g., cow, horse, pig) or pet (e.g., dog, cat). More preferably, the subject is a human. Subjects can be selected from various age groups. For example, subjects can be children, adults, or the elderly. Adult humans are subjects having an age of 18 or more, while human children have an age of 17 or less. Human subjects can be selected from any age group. For example, subjects can have an age that ranges from 1 to 100, or any age there between. [00035] In some exemplary embodiments, the subject is a leucine conversion-compromised human. The phrase "leucine conversion-compromised" as used herein refers to a human with reduced leucine conversion efficiency, such as females, subjects experiencing conditions of muscle loss, and subjects experiencing conditions of muscle atrophy.

[00036] In some exemplary embodiments, the subject is a human undergoing treatment for diabetes. The treatment may include known diabetic treatments, such as metformin or other drugs, which is a drug known to stimulate GLUT4 translocation to improve insulin resistance. Such diabetic treatment may give rise to leucine conversion-compromised subjects, whereby even if HMB is effectively converted in the diabetic subject, the HMB does not act on the target tissue.

[00037] The term HMB, which is also referred to as beta-hydroxy-beta-methylbutyric acid, or beta-hydroxy-isovaleric acid, can be represented in its free acid form as (CH ₃ ) ₂ (OH)CCH ₂ COOH. HMB is a metabolite of leucine formed by transamination to alpha- ketoisocaproate (KIC) in muscle followed by oxidation of the KIC in the cytosol of the liver to give HMB.

[00038] The term "nutritional composition," as used herein, unless otherwise specified, refers to nutritional products in various forms including, but not limited to, liquids, solids, powders, semisolids, semi-liquids, nutritional supplements, and any other nutritional food product known in the art. A nutritional composition in powder form may often be reconstituted to form a nutritional composition in liquid form. The nutritional compositions disclosed herein are generally suitable for oral consumption by a human.

[00039] The term "liquid nutritional composition" as used herein, unless otherwise specified, refers to nutritional compositions in ready-to-drink liquid form, concentrated liquid form, and nutritional liquids made by reconstituting nutritional powders described herein prior to use. The liquid nutritional composition may also be formulated as a suspension, an emulsion, a solution, and the like.

[00040] The term "nutritional powder" or "reconstitutable powder" as used herein, unless otherwise specified, refers to nutritional compositions in a solid flowable or scoopable form that can be reconstituted with water or another liquid prior to consumption and includes spray-dried powders, dry-mixed, or dry-blended powders, and the like. [00041] The term "nutritional semi-solid" as used herein, unless otherwise specified, refers to nutritional products that are intermediate in properties, such as rigidity, between solids and liquids. Some semi-solid examples include puddings, yogurts, gels, gelatins, doughs, and the like.

[00042] The term "nutritional semi-liquid" as used herein, unless otherwise specified, refers to nutritional compositions that are intermediate in properties, such as flow properties, between liquids and solids. Some semi-liquid examples include thick shakes, liquid yogurts, liquid gels, and the like.

[00043] The term "serving," unless otherwise indicated herein, generally refers to an amount of nutritional composition that is consumed in one sitting, which may last up to one or up to two hours.

[00044] The terms "administer," "administering," "administered," or "administration" as used herein, unless otherwise specified, should be understood to include providing the composition (e.g., in the form of a nutritional product) to a subject, the act of consuming the nutritional product by the subject, and combinations thereof. In addition, it should be understood that the methods disclosed herein (e.g., administering the composition) may be practiced with or without doctor supervision or other medical direction.

[00045] The term "lean body mass," as used herein, unless otherwise specified, refers to the amount or size of muscle or muscle groups, as expressed by muscle weight, mass, area, or volume. Muscle mass may also be expressed as total muscle mass, lean body mass of a body compartment such as a leg or arm, or cross-sectional area of a leg or arm compartment. The volume or mass of the muscle can be determined using any known or otherwise effective technique that provides muscle area, volume, or mass, such as DEXA, or by using visual or imaging techniques such as MRI or CT scans.

[00046] The term "muscle atrophy," as used herein, unless otherwise specified, refers to the loss of muscle mass (also known as muscle wasting). Muscle atrophy may be caused by normal aging (e.g., sarcopenia), inactivity (e.g., muscle disuse or immobility), or disease-related disorders (e.g., cachexia), or drug/medication induced atrophy.

[00047] The general inventive concepts relate to methods of increasing or maintaining lean body mass in a leucine conversion-comprised subject. The leucine conversion inefficiency in the subject may be due, for example, to gender, existing muscle atrophy, or treatment for diabetes. Accordingly, the general inventive concepts relate to methods of increasing or maintaining lean body mass in human females and humans (male or female) suffering from muscle atrophy. The general inventive concepts further relate to methods of increasing or maintaining lean body mass in subjects undergoing treatment for diabetes. The methods include administering an effective amount of HMB, or an acceptable salt thereof, to the subject in order to increase or maintain the subject's lean body mass. In certain exemplary embodiments, the methods include administering the HMB as part of a nutritional composition.

[00048] HMB is a metabolite of the branched chain amino acid leucine. HMB has been shown to increase protein synthesis and decrease protein degradation by decreasing the ubiquitin-proteasome proteolytic pathway. Decreasing protein degradation allows for faster strength increase, since the body has less muscle damage that needs to be repaired.

[00049] Traditionally, HMB has been administered through the delivery of leucine, which is then converted to HMB in the body. As an essential amino acid, leucine cannot be synthesized by the human body and must be ingested. The conversion of leucine to HMB is a 2-step enzymatic process. The first step involves the conversion of leucine to keto-iso caproate (KIC) in the muscle catalyzed by branched chain amino acid transferase (BCAT). KIC is then converted to HMB in the cytosol of the liver catalyzed by KIC dioxygenase. However, it has been discovered that certain subjects have lower leucine conversion efficiencies than other subjects, thereby receiving less of a benefit from the administration of leucine. Such subjects include those experiencing conditions of muscle atrophy, females, and those undergoing certain diabetic therapies. Therefore, methods for maintaining or increasing lean body mass in leucine conversion-compromised subjects by administering an effective amount of HMB, or an acceptable salt thereof, are provided.

[00050] In one exemplary embodiment of the present invention, the leucine conversion- compromised subjects are those experiencing conditions of muscle atrophy, such as during glucocorticoid treatment. It has been discovered that subjects experiencing conditions of muscle atrophy, such as subjects undergoing glucocorticoid treatment, do not efficiently convert leucine to HMB. Therefore, exemplary embodiments include at least partially replacing dosages of leucine with an effective amount of HMB, for maintaining or increasing lean body mass in subjects experiencing conditions of muscle atrophy. Providing an effective amount of HMB or an acceptable salt thereof to a subject receiving glucocorticoid treatment may effectively stop or reverse muscle atrophy resulting from the glucocorticoid treatment.

[00051] Glucocorticoids are a class of steroid hormones that bind to the glucocorticoid receptor. Examples of glucocorticoids include Cortisol (i.e., hydrocortisone), cortisone, prednisone, prednisolone, methylprednisolone, dexamethasone, betamethasone, triamcinolone, and beclometasone. A commonly administered glucocorticoid is dexamethasone, which has the chemical name (8S,9R,10S,1 lS,13S,14S,16R,17R)-9- Fluoro-1 l,17-dihydroxy-17-(2- hydroxyacetyl)-10,13,16- trimethyl-6,7,8,9,10,11,12,13, 14,15, 16,17-dodecahydro-3H- cyclopenta[a]phenanthren-3-one. Glucocorticoids differ from one another in terms of both pharmacokinetics (e.g., absorption factor, half-life, volume of distribution, clearance) and pharmacodynamics (e.g., mineralocorticoid activity and renal physiology). Glucocorticoids are primarily provided orally, but can also be provided by other methods, such as by inhalation, topically or parenterally.

[00052] Glucocorticoids are commonly used to treat a wide variety of chronic inflammatory conditions such as systemic lupus erythematosus, sarcoidosis, rheumatoid arthritis, and bronchial asthma. Glucocorticoids may also be used in low doses to treat adrenal insufficiency. They are also administered as post-transplantory immunosuppressants to prevent the acute transplant rejection and graft- versus-host disease. Cancer patients may also be administered glucocorticoids both for an anticancer effect, and to counteract some of the side-effects of chemotherapy. However, administration of high doses of glucocorticoids have a number of side effects, including immunodeficiency, hyperglycemia, weight gain, and osteoporosis. In particular, glucocorticoid administration causes muscle atrophy in humans and animals. Dardavet et al., J. Clin. Invest. 96, 2113-2119 (1995). The combination of clinical problems produced by prolonged administration or endogenous overproduction of glucocorticoids is termed Cushing's syndrome. Studies have suggested that glucocorticoids inhibit protein synthesis and simulate protein degradation in skeletal muscle, and that this may be mediated, at least in part, by an increase in myostatin expression. Ma et al., Am. J. Physiol Endocrinol Metab, 285, E363-E371 (2003).

[00053] Glucocorticoids can be given in a dose that provides approximately the same glucocorticoid effects as normal physiological Cortisol production. This dose is referred to as physiologic, replacement, or maintenance dosing, and is approximately 6-12 mg/m ² /day (m ² refers to body surface area (BSA), and is a measure of body size; an average man's BSA is 1.7 m ² ). The dose for a given glucocorticoid varies based on its potency. The potency of known glucocorticoids are known to those skilled in the art. A high dose of glucocorticoid is an amount above the physiologic or replacement dose, whereas a low dose of glucocorticoid is an amount of less than or equal to the physiologic or replacement dose. Because high doses of glucocorticoids may result in a higher level of negative effects on muscle, in some embodiments, HMB is provided to a subject who is receiving high doses of glucocorticoids as defined herein.

[00054] HMB or an acceptable salt thereof may be administered before, after, or concurrent with administration of the glucocorticoids. Alternatively, the HMB may be provided both before and concurrent with, both concurrent with and after, both before and after, or before, concurrent with and after administration of the glucocorticoids. Concurrent administration, as used herein, includes providing the HMB and glucocorticoids within about 10 minutes or less of one another, within about 5 minutes or less of one another, or in some embodiments essentially simultaneously. Concurrent administration may be most feasible when the glucocorticoids and HMB are administered to a subject by the same route (e.g., both orally). When the HMB is provided before or after administration of the glucocorticoids, administration should still occur during a time when the effects of the HMB and the glucocorticoids will overlap. For example, when the HMB is provided after glucocorticoid administration, the HMB should be provided while the negative effect of the glucocorticoids on muscle are either developing or manifest. For example, the HMB may be provided within 1 hour, 12 hours, 24 hours, or one week or more after glucocorticoid administration and may continue on a daily basis for multiple days after glucocorticoid treatment (e.g., 1-14 days, 1-7 days or 7-14 days). Alternately, when HMB is provided before glucocorticoid administration, the HMB should be provided at a time such that the beneficial effect of HMB on muscle will be able to prevent or decrease the negative effects of the glucocorticoid administration. For example, HMB can be provided within 1 hour, 12 hours, 24 hours, or one week or more before glucocorticoid administration. When the glucocorticoids are administered as part of a chemotherapy regimen, it may be especially beneficial to administer the HMB before, after or both before and after, but not concurrently with the glucocorticoids (especially when the glucocorticoids are administered immediately prior to the chemotherapy regimen when it may be difficult for the subject to tolerate concurrent oral administration of the HMB). [00055] It has further been discovered that there is an unexpected difference in conversion efficiency of leucine to HMB between male and female subjects. Particularly, males are believed to be more efficient in leucine-to-HMB conversion than females. Therefore, providing HMB to female subjects, rather than leucine, may allow female subjects to incur the same benefits of HMB as males without having to account for the conversion inefficiency of leucine to HMB.

[00056] Accordingly, in another exemplary embodiment, the leucine conversion- compromised subjects include females. Therefore, methods for administering an effective amount of HMB are provided for maintaining or increasing lean body mass in a female subject.

[00057] It has additionally been discovered that diabetic subjects accumulate significantly higher levels of HMB when compared to non-diabetic subjects, which indicates that, although diabetic subjects may effectively convert leucine to HMB, the HMB may not be acting on the target tissue. Such a HMB-resistant state may be related to the insulin resistance of diabetic subjects. Diabetic treatments often includes controlling blood sugar by improving insulin resistance. For example, metformin is a drug often used as a first-line treatment for diabetes and is known to stimulate GLUT4 translocation thus improving insulin resistance. Thus, metformin increases insulin sensitivity to provide a more effective use of insulin. Therefore, by not intending to be limited by any theory, it is believed that HMB efficacy may also be improved by co-administering HMB with a diabetic treatment, such as metformin. Although exemplary embodiments may be described as co-administration of HMB with metformin, it should be understood that any of the medications and therapies that are used by medical professionals to treat diabetes (e.g., improve insulin resistance) may be suitable for use in the methods described herein.

[00058] Accordingly, yet another exemplary embodiment of the present invention is directed to a method for administering an effective amount of HMB, or an acceptable salt thereof, to a subject receiving a diabetic treatment in order maintain or increase lean body mass. As the diabetic treatment increases insulin sensitivity, it is provided that the co -administration of HMB with the diabetic treatment may also increase a subject's HMB sensitivity to allow the HMB to act on the target tissue thereby increasing or maintaining a subject's lean body mass.

[00059] The increase in the amount of lean body mass can be expressed as a percentage of the subject's overall lean body mass. Accordingly, in various embodiments, the increase in the amount of lean body mass is about 5%, about 10%, about 20%, about 30%>, about 40%>, about 50%, about 60%, about 70%, about 80%, about 90%, about 95%, or about 100%. In the case where about 100% of the subject's lean body mass in increased, the HMB is effective to double the subject's lean body mass. In some embodiments, the amount of HMB provided can be varied to affect the increase or maintenance in a subject's lean body mass.

[00060] The increase or maintenance in lean body mass achieved by the methods disclosed herein can also be viewed as prevention of loss of lean body mass. Accordingly, in various embodiments, administration of HMB can be used to maintain 100% of the lean body mass of a leucine-conversion compromised subject and/or a diabetic subject.

[00061] Glucocorticoids can have a variety of different negative effects on muscle. Different embodiments of the methods disclosed herein include decreasing one or more of these different negative effects. In some embodiments, HMB is provided to increase the lean body mass in subjects undergoing administration of glucocorticoids. Glucocorticoid administration can also result in muscle atrophy, which can be prevented by providing HMB to the subject. Muscle atrophy corresponds to a decrease in lean body mass. Lean body mass can be evaluated using a variety of different techniques known to those skilled in the art, including determination of muscle weight or muscle fiber diameter. In various embodiments, muscle atrophy can be prevented by about 5%, about 10%>, about 20%>, about 30%>, about 40%>, about 50%>, about 60%>, about 70%), about 80%>, about 90%>, about 95%, or about 100%. Glucocorticoid administration can also have a negative effect on various other aspects of muscle, such as decreasing protein content or increasing susceptibility to fatigue, which can be ameliorated by providing HMB to the subject.

[00062] The decrease in muscle atrophy achieved by the methods disclosed herein can also be viewed as maintenance of lean body mass. Maintenance of lean body mass, as used herein, refers to retaining an amount of lean body mass that corresponds to that of a subject who is not receiving glucocorticoid treatment, or a percentage thereof (or that corresponds to the lean body mass of the subject prior to undergoing glucocorticoid treatment). Accordingly, in various embodiments, administration of HMB or lower doses of HMB and leucine can be used to maintain 100% of the lean body mass of a subject receiving glucocorticoids, or in other embodiments lesser amounts. For example, embodiments of the method can maintain about 50% lean body mass, about 60%> lean body mass, about 70%> lean body mass, about 80%> lean body mass, about 90% lean body mass, about 95% lean body mass, or any amounts ranging from about 50% to about 100%.

[00063] HMB can be provided to a variety of subjects, such as females, those who are receiving glucocorticoids, and diabetic subjects who are receiving a diabetic treatment. The subjects receiving glucocorticoids or diabetic treatment may be in a home setting, or the subjects may be currently hospitalized. In certain such embodiments, the subject is administered a nutritional composition containing HMB concurrent with receiving the glucocorticoid or diabetic treatment. In other such embodiments, the subject is administered the nutritional composition containing HMB subsequent to receiving the glucocorticoid or diabetic treatment. In yet other such embodiments, the subject is administered the nutritional composition containing HMB concurrent with and subsequent to receiving the glucocorticoid or diabetic treatment.

[00064] The methods of maintaining or increasing lean body mass in subjects described herein include providing an effective amount of HMB or an acceptable salt thereof. As used herein, the abbreviation HMB is meant to encompass both HMB and its acceptable salts, unless indicated otherwise. A preferred form of HMB is the calcium salt of HMB, also designated as Ca-HMB, which is most typically the monohydrate calcium salt. The HMB used in the methods disclosed herein can come from virtually any source. Calcium HMB monohydrate is commercially available from Technical Sourcing International (TSI) of Salt Lake City, Utah. Note that the amounts of HMB described in the exemplary embodiments presented herein are based on use of Ca-HMB.

[00065] Although the calcium monohydrate salt is one preferred form of HMB for use herein, other suitable sources include HMB as a free acid, a salt, an anhydrous salt, an ester, a lactone, or other product forms that provide a bioavailable form of HMB suitable for administration. Non- limiting examples of suitable salts of HMB (hydrated or anhydrous) for use herein include sodium, potassium, chromium, calcium, and other non-toxic salt forms.

[00066] The HMB may be formulated in a suitable composition (e.g., a nutritional composition) and then, in accordance with the methods disclosed herein, administered to a subject in a form adapted to the chosen route of administration. Examples of nutritional compositions include nutritional liquids, nutritional powders, nutritional semi-solids, and nutritional semi-liquids. While certain portions of the remainder of the formulation description explicitly refers to HMB alone, it should be understood that the formulations described are also intended to encompass the combination of lower doses of HMB and a branched chain amino acid such as leucine, unless it is clear from the context that the formulation is intended to contain HMB alone.

[00067] Formulations suitable for administration of HMB (or HMB in combination with a branched chain amino acid, and/or a diabetic treatment) include, but are not limited to, those suitable for oral administration, enteral administration, parental administration, injectable administration (including subcutaneous and intramuscular) and intravenous administration. Oral administration, as defined herein, includes any form of administration in which the HMB, or HMB in combination with the branched chain amino acid, or diabetic treatment, passes through the esophagus of the subject. For example, oral administration includes nasogastric intubation, in which a tube is run from through the nose to the stomach of the subject to administer food or drugs. Enteral administration includes administration through a gastric tube or a jejunal tube.

[00068] Oral formulations include liquid, powder including reconditionable powder, semisolid, and semi-liquid compositions, provided that such formulations allow for the safe and effective oral delivery of HMB and optional nutritive components. In certain exemplary embodiments, the oral formulation is a nutritional composition. A nutritional composition is a composition that is edible and includes additional nutrients beyond HMB, such as proteins, carbohydrates, fats, vitamins, and minerals. Formulations suitable for oral administration may be presented as discrete units such as tablets, troches, capsules, lozenges, wafers, or cachets, each containing a predetermined amount of the HMB as a powder or granules or as a solution or suspension in an aqueous liquor or non-aqueous liquid such as a syrup, an elixir, an emulsion, or a draught.

[00069] In certain exemplary embodiments, the nutritional composition used in the methods includes HMB; HMB in combination with a branched chain amino acid (e.g., leucine, isoleucine, valine, and combinations thereof); HMB in combination with a metabolite of a branched chain amino acid (e.g., keto isocaproic acid, alpha hydroxyl isocaproic acid, and combinations thereof); HMB in combination with a branched chain amino acid (e.g., leucine, isoleucine, valine, and combinations thereof) and a metabolite of a branched chain amino acid (e.g., keto isocaproic acid, alpha hydroxy isocaproic acid, and combinations thereof), or HMB in combination with a diabetic treatment. [00070] In certain exemplary embodiments the nutritional composition used in the methods includes HMB (or HMB in combination with a branched chain amino acid (e.g., leucine, isoleucine, valine, or combinations thereof) or a metabolite thereof, and vitamins and minerals. In certain such embodiments, the vitamins and minerals that are present are the 24 "essential" vitamins and minerals. The addition of vitamins and minerals to the nutritional composition used in the methods may result in an additional improvement to the individual who consumes the nutritional composition.

[00071] The term "nutritional liquid" as used herein, unless otherwise specified, refers to nutritional compositions in ready-to-drink liquid form, concentrated form, and nutritional liquids made by reconstituting the nutritional powders described herein prior to use. The nutritional liquid may also be formulated as a suspension, an emulsion, a solution, and so forth.

[00072] The nutritional composition administered to the subject can provide from 0.1 g/day to 10 g/day of HMB. Alternately, the composition can provide 0.5 g/day to 10 g/day, 1.5 g/day to 10 g/day, 0.5 g/day to 5 g/day, or 1.5 g/day to 4 g/day, of HMB. Subjects may be administered one serving per day, two servings per day, three servings per day, or four or more servings per day to receive the desired amount of HMB from the nutritional composition. In some exemplary embodiments, each serving of a nutritional composition including HMB provides about 0.5 to 3 grams HMB, or about 1.5 grams of HMB. In addition, in further embodiments, the HMB can be provided to the subject for a period of 3-7 days (e.g., the HMB is provided twice daily for 3 days, 4 days, 5 days, 6 days or 7 days). In other exemplary embodiments, the HMB is provided to the subject during a period of hospitalization, which typically lasts 3-5 days. However, to offset the poor conversion of leucine to HMB seen with glucocorticoid treatment, female sex, or the HMB- resistance of diabetes, much longer duration of treatment (up to years) would be envisaged.

[00073] In certain exemplary embodiments, the nutritional composition includes one or more ingredients that help satisfy the subjects nutritional requirements, in addition to providing a useful formulation for the HMB. For example, the nutritional composition can include a protein, a carbohydrate, and/or a fat. In certain embodiments, the nutritional composition includes at least one source of protein, at least one source of carbohydrate, and at least one source of fat. In some exemplary embodiments, the nutritional composition can be formulated to provide a specialized nutritional product for use in subjects afflicted with specific diseases or conditions. Many different sources and types of proteins, carbohydrates, and fats are known and can be used in nutritional compositions including HMB. In certain exemplary embodiments, the nutritional composition is in the form of a powder suitable for reconstitution to a liquid, a ready-to-drink liquid or a bar.

[00074] In certain exemplary embodiments, the nutritional composition may be a solid nutritional product. Non-limiting examples of solid nutritional products include snack and meal replacement products, including those formulated as bars; sticks; cookies; breads; cakes; other baked goods; frozen liquids; candy; breakfast cereals; powders; granulated solids; other particulates; snack chips; bites; frozen or retorted entrees; and so forth. In certain exemplary embodiments, when the nutritional composition is a solid product, the serving may be 25 grams to 150 grams.

[00075] In certain exemplary embodiments, the nutritional composition may be a nutritional liquid. Non-limiting examples of nutritional liquids include snack and meal replacement products; hot or cold beverages; carbonated or non carbonated beverages; juices or other acidified beverages; milk or soy-based beverages; shakes; coffees; teas; enteral feeding compositions; and so forth. Generally, the nutritional liquids are formulated as suspensions or emulsions, but the nutritional liquids can also be formulated in any other suitable forms such as clear liquids, solutions, liquid gels, liquid yogurts, and so forth.

[00076] Where the nutritional composition is a liquid nutritional product, a serving thereof may be within a range of 30 milliliters to 500 milliliters (~1 fl. oz. to ~17 fl. oz.). In certain exemplary embodiments, the serving is 237 milliliters (~8 fl. oz.). In certain exemplary embodiments, the serving is 125 milliliters (~4 fl. oz.). In certain exemplary embodiments, the serving is 177 milliliters to 417 milliliters (~6 fl. oz. to ~14 fl. oz.). In other exemplary embodiments, the serving is 207 milliliters to 266 milliliters (~7 fl. oz. to ~9 fl. oz.). In other exemplary embodiments, the serving is 30 milliliters to 75 milliliters (~1 fl. oz. to ~ 2.5 fl. oz.). In certain exemplary embodiments, one serving to 14 servings of the nutritional composition is administered to the subject per week.

[00077] The liquid products may be and typically are shelf stable. The liquid products typically contain up to about 95% by weight of water. In certain exemplary embodiments, the liquid nutritional products contain from about 50% to about 95%, about 60% to about 90%, and about 70%) to about 85% of water by weight. [00078] The liquid products may have a low pH ranging from about 2 to 5. In accordance with certain embodiments, the pH of the liquid nutritional composition may be from 2.5 to 4.6, including a pH of 3 to 3.5.

[00079] In yet other exemplary embodiments, the nutritional composition may be formulated as semi-solid or semi-liquid compositions (e.g., puddings, gels, yogurts), as well as conventional product forms such as capsules, tablets, caplets, pills, and so forth. In other exemplary embodiments, the nutritional composition may be in the form of lozenges or tablets (e.g., chewable, coated).

[00080] In yet other exemplary embodiments, the nutritional composition may be formulated as a nutritional powder. The nutritional powders (including pressed or solid nutritional powders including tablet forms) may be reconstituted by the intended user with a suitable aqueous liquid, typically water, to form a nutritional liquid for immediate oral or enteral use. In this context, "immediate" use generally means within about 48 hours, more typically within about 24 hours, most typically right after or within 20 minutes of reconstitution.

[00081] The exemplary nutritional powders may include spray dried powders, dry mixed powders, agglomerated powders, and combinations thereof, as well as powders prepared by other suitable methods.

[00082] The nutritional compositions disclosed herein are useful to provide sole, primary, or supplemental sources of nutrition, as well as providing one or more of the benefits as described herein. Accordingly, the nutritional compositions disclosed herein may include one or more macronutrients. For example, in certain exemplary embodiments, the nutritional compositions comprise at least one source of protein, at least one source of carbohydrate, and at least one source of fat. In other exemplary embodiments, the nutritional compositions comprise at least one source of protein, at least one source of carbohydrate, but no source of fat. In certain exemplary embodiments, the nutritional compositions provide up to 1000 kcal of energy per serving or dose. In certain exemplary embodiments, the nutritional compositions provide from 20 kcal to 900 kcal, from 25 kcal to 700 kcal, from 50 kcal to 500 kcal, from 100 kcal to 450 kcal, or from 150 kcal to 400 kcal per serving.

[00083] In certain exemplary embodiments, the nutritional composition may comprise 8 grams to 100 grams of protein per serving or 10 grams to 100 grams of protein per serving. In other exemplary embodiments, the nutritional composition may comprise 10 grams to 50 grams of protein per serving. In still other exemplary embodiments, the nutritional composition may comprise 10 grams to 25 grams of protein per serving. In certain other exemplary embodiments, the at least one source of protein comprises from 5% to 40%, from 5% to 30%, or from 15% to 25% by weight of the nutritional composition.

[00084] Virtually any source of protein may be used so long as it is suitable for use in oral nutritional compositions and is otherwise compatible with any other selected ingredients or features in the nutritional composition.

[00085] The source of protein may include, but is not limited to, intact, hydrolyzed, and partially hydrolyzed protein, which may be derived from any known or otherwise suitable source such as milk (e.g., casein, whey), animal (e.g., meat, fish), cereal (e.g., rice, corn), vegetable (e.g., soy, pea), insect (e.g., cricket, locust), and combinations thereof. Non-limiting examples of the source of protein include a whey protein concentrate, a whey protein isolate, a whey protein hydrolysate, an acid casein, a sodium caseinate, a calcium caseinate, a potassium caseinate, a casein hydrolysate, a milk protein concentrate, a milk protein isolate, a milk protein hydrolysate, nonfat dry milk, condensed skim milk, skim milk powder, a soy protein concentrate, a soy protein isolate, a soy protein hydrolysate, a pea protein concentrate, a pea protein isolate, a pea protein hydrolysate, a collagen protein, collagen protein isolates, potato protein, insect protein isolates, earthworm protein, rice protein, quinoa protein, legume protein, grain protein, and combinations thereof.

[00086] In some instances, the at least one source of protein provides 6 grams to 50 grams of protein per serving and comprises any one or more sources of protein provided in the non- limiting list presented above.

[00087] As previously discussed, in various exemplary embodiments, the nutritional composition further comprises at least one source of carbohydrate. In some exemplary embodiments, where the nutritional composition contains at least one source of carbohydrate, the at least one source of carbohydrate comprises from 10% to 70%, including from 30%> to 60%> and from 45% to 58%, by weight of the nutritional composition. In other exemplary embodiments, the nutritional composition comprises 15 grams to 1 10 grams of at least one source of carbohydrate per serving. In some exemplary embodiments, the nutritional composition comprises 25 grams to 90 grams, including 40 grams to 65 and 45 grams to 55 grams of at least one source of carbohydrate per serving. [00088] The at least one source of carbohydrate suitable for use in various exemplary embodiments of the nutritional compositions may be simple, complex, or variations or combinations thereof. Generally, any source of carbohydrate may be used so long as it is suitable for use in oral nutritional compositions and is otherwise compatible with any other selected ingredients or features present in the nutritional compositions. Non-limiting examples of a source of carbohydrate suitable for use in the nutritional compositions described herein include maltodextrin; hydrolyzed or modified starch or cornstarch; glucose polymers; corn syrup, corn syrup solids; rice-derived carbohydrate; sucrose; glucose; fructose; lactose; high fructose corn syrup; honey; sugar alcohols (e.g., maltitol, erythritol, sorbitol); isomaltulose; sucromalt; pullulan; potato starch; and other slowly-digested carbohydrates; dietary fibers including, but not limited to, fructooligosaccharides (FOS), galactooligosaccharides (GOS), oat fiber, soy fiber, gum arabic, sodium carboxymethylcellulose, methylcellulose, guar gum, gellan gum, locust bean gum, konjac flour, hydroxypropyl methylcellulose, tragacanth gum, karaya gum, gum acacia, chitosan, arabinoglactins, glucomannan, xanthan gum, alginate, pectin, low and high methoxy pectin, cereal beta-glucans (e.g., oat beta-glucan, barley beta-glucan), carrageenan and psyllium, Fibersol™; other resistant starches; and combinations thereof.

[00089] Additionally, in certain exemplary embodiments, the nutritional composition further comprises at least one source of fat. However, in other exemplary embodiments, the nutritional composition may be free of any fat content, or essentially free of any fat content (i.e., having less than 0.5 grams of fat per serving). In some exemplary embodiments, where the nutritional composition contains fat, the nutritional composition comprises from 0.5 grams to 45 grams of at least one source of fat per serving. In other exemplary embodiments, the nutritional composition comprises from 5 grams to 35 grams, from 8 grams to 25 grams, from 10 grams to 20 grams of at least one source of fat per serving. In certain exemplary embodiments, the at least one source of fat comprises from 20% to 65%, from 25% to 45% by weight of the nutritional composition. In other exemplary embodiments, the at least one source of fat comprises from 1% to 30% or from 3% to 15% by weight of the nutritional composition.

[00090] In general, any source of fat may be used so long as it is suitable for use in oral nutritional compositions and is otherwise compatible with any other selected ingredients or features present in the nutritional compositions. The source of fat may be derived from plants, animals, and combinations thereof. Non-limiting examples of suitable sources of fat for use in the nutritional compositions described herein include coconut oil; fractionated coconut oil; soy oil; corn oil; olive oil; safflower oil; high oleic safflower oil; high gamma-linolenic acid (GLA) safflower oil; MCT (medium chain triglycerides) oil; sunflower oil; high oleic sunflower oil; palm and palm kernel oils; palm olein; canola oil; marine oils; cottonseed oils; eicosapentaenoic acid; docosahexaenoic acid; gamma-linolenic acid; rice bran oil; wheat germ oil; algal oil; nut oil; conjugated linolenic acid from any source; and combinations thereof.

[00091] The amount or concentration of the at least one source of protein, at least one source of carbohydrate; and at least one source of fat present in certain embodiments of the nutritional compositions described herein may vary widely depending on the product formulation of the nutritional composition (e.g., solid, liquid, etc.). In addition, the amount or concentration of the at least one source of protein, at least one source of carbohydrate, and at least one source of fat present in certain embodiments of the nutritional compositions described herein may be characterized based upon: (i) a percentage of the total calories per serving in the nutritional composition; or (ii) the total weight of each ingredient present in a serving of the nutritional composition; or both (i) and (ii). For example, in certain exemplary embodiments, the amount or concentration of the at least one source of protein, the at least one source of carbohydrate, and at least one source of fat present in the nutritional composition can be within the ranges shown in the examples provided in Tables I and II below. In other exemplary embodiments, the nutritional composition contains no fat or essentially no fat (i.e., less than 0.5 grams per serving). TABLE I

TABLE II

[00092] In certain exemplary embodiments, the nutritional composition comprises at least one source of carbohydrate and at least one source of fat, and the at least one source of carbohydrate provides 5 percent to 80 percent of the caloric density per serving and the at least one source of fat provides 10 percent to 85 percent of the caloric density per serving. In other exemplary embodiments, the nutritional composition comprises at least one source of carbohydrate and at least one source of fat, and the at least one source of carbohydrate provides 20 percent to 75 percent of the caloric density per serving and the at least one source of fat provides 10 percent to 65 percent of the caloric density per serving. In yet other exemplary embodiments, the nutritional composition comprises at least one source of carbohydrate and at least one source of fat, and the at least one source of carbohydrate provides 30 percent to 50 percent of the caloric density per serving and the at least one source of fat provides 30 percent to 50 percent of the caloric density per serving. Such embodiments provide flexibility in formulating calorie dense nutritional compositions with various other ingredients.

[00093] The HMB -containing nutritional composition may include additional ingredients that may modify the physical, nutritional, chemical, hedonic, or processing characteristics of the product or serve as pharmaceutical or additional nutritional components. Non-limiting examples of such optional ingredients include preservatives; antioxidants; emulsifying agents; buffers; fructooligosaccharides; chromium picolinate; pharmaceutical additives; colorants; flavors or masking agents; thickening agents and stabilizers; artificial sweeteners; hydrocolloids such as guar gum, xanthan gum, carrageenan, gellan gum, gum acacia, and so forth.

[00094] The HMB-containing nutritional composition may also include vitamins, minerals, and combinations thereof. Exemplary vitamins include, but are not limited to, vitamin A; vitamin E; vitamin D2; vitamin D3, including cholecalciferol, 1 ,25-dihydroxycholecalciferol, 24,25-dihydroxycholecalciferol, 1 -hydroxycholecalciferol, and 25 -hydroxy cholecalciferol; vitamin A palmitate; vitamin E acetate; vitamin C palmitate (ascorbyl palmitate); vitamin K; thiamine; riboflavin; pyridoxine; vitamin B12; carotenoids (e.g., beta-carotene, zeaxanthin, lutein, lycopene); niacin; folic acid; pantothenic acid; biotin; vitamin C; choline; inositol; salts and derivatives thereof, and combinations thereof. Exemplary minerals include, but are not limited to, calcium; selenium; potassium; iodine; phosphorus; magnesium; iron; zinc; manganese; copper; sodium; molybdenum; chromium; chloride; and combinations thereof. [00095] In a specific exemplary embodiments of the methods disclosed herein, the nutritional composition includes water; corn syrup; sucrose; milk protein concentrate; sodium caseinate; canola oil; corn oil; fructooligosaccharides; soy protein isolate; calcium beta-hydroxy-beta- methylbutyrate (Ca-HMB); whey protein concentrate; potassium citrate; natural and artificial flavors; potassium phosphate; lecithin; cellulose gel; magnesium hydroxide; calcium carbonate; ascorbic acid; calcium phosphate; choline chloride; sodium chloride; sodium phosphate; potassium hydroxide; zinc sulfate; cellulose gum; L-carnitine; carrageenan; DL-alpha-tocopherol acetate; dextrose; ferrous sulfate; maltodextrin; niacinamide; gellan gum; calcium pantothenate; citric acid; cupric sulfate; manganese sulfate; chromium chloride; thiamine chloride hydrochloride; coconut oil; vitamin A; palmitate pyridoxine hydrochloride; riboflavin; folic acid; biotin; sodium selenate; sodium molybdate; potassium iodide; phylloquinone; cyanocobalamin; and vitamin D3.

[00096] An exemplary liquid nutritional composition containing HMB and formulated for use with diabetic subjects is provided in Table III below, with the specific ingredients provided immediately thereafter. While the formulation as provided in Table III contains only HMB and no leucine, isoleucine, or valine, it should be understood that any one or more of these branched chain amino acids could be added to the formulation in the amounts previously discussed.

Table III: Diabetic Formulation Information

Molybdenum Meg 38

[00097] The exemplary liquid nutritional composition described in Table III includes water, corn maltodextrin, sodium & calcium casemates; maltitol syrup; high oleic safflower oil; fructose; soy protein isolate; soy fiber; short-chain fructooligosaccharides; canola oil;calcium phosphate; magnesium chloride; soy lecithin; artificial fiavor; sodium citrate; magnesium phosphate; potassium citrate; potassium chloride; potassium phosphate; ascorbic acid; choline chloride; DL-alpha-tocopheryl acetate; gellan gum; acesulfame potassium; ferrous sulfate; zinc sulfate; niacinamide; manganese sulfate; calcium pantothenate; cupric sulfate; sucralose; pyridoxine hydrochloride; thiamine chloride hydrochloride; vitamin A palmitate; riboflavin; chromium chloride; beta-carotene; folic acid; biotin; sodium molybdate; potassium iodide; sodium selenate; phylloquinone; cyanocobalamin; and vitamin D3. [00098] An exemplary liquid nutritional composition containing essentially no fat, having a clear, juice-like appearance, and having an acidic pH is provided in Table IV below, with the specific ingredients provided immediately thereafter. While the composition as provided in Table IV contains only HMB and no leucine, isoleucine, or valine, it should be understood that any one or more of these branched chain amino acids could be added to the formulation in the amounts previously discussed.

Table IV: Clear Formulation Information

Iodine % DV 35

Selenium % DV 20

Chromium % DV 15

Molybdenum % DV 50

[00099] The liquid nutritional composition described in Table IV includes water, corn syrup solids, sugar, whey protein isolate and less than 0.5% of the following: citric acid, natural and artificial flavor, phosphoric acid, ascorbic acid, acesulfame potassium, sucralose, zinc sulfate, DL-alpha-tocopheryl acetate, ferrous sulfate, niacinamide, manganese sulfate, calcium pantothenate, cupric sulfate, fd&c yellow #6, vitamin A palmitate, thiamine chloride hydrochloride, pyridoxine hydrochloride, fd&c red #40, riboflavin, folic acid, chromium chloride, sodium molybdate, biotin, potassium iodide, sodium selenate, phylloquinone, vitamin D3, and cyanocobalamin.

[000100] The various exemplary embodiments of the nutritional compositions disclosed or otherwise suggested herein may be prepared by any process or suitable method (now known or known in the future) for making a selected product form, such as a nutritional solid, a nutritional liquid, or a nutritional powder. Many such techniques are known for any given product form such as nutritional liquids or nutritional powders and can easily be applied by one of ordinary skill in the art to the various embodiments of the nutritional composition according to the exemplary embodiments disclosed herein.

[000101] Liquid nutritional compositions can be manufactured by any process or suitable method for making nutritional emulsions. In one suitable manufacturing process, at least three separate slurries are prepared. These slurries include: a protein-in-fat (PIF) slurry, a carbohydrate-mineral (CHO-MIN) slurry and a protein-in-water (PIW) slurry. The PIF slurry is formed by heating and mixing any oils that are selected for the fat component (when present) and then adding an emulsifier (e.g., lecithin), fat-soluble vitamins and a portion of the total protein (preferably about half of the milk protein concentrate) with continued heat and agitation. The CHO-MIN slurry is formed by adding to water (with heat and agitation), minerals (e.g., potassium citrate, dipotassium phosphate, sodium citrate), trace and ultra trace minerals (often as pre-mix(es)), thickening-type or suspending agents (e.g., Avicel, gellan, carragenan) and any HMB source. The CHO-MIN slurry that results is held for 10 minutes with continued heat and agitation and then additional minerals may be added (e.g., potassium chloride, magnesium carbonate, potassium iodide) and/or carbohydrates (e.g., fructooligosaccharides, sucrose, corn syrup). The PIW slurry is formed by mixing the remaining protein (i.e., sodium caseinate, soy protein, why protein) into water.

[000102] The three slurries are blended together with heat and agitation and the pH is adjusted to the desired range (typically near neutral, around 6.6-7), after which the composition is subjected to high-temperature short-time (HTST) processing during which time the composition is heat treated, emulsified, and homogenized and allowed to cool. Water soluble vitamins and ascorbic acid are added (if applicable), the pH is again adjusted (if necessary), flavors are added and any additional water can be added to adjust the solids content to the desired range.

[000103] A nutritional solid, such as a spray dried nutritional powder or drymixed nutritional powder, may be prepared by any known or otherwise effective techniques, suitable for making and formulating a nutritional powder.

[000104] For example, when the nutritional powder is a spray dried nutritional powder, the spray drying step may likewise include any spray drying technique that is known for or otherwise suitable for use in the production of nutritional powders. Many different spray drying methods and techniques are known for use in the nutrition field, all of which are suitable for use in the manufacture of the spray dried nutritional powders herein.

[000105] One method of preparing the spray dried nutritional powder comprises forming and homogenizing an aqueous slurry or liquid comprising predigested fat, and optionally protein, carbohydrate, and other sources of fat, and then spray drying the slurry or liquid to produce a spray dried nutritional powder. The method may further comprise the step of spray drying, drymixing, or otherwise adding additional nutritional ingredients, including any one or more of the ingredients described herein, to the spray dried nutritional powder.

[000106] Other suitable methods for making nutritional products are described, for example, in U.S. Pat. No. 6,365,218 (Borschel et al); U.S. Pat. No. 6,589,576 (Borschel et al); U.S. Pat. No. 6,306,908 (Carlson et al); and U.S. Pat. Appl. No. 20030118703 Al (Nguyen et al); which descriptions are incorporated herein by reference in their entireties to the extent that they are consistent herewith.

[000107] The following examples are included for purposes of illustration and are not intended to limit the scope of the methods described herein. EXAMPLES

Example 1: Leucine-to-HMB Conversion in Normal vs Dexamethasone-treated Rats

[000108] Twenty 10-11 week old rats were divided into 2 groups of 10 animals. Group 1 received 0.25 mg/kg dexamethasone for a period of 21 days. Group 2 rats were given saline. After 21 days, each Group received a single bolus dose of leucine at 1.0 g/kg body weight. Blood samples were collected at 0 hr., 30 min., 1 hr., 2 hr., 3 hr., 4 hr., 6 hr., 8 hr., and 10 hr., post-dosing. As illustrated in Figures 1 and 2, Group 1 dexamethasone-treated rats exhibited a significant reduction in HMB concentration over a 10 hour period compared to the untreated Group 2.

[000109] The Group 1 and Group 2 rats were then tested for lean body mass. As illustrated in Figure 3, the Group 2 rats, given only saline had approximately 100 grams of lean mass more than the rats from Group 1, treated with dexamethasone. It is believed that the difference in lean mass is related to the lack of leucine to HMB conversion happening in the rats undergoing treatment with dexamethasone.

Example 2: Leucine-to-HMB Conversion in Females vs. Male Rats

[000110] Sixteen 6-7 week old rats were divided into two groups based on sex, with eight rats in each group. Each group received 1 gm/kg (10 ml/kg) body weight leucine and blood samples were taken from each rat at 0 hr., 15 min., 30 min., 1 hr., 2 hr., 3 hr., 4 hr., 6 hr., 8 hr., and 10 hr., post-dosing. As illustrated in Figure 4, male rats exhibited a higher concentration of HMB as compared to females after 30 minutes. This indicates that male rats have a higher efficiency in the conversion of leucine to HMB, as compared to female rats.

[000111] Figure 5 illustrates the difference in the HMB AUC of males and females after administration of 1 g/kg (10 ml)/kg body weight of leucine. The AUC (area under the curve) represents the total amount of HMB produced endogenously by the body. The female rats demonstrated an average AUC of about 1750 hr*ng/ml, while the male rats demonstrated an AUC of about 2200 hr*ng/ml.

Example 3: Leucine-to-HMB Conversion in Diabetic Rats

[000112] Zucker diabetic fatty male rats and Zucker lean male rats (11 weeks old) were divided into two groups, with eight animals in each group. Each group received 1.0 g/kg body weight leucine and blood samples were taken from each rat at 0 hr., 15 min., 30 min., 1 hr., 2 hr., 3 hr., 4 hr., 6 hr., 8 hr., and 10 hr., post-dosing. [000113] As illustrated in Figure 6, for a period of six hours after the point of administration, diabetic rats illustrated a higher efficiency for converting leucine to HMB. For instance, at 2 hours post-dosing, diabetic rats exhibited an HMB concentration of about 1300 ng/ml, compared to lean rats, which demonstrated an HMB concentration of about 800 ng/ml.

[000114] Figure 7 illustrates the difference in HMB AUC of Zucker diabetic fatty male rats and Zucker lean male rats after administration of 1.0 g/kg body weight of leucine. The AUC represents the total amount of HMB absorbed by the body. The Zucker diabetic fatty rats demonstrated an average AUC of about 5000 hr*ng/ml, while the Zucker lean rats demonstrated an AUC of about 4100 hr*ng/ml.

[000115] The complete disclosure of all patents, patent applications, and publications, and electronically available material cited herein are incorporated by reference. The foregoing detailed description and examples have been given for clarity of understanding only. No unnecessary limitations are to be understood therefrom. The invention is not limited to the exact details shown and described, for variations obvious to one skilled in the art will be included within the invention defined by the claims.