# Patent

## Title
Methods for enhancing the quality of life of a senior animal

## Abstract

Methods for enhancing the quality of life of a senior or super senior animal by feeding the animal a composition comprising at least one omega-3 polyunsaturated fatty acid and various combinations of amino acids, minerals, and antioxidants in amounts effective to enhance alertness, improve vitality, protect cartilage, maintain muscle mass, enhance digestibility, and improve skin and pelage quality. Changes in expression of genes associated with several biological pathways induced in an animal by feeding it said composition are consistent with an enhanced quality of life.

## Background
CROSS-REFERENCE TO RELATED APPLICATIONS
This application is a continuation-in-part of pending U.S. patent application Ser. No. 11/813,276, filed Mar. 28, 2008, which is a US national stage entry under 35 U.S.C. §371 of International Application No. PCT US 2005/047461 filed Dec. 30, 2005, publication No. WO 2006/074089, which claims priority to U.S. Provisional Application Ser. No. 60/640,890, filed Dec. 30, 2004, each of which is incorporated by reference in its entirety for all purposes.


FIELD OF THE INVENTION
The present invention relates generally to methods for enhancing the quality of life of an animal and particularly to using food compositions containing omega-3 polyunsaturated fatty acids for enhancing the quality of life of a senior or super senior animal.
BACKGROUND OF THE INVENTION
Companion animals such as dogs and cats frequently require differing diets depending on their life stage (age), size, body composition, and breed. Both dog and cat nutrient requirements can be separated into three different life-stages, based on age: growing dogs (or cats), adult dogs (or cats), and senior dogs (or cats). The latter category, senior dogs (or cats), can be further separated into two stages, which include senior (or mature adult) and super senior (or geriatric). Dogs are further separated into different categories for regular breed dogs versus large-breed dogs.
Essential fatty acids, consisting of omega-3 and omega-6 polyunsaturated fatty acids, are critical nutrients for the health of an animal. These nutrients, however, either cannot be made by animals or cannot be made in sufficient amounts to elicit benefits and therefore must be consumed in an animal's diet. See, e.g., Hornstra, G., et al., “Essential fatty acids in pregnancy and early human development”, Eur. J. Obs. & Gyn. and Reprod. Biology, 61:57-62 (1995). It has previously been postulated that Docosahexaenoic Acid (“DHA”), an omega-3 polyunsaturated fatty acid, is effective in increasing the maze-learning ability and brain functions in aged mice. See, Lim, S.-Y., “Intakes of dietary docosahexaenoic acid ethyl ester and egg phosphatidylcholine improve maze-learning ability in young and old mice”, J. Nutr., 130:1629-1632 (2000).
Rogers discusses the theory of the potential use of antioxidants to slow the deterioration of cognitive function, particularly in the elderly. See Rogers, P., “A healthy body, a healthy mind: long-term impact of diet on mood and cognitive function”, Proceedings of the Nutrition Society, 60:135-143 (2001).
Despite the studies and developments relating to improving cognitive abilities, there continues to be a need for methods for enhancing the quality of life of senior animals, as measured by, e.g., enhanced alertness, improved vitality, cartilage protection, maintenance of muscle mass, enhanced digestibility, and improved skin and pelage quality in senior and super senior animals. As previously reported, the super senior pet food composition described herein may be administered to achieve this result. Additionally, we now report herein our surprising discovery that the enhanced quality of life of senior and super senior animals achieved by the administration of the pet food compositions disclosed herein is reflected at the genomic level. Specifically, as described in detail in the Examples below, gene chip data indicate that the expression of genes that encode proteins associated with several biological pathways such as blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport are modified, i.e., in general, the majority are beneficially altered through administration to the animal of the super senior pet food compositions described herein.
SUMMARY OF THE INVENTION
The invention provides methods for improving the quality of life of senior and super senior animals by feeding the animal a composition comprising at least about 9% by weight protein, at least about 5% by weight fat, and at least about 0.05% by weight of at least one omega-3 polyunsaturated fatty acid.
In one embodiment, the method comprises feeding the animal an amount of a composition effective to enhance the animal's quality of life, wherein enhanced quality of life is evidenced by improvement in one or more characteristics selected from the group consisting of alertness, vitality, cartilage protection, muscle mass maintenance, digestibility, and skin and pelage quality.
In another embodiment, the method comprises feeding the animal a composition comprising at least one omega-3 polyunsaturated fatty acid selected from the group consisting of docosahexaenoic acid (“DHA”) and eicosapentaenoic acid (“EPA”). In an additional embodiment, the method comprises feeding the animal a composition further comprising at least one antioxidant and at least one nutrient selected from the group consisting of choline, manganese, methionine, cysteine, L-carnitine, lysine, and mixtures thereof.
In one embodiment, the method comprises feeding the animal an amount of a composition effective to improve or enhance the animal's quality of life, wherein enhanced quality of life is evidenced by improvement in one or more biological pathways selected from the group consisting of blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport.
In another embodiment, the method comprises feeding the animal an amount of a composition effective to enhance the animal's quality of life, wherein enhanced quality of life is evidenced by a change in expression of one or more genes which encode proteins associated with or related to biological pathways selected from the group consisting of blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport.
In yet another embodiment, the invention relates to a method to treat an animal suffering from a disorder or disease associated with or related to a biological pathway selected from the group consisting of blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport comprising administering to said animal a composition disclosed herein. In one embodiment, said composition comprises at least about 9% by weight protein, at least about 5% by weight fat, and at least about 0.05% by weight of at least one omega-3 polyunsaturated fatty acid. In a further embodiment said composition comprises at least one omega-3 polyunsaturated fatty acid selected from the group consisting of docosahexaenoic acid (“DHA”) and eicosapentaenoic acid (“EPA”). In yet an additional embodiment, the composition further comprises at least one antioxidant and at least one nutrient selected from the group consisting of choline, manganese, methionine, cysteine, L-carnitine, lysine, and mixtures thereof.
In another embodiment, the invention relates to methods of measuring or characterizing the enhancement in the quality of life of an animal, particularly a senior or super senior animal, fed a composition described herein by quantitating the gene expression levels of one or more genes selected from a group consisting of those disclosed in Tables 5-14 in said animal and comparing said levels in the animal to levels in the animal prior to administration of the feed composition.
In a further embodiment, the invention relates to methods to enhance the quality of life of an animal by modulating the expression level of one or more genes listed on Tables 5-14 (i.e., up or down regulation as indicated therein) in an animal in order to mimic the pattern of expression seen in vivo after administration of the pet food compositions of the present invention. It is also contemplated herein that modulating the expression levels of these genes may have therapeutic value with regard to the treatment of diseases or disorders associated with the various biological pathways.
The invention also relates to methods to identify an animal that might benefit from feeding a composition as disclosed herein comprising measuring the gene expression levels of any one or more genes listed in Tables 5-14 in said animal and comparing said levels to the gene expression levels seen in Tables 5-14 wherein an animal with levels different than those seen in Tables 5-14 would be identified as potentially benefiting from feeding a composition of the present invention.
In yet another aspect of the present invention there are provided assay methods and kits comprising the components necessary to detect expression of polynucleotides encoding the genes disclosed herein, or levels of encoded protein, or fragments thereof, in body tissue samples derived from an animal, such kits comprising, e.g., antibodies that bind to said polypeptides, or to fragments thereof, or oligonucleotide probes that hybridize with said polynucleotides. In a preferred embodiment, such kits also comprise instructions detailing the procedures by which the kit components are to be used.
Other and further objects, features, and advantages of the present invention will be readily apparent to those skilled in the art.


DETAILED DESCRIPTION OF THE INVENTION
Definitions
It is contemplated that the invention described herein is not limited to the particular methodology, protocols, and reagents described as these may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention in any way.
Unless defined otherwise, all technical and scientific terms used herein have the same meanings as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods, devices and materials are now described. All publications mentioned herein are incorporated by reference for the purpose of describing and disclosing the materials and methodologies that are reported in the publication which might be used in connection with the invention.
In practicing the present invention, many conventional techniques in molecular biology may be used. These techniques are well known and are explained in, for example, Current Protocols in Molecular Biology, Volumes I, II, and III, 1997 (F. M. Ausubel ed.); Sambrook et al., 1989, Molecular Cloning: A Laboratory Manual, Second Edition, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y.; DNA Cloning: A Practical Approach, Volumes I and II, 1985 (D. N. Glover ed.); Oligonucleotide Synthesis, 1984 (M. L. Gait ed.); Nucleic Acid Hybridization, 1985, (Hames and Higgins); Transcription and Translation, 1984 (Hames and Higgins eds.); Animal Cell Culture, 1986 (R. I. Freshney ed.); Immobilized Cells and Enzymes, 1986 (IRL Press); Perbal, 1984, A Practical Guide to Molecular Cloning; the series, Methods in Enzymology (Academic Press, Inc.); Gene Transfer Vectors for Mammalian Cells, 1987 (J. H. Miller and M. P. Calos eds., Cold Spring Harbor Laboratory); and Methods in Enzymology Vol. 154 and Vol. 155 (Wu and Grossman, and Wu, eds., respectively).
As used herein and in the appended claims, the singular forms “a”, “an”, and “the” include plural reference unless the context clearly dictates otherwise.
The terms “senior” or “mature adult” refers to the life-stage of an animal. For small or regular breed canines, the “senior” life stage is from about 7 to about 10 years of age. For felines, the “senior” life stage is from about 7 to about 12 years of age. For large breed canines, over 5 years of age represents “super senior” as described below.
The terms “super senior” or “geriatric” refers to a specific life-stage of an animal. For small or regular breed canines, the super senior stage is any age greater than 10 years of age. For large breed canines, the super senior stage is any age greater than 5 years of age. For felines, the super senior stage is any age greater than 12 years of age.
The term “large breed” canine means a canine that weighs more than 55 pounds when an adult.
The term “regular breed” canine means a canine that weighs less than 55 pounds when an adult.
The term “small breed” canine means a canine that weighs less than 20 pounds when an adult.
The term “super senior pet food composition” refers to any and all of the pet food compositions disclosed herein.
The term “carbohydrate” as used herein includes polysaccharides (e.g., starches and dextrins) and sugars (e.g. sucrose, lactose, maltose, glucose, and fructose) that are metabolized for energy when hydrolyzed. Examples of carbohydrates suitable for inclusion in the compositions disclosed herein include, but are not limited to, corn, grain sorghum, wheat, barley, and rice.
The term “antioxidant” means a substance that is capable of reacting with free radicals and neutralizing them. Illustrative examples of such substances include beta-carotene, selenium, coenzyme Q10 (ubiquinone), luetin, tocotrienols, soy isoflavones, S-adenosylmethionine, glutathione, taurine, N-acetylcysteine, vitamin E, vitamin C, lipoic acid and L-carnitine. Examples of foods containing useful levels of one or more antioxidants include but are not limited to ginkgo biloba, green tea, broccoli, citrus pulp, grape pomace, tomato pomace, carrot spinach, and a wide variety of fruit meals and vegetable meals. It will be understood by one of skill in the art that while units of antioxidants may be provided herein as “ppm”, appropriate amounts of antioxidants may also be provided as “IU/kg” where appropriate and customary for a given antioxidant such as, e.g., Vitamin E
The terms “beneficial change” in gene expression, or gene expression may be “beneficially altered” and like terms refer to a modification in gene expression (e.g., up or down regulation of mRNA levels) such that levels of proteins encoded by the genes may be correspondingly modified such that an associated biological pathway may be more likely to function normally and with less tendency to reflect pathological changes in the pathway that, e.g., may be typical of a super senior animal. Generally, beneficial changes in gene expression relate to improved health and/or reduced propensity for disease in an animal. As used herein, measuring differences in gene expression “levels” and like terms refer to, e.g., characterizing whether expression of a gene is up or down regulated in an animal compared to a control level.
As used herein, “improving” or “enhancing” the quality of life of an animal refers to as an improvement or enhancement in one or more characteristics selected from a group consisting of alertness, vitality, protection of cartilage, maintenance of muscle mass, digestibility, and skin and pelage quality. Additionally, improvement/enhancement in blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport are also contemplated.
An “improvement” or an “enhancement” in a characteristic or biological pathway refers to a modification in said characteristic or biological pathway such that there is a tendency for the characteristic or pathway to appear and/or function normally and with less tendency to reflect pathological changes in the characteristic or pathway that, e.g., may be typical of a super senior animal.
As used herein, methods to “treat” an animal suffering from a disease or disorder is also meant to encompass methods to prevent and/or to ameliorate the disease or disorder as well.
The Invention
The present invention provides methods for improving or enhancing the quality of life of a senior or super senior animal. The methods comprise feeding the animal a composition comprising at least about 9% by weight protein, at least about 5% by weight fat, and at least about 0.05% by weight omega-3 polyunsaturated fatty acid. The methods are useful for enhancing alertness, improving vitality, protecting cartilage, maintaining muscle mass, enhancing digestibility, and improving skin and pelage quality in a senior or super senior animal. The methods are also useful for improving in an animal one or more biological pathways selected from the group consisting of blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and the electron transport pathway, such improvements also being reflected in overall beneficial changes at the genomic level. Methods for treating animals suffering from disorders or diseases associated with or related to these biological pathways comprising administering the compositions of the present invention are also contemplated herein.
Without being bound by theory, the benefits of the invention may be the result of physiological effects from the addition of omega-3 polyunsaturated fatty acids to a senior or super senior animal's diet. Similarly, the antioxidants, choline, and other nutrients may play a role in enhancing a senior or super senior animal's quality of life.
Although the methods of the present invention may improve an animal's quality of life by enhancing all of the above described characteristics or improving all of the described biological pathways, it is not necessary to demonstrate substantial improvements in each of the characteristics or pathways to achieve the “enhanced quality of life” as defined herein.
When the compositions are administered to a senior or super senior animal, the animal experiences an enhanced quality of life, e.g., exhibits or experiences one or more of enhanced alertness, improved vitality, protected cartilage, maintained muscle mass, enhanced digestibility, improved skin and pelage quality, as well as improvements in e.g., blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and the electron transport pathway as indicated by overall beneficial changes at the genomic level. Methods for determining these measurements of quality of life are known to skilled artisans. For example, alertness can be measured by various means, including an analysis of metabolism and antioxidant markers, as well as through clinical studies with follow-up questions to participating pet owners. Potential metabolism markers may include ghrelin, GLP-1, thyroid hormone, and/or growth hormone. Potential markers of antioxidant status may include serum vitamin E, ORAC, glutathione peroxidase, alkanels, and/or cell damage indicators. Further, vitality can be measured by various means, including an analysis of metabolism and antioxidant markers, as well as through clinical studies with follow-up questions to participating pet owners. Similarly, cartilage protection can be measured by various means, including an analysis of arthritis biomarkers. Potential arthritis biomarkers may include type TI collagen synthesis, matrix metaloproteinase, osteocalcin, alkaline phosphatase activity, COMP, and fragments of cartilage damage. Muscle mass maintenance can be measured by various means, including an analysis of body composition and digestibility can be measured by various means, including clinical studies with follow-up questions to participating pet owners and animal feeding to determine the percentage of nutrients digested. Skin and pelage quality can be measured by various means, including clinical studies with follow-up questions to participating pet owners. Additionally, as discussed above, improvements in quality of life is also reflected at the genomic level, as evidenced by gene chip data which indicate beneficial changes on the expression of a majority of genes associated with various important biological pathways including blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and protection and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and the electron transport pathway. The identities of these genes are provided in the Examples below.
The methods of the invention are useful for enhancing the quality of life of humans and animals, including primates (e.g., monkeys, chimpanzees, etc.), companion animals (e.g., dogs, cats, horses, etc.), farm animals (e.g., goats, sheep, swine, cattle, etc.), laboratory animals (e.g., mice, rats, etc.), birds (e.g., domestic birds such as canaries, parrots, etc. and commercial birds such as chickens, ducks, turkeys, etc.), rodents (e.g., hamsters, guinea pigs, gerbils, rabbits, hedgehogs, ferrets, chinchillas, etc.), and wild, exotic, and zoo animals (e.g., wolves, bears, deer, etc.). In various embodiments, the animal is a cat, a dog, or a horse.
The compositions of the present invention are designed to enhance digestibility and improve chewability. Canine and feline foods are typically formulated based on life stage (age), size, body composition, and breed. Thus, some embodiments of the present invention include compositions that are formulated to address specific nutritional differences between regular or small breed dogs, large breed dogs, and cats.
The invention provides methods utilizing a variety of compositions containing at least one omega-3 polyunsaturated fatty acid. The compositions include foods, supplements, treats, and toys (typically chewable and consumable toys). The methods also provide the compositions to the designated animals over a period of time that is long enough to effectuate the improved quality of life. In one embodiment, the method provides the animal with a composition for at least thirty days.
The compositions for use in the methods of the present invention generally have an omega-3 polyunsaturated fatty acid content of at least about 0.02% (or from about 0.05% to about 10%, or from about 0.1% to about 6%) by weight on a dry matter basis. In some embodiments, the omega-3 polyunsaturated fatty acid is DHA. In other embodiments, the omega-3 polyunsaturated fatty acid is EPA. In still other embodiments, the omega-3 polyunsaturated fatty acid comprises a mixture of DHA and EPA.
In some embodiments, the composition containing omega-3 polyunsaturated fatty acid is a food. Although both liquid and solid foods are provided, solid foods are typically preferred. Foods include both dry foods and wet foods. Some of the non-polyunsaturated fatty acid components of the food, and their preferred proportions, include those listed in Table 1.








TABLE 1






Proportion of the composition (% of dry weight of


Component
composition or parts per million)







Protein
from about 9% to about 55%, or from about 18% to about



30%, or from about 33% to about 55% or from about 18%



to about 20% or from about 33% to about 36%


Fat
from about 7% to about 35%, or from about 18% to about



35%, or from about 7% to about 24%, or from about 14%



to about 24%, or from about 14% to about 16% or from



about 18% to about 24%


Antioxidant
from about 0 ppm to about 7500 ppm, or from about 0.05



ppm to about 3600 ppm, or from about 250 to about 3600,



or from about 250 ppm to about 1650 ppm, or from about



5 ppm to about 225 ppm, or from about 0.05 ppm to about



2.4 ppm









In one embodiment, the methods of this invention comprise feeding a super senior animal a composition in an amount effective to enhance the animal's quality of life. Such compositions generally comprise:

    
    
        (a) 0.02% (or from about 0.05% to about 10%, or from about 0.1% to about 6%) at least one omega-3 polyunsaturated fatty acid, and
        (b) at least one of the following:
        
            (i) from about 10% to about 55% (or from about 18% to about 30%, or from about 33% to about 55% or from about 18% to about 20% or from about 33% to about 36%) protein,
            (ii) from about 7% to about 35% (or from about 18% to about 35%, or from about 7% to about 24%, or from about 14% to about 24%, or from about 14% to about 16% or from about 18% to about 24%) fat, and
            (iii) at least about 0.05 (or from about 0.05 ppm or IU/kg to about 7500 ppm or IU/kg, or from about 250 ppm or IU/kg to about 3600 ppm or IU/kg, or from about 250 ppm or IU/kg to about 1650 ppm or IU/kg, or from about 5 ppm or IU/kg to about 225 ppm or IU/kg, or from about 0.05 ppm or IU/kg to about 2.4 ppm or IU/kg) antioxidant.
        
        
    
    


In another embodiment, the methods of this invention comprise feeding a super senior regular or small breed canine a composition in an amount effective to enhance the canine's quality of life. The composition generally comprises:

    
    
        (a) at least one of the following:
        
            (i) at least about 0.02% (or from about 0.02% to about 0.3%, or from about 0.05% to about 0.3%, or from about 0.05% to about 0.2%) DHA, and
            (ii) at least about 0.1% (or from about 0.1% to about 0.5%, or from about 0.2% to about 0.5%, or from about 0.2% to about 0.3%) EPA,
        
        
        (b) at least about 9% (or from about 9% to about 30%, or from about 18% to about 30%, or from about 18% to about 20%) protein,
        (c) at least about 7% (or from about 7% to about 24%, or from about 14% to about 24%, or from about 14% to about 16%) fat, and
        (d) at least one of the following:
        
            (i) at least about 250 IU/kg (or from about 250 IU/kg to about 1500 IU/kg, or from about 500 IU/kg to about 1500 IU/kg, or from about 500 IU/kg to about 1000 IU/kg) vitamin E,
            (iv) at least about 50 ppm (or from about 50 ppm to about 500 ppm, or from about 100 ppm to about 500 ppm, or from about 100 ppm to about 301 ppm) vitamin C,
            (v) at least about 600 ppm (or from about 600 ppm to about 2400 ppm, or from about 1260 ppm to about 2400 ppm, or from about 1260 ppm to about 1545 ppm) taurine,
            (vi) at least about 50 ppm (or from about 50 ppm to about 200 ppm, or from about 100 to about 160, or from about 100 to about 155) lipoic acid, and
            (vii) at least about 50 ppm (or from about 50 ppm to about 500 ppm, or from about 200 ppm to about 500 ppm, or from about 200 ppm to about 350 ppm) carnitine.
        
        
    
    


In another embodiment, the methods of this invention comprise feeding a super senior large breed canine a composition in an amount effective to enhance the canine's quality of life. The compositions generally comprise:

    
    
        (a) at least one of the following:
        
            (i) at least about 0.02% (or from about 0.02% to about 0.3%, or from about 0.05% to about 0.3%, or from about 0.05% to about 0.2%) DHA, and
            (ii) at least about 0.1% (or from about 0.1% to about 0.5%, or from about 0.2% to about 0.5%, or from about 0.2% to about 0.3%) EPA,
        
        
        (b) at least about 9% (or from about 9% to about 30%, or from about 18% to about 30%, or from about 18% to about 20%) protein,
        (c) at least about 7% (or from about 7% to about 24%, or from about 14% to about 24%, or from about 14% to about 16%) fat, and
        (d) at least one of the following:
        
            (i) at least about 250 IU/kg (or from about 250 IU/kg to about 1500 IU/kg, or from about 500 IU/kg to about 1500 IU/kg, or from about 500 IU/kg to about 1000 IU/kg) vitamin E,
            (viii) at least about 50 ppm (or from about 50 ppm to about 500 ppm, or from about 100 ppm to about 500 ppm, or from about 100 ppm to about 301 ppm) vitamin C,
            (ix) at least about 600 ppm (or from about 600 ppm to about 2400 ppm, or from about 1260 ppm to about 2400 ppm, or from about 1260 ppm to about 1575 ppm) taurine, and
            (x) at least about 50 ppm (or from about 50 ppm to about 200 ppm, or from about 100 to about 160, or from about 100 to about 155) lipoic acid, and
            (xi) at least about 50 ppm (or from about 50 ppm to about 500 ppm, or from about 200 ppm to about 500 ppm, or from about 200 ppm to about 350 ppm) carnitine.
        
        
    
    


In another embodiment, the methods of this invention comprise feeding a super senior feline a composition in an amount effective to enhance the feline's quality of life. The compositions generally comprise:

    
    
        (a) at least one of the following:
        
            (i) at least about 0.05% (or from about 0.05% to about 0.30%, or from about 0.1% to about 0.30%, or from about 0.1% to about 0.2%) DHA, and
            (ii) at least about 0.1% (or from about 0.1% to about 0.5%, or from about 0.2% to about 0.5%, or from about 0.2% to about 0.3%) EPA,
        
        
        (b) at least about 15% (or from about 15% to about 55%, or from about 30% to about 55%, or from about 33% to about 36%) protein,
        (c) at least about 9% (or from about 9% to about 35%, or from about 18% to about 35%, or from about 18% to about 24%) fat, and
        (d) at least one of the following:
        
            (i) at least about 250 IU/kg (or from about 250 IU/kg to about 1500 IU/kg, or from about 500 IU/kg to about 1500 IU/kg, or from about 500 IU/kg to about 1100 IU/kg) vitamin E,
            (xii) at least about 50 ppm (or from about 50 ppm to about 300 ppm, or from about 100 ppm to about 300 ppm, or from about 100 ppm to about 200 ppm) vitamin C,
            (xiii) at least about 1100 ppm (or from about 1100 ppm to about 3500 ppm, or from about 2300 ppm to about 3500 ppm, or from about 2300 ppm to about 2350 ppm) taurine, and
            (xiv) at least about 200 ppm (or from about 200 to about 750 ppm, or from about 400 ppm to about 750 ppm, or from about 400 to about 525 ppm) carnitine, and
            (xv) at least about 0.05% (or from about 0.05% to about 0.6%, or from about 0.1% to about 0.6%, or from about 0.1% to about 0.4%) cystine.
        
        
    
    


In another embodiment, the methods of this invention comprise feeding a super senior animal a composition in an amount effective to enhance the animal's alertness and vitality. The composition generally comprises:

    
    
        (a) 0.02% (or from about 0.05% to about 10%, or from about 0.1% to about 6%) at least one omega-3 polyunsaturated fatty acid, and
        (b) at least one of the following:
        
            (xvi) from about 10% to about 55% (or from about 18% to about 30%, or from about 33% to about 55% or from about 18% to about 20% or from about 33% to about 36%) protein,
            (xvii) from about 7% to about 35% (or from about 18% to about 35%, or from about 7% to about 24%, or from about 14% to about 24%, or from about 14% to about 16% or from about 18% to about 24%) fat,
            (xviii) at least about 0.05 (or from about 0.05 ppm to about 7500 ppm, or from about 250 to about 3600, or from about 250 ppm to about 1650 ppm, or from about 5 ppm to about 225 ppm, or from about 0.05 ppm to about 2.4 ppm) antioxidant, and
            (xix) at least about 1000 ppm (or from about 1000 ppm to about 5000 ppm, from about 3300 ppm to about 5000 ppm, or from about 2000 ppm to about 3000 ppm, or from about 3000 ppm to about 4000 ppm) choline.
        
        
    
    


In another embodiment, the methods of this invention comprise feeding a super senior regular or small breed canine a composition in an amount effective to enhance the canine's alertness and vitality. The composition generally comprises:

    
    
        (a) at least one of the following:
        
            (i) at least about 0.02% (or from about 0.02% to about 0.3%, or from about 0.05% to about 0.3%, or from about 0.05% to about 0.2%) DHA, and (ii) at least about 0.1% (or from about 0.1% to about 0.5%, or from about 0.2% to about 0.5%, or from about 0.2% to about 0.3%) EPA,
        
        
        (b) at least about 9% (or from about 9% to about 30%, or from about 18% to about 30%, or from about 18% to about 20%) protein,
        (c) at least about 7% (or from about 7% to about 24%, or from about 14% to about 24%, or from about 14% to about 16%) fat,
        (d) at least one of the following:
        
            (i) at least about 250 IU/kg (or from about 250 IU/kg to about 1500 IU/kg, or from about 500 IU/kg to about 1500 IU/kg, or from about 500 IU/kg to about 1000 IU/kg) vitamin E,
            (xx) at least about 50 ppm (or from about 50 ppm to about 500 ppm, or from about 100 ppm to about 500 ppm, or from about 100 ppm to about 301 ppm) vitamin C,
            (xxi) at least about 600 ppm (or from about 600 ppm to about 2400 ppm, or from about 1260 ppm to about 2400 ppm, or from about 1260 ppm to about 1545 ppm) taurine, and
            (xxii) at least about 50 ppm (or from about 50 ppm to about 200 ppm, or from about 100 to about 160, or from about 100 to about 155) lipoic acid, and
            (xxiii) at least about 50 ppm (or from about 50 ppm to about 500 ppm, or from about 200 ppm to about 500 ppm, or from about 200 ppm to about 350 ppm) carnitine,
        
        
        (e) at least about 1000 ppm (or from about 1000 ppm to about 3200 ppm, or from about 2000 ppm to about 3200 ppm, or from about 2000 ppm to about 2500 ppm) choline,
        (f) at least about 50 ppm (or from about 50 ppm to about 150 ppm, or from about 100 ppm to about 150 ppm, or from about 100 ppm to about 110 ppm) manganese, and
        (g) at least about 0.4% (or from about 0.4% to about 2%, or from about 0.9% to about 2%, or from about 0.9% to about 1.2%) lysine, and
        (h) at least about 0.4% to about 1.5% methionine.
    
    


In another embodiment, the methods of this invention comprise feeding a super senior large breed canine a composition in an amount effective to enhance the canine's alertness and vitality. The composition generally comprises:

    
    
        (a) at least one of the following:
        
            (i) at least about 0.02% (or from about 0.02% to about 0.3%, or from about 0.05% to about 0.3%, or from about 0.05% to about 0.2%) DHA, and
            (ii) at least about 0.1% (or from about 0.1% to about 0.5%, or from about 0.2% to about 0.5%, or from about 0.2% to about 0.3%) EPA,
        
        
        (b) at least about 9% (or from about 9% to about 30%, or from about 18% to about 30%, or from about 18% to about 20%) protein,
        (c) at least about 7% (or from about 7% to about 24%, or from about 14% to about 24%, or from about 14% to about 16%) fat,
        (d) at least one of the following:
        
            (i) at least about 250 IU/kg (or from about 250 IU/kg to about 1500 IU/kg, or from about 500 IU/kg to about 1500 IU/kg, or from about 500 IU/kg to about 1000 IU/kg) vitamin E,
            (xxiv) at least about 50 ppm (or from about 50 ppm to about 500 ppm, or from about 100 ppm to about 500 ppm, or from about 100 ppm to about 301 ppm) vitamin C,
            (xxv) at least about 600 ppm (or from about 600 ppm to about 2400 ppm, or from about 1260 ppm to about 2400 ppm, or from about 1260 ppm to about 1575 ppm) taurine, and
            (xxvi) at least about 50 ppm (or from about 50 ppm to about 200 ppm, or from about 100 to about 160, or from about 100 to about 155) lipoic acid, and
            (xxvii) at least about 50 ppm (or from about 50 ppm to about 500 ppm, or from about 200 ppm to about 500 ppm, or from about 200 ppm to about 350 ppm) carnitine,
        
        
        (e) at least about 1000 ppm (or from about 1000 ppm to about 3200 ppm, or from about 2000 ppm to about 3200 ppm, or from about 2000 ppm to about 2500 ppm) choline,
        (f) at least about 50 ppm (or from about 50 ppm to about 150 ppm, or from about 100 ppm to about 150 ppm, or from about 100 ppm to about 110 ppm) manganese, and
        (g) at least about 0.4% (or from about 0.4% to about 2%, or from about 0.9% to about 2%, or from about 0.9% to about 1.2%) lysine, and
        (h) at least about 0.4% to about 1.5% methionine.
    
    


In another embodiment, the methods of this invention comprise feeding a super senior feline a composition in an amount effective to enhance the feline's alertness and vitality. The composition generally comprises:

    
    
        (a) at least one of the following:
        
            (i) at least about 0.05% (or from about 0.05% to about 0.30%, or from about 0.1% to about 0.30%, or from about 0.1% to about 0.2%) DHA, and
            (ii) at least about 0.1% (or from about 0.1% to about 0.5%, or from about 0.2% to about 0.5%, or from about 0.2% to about 0.3%) EPA,
        
        
        (b) at least about 15% (or from about 15% to about 55%, or from about 30% to about 55%, or from about 33% to about 36%) protein,
        (c) at least about 9% (or from about 9% to about 35%, or from about 18% to about 35%, or from about 18% to about 24%) fat,
        (d) at least one of the following:
        
            (i) at least about 250 IU/kg (or from about 250 IU/kg to about 1500 IU/kg, or from about 500 IU/kg to about 1500 IU/kg, or from about 500 IU/kg to about 1100 IU/kg) vitamin E,
            (xxviii) at least about 50 ppm (or from about 50 ppm to about 300 ppm, or from about 100 ppm to about 300 ppm, or from about 100 ppm to about 200 ppm) vitamin C,
            (xxix) at least about 1100 ppm (or from about 1100 ppm to about 3500 ppm, or from about 2300 ppm to about 3500 ppm, or from about 2300 ppm to about 2350 ppm) taurine, and
            (xxx) at least about 200 ppm (or from about 200 to about 750 ppm, or from about 400 ppm to about 750 ppm, or from about 400 to about 525 ppm) carnitine, and
            (xxxi) at least about 0.05% (or from about 0.05% to about 0.6%, or from about 0.1% to about 0.6%, or from about 0.1% to about 0.4%) cystine.
        
        
        (e) at least about 1600 ppm (or from about 1600 ppm to about 5000 ppm, or from about 3300 ppm to about 5000 ppm, or from about 3300 ppm to about 3400 ppm) choline,
        (f) at least about 50 ppm (or from about 50 ppm to about 150 ppm, or from about 100 ppm to about 150 ppm, or from about 100 ppm to about 110 ppm) manganese, and
        (g) at least about 0.7% (or from about 0.7% to about 3%, or from about 1.4% to about 3%, or from about 1.4% to about 1.7%) lysine, and
        (h) at least about 0.4% to about 1.5% methionine.
    
    


In another embodiment, this invention provides a method for improving the quality of life of a senior or super senior small or regular breed canine. The method comprises feeding the canine a composition comprising:

    
    
        from about 60% to about 70% by weight carbohydrate;
        from about 15% to about 25% by weight protein selected from the group consisting of animal protein and vegetable protein;
        from about 5% to about 7% by weight fat selected from the group consisting of animal fat and vegetable fat;
        from about 2.5% to about 4% by weight of at least one omega-3 polyunsaturated fatty acids;
        from about 1% to about 4% by weight fiber;
        from about 1% to about 2% by weight minerals; and
        from about 0.5 to about 1.5% by weight vitamins.
    
    


In another embodiment, this invention provides a method for improving the quality of life of a senior or super senior large breed canine. The method comprises feeding the canine a composition comprising:

    
    
        from about 60% to about 70% by weight carbohydrate;
        from about 15% to about 25% by weight protein selected from the group consisting of animal protein and vegetable protein;
        from about 5% to 10% by weight fat selected from the group consisting of animal fat and vegetable fat;
        from about 3% to about 5% by weight of at least one omega-3 polyunsaturated fatty acids;
        from about 1% to about 4% by weight fiber;
        from about 0.5% to about 1% by weight minerals; and
        from about 0.75 to about 1.25% by weight vitamins.
    
    


In another embodiment, this invention provides a method for improving the quality of life of a senior or super senior feline. The method comprises feeding the feline a composition comprising:

    
    
        from about 30% to about 35% by weight carbohydrate;
        from about 35% to about 50% by weight protein selected from the group consisting of animal protein and vegetable protein;
        from about 12% to about 15% by weight fat selected from the group consisting of animal fat and vegetable fat;
        from about 1% to about 2% by weight of at least one omega-3 polyunsaturated fatty acids;
        from about 1% to about 5% by weight fiber;
        from about 1% to about 2% by weight minerals; and
        from about 1% to about 2% by weight vitamins.
    
    


In a further embodiment, this invention provides a method for improving the quality of life of a senior or super senior animal comprising feeding the animal (e.g., small, regular or large breed canine or feline, as the case may be) a composition comprising the components as indicated in Table 1A below:







TABLE 1A







Chemical composition of Super Senior Foods











Small/Regular
Large




Breed
Breed



Nutrient Component
Canine
Canine
Feline













Crude Protein, %
20.1
19.34
35.73


Fat, %
16.45
16.92
22.47


Calcium, %
0.71
0.73
0.94


Phosphorus, %
0.61
0.68
0.77


EPA, %
0.32
0.32
0.23


DHA, %
0.22
0.22
0.32


Linoleic Acid, %
3.96
4.04
5.05


Total N-3 fatty acids, %
1.3
2.24
1.14


Total N-6 fatty acids, %
3.96
3.99
5.09


Taurine, ppm
1400
15.25
2100


Carnitine, ppm
314
337
367


Methioinine, %
1
1.19
1.32


Cystine, %
0.25
0.24
0.47


Manganese, ppm
87
100
104


Vitamin E, IU/kg
1492
1525
1292


Vitamin C, ppm
127
261
141


Lipoic Acid, ppm*
101
135





*Lipoic acid based on formulated, not analyzed values.






The compositions for use in the methods of this invention further comprise at least one nutrient selected from the group consisting of manganese, methionine, cysteine, mixtures of methionine and cysteine, L-carnitine, lysine, and arginine. Specific preferred amounts for each component in a composition will depend on a variety of factors including, for example, the species of animal consuming the composition; the particular components included in the composition; the age, weight, general health, sex, and diet of the animal; the animal's consumption rate, and the like. Thus, the component amounts may vary widely, and may even deviate from the proportions given herein.
The omega-3 fatty acids may be obtained from a variety of sources. One convenient source is fish oils from, for example, menhaden, mackerel, herring, anchovy, and salmon. DHA and EPA are typical fatty acids present in such fish oils, and, together often make up a significant portion of the oil, such as from about 25% to about 38% of the oil.
When the composition is an animal food, vitamins and minerals preferably are included in amounts required to avoid deficiency and maintain health. These amounts are readily available in the art. The National Research Council (NRC), for example, provides recommended amounts of such ingredients for farm animals. See, e.g., Nutrient Requirements of Swine (10th Rev. Ed., Nat'l Academy Press, Wash. D.C., 197298), Nutrient Requirements of Poultry (9th Rev. Ed., Nat'l Academy Press, Wash. D.C., 1994), Nutrient Requirements of Horses (Fifth Rev. Ed., Nat'l Academy Press, Wash. D.C., 1989), Nutrient Requirements of Dogs and Cats (Nat'l Academy Press, Wash. D.C., 2006). The American Feed Control Officials (AAFCO), for example, provides recommended amounts of such ingredients for dogs and cats. See American Feed Control Officials, Inc., Official publication, pp. 126-140 (2003). Examples of vitamins useful as food additives include vitamin A, B1, B2, B6, B12, C, D, E, K, H (biotin), K, folic acid, inositol, niacin, and pantothenic acid. Examples of minerals and trace elements useful as food additives include calcium, phosphorus, sodium, potassium, magnesium, copper, zinc, chloride, and iron salts.
The methods of the present invention include compositions that may further contain other additives known in the art. Preferably, such additives are present in amounts that do not impair the purpose and effect provided by the invention. Examples of additives include, for example, substances with a stabilizing effect, processing aids, substances that enhance palatability, coloring substances, and substances that provide nutritional benefits.
Stabilizing substances include, for example, substances that tend to increase the shelf life of the composition. Potentially suitable examples of such substances include, for example, preservatives, antioxidants, synergists and sequestrants, packaging gases, stabilizers, emulsifiers, thickeners, gelling agents, and humectants. Examples of emulsifiers and/or thickening agents include, for example, gelatin, cellulose ethers, starch, starch esters, starch ethers, and modified starches.
Additives for coloring, palatability (“pal enhancers”), and nutritional purposes include, for example, colorants (e.g., iron oxide, such as the red, yellow, or brown forms); sodium chloride, potassium citrate, potassium chloride, and other edible salts; vitamins; minerals; and flavoring. Such additives are known in the art. See, e.g., U.S. Pat. No. 3,202,514. See also, U.S. Pat. No. 4,997,671. Flavorants include, for example, dairy product flavorants (e.g., milk or cheese), meat flavorants (e.g., bacon, liver, beef, poultry, or fish), oleoresin, pinacol, and the various flavorants identified in the trade by a FEMA (Flavor Extract Manufacturers Association) number. Flavorants help provide additional palatability, and are known in the art. See, e.g., U.S. Pat. No. 4,997,672. See also, U.S. Pat. No. 5,004,624. See also, U.S. Pat. No. 5,114,704. See also, U.S. Pat. No. 5,532,010. See also, U.S. Pat. No. 6,379,727. The concentration of such additives in the composition typically may be up to about 5% by weight. In some embodiments, the concentration of such additives (particularly where such additives are primarily nutritional balancing agents, such as vitamins and minerals) is from about 0% to about 2.0% by weight. In some embodiments, the concentration of such additives (again, particularly where such additives are primarily nutritional balancing agents) is from about 0% to about 1.0% by weight.
Supplements include, for example, a feed used with another feed to improve the nutritive balance or performance of the total. Supplements include compositions that are fed undiluted as a supplement to other feeds, offered free choice with other parts of an animal's ration that are separately available, or diluted and mixed with an animal's regular feed to produce a complete feed. The AAFCO, for example, provides a discussion relating to supplements in the American Feed Control Officials, Inc. Official Publication, p. 220 (2003). Supplements may be in various forms including, for example, powders, liquids, syrups, pills, encapsulated compositions, and the like.
Treats include, for example, compositions that are given to an animal to entice the animal to eat during a non-meal time. Treats for canines include, for example, dog bones. Treats may be nutritional, wherein the composition comprises one or more nutrients, and may, for example, have a composition as described above for food. Non-nutritional treats encompass any other treats that are non-toxic.
Toys include, for example, chewable toys. Toys for dogs include, for example, artificial bones. There is a wide range of suitable toys currently marketed. See, e.g., U.S. Pat. No. 5,339,771 (and references disclosed in U.S. Pat. No. 5,339,771). See also, e.g., U.S. Pat. No. 5,419,283 (and references disclosed in U.S. Pat. No. 5,419,283). The invention provides both partially consumable toys (e.g., toys comprising plastic components) and fully consumable toys (e.g., rawhides and various artificial bones). It should be further recognized that this invention provides toys for both human and non-human use, particularly for companion, farm, and zoo animal use, and particularly for dog, cat, or bird use.
A “food” is a nutritionally complete diet for the intended recipient animal (e.g., domestic cat or domestic dog). A “nutritionally complete diet” is a diet that includes sufficient nutrients for maintenance of normal health of a healthy animal on the diet. The methods of this invention utilize compositions that are not intended to be restricted by any specific listing of proteinaceous or fat ingredients or product form. The compositions can be prepared in, for example, a dry, canned, wet, or intermediate moisture form using conventional pet food processes. In some embodiments, the moisture content is from about 10% to about 90% of the total weight of the composition. In other embodiments, the moisture content is from about 65% to about 75% of the total weight of the composition.
In preparing a composition for use with the methods of the present invention, any ingredient (e.g., fish oil) generally may, for example, be incorporated into the composition during the processing of the formulation, such as during and/or after mixing of other components of the composition. Distribution of these components into the composition can be accomplished by conventional means. In one embodiment, ground animal and poultry proteinaceous tissues are mixed with the other ingredients, including fish oils, cereal grains, other nutritionally balancing ingredients, special-purpose additives (e.g., vitamin and mineral mixtures, inorganic salts, cellulose and beet pulp, bulking agents, and the like); and water that is sufficient for processing is also added. These ingredients preferably are mixed in a vessel suitable for heating while blending the components. Heating of the mixture may be effected using any suitable manner, such as, for example, by direct steam injection or by using a vessel fitted with a heat exchanger. Following the addition of the last ingredient, the mixture is heated to a temperature range of from about 50° F. (10° C.) to about 212° F. (100° C.). In some embodiments, the mixture is heated to a temperature range of from about 70° F. (21° C.) to about 140° F. (60° C.). Temperatures outside these ranges are generally acceptable, but may be commercially impractical without use of other processing aids. When heated to the appropriate temperature, the material will typically be in the form of a thick liquid. The thick liquid is filled into cans. A lid is applied, and the container is hermetically sealed. The sealed can is then placed into conventional equipment designed to sterilize the contents. This is usually accomplished by heating to temperatures of greater than about 230° F. (110° C.) for an appropriate time, which is dependent on, for example, the temperature used and the composition.
Methods of the present invention include utilizing compositions that can be prepared in a dry form using conventional processes. In one embodiment, dry ingredients, including, for example, animal protein sources, plant protein sources, grains, etc., are ground and mixed together. Moist or liquid ingredients, including fats, oils, animal protein sources, water, etc., are then added to and mixed with the dry mix. The mixture is then processed into kibbles or similar dry pieces. Kibble is often formed using an extrusion process in which the mixture of dry and wet ingredients is subjected to mechanical work at a high pressure and temperature, and forced through small openings and cut off into kibble by a rotating knife. The wet kibble is then dried and optionally coated with one or more topical coatings which may include, for example, flavors, fats, oils, powders, and the like. Kibble also can be made from the dough using a baking process, rather than extrusion, wherein the dough is placed into a mold before dry-heat processing.
The compositions are also designed to be easier to chew. Canine and feline foods are typically formulated based on life stage (age), size, body composition, and breed. In the methods of this invention, some embodiments of the compositions address specific nutritional differences between super senior regular or small breed dogs, large breed dogs, and cats.
All percentages expressed herein are on a weight by dry matter basis unless specifically stated otherwise.
As noted previously, this invention is directed, in part, to a method for enhancing the quality of life of an animal. The method comprises feeding a senior or super senior animal a composition in an amount effective to enhance alertness, improve vitality, protect cartilage, maintain muscle mass, enhance digestibility, and improve skin and pelage quality. Additionally, we now report herein our surprising discovery that the enhanced quality of life of an animal achieved by administration of the compositions of the present invention is reflected at the genomic level. While it may be that a change in expression of any one gene disclosed in the tables presented below may result in beneficial or deleterious biological effects, the data presented herein indicate that, overall, the observed expression profiles are consistent with the beneficial biological effects seen in vivo after administration of the diets disclosed herein. Specifically, gene chip data indicate that the expression of genes that encode proteins associated with or related to several biological pathways such as blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport are, for the most part, beneficially altered through administration to the animal of compositions described herein. Thus, the invention also relates to methods of measuring or characterizing the enhancement in the quality of life of an animal, particularly a senior or super senior animal, fed a composition described herein by quantitating the gene expression levels of one or more genes selected from a group consisting of those disclosed in Tables 5-14 in said animal and comparing said levels in the animal to levels in the animal prior to administration of the feed composition. Quantitation of gene expression may be carried out in numerous ways familiar to one of skill in the art and include such techniques as RT PCR as well as gene chip assays and Northern blotting. Thus, it is contemplated herein that the expression levels detected may be used, for example, in methods to measure enhancement in the quality of life of an animal as disclosed herein.
In another aspect, the present invention relates to kits which comprise:
(a) a polynucleotide of a gene disclosed herein or a fragment thereof,
(b) a nucleotide sequence complementary to that of (a);
(c) a polypeptide encoded by a gene disclosed herein, or a fragment thereof, or
(d) an antibody to a polypeptide encoded by a gene disclosed herein, or a fragment thereof.
It will be appreciated that in any such kit, (a), (b), (c) or (d) may comprise a substantial component. The manufacture of kits as described herein and components thereof (e.g., antibody production) may be achieved according to conventional methods.
It is contemplated herein that modulating the expression levels of the genes disclosed herein may have therapeutic value with regard to the treatment of diseases or disorders associated with the various biological pathways. Such determination may be made on a gene by gene basis without undue experimentation, for example, by assessing expression levels in tissues as well as in blood samples, or by assaying expression levels in vitro in cells or cell lines relevant to particular disease states and suitable for such experimentation. In vivo models of disease might also be utilized in such experimentation. The nature of these and other suitable additional assays would be familiar to one of skill in the art. Thus, based on the genomic data disclosed herein, the invention also relates to methods to enhance the quality of life of an animal by modulating the expression level of one or more genes listed on Tables 5-14 (i.e. up or down regulation as indicated therein) in an animal in order to mimic the pattern of expression seen in vivo after administration of the pet food compositions of the present invention.
Modulation of gene expression levels may be achieved through the use of known modulators of gene expression suitable for administration in vivo, including, but not limited to, ribozymes, antisense oligonucleotides, triple helix DNA, RNA aptamers and/or double stranded RNA directed to an appropriate nucleotide sequence of a gene of interest. These inhibitory molecules may be created using conventional techniques by one of skill in the art without undue burden or experimentation. For example, modification (e.g. inhibition) of gene expression may be obtained by designing antisense molecules, DNA or RNA, to the control regions of the genes discussed herein, i.e. to promoters, enhancers, and introns. For example, oligonucleotides derived from the transcription initiation site, e.g., between positions −10 and +10 from the start site may be used. Notwithstanding, all regions of the gene may be used to design an antisense molecule in order to create those which gives strongest hybridization to the mRNA and such suitable antisense oligonucleotides may be produced and identified by standard assay procedures familiar to one of skill in the art.
Similarly, inhibition of gene expression may be achieved using “triple helix” base-pairing methodology. Triple helix pairing is useful because it causes inhibition of the ability of the double helix to open sufficiently for the binding of polymerases, transcription factors, or regulatory molecules. Recent therapeutic advances using triplex DNA have been described in the literature (Gee, J. E. et al. (1994) In: Huber, B. E. and B. I. Carr, Molecular and Immunologic Approaches, Futura Publishing Co., Mt. Kisco, N.Y.). These molecules may also be designed to block translation of mRNA by preventing the transcript from binding to ribosomes.
Ribozymes, enzymatic RNA molecules, may also be used to modulate gene expression by catalyzing the specific cleavage of RNA. The mechanism of ribozyme action involves sequence-specific hybridization of the ribozyme molecule to complementary target RNA, followed by endonucleolytic cleavage. Examples which may be used include engineered “hammerhead” or “hairpin” motif ribozyme molecules that can be designed to specifically and efficiently catalyze endonucleolytic cleavage of gene sequences.
Specific ribozyme cleavage sites within any potential RNA target are initially identified by scanning the target molecule for ribozyme cleavage sites which include the following sequences: GUA, GUU and GUC. Once identified, short RNA sequences of between 15 and 20 ribonucleotides corresponding to the region of the target gene containing the cleavage site may be evaluated for secondary structural features which may render the oligonucleotide inoperable. The suitability of candidate targets may also be evaluated by testing accessibility to hybridization with complementary oligonucleotides using ribonuclease protection assays.
Ribozyme methods include exposing a cell to ribozymes or inducing expression in a cell of such small RNA ribozyme molecules (Grassi and Marini, 1996, Annals of Medicine 28: 499-510; Gibson, 1996, Cancer and Metastasis Reviews 15: 287-299). Intracellular expression of hammerhead and hairpin ribozymes targeted to mRNA corresponding to at least one of the genes discussed herein can be utilized to inhibit protein encoded by the gene.
Ribozymes can either be delivered directly to cells, in the form of RNA oligonucleotides incorporating ribozyme sequences, or introduced into the cell as an expression vector encoding the desired ribozymal RNA. Ribozymes can be routinely expressed in vivo in sufficient number to be catalytically effective in cleaving mRNA, and thereby modifying mRNA abundance in a cell (Cotten et al., 1989 EMBO J. 8:3861-3866). In particular, a ribozyme coding DNA sequence, designed according to conventional, well known rules and synthesized, for example, by standard phosphoramidite chemistry, can be ligated into a restriction enzyme site in the anticodon stem and loop of a gene encoding a tRNA, which can then be transformed into and expressed in a cell of interest by methods routine in the art. Preferably, an inducible promoter (e.g., a glucocorticoid or a tetracycline response element) is also introduced into this construct so that ribozyme expression can be selectively controlled. For saturating use, a highly and constituently active promoter can be used. tDNA genes (i.e., genes encoding tRNAs) are useful in this application because of their small size, high rate of transcription, and ubiquitous expression in different kinds of tissues. Therefore, ribozymes can be routinely designed to cleave virtually any mRNA sequence, and a cell can be routinely transformed with DNA coding for such ribozyme sequences such that a controllable and catalytically effective amount of the ribozyme is expressed. Accordingly the abundance of virtually any RNA species in a cell can be modified or perturbed.
Ribozyme sequences can be modified in essentially the same manner as described for antisense nucleotides, e.g., the ribozyme sequence can comprise a modified base moiety.
RNA aptamers can also be introduced into or expressed in a cell to modify RNA abundance or activity. RNA aptamers are specific RNA ligands for proteins, such as for Tat and Rev RNA (Good et al., 1997, Gene Therapy 4: 45-54) that can specifically inhibit their translation.
Gene specific inhibition of gene expression may also be achieved using conventional RNAi technologies. Numerous references describing such technologies exist and include, for example, WO 99/32619; Miller et al. Cell Mol Neurobiol 25:1195-207 (2005); Lu et al. Adv Genet 54:117-42 (2005).
Antisense molecules, triple helix DNA, RNA aptamers and ribozymes of the present invention may be prepared by any method known in the art for the synthesis of nucleic acid molecules. These include techniques for chemically synthesizing oligonucleotides such as solid phase phosphoramidite chemical synthesis. Alternatively, RNA molecules may be generated by in vitro and in vivo transcription of DNA sequences encoding the genes discussed herein. Such DNA sequences may be incorporated into a wide variety of vectors with suitable RNA polymerase promoters such as T7 or SP6 according to conventional methods. Alternatively, cDNA constructs that synthesize antisense RNA constitutively or inducibly can be introduced into cell lines, cells, or tissues using methods familiar to one of skill in the art. Vectors may be introduced into cells or tissues by many available means, and may be used in vivo, in vitro or ex vivo. For ex vivo therapy, vectors may be introduced into stem cells taken from an animal and clonally propagated for autologous transplant back into that same animal. Delivery by transfection and by liposome injections may be achieved using methods that are well known in the art.
The instant invention also includes a method to identify an animal that might benefit from feeding a composition as disclosed herein comprising measuring the gene expression levels of any one or more genes listed in Tables 5-14 in said animal and comparing said levels to the gene expression levels seen in Tables 5-14 wherein an animal with levels different than those seen in Tables 5-14 (e.g., up regulated versus down regulated) would be identified as potentially benefiting from feeding a composition of the present invention.
It is also contemplated herein that the invention relates to methods for treating an animal suffering from disorders or disease associated with or relating to any one of more of the following biological pathways: blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport comprising administering to the animal a composition of the present invention.
This invention is not limited to the particular methodology, protocols, and reagents described herein because they may vary. Further, the terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention. The terms “comprise”, “comprises”, and “comprising” are to be interpreted inclusively rather than exclusively.
Unless defined otherwise, all technical and scientific terms and any acronyms used herein have the same meanings as commonly understood by one of ordinary skill in the art in the field of the invention. Although any methods and materials similar or equivalent to those described herein can be used in the practice of the present invention, the preferred methods, devices, and materials are described herein.
All patents, patent applications, and publications mentioned herein are incorporated herein by reference to the extent allowed by law for the purpose of describing and disclosing the compositions, compounds, methods, and similar information reported therein that might be used with the present invention. However, nothing herein is to be construed as an admission that the invention is not entitled to antedate such disclosure by virtue of prior invention.
In the specification there have been disclosed typical preferred embodiments of the invention and, although specific terms are employed, they are used in a generic and descriptive sense only and not for purposes of limitation, the scope of the invention being set forth in the following claims. Many modifications and variations of the present invention are possible in light of the above teachings. It is therefore to be understood that within the scope of the appended claims the invention may be practiced otherwise than as specifically described.
EXAMPLES
This invention can be further illustrated by the following examples of preferred embodiments thereof, although it will be understood that these examples are included merely for purposes of illustration and are not intended to limit the scope of the invention unless otherwise specifically indicated.
Example 1
A composition formulated for senior or super senior regular or small breed canines is described in Table 2.







TABLE 2







Ingredient Composition for Canine Regular


or Small Breed Super Senior










Ingredient
% of composition













Carbohydrate
65.83



Animal Protein
14.31



Vegetable Protein
6.05



Animal/Vegetable Fat
6.60



Omega Fat
3.38



Fiber
1.42



Minerals
1.63



Vitamins
0.78









Example 2
A composition formulated for senior or super senior large breed canines is described in Table 3.







TABLE 3







Ingredient Composition for Canine Large Breed Super Senior










Ingredient
% of composition













Carbohydrate
65.15



Animal Protein
14.79



Vegetable Protein
6.45



Animal/Vegetable Fat
6.23



Omega Fat
4.12



Fiber
1.30



Minerals
0.91



Vitamins
1.05









Example 3
A composition formulated for senior or super senior felines is described in Table 4.







TABLE 4







Ingredient Composition for Feline Super Senior










Ingredient
% of composition













Carbohydrate
31.47



Animal Protein
25.57



Vegetable Protein
20.14



Animal/Vegetable Fat
13.31



Omega Fat
1.61



Fiber
4.80



Minerals
1.77



Vitamins
1.34









Example 4
Genomic Analysis of Control vs. Super Senior Pet Food
To further characterize the nutritional benefits of the super senior pet food compositions of the present invention, gene expression profiles from animals fed the compositions compared to control animals are assayed and the results are described in detail below.
Materials and Methods:
Study Design:
Blood samples are drawn from 9 Beagles according to conventional methods before and after feeding for 14 days on Super Senior K9 diet (a total of 18 samples). Each sample taken after the 14 day trial is compared to its own control.
Isolation of Lymphocytes from Canine Blood
Reagents:
4 ml canine blood, heparin or EDTA tubes, Hank's Balanced Salt Solution (Gibco 14175-095), HEPES buffer (Gibco 15630-080), Accu-Paque (Accurate Chemical & Scientific Corp AN3100).
Materials/Equipment:
Transfer pipettes (VWR 14670-147), 14 ml centrifuge tubes w/caps, 9′′ Pasteur pipettes, 1.5 ml microcentrifuge tubes (VWR 20170-038), centrifuge tube racks, microcentrifuge tube rack, waste container, Beckman Coulter Allegra 25R Centrifuge, SN AJC01J015Eppendorf Centrifuge, 5417C.
Solutions:
Hank's Balanced Salt Solution (HBSS) w/25 mM HEPES buffer solution is made by adding 12.8 ml of HEPES buffer solution to a 500 ml bottle of HBSS. Hank's Balanced Salt Solution and Accu-Paque need to be removed from the refrigerator and placed at room temperature at least 30 minutes before beginning the lymphocyte isolation. Both solutions should be place back in the refrigerator (4° C.) immediately following their use. Procedure:

    
    
        1. Measure 4 ml of HBSS w/HEPES into the correct number of 14 ml centrifuge tubes (one tube for each 4 ml draw of blood)
        2. Using a transfer pipette, transfer 4 ml blood from the Vacutainer® tubes to the 14 ml centrifuge tube containing the HBSS w/HEPES.
        3. Mix the sample well using the transfer pipette to pipette up and down for 30 seconds.
        4. Insert a 9′′ Pasteur pipette into each of the 14 ml centrifuge tubes. Make sure the bottom tip of the Pasteur pipette touches the bottom of the tube.
        5. Using a transfer pipette, slowly add 4 ml of Accu-Paque by running the liquid down the inside of the Pasteur pipette allowing gravity to layer the Accu-Paque under the diluted blood sample.
        6. Plug the top of the Pasteur pipette using your finger and gently remove the pipette.
        7. Centrifuge the tubes at 800×g for 20 minutes at room temperature. For puppy blood a longer centrifugation of 45 minutes is necessary to allow for a good separation of RBC's from WBC's.
        8. Using a transfer pipette, carefully remove the top layer to within 0.5 cm of the middle opaque layer and discard.
        9. Using a new transfer pipette, carefully remove the middle opaque layer and transfer to a 1.5 ml microcentrifuge tube. Be careful not to transfer any of the bottom layers.
        10. Centrifuge the microcentrifuge tubes at 13,200 rpm for 3.5 minutes at room temperature.
        11. Carefully remove the supernatant and flash freeze the remaining pellet (lymphocytes) in liquid nitrogen. Store the final samples at −80° C.

RNA Isolation:

Reagents:

    
    


Deionized H2O, Absolute ethanol (Sigma E7023), RNA Storage Solution (Ambion 7000), RNase Zap® (Ambion 9780), Buffer RLT, Buffer RW1 and Buffer RPE (provided in the RNeasy Mini Kit).
Equipment/Materials:
RNeasy Mini Kit (Qiagen 74104), QIAshredder spin columns (Qiagen 79656), P1000 Pipetman pipette (Rainin), P200 Pipetman pipette (Rainin), 100-100 μl filtered pipette tips (USA Scientific 1126-7810), 1-200 μl filtered pipette tips (USA Scientific 1120-8810), sterile transfer pipettes (VWR 14670-147), 55 ml sterile solution basin (VWR 21007-974), 2 waste containers (one for liquid, one for tips/pipettes), 1.5 ml sterile microcentrifuge tubes (VWR 20170-038), Microcentrifuge tube rack, permanent marker, Eppendorf Microcentrifuge, model #5417C.
Procedure:

    
    
        1. Loosen the pellet in the microcentrifuge tubes by thawing slightly and then flick the tube to dislodge the pellet.
        2. Add the appropriate volume of Buffer RLT (in this case use 600 μl). Vortex or pipette to mix.
        3. Transfer sample to a QIAshredder tube to homogenize the sample. Centrifuge for 2 minutes at 14,000 rpm. Discard spin column but keep the collection tube and its contents.
        4. Add one volume (600 μl) of 70% ethanol to the homogenized lysate and mix by pipetting.
        5. Apply a 600 μl aliquot of the sample to an RNeasy mini column placed in a 2 ml collection tube. Close tube gently and centrifuge for 15 sec at 14,000 rpm. Discard the flow-through. Add the second 600 μl aliquot of the cell lysate to the same spin column and repeat. Discard flow-through.
        6. Reuse the collection tube from step 5. Add 700 μl Buffer RW1 to the column.
    
    


Centrifuge for 15 sec at 14,000 rpm. Discard the flow-through and collection tube.

    
    
        7. Transfer the column to a new 2 ml collection tube and pipette 500 μl Buffer RPE onto the column. Centrifuge for 15 sec at 14,000 rpm to wash the column. Discard the flow-through but save the collection tube for step 8.
        8. Add another 500 ml Buffer RPE to the column. Centrifuge for 2 min at 14,000 rpm to dry the membrane.
        9. Transfer the column to a new 1.5 ml collection tube. Pipette 10 μl of RNA Storage Solution directly onto the membrane. Centrifuge for 1 min at 14,000 rpm to elute the RNA. Add a second volume of 5 μl of RNA Storage Solution directly to the membrane and spin for an additional minute. Store the final elution of RNA at −80° C.

RNA Probe Preparation and Hybridization.

Reagent:

    
    


Ovation TM Biotin System v1.0 for probe preps.
Protocol:
User Guide (Cat#D01002, version Oct. 27, 2004, NuGEN Technologies, Inc). The experimental procedure is followed as described in the user guide. All probe preparation starts with 50 ng of total RNA.
Genechip Procedures:
The Genechips used for the test is the Canine Genome 2.0 Array (Affymetrix). This Genechip contains 44,000 probe sets. Detailed sequence information for each unique probe identification number is available from the manufacturer.
Gene Expression Analysis:
Normalization is performed using MAS 5 provided in GCOS Affymetrix software (version 1.2). Expression levels for the genes analyzed are indicated on the tables included in the examples below, where an upward facing arrow refers to “up regulation” or increase and a downward facing arrow indicates “down regulation” in gene expression. Similarly, in some tables, upward or downward facing arrows also indicate increases or decreases in activity of certain proteins involved in a particular pathway, and are otherwise self explanatory.
Gene List Selection:
15,411 genes are selected for further analysis based on their “present” calls in at least 9 out of 18 samples.
Results of the gene chip analysis indicate that 1088 genes are differentially expressed between the control and Super Senior diet treated groups. The expression levels of these 1088 genes are statistically significant when grouped by ‘diet’; using a parametric test where the variances is not assumed to be equal (Welch t-test). The p-value cutoff is 0.01 with no multiple testing correction. Under those selection criteria only about 154 genes would be expected to pass the restriction by chance. The genomic data is discussed in detail below.
Results:
Effect of Nutrition on Genes Associated with Pain and Inflammation
Based on an analysis of the gene chip data, at the P<0.01 level, 1,088 genes changed compared to control expression levels (10 were up regulated and the rest down regulated). At the P<0.001 level, data indicate that 35 genes are down regulated in beagles fed the super senior food. Nine of these down regulated genes are identified as related to the inflammatory and pain response. Down regulation of these genes may be predicted to result in pain relief, cartilage protection (less damage) and reduction in inflammatory responses. The compositions disclosed herein may be part of a therapeutic regimen to treat animals suffering from pain and/or inflammatory diseases. These genes and their putative role in inflammation and pain response are provided below in Tables 5-6.







TABLE 5







Genes involved in inflammation and pain response (P < 0.001)

















Best
% match



Se-



Current
of probe



quence

Also

BLAST
sequence to
Probe


ID No.
Genes
Known As
Probe
Annotation
BLAST hit
Target Sequence
















1
Phospho-
IPLA2GAMMA,
CfaAffx.6431.1.S1_s_at
PREDICTED: Canis
100
GGAGCCATGCATTTATG



lipase A2
IPLA2-2

familiaris

ACAGTCAAACGTGGGAA






similar to

AATATTCTTAAGGACAG






intracellular

AATGGGATCCTCGCTAA






membrane-

TGATTGAAACAGCAAGA






associated

AACCCTTCATGTCCTAA






calcium-indepen-

GGATGGAGGTTTGCTTC






dent phospholi-

TGAATAACCCTTCAGCG






pase A2 gamma;

CTAGCAATGCACGAGTG






transcript

CAAATGTCTTTGGCCTG






variant 3

ACGTCCCATTAGAGTGC






(LOC475880);

ATTGTGTCCCTGGGCAC






mRNA

CGGGCGTTATGAGAGTG








ATGTGAGAAACTCTGTG








ACATCTACAAGCTTGAA








AACCAAACTGTCTAATG








TCATTAACAGTGCTACA








GATACAGAAGAAGTCCA








CGTAATGCTTGATGGTC








TTTTACCTCCTGACACC








TATTTTAGAT


 


2
Dipeptidase
Putative
CfaAffx.31124.1.S1_at
PREDICTED: Canis
82.197
GTGCTGCAATGCAACCT



2
dipeptidase

familiaris

GTTAGCTAACGTGTCCA






similar to

CTGTGGCAGTTCCCACG






dipeptidase 2

CATCCCTGCCCTGGAAG






(LOC611083);

CCCCACAGTGCTGACTC






mRNA

TCCATCCCTCAGATCAC








TTTGACTACATCAGGGC








AGTCATTGGATCCAAGT








TCATTGGAATTGGTGGA








GATTATGATGGGGCCAG








ACGTTTCCCTCAGGGGC








TGGAGGATGTGTCCACA








TACCCAGTTCTGATAGA








GGAGTTGCTGAGGCGTG








GCTGGAGTAGGGAAGAG








CTCCAGGGTGTCCTTCG








AGGAAACCTACTGCGGG








TCTTTGGACAGGTGGAA








CAGGTACGGGAGGCAAG








CAAGGGGCAAAGGCCCT








TGGAGGATGAGTTCCCG








GATGAGCAGCTGAGCAG








CTCTTGCCGCTCCGTTC








TCTCACGTCTGCATCAG








ACACAGTACCCTGCTCC








ATACCAGAAACTAACTG








AGATTTCACCTGAGTGG








TCCCCTAAACAGTCATT








GTCAAAATCTCTCCCCA








TCATGGCCCCAGGCCTC








ATAGTTATTGCTGCTTG








T


 


3
Thromboxane
Thromboxane A
CfaAffx.6939.1.S1_s_at
PREDICTED: Canis
100
ATCGCTGGCTATGAGAT



synthase
synthase 1,

familiaris

CATCACCAACACGCTCT




Thromboxane A

similar to

CTTTTGCCACCTACCTC




synthase,

Thromboxane-A

CTGGCCACCAACCCTGA




Platelet,

synthase (TXA

CTGCCAAGAGAAGCTTC




Cytochrome

synthase) (TXS)

TGGCAGAGGTGGACAGC




P450

(LOC482771);

TTTAAGGAGAAATATAC




subfamily V,

mRNA

GGCCCTTGACTACTGCA




CYP5, CYP5A1,



GCCTCCAGGAAGGCCTG




Thromboxane



CCCTACCTGGACATGGT




synthatase,



GATTGCGGAGACCTTGA




TXA synthase,



GGATCTACCCCCCGGCT




TXS



TTCAGGTTCACACGGGA








GGCGGCGCGGGACTGCG








AGGTGCGGGGACAGCGC








ATCCCCGCGGGCGCCGT








GGTGGAGGTGGCCGTGG








GCGCCCTGCACCGTGAC








CCTGAGTACTGGCCACA








ACCGGAGACCTTCAACC








CCGAGAGGTTCAAGGCC








GAGGCGCAGCGACGACA








GCAACCCTTCACCTACC








TGCCGTTCGGCGCGGGC








CCCCGGAGCTGCCTCGG








GGTGCGGCTGGGGCTGC








TGGAGGTCAAGCTGACG








CTGCTGCAGGTCCTGCA








CCAGTTCCGGTTCGAGG








CCTGCCCGGAGACGCAG








GTACCACTGCAGCTAGA








CTCCAAATCTGCCCTAG








GTCCAAAGAATGGCATC








TACATCAAGATTGTCTC








CCGCT


 


4
Ubiquitin
Ubiquitin
CfaAffx.275.1.S1_s_at
PREDICTED: Pan
97.19626
GATTTGGCCCGTGACCC



conjugating
protein 

troglodytes

TCCAGCACAATGTTCTG



enzyme E2D
ligase, 

LOC461941

CAGGTCCTGTTTGGGAT



3
Ubiquitin

(LOC461941);

GATATGTTTCATTGGCA




carrier pro-

mRNA

AGCCACAATTATAGGAC




tein, E2 (17) 



CTAATGACAGCCCATAT




KB 3,



CAAGG




Ubiquitin








conjugating








enzyme E2-17








kDa 3, UBC4/5,








UBCH5C






 


5
NEDD8
Neural pre-
Cfa.12556.1.A1_s_at
PREDICTED: Canis
99.12473
GGAATGGGCTACTCTAC



ultimate
cursor cell

familiaris

TCATGCAGNCAAGCAGG



buster-1
expressed,

similar to NEDD8

NCCTGCATCAGGCCAGT




developmen-

ultimate buster-

GGGAACCTGGACGAAGC




tally down

1 (NY-REN-18

CCTGAAGATTCTTCTCA




regulated 8,

antigen)

GCAATCCTCAGATGTGG




Ubiquitin

(LOC475542);

TGGTTAAATGATTCAGA




like protein

mRNA

TCCTGAAACGANCAACC




NEDD8



AGCAAGAAAGTCCTTCC








CAGGAAAACATTGACCA








ACTGGTGTACATGGGCT








TCGACGCTGTGGTGGCT








GATGCTGCCTTGAGAGT








GTTCAGGGGAAACGTGC








AGCTGGCAGCTCAGNCC








CTCGCCCACAACGGAGG








AACTCTTCCTCCTGACC








TGCAGCTCTTGGTGGAA








GACTCTTCATCAACGCC








ATCCACGTCCCCTTCCG








ACTCCGCAGGTACCTCT








AGTGCCTCAACAGATGA








AGATATGGAAACCGAAG








CTGTCAATGAAATACTG








GAAGATATTCCAGAACA








TGAAGAAGATTATCTTG








ACTCAACACTGGAAG


 


6
Mitogen-
p38, Mitogen
CfaAffx.2947.1.S1_at
Homo sapiens
97.84946
GAGATGGAGTCCTGAGC



activated
activated

mitogen-acti-

ACCTGGTTTCTGTTTTG



protein
protein kinase

vated protein

TTGATCCCACTTCACTG



kinase
14, Cytokine

kinase 14,

TGAGGGGAAGGCCTTTT



14 (p38)
suppressive

transcript

CATGGGAACTCTCCAAA




antiinflam-

variant 2; mRNA

TATCATTC




matory drug

(cDNA clone MGC:






binding pro-

34610 IMAGE:






tein 1, CSBP1,

5181064);






CSAID binding

complete cds






protein 1,








Stress acti-








vated protein








kinase 2A,








SAPK2A, p38








MAP kinase,








p38 alpha, RK,








MXI2, Cytokine








suppressive








antiinflam-








matory drug








binding pro-








tein 2, CSBP2,








CSAID binding








protein 2






 


7
Matrix
MMP 19
Cfa.4573.1.A1_at
Homo sapiens
48.93048
GTAGTTGATTCCTGGTT



metallo-


cDNA FLJ38021

CGCCTTTCCTCTTGGGT



proteinase


fis; clone

CCCATAGGTTCGAATCC



19 (MMP-19)


CTONG2012847

CCTTCTACCTCAGTCGG








GAGTACTGTCCTCCATG








GTGCTTCCCTTCCTCTC








CTTAATGTGGGGAAGAC








CATGGGGCAATGCATGG








CGCAGGACCTGCCTCCC








CCAAAAGCAGTCTACTT








GCTCCACGGAGAGAGAA








CTGGGTCCACGTGCCAG








AGTCTTGCCCTTTGGCC








CAGAGTAGCCTGGTCTT








CATGGCTGTATGGGAGA








CAAGTGCCTTCTCTGCT








TCTTGTTGTAGGTGATG








CTAATCTCCTTAACCAA








ACCTTTGTCCCAGCCGC








TAATCTGTTCTAACTCT








CCCTCCTCNTGATTCTC








CTGCTCAAAGTCTGTTC


 


8
Tissue
TIMP-1
Cfa.3680.1.S1_s_at
Canis familiaris
99.4
AGATGTTCAAGGGTTTC



Inhibitor


TIMP metallo-

AGCGCCTTGGGGAATGC



of


peptidase

CTCGGACATCCGCTTCG



metallo-


inhibitor 1

TCGACACCCCCGCCCTG



proteinases


(TIMP1); mRNA

GAGAGCGTCTGCGGATA



(TIMP-1)




CTTGCACAGGTCCCAGA








ACCGCAGCGAGGAGTTT








CTGGTCGCCGGAAACCT








GCGGGACGGACACTTGC








AGATCAACACCTGCAGT








TTCGTGGCCCCGTGGAG








CAGCCTGAGTACCGCTC








AGCGCCGGGGCTTCACC








AAGACCTATGCTGCTGG








CTGTGAGGGGTGCACAG








TGTTTACCTGTTCATCC








ATCCCCTGCAAACTGCA








GAGTGACACTCACTGCT








TGTGGACGGACCAGTTC








CTCACAGGCTCTGACAA








GGGTTTCCAGAGCCGCC








ACCTGGCCTGCCTGCCA








AGAGAGCCAGGGATATG








CACCTGGCAGTCCCTGC








GGCCCCGGATGGCCTAA








ATCCTACTCCCCGTGGA








AGCCAAAGCCTGCACAG








TGTTCACCCCACTTCCC








ACTCCTGTCTTTCTTTA








TCCAAAA


 


9
Fatty
Oleamide
CfaAffx.7308.1.S1_x_at
PREDICTED: Canis
63.33333
GAAGTGGAGTAGGTGCC



acid
hydrolase

familiaris

GCTGTTGCTGCTGGTGT



amide
Anandamide

similar to

TGAATTCAGAACTGTAG



hydrolase
amido-

Ubiquinol-

CGGGACATGGGGCTGGA



(FAAH)
hydrolase

cytochrome c

GGACGAGCAAAAGATGC




FAAH

reductase com-

TGACCGGGTCCGGAGAT






plex 11 kDa pro-

CCCAAGGAGGATCCCCT






tein; mitochon-

AACAACAGTGAGAGAGC






drial precursor

AATGCGAGCAGCTGGAG






(Mitochondrial

AAATGTGTAAAGGCTCG






hinge protein)

GGAGCGGCTAGAGCTCT






(Cytochrome C1;

GTGACCAGCGTGTATCC






nonheme 11 kDa

TCCAGGTCACAGACAGA






protein) (Com-

GGAGGATTGCACAGAGG






plex III subunit

AGCTCTTTGACTTCCTG






VIII); tran-

CATGCAAGGGACCACTG






script variant

TGTGGCCCACAAACTCT






2 (LOC608530);

TTAACAGCTTG






mRNA
















TABLE 6







Summary of down-regulated enzyme roles involved


in the eicosanoid pathway (inflammatory response)











Gene 





Expression





Compared 




Gene
to Control
Results in
Role





Phospholipase A2
↓
↓ in arachidonic
↓ in 2-series 




release from
inflammatory response




phospholipids



Thromboxane 
↓
↓ Thromboxane A2
↓ platelet aggregation,


synthase


vasoconstriction, 





lymphocyte 





proliferation and 





bronchoconstriction



↓
↓ Thromboxane B2
↓ vasoconstriction


Dipeptidase 2
↓
↓ Leukotriene E4
↓ component of slow-





reactive substance of 





anaphylaxis, micro-





vascular vaso-





constrictor and 





bronchoconstriction


Ubiquitin 
↓
↓ ubiquination or
↓ MMP Production


conjugating

activation of 



enzyme E2D 3

TAK1, IRAK 



(and NEDD8 

and TRAF



ultimate buster-1)





Mitogen activated
↓
↓ in c-Jun 
↓ MMP Production


protein kinase 

promotor



14 (p38)





MMP-19
↓
↓ MMP-19
↓ in T-cell derived 





MMP-19 which has 





been implicated in 





rheumatoid arthritis


TIMP-1
↓
↓ TIMP-1
Deactivates MMP's 





concentration is directly





related to MMP 





concentration


Fatty acid amide
↓
↑ anandmide
↓ pain response


hydrolase









Effect of Nutrition on Genes Involved in Heart Health and Blood Coagulation

At the P<0.001 and P<0.01 level, 12 genes are identified to be related to heart health through regulation of the eicosanoid pathway and blood coagulation pathway. The genes are responsible for blood coagulation through platelet activation and aggregation. The down regulation of these genes through nutrition can prevent inappropriate blood clotting which may result in heart or brain related disorders. The compositions of the present invention may be part of a therapeutic regimen to treat animals suffering from disorders or diseases of the blood, heart or brain. These genes and their putative role in vivo are described in Tables 7 and 8 below.







TABLE 7







Genes involved in heart health and blood coagulation

















Best
% match



Se-



current
of probe



quence


P-
BLAST
sequence to



ID No.
Gene
Probe
value
annotation
BLAST hit
Probe Target Seq.
















10
Glycoprotein
Cfa.3503.1.S1_at
<0.01
Canis familiaris
98.57143
TGTGGGTCCGAGCTAACAGCT



Ib


glycoprotein Ib

ACGTGGGGCCTCTGATGGCAG






mRNA; complete

GACGGCGGCCCTCTGCCCTGA






cds

GCCTGGGTCGTGGGCAGGACC








TGCTAGGTACGGTGGGCGTTA








GGTACTCCAGCCACAGCCTCT








GAGGCGACGGTGGGCAGTTTG








GGGACCTTGAGAGGCTGTGAT








GGGCCCTCCTATCAGGATCTT








GCTGGGGGTGGGTGGGCAGGG








AGCACAGGATTGGGGGGAGGC








CTTAAGCACCTTTTCTGGGTC








AGAAGCCTCCTCTCCGCATTG








CATGTGCAACCTCAGTGAAGC








AGCATGGGCAGGGGAGCCGGA








CGGGCCACCCAACAGAGCTCC








TTATGCTGCAGGAGGGGTTCA








CAGACCACTCGGACATCACCA








TCACCTTGGGGGGGGTGCTTG








AGGGAAAAGCAAATTGAACAG








AGCGTGATTCTCACGTGCAGG








TACCTAAGGGAACTGGGGAAG








AGATGCACCAAGACGAGAGCC








CTCGTCATCCCTGGGGAGCCC








AAGCCTAGGGGTTTTCTTCCT








CTTCCCGTTTAGCATTTTCCA








CCATCGTATGTTAC


 


11
Platelet
CfaAffx.4809.1.S1_at
<0.01
PREDICTED: Canis
50
AGTTTTGACCAATTCGCTCTG



glycoprotein


familiaris

TACAAGGAGGGGGACACTGAG



VI


similar to

CCCCACAAGCAATCTGCAGAA






glycoprotein VI

CAGTACTGGGCCAATTTCCCC






(platelet)

ATCACCGCAGTGACTGTTGCC






(LOC484303);

CACAGTGGGATCTACCGATGC






mRNA

TATAGCTTTTCCAGCAAGTTC








CCGTACCTGTGGTCAGCCCCC








AGCGACCCCCTGGAGCTTGTG








GTAACAGGTGAGGGAGATGCA








GTCCAAGCCTTTCTTCTTCAG








CTCTTGCATACTCTGGTGGAA








GTTCCAGGGGAGGGGCCAACA








GTGCCTTCTAGGACTATCACT








GTCTCTCCAAAGGGGTCAGAC








TCTCCAACTGGTCTTGCTCAC








CAGCACTACACCAAGGGCAAT








CTGGTCCGGATATGCCTTGGA








GCTGTGATTCTAATACTCCTG








GTGGGAATTCTGGCAGAAGAT








TGGCACAGCAGAAAGAAACCC








CTGTTGCTCCGGGTCAGAGCT








GTCCACAGGCCACTCCCACCC








CTCCCACAGACCCAGAAACCA








CACAGTCATCAGGATGGGGGT








CGACCAGATGGCCATAACCAT


 


12
Platelet
CfaAffx.7430.1.S1_at
<0.01
PREDICTED: Canis
100
TCTGGGCTGCCACGGAGGCCA



glycoprotein


familiaris

CCAACGACTGCCCCGCAGAGT



IX precursor


similar to

GCACCTGCCAGACCCTGGAGA






Platelet glyco-

CCATGGGGCTGTGGGTGGACT






protein IX pre-

GCAGGGGGCGGGGACTCAAGG






cursor (GPIX)

CCCTGCCCGCCCTGCCGGTCC






(CD42A)

ACACCCGCCACCTCCTGCTGG






(LOC609630);

CCAATAACAGCCTCCGCTCCG






mRNA

TGCCCCCTGGTGCCTTCGACC








ACCTGCCTGGGCTGCAGATCC








TCGACGTGATGCACAACCCCT








GGCACTGTGACTGCAGCCTCA








CCTACCTGCGTCTCTGGCTGG








AGGACCACACGCCCGAGGCCT








TGCTGCAGGTCCGCTGTGCCA








GCCCCGCGCTGGCCACCACCC








GGCCGCTGGGCTGGCTGACGG








GCTACGAGCTGGGCAGCTGCG








GCTGGCAGCTACAGGCACCCT








GGACCTA


 


13
Coagulation
CfaAffx.14964.1.S1_s_at
<0.01
PREDICTED: Canis
99.6008
ATCTCTCAGGCAACATCGTCT



factor XIII


familiaris

TCTACACCGGGGTCTCCAAGA



A chain


similar to

CGGAATTCAAGAAGGAGACAT



precursor


Coagulation

TTGAAGTGACACTGGAGCCCT






factor XIII A

TGTCTTTCAAGAGAGAGGAGG






chain precursor

TGCTGATCAGAGCGGGCGAGT






(Coagulation

ACATGGGCCAGCTGCTAGAGC






factor XIIIa)

AAGCATACCTGCACTTCTTTG






(Protein-gluta-

TCACAGCGCGTGTCAATGAGT






mine gamma-

CCAAGGATATTCTGGCCAAGC






glutamyltrans-

AGAAGTCCACCGTGCTGACGA






ferase A chain)

TCCCCCAGCTCATCATCAAGG






(Transgluta-

TCCGTGGCGCCAAGATGGTTG






minase A chain);

GTTCTGACATGGTGGTGACAG






transcript

TTGAGTTCACCAATCCCCTGA






variant 1

AAGAAACTCTGCGGAATGTGT






(LOC478711);

GGATACACCTGGATGGTCCTG






mRNA

GAGTGATAAAGCCAATGAGGA








AGATGTTCCGTGAAATCCAGC








CCANTGCCACCATACAATGGG








AAGAAGTGTGTCGACCCTGGG








TGTCTGGCCCTCGGAAGCTGA








TAGCCAGCATGACGAGTGACT








CCCTGAGACACGTGTATG


 


 3
Thromboxane
CfaAffx.6939.1.S1_s_at
<0.001
PREDICTED: Canis
100
ATCGCTGGCTATGAGATCATC



synthase


familiaris

ACCAACACGCTCTCTTTTGCC






similar to

ACCTACCTCCTGGCCACCAAC






Thromboxane-A

CCTGACTGCCAAGAGAAGCTT






syhthase (TXA

CTGGCAGAGGTGGACAGCTTT






synthase) (TXS)

AAGGAGAAATATACGGCCCTT






(LOC482771);

GACTACTGCAGCCTCCAGGAA






mRNA

GGCCTGCCCTACCTGGACATG








GTGATTGCGGAGACCTTGAGG








ATCTACCCCCCGGCTTTCAGG








TTCACACGGGAGGCGGCGCGG








GACTGCGAGGTGCGGGGACAG








CGCATCCCCGCGGGCGCCGTG








GTGGAGGTGGCCGTGGGCGCC








CTGCACCGTGACCCTGAGTAC








TGGCCACAACCGGAGACCTTC








AACCCCGAGAGGTTCAAGGCC








GAGGCGCAGCGACGACAGCAA








CCCTTCACCTACCTGCCGTTC








GGCGCGGGCCCCCGGAGCTGC








CTCGGGGTGCGGCTGGGGCTG








CTGGAGGTCAAGCTGACGCTG








CTGCAGGTCCTGCACCAGTTC








CGGTTCGAGGCCTGCCCGGAG








ACGCAGGTACCACTGCAGCTA








GACTCCAAATCTGCCCTAGGT








CCAAAGAATGGCATCTACATC








AAGATTGTCTCCCGCT


 


14
Dystrobrevin
CfaAffx.15541.1.S1_s_at
<0.01
PREDICTED: Canis
99.65986
GGCAACATGTCGTCCATGGAG



binding


familiaris

GTCAACATCGACATGCTGGAG



protein 1


similar to

CAGATGGACCTGATGGACATC



isoform a


dystrobrevin

TCTGACCAGGAGGCCCTGGAC






binding protein

GTCTTCCTGAACTCCGGCGCT






1 isoform a

GAAGACAACACGGTGCCGTCT






(LOC610315);

CCGGTCTCAGGGCCTGGCTCG






mRNA

GGGGACAGTCGGCAGGAAATC








ACGCTCCGGGTTCCAGATCCC








GCCGAATCGCAAGCTGAGCCT








CCTCCCTCGCCGTGTGCCTGT








CCTGAGCTGGCCGCCCCGGCC








CCCGGCGACGGTGAGGCCCCC








GTGGTCCAGTCTGACGAGGAG


 


15
Integrin
Cfa.11961.1.A1_s_at
<0.01
PREDICTED: Canis
99.0909
ATTACAACGTGACTCTGGCTT



beta-7


familiaris

TGGTCCCTGTCCTGGATGACG



precursor


similar to

GCTGGTGCAAAGAGAGGACCC






Integrin beta-7

TAGACNAACCAGCTGCTGTTC






precursor

TTCCTGGTGGAGGAGGAANCC






(LOC477598);

GGAGGCATGGTTGTGTTGACA






mRNA

GTGAGACCCCAAGAGAGAGGC








GCGGATCACACCCAGGCCATC








GTGCTGGGCTGTGTAGGGGGC








ATCGTGGCAGTGGGGCTGGGG








CTGGTCCTGGCTTACCGGCTC








TCTGTGGAAATCTACGNCCGC








CGAGAATTTAGCCGCTTTGAG








AAGGAGCAGAAGCACCTCAAC








TGGAAGCAGGAAAACAATCCT








CTCTACAGAAGCGCC


 


16
integrin-
Cfa.465.1.S1_s_at
<0.01
PREDICTED: Canis
100
TGGGCGCATGTATGCACCTGC



linked


familiaris

CTGGGTGGCCCCTGAAGCTCT



kinase


similar to

GCAGAAGAAGCCTGAAGATAC






integrin linked

AAACAGACGCTCAGCAGATAT






kinase; tran-

GTGGAGTTTTGCAGTGCTTCT






script variant

GTGGGAACTGGTGACGAGGGA






1 (LOC476838);

GGTACCCTTTGCTGACCTCTC






mRNA

CAACATGGAGATTGGAATGAA








GGTGGCACTGGAAGGCCTTCG








GCCTACTATCCCACCAGGCAT








TTCCCCCCATGTGTGTAAGCT








CATGAAGATCTGCATGAATGA








AGACCCTGCTAAGCGGCCCAA








GTTTGACATGATTGTGCCTAT








CCTGGAGAAGATGCAGGACAA








GTAGAGCTGGAAAGCCCTTGC








CTAAACTCCAGAGGTGTCAGG








ACACGGTTAGGGGAGTGTGTC








TCCCCAAAGCAGCAGGC


 


17
Thrombo-
Cfa.21204.1.S1_at
<0.01
PREDICTED: Canis
54.83871
ATACGAATGCAGAGATTCCTA



spondin 1


familiaris

ATCAAACTGTTGATCAAAAGA






similar to

CTGATCCTAACCAATGCTGGT






thrombospondin

GTTGCACCTTCTGGAACCACG






1 precursor

GGCTTAAGAAAACCCCCAGGA






(LOC487486);

TCACTCCTCCCTGCCTTTTCT






mRNA

CTGCTTGCATATCATTGTGGA








CACCTAGAATACGGGACTTGC








CTCGAGACCATGCNNNNNTCC








AAATCAGACTNNNNNNGTAGC








CTCTGAACGCGAAGAGAATCT








TCCAAGAGCATGAACAG


 


18
Thrombo-
CfaAffx.18675.1.S1_s_at
<0.01
PREDICTED: Canis
100
GAAGCCCTTGATGGATACTGT



spondin


familiaris

GAACGGGAACAGGCTATAAAG



repeat


similar to

ACCCACCACCACTCCTGTTGC



containing


extracellular

CACCACCCTCCTAGCCCTGCC



1


matrix protein

CGCGATGAGTGCTTTGCCCGT






1 isoform 1

CAGGCGCCATACCCCAACTAT






precursor

GACCGGGACATCCTGACCCTT






(LOC808791);

GATTTCAGCCAAGTTACCCCC






mRNA

AACCTCATGCAACATCTCTGT








GGAAATGGAAGACTTCTCACC








AAGCATAAACAGATTCCTGGG








CTGATCCGGAACATGACTGCC








CACTGCTGTGACCTGCCATTT








CCAGAGCAGGCCTGCTGTGCT








GAGGAGGAGAAATCGGCCTTC








ATTGCAGACTTGTGTGGTTCC








CGACGTAACTTCTGGCGAGAC








TCTGCCCTCTGCTGTAACCTG








AATCCTGGAGATGAACAGACC








AACTGCTTCAACACTTATTAT








CTGAGGAATGTGGCTCTAGTG








GCTGGAGACAAT


 


19
Thrombo-
CfaAffx.16694.1.S1_at
<0.01
PREDICTED: Canis
98.13084
TGGTTGTAGCTCCTCACTTGT



spondin


familiaris

CCAAGACCGAAGCAGCAACCA



type 1


similar to lines

AACTGAACTTAGCCTTTGGGC



motif, 17


homolog 1

TGCTCTTGGTAGTCACAGAAA






isoform 1

TGCCCACGCTTCAGTCCCCTG






(LOC607902);

GGCTTCCAATGCTTCTGGACC






mRNA

TCTGAACCAGCCTGTGATGTC








CAAGGAACCCCACGTCACGCT








CCAGGCTGCTGCTGGTCTGTC








TCCCCCACAAGCTTCTCAAAG








TCTGGTAGATTATGACAGCTC








TGATGATTCTGAAGTAGAAGT








CACAGACCAGCACTCAACAAA








CAGTAAACAAACATCTTTACA








GCAAGAAGCAAAGAAGAAATT








TCAGGACACAGTTAGAACAGG








TCCAGATGAAAAAGAACTTAG








CATGGAGCCTCAATCAAGGCC








TCTGGTTCCAGAACAATCTAA








TATTAATATTCCCTTCTCTGT








TGACTGTGACATCTCCAAAGT








AGGAATATCTTACAGGACACT








GAAGTGCTTTCAGGAGCTACA








GGGTGCCATTTACCGTTTGCA








GAAAAAAAATCTTTTCCCCTA








TAATGCCACA


 


20
Angio-
Cfa.8616.1.A1_s_at
<0.001
Canis familiaris
64.77273
GCGGACTGTGTTCCAACCCCT



associated


angio-associated

TCAGCCGACTTGCCCCCTCCG



migratory


migratory cell

TCCCTTCTCTTAAGAGACCCA



cell


protein (AAMP)

TCCCTTGGCCCCCCACCCCAC



protein


gene; complete

CCTCACCCAGACCTGCGGGTC



(AAMP)


cds

CCTCAGAGGGGGGTCAGGCCT








CTTTCTCTTTCACCTTCATTT








GCTGGCGTGAGCTGCGGGGGT








GTGTGTTTGTATGTGGGGAGT








AGGTGTTTGAGGTTCCCGTTC








TTTCCCTTCCCAAGTCTCTGG








GGGTGGAAAGGAGGAAGAGAT








ATTAGTTACAGA
















TABLE 8







Summary of down regulated enzyme roles


involved in heart health and blood coagulation










Gene




Expression




compared



Gene
to Control
Role





Glycoprotein Ib
↓
GP-Ib, a surface membrane 




protein of platelets, participates in 




the formation of platelet plugs by 




binding to the A1 domain of von




Willebrand factor, which is




already bound to the




subendothelium.


Platelet glycoprotein VI
↓
Collagen receptor belonging to 




the immunoglobulin-like protein 




family that is essential for platelet




interactions with collagen


Platelet glycoprotein IX
↓
The GPIb-V-IX complex


precursor

functions as the von Willebrand 




factor receptor and mediates von




Willebrand factor-dependent 




platelet adhesion to blood vessels. 




The adhesion of platelets to




injured vascular surfaces in the 




arterial circulation is a critical 




initiating event in hemostasis


Coagulation factor XIII A
↓
Factor XIII is activated by


chain precursor

thrombin and calcium ion to




a transglutaminase that catalyzes




the formation of gamma-




glutamyl-epsilon-lysine cross-




links between fibrin chains, thus




stabilizing the fibrin clot.


Thromboxane synthase
↓
↓ platelet aggregation,




vasoconstriction,




lymphocyte proliferation




and bronchoconstriction


Angio-associated migratory
↓
contains a heparin-binding


cell protein (AAMP)

domain (dissociation constant, 




14 pmol) and mediates heparin-




sensitive cell adhesion


Dystrobrevin binding
↓
Plays a role in the biogenesis 


protein 1 isoform a

of lysosome-related organelles 




such as platelet dense granule 




and melanosomes


Thrombospondin 1
↓
Adhesive glycoprotein that




mediates cell-to-cell and cell-




to-matrix interactions. Can bind 




to fibrinogen, fibronectin, 




laminin, type V collagen and 




integrins alpha-V/beta-1, alpha-




V/beta-3 and alpha-IIb/beta-3.


Thrombospondin type 1
↓
Metalloprotease activity


motif, 17




Thrombospondin repeat
↓



containing 1




Integrin beta-7 precursor
↓
Integrin alpha-4/beta-7




(Peyer's patches-specific




homing receptor LPAM-1)




is expected to play a role in




adhesive interactions of




leukocytes. It is a receptor for 




fibronectin and recognizes one 




or more domains within the




alternatively spliced CS-1




region of fibronectin. Integrin 




alpha-4/beta-7 is also a receptor 




for MADCAM1 and VCAM1.




It recognizes the sequence




L-D-T in MADCAM1.




Integrin alpha-E/beta-7 (HML-1) 




is a receptor for E-cadherin.


Integrin linked kinase
↓
Receptor-proximal protein




kinase regulating integrin-




mediated signal transduction. 




May act as a mediator of inside-




out integrin signaling. Focal




adhesion protein part of the




complex ILK-PINCH. This




complex is considered to be one 




of the convergence points of 




integrin- and growth factor-




signaling pathway. Could be




implicated in mediating cell




architecture, adhesion to integrin 




substrates and anchorage-




dependent growth in epithelial 




cells. Phosphorylates beta-1 and




beta-3 integrin subunit on




serine and threonine residues, 




but also AKT1 and GSK3B.









Effect of Nutrition on Genes Involved with Muscle and Bone Regulation

Ten down regulated genes are identified as related to body composition through regulation of bone and muscle. The genes spare muscle and bone deterioration by reducing nitric oxide production and glucocorticoid degradation of muscle. Down regulation of these genes results in a decrease in nitric oxide production and glucocorticoid response. The compositions disclosed herein may be part of a therapeutic regimen to treat animals suffering from diseases or disorders associated with or relating to muscle or bone. These genes and their putative role in muscle and bone regulation are detailed in Tables 9 and 10 below.







TABLE 9







Genes involved in muscle and bone regulation

















Best
% match



Se-



current
of probe



quence


P-
BLAST
sequence to



ID No.
Gene
Probe
value
annotation
BLAST hit
Probe Target Sequence
















21
Capping
Cfa.1044.1.S1_at
0.001
PREDICTED: Canis
44.87179
AGGTCCCGTAACACCGGCATCGCG



Protein


familiaris

ACCGCACAGCGCCATCTCCCCAGA






similar to F-

ATAAAGCCCAGTAAACACCCCTGN






actin capping

NNNNNANNNNNANNNNNCACCACG






protein beta

TTTTGCTATCAGAACTCTCCTTGT






subunit

TTCCAGAGCCCGTGTGCTTTTGTT






(LOC478209);

TGCCCCAGCCCC






mRNA




 


22
Calmodulin
Cfa.4168.1.S1_at
0.01
PREDICTED: Canis
52.54237
CCACCCATGGTGACGATGACACAC






familiaris

ATCCTGGTGGCATGCGTGTGTTGG






similar to

TTTAGCGTTGTCTGCGTTGTACTA






calmodulin 1;

GAGCGAAAATGGGTGTCAGGCTTG






transcript

TCACCATTCACACAGAAATTTAAA






variant 3

AAAAAAAAAAAAANNNNGANAAAA






(LOC480416);

AACCTTTACCAAGGGAGCATCTTT






mRNA

GGACTCTCTGTTTTTAAAACCTCC








TGAACCATGACTTGGAGCCAGCAG








ATTAGGCTGTGGCTGTGGACTTCA








GCACAACCATCAACATTGCTGATC








AAGAAATTACAATATACGTCCATT








CCAAGTT


 


23
Dynein
Cfa.4942.1.A1_s_at
0.001
PREDICTED: Canis
99.6016
ATACCTCAGAGGTCTCGTAGCTCG






familiaris

TGCCCTTGCCATCCAGAGCTGGGT






similar to

GGNAGAGAGCTGAGAAGCAGGCTC






dynein; cyto-

TTTTCTCTGATACACTCGACCTGT






plasmic; heavy

CAGAACTCTTCCACCCAGACACAT






polypeptide 2;

TTCTCAATGCTCTTCGCCAGGAAA






transcript

CAGCAAGGGTGATGGGCTGCTCTG






variant 2

TGGATAGCCTTAAGTTTGTAGCTT






(LOC479461);

CGTGGAAAGGTCGGCTGCAAGAAG






mRNA

CAAAGCTGCAGATCAAGATGGGCG








GCTTGCTTCTGGAAGGCTGCAGTT








TTGACGGGAGCCGGCTCTCTGAAA








ACCACCACGATTCTCCAAGTGTGT








CACCAGTTCTCCCTTGCTGTGTTG








GCTGGATTCCCCAGGGTGCATATG








GTCCCTATTCTCCTGACGAGTGCA








TATCTCTGCCCGTGTACACGAGCG








CTGAGAGGGATCGTGTGGTAGCCA








ACATCGACGTCCCGTGTGGGGGCA








NCCAAGACCAGTGGATTCAGTGTG








GAGCCGCTCTGTTTCTAAAAAA


 


24
Dynactin
Cfa.1807.1.S1_at
0.01
PREDICTED: Canis
100
AGGACGACAAGGCTCAGGACGCAA






familiaris

AGTGTGAAACTGCCTTTGTAACAG






similar to

GGCAGAAGCAGCTCTGTATTGGAT






dynactin 3 iso-

TCACAACCTACCTATCTGCATTCA






form 2; tran-

GGTGGGGCTCGGAGGTCAGAGGTC






script variant 1

TGGCTACTTGAGGTTTGCTGTTTG






(LOC474750);

CAC






mRNA




 


25
Kinesin
Cfa.10496.1.S1_s_at
0.01
PREDICTED: Canis
99.73046
AGCCACAGCATTTCCTTTTAACTT






familiaris

GGTTCAATTTTTGTAGCAAGACTG






similar to

AGCAGTTCTAAATCCTTTGCGTGC






Kinesin-like

ATGCATACCTCATCAGTGNACTGT






KIF2 (Kinesin-

ACATACCTTGCCCTCTCCCAGAGA






2) (HK2); tran-

CAGCTGTGCTCACCTCTTCCTGCT






script variant 5

TTGTGCCTTGACTAAGGCTTTTGA






(LOC478071);

CCCTAAATTTCTGAAGCACAGCCA






mRNA

AGATAAAGTACATTCCTTAATTGT








CAGTGTAAATTACCTTTATTGTGT








GTACATTTTTACTGTACTTGAGAC








ATTTTTTGTGTGTGACTAGTTAAT








TTTGCAGGATGTGCCATATCATTG








AATGGAACTAAAGTCTGTGACAGT








GGACATAGCTGCTGGACCATTCCA








TCTTACATGTA


 


26
Heat
CfaAffx.11022.1.S1_s_at
0.01
PREDICTED: Canis
100
GGTGCTACTGTTTGAAACAGCTCT



Shock


familiaris

ACTCTCCTCCGGCTTCTCACTGGA



Protein 1


similar to

GGATCCCCAGACTCACTCCAACCG



(HSP90)


Heat shock pro-

CATTTACCGCATGATAAAGCTAGG






protein HSP 90-

CCTGGGCATCGATGAAGATGAAGT






beta (HSP 84)

GGCAGCGGAGGAACCCAGTGCTGC






(Tumor specific

TGTTCCTGATGAGATCCCTCCACT






transplantation

TGAGGGTGATGAGGATGCCTCTCG






84 kDa antigen)

CATGGAAGAAGTC






(TSTA)








(LOC611252);








mRNA




 


27
PPlase
CfaAffx.1740.1.S1_at
0.01
PREDICTED: Canis
100
GACATCACCAGTGGAGACGGCACC






familiaris

GGCGGTATAAGCATTTATGGTGAG






similar to

ACGTTTCCAGATGAAAACTTCAAA






Peptidyl-prolyl

CTGAAGCATTATGGCATTGGTTGG






cis-trans iso-

GTCAGCATGGCCAACGCTGGGCCT






merase C

GACACCAACGGCTCTCAGTTCTTT






(PPLASE)

ATCACCTTGACCAAGCCCACTTGG






(Rotamase)

TTGGATGGCAAACATGTGGTATTT






(Cyclophilin C)

GGAAAAGTCCTTGATGGAATGACT






(LOC481480);

GTGGTCCACTCCATAGAACTTCAG






mRNA

GCAACCGATGGGCACG


 


28
Calcinuerin
Cfa.19761.1.S1_at
0.001
PREDICTED: Canis
98.83382
GAATTAACAATCTGCTTGAGCCCC






familiaris

AAAACACTACTTATGCACTTCACT






similar to

TGCCAAAAGATTTGNGCAAGGTTT






protein phospha-

TGTACCCTGGTAAATGATGCCAAA






tase 3 (formerly

GTTTGTTTTCTGTGGTGTTTGTCA






2B); catalytic

AATGTTCTATGTATAATTGACTGT






subunit; beta

CTGTAACATGCTGTTTNCTTCCTC






isoform

TGCAGATGTAGCTGCTTTCCTAAA






(calcineurin A

TCTGTCTGTCTTTCTTTAGGTTAG






beta); tran-

CTGTATGTCTGTAAAAGTATGTTA






script variant 5

AATTAAATTACTCTATCAGACGCT






(LOC479248);

TGTCTGTCTTTTGATGTAGAAGCA






mRNA

ACTTTGTAGCACCTTGTTTTGAGG








TNNGCTGCATTTGTTGCTGTACTT








TGTGCAT


 


29
Protein
CfaAffx.408.1.S1_s_at
0.01
PREDICTED: Canis
99.64664
TTCAGTTCCTGTCTCATGGCCGCT



kinase C


familiaris

CCCGGGACCATGCCATCGCCGCCA






similar to

CTGCCTTCTCCTGCATCGCTTGTG






myeloid-associ-

TGGCTTATGCCACCGAAGTGGCCT






ated differen-

GGACCCGGGCCCGTCCCGGAGAGA






marker

TCACCGGCTACATGGCCANTGTGC






(LOC611521);

CGGGCCTGCTCAAGGTGCTGGAGA






mRNA

CCTTTGTGGCCTGCATCATCTTCG








CCTTCATCAGCAACCCCTCCCTGT








ACCAGCACCAGCCGGCCCTGGAGT








GGTGTGTGGCCGTCTACTCCATCT








GTTTCATCCTGGCGGCTGTGGCCA








TCCTACTGAACCTGGGGGACTGCA








CCAACATGCTGCCCATCTCCTTCC








CCAGTTTCCTGTCGGGCCTGGCCC








TGCTCTCCGTCCTGCTGTATGCCA








CGGCTCTGGNTCTCTGGCCGCTCT








ACCAGTTCAACGAGAAGTATGGTG








GCCAGCCCCGTCGGTCGAGGGATG








TTAGCTGCGCCGACAGGCACACCT








ACTACGTGTGTACCTGGGACCGCC








GCCTGGCTGTGGCCATCCTGACAG








CCATCAACCTGCTGGCTTACGTGG








CTGACCTGGTGTAC


 


30
Protein
Cfa.15485.1.A1_s_at
0.01
PREDICTED: Canis
100
GGAGCAGTCAGAACTAAGACATGG



Kinase C


familiaris

TCCGTTTTACTATATGAAGCAGCC



Binding


similar to

ACTCACCACAGACCCTGTTGATGT



Protein


protein kinase C

TGTACCGCAGGATGGACGGAA






binding protein








1 isoform b;








transcript








variant 11








(LOC477252);








mRNA
















TABLE 10







Summary of genes affecting glucocorticoid receptors


and nitric oxide production










Gene




Expression




Compared



Gene
to Control
Role





Kinesin
↓
Transport of organelles from the 




(−) to (+) ends.




Binds microtubules.




ATPase activity


Capping Protein
↓
Part of dynactin-dynein




hetero-complex


Calmodulin
↓
Directly influences calcium




dependent dynein activity.




Binds to nitric oxide synthase and 




up regulates the production of 




nitric oxide


Dynein
↓
Transport of organelles from the 




(+) to (−) ends.




Binds microtubules.




ATPase activity and force




production


Dynactin
↓
Cytoplasmic dynein activator. 




Binds mirotubules and ↑average




length of dyein movements.


Heat Shock Protein 1 beta
↓
Necessary for glucocorticoid 


(HSP90)

receptor binding and fast trans-




port of dynein complex to 




nucleus.




Calcinuerin activity.




Enhances the nitric oxide




production by binding to




nitric oxide synthase


PPIase
↓
Necessary for 




dynein/glucocorticoid




interaction and movement


Calcinuerin
↓
Part of dynactin-dynein




hetero-complex. Catalyzes




the conversion of arginine




to citrulline and nitric oxide


Protein kinase C
↓
Calcium-activated,




phospholipid-dependent, serine- 




and threonine-specific enzyme.


Protein Kinase C Binding
↓
Associated with protein


Protein

kinase C









Effect of Nutrition on Genes Involved with DNA Damage/Protection and Neural Function

Eleven genes are identified that are related to DNA damage/protection and neural function. With regard to the latter, the genes identified are important for rebound potentiation; they are believed to have a potential role in motor learning. Interestingly, of these genes, all were down regulated except for of gamma-aminobutyric acid (GABA) A receptor, gamma 2 which was up regulated. The compositions disclosed herein may be part of a therapeutic regimen to treat animals suffering from diseases or disorders associated with or relating to DNA damage/protection and neural function. The identity of these genes and their putative role in DNA damage/protection and neural function are described in Tables 11 and 12 below.







TABLE 11







Genes involved in DNA damage/protection and neural function

















Best
% match







current
of probe



Sequence


P-
BLAST
sequence to



ID No.
Gene
Probe
value
annotation
BLAST hit
Probe Target Sequence
















31
Gamma-
CfaAffx.26362.1.S1_at
<0.01
Homo sapiens
100
CCTCTTCTTCGGATGTTTTCCT



aminobutyric


gamma-amino-

TCAAGGCCCCTACCATTGAT



acid (GABA)


butyric acid





A receptor,


(GABA) A





gamma 2


receptor; gamma








2 (GABRG2);








transcript








variant 1; mRNA




 


22
Calmodulin
Cfa.4168.1.S1_at
<0.01
PREDICTED: Canis
52.54237
CCACCCATGGTGACGATGACAC






familiaris

ACATCCTGGTGGCATGCGTGTG






similar to

TTGGTTTAGCGTTGTCTGCGTT






calmodulin 1;

GTACTAGAGCGAAAATGGGTGT






transcript

CAGGCTTGTCACCATTCACACA






variant 3

GAAATTTAAAAAAAAAAAAAAA






(LOC480416);

ANNNNGANAAAAAACCTTTACC






mRNA

AAGGGAGCATCTTTGGACTCTC








TGTTTTTAAAACCTCCTGAACC








ATGACTTGGAGCCAGCAGATTA








GGCTGTGGCTGTGGACTTCAGC








ACAACCATCAACATTGCTGATC








AAGAAATTACAATATACGTCCA








TTCCAAGTT


 


28
Calcinuerin
Cfa.19761.1.S1_at
<0.001
PREDICTED: Canis
98.83382
GAATTAACAATCTGCTTGAGCC






familiaris

CCAAAACACTACTTATGCACTT






similar to

CACTTGCCAAAAGATTTGNGCA






protein phospha-

AGGTTTTGTACCCTGGTAAATG






tase 3 (formerly

ATGCCAAAGTTTGTTTTCTGTG






2B); catalytic

GTGTTTGTCAAATGTTCTATGT






subunit; beta

ATAATTGACTGTCTGTAACATG






isoform

CTGTTTNCTTCCTCTGCAGATG






(calcineurin A

TAGCTGCTTTCCTAAATCTGTC






beta); tran-

TGTCTTTCTTTAGGTTAGCTGT






script variant 5

ATGTCTGTAAAAGTATGTTAAA






(LOC479248);

TTAAATTACTCTATCAGACGCT






mRNA

TGTCTGTCTTTTGATGTAGAAG








CAACTTTGTAGCACCTTGTTTT








GAGGTNNGCTGCATTTGTTGCT








GTACTTTGTGCAT


 


32
Calcium/
Cfa.3884.1.S1_at
<0.01
Homo sapiens
24.10714
GGTGCTGTTCACCACAGTAAGT



calmodulin-


PTEn induced

GGCCTCTCAGTGTTGCTGACCA



dependent


putative kinase

AAGTGTGAAATCCTAGAGCTTC



protein


1 (PINK1); mRNA

AGGGGAGAGGACGTGGGGGAAA



kinase II




TCCGGGGCTTGACTTTATAATA








GGATTATAGAGATGAAAAGTAC








ACCTTGCTTTAGGCAACAGTTG








GGATTCCTAAGACGCATGTGTA








AGAGCATATGTGAAATCCCTTC








CCCATTGTTGATCTCTACTCAC








AGAATTTTGTCTTTATTATGGT








GTAAGAATCACTCTTAAAGCCA








CATATTCAATTCAAAGCAAATA








CGTGTTCTGCAGTTGCAAATGT








GTATTTAATTCTTCACAATTCC








TGTAAG


 


33
Adenylate
CfaAffx.5462.1.S1_s_at
<0.01
PREDICTED: Canis
100
GAAACTCGGTCTGGTGTTCGAT



cyclase-


familiaris

GACGTCGTGGGCATTGTGGAGA



associated


similar to

TAATCAATAGTAGGGATGTCAA



protein 1


Adenylyl

AGTTCAGGTAATGGGTAAAGTG






cyclase-

CCAACCATTTCCATCAACAAAA






associated pro-

CAGATGGCTGCCATGTTTACCT






tein 1 (CAP 1);

GAGCAAGAATTCCCTGGATTGC






transcript

GAAATAGTCAGTGCCAAATCTT






variant 1

CTGAGATGAATGTCCTCATTCC






(LOC475317);

TACTGAAGGCGGTGACTATAAT






mRNA

GAATTCCCAGTCCCTGAGCAGT








TCAAGACCCTATGGAATGGGCA








GAAGTTGGTCACCACAGTGACA








GAAATTGCTGGATAAGCGAAGT








GCCACTGGGTTCTTTGCCCTCC








CCCTCACACCATGGGATAAATC








TATCAGGACGGTTCTTTTCTAG








ATTTCCTTTACCTTTCTGCTCT








TAAACTGCTT


 


34
Protein
Cfa.6174.1.A1_at
<0.01
PREDICTED: Canis
100
AAATCTTACGAAGCCCAATATG



Phosphatase


familiaris

CAGGGAGTTAACTGAAAACTAT



I


similar to

CTTGGCAGTGAGGTTGGCACTG






protein phospha-

TTGATAAAGCTGGTCCCTTCCT






tase 1A isoform

TTAACTGTCTTTTAGGTTGTTC






1; transcript

TTGCCTTGTTGCCAGGAGTATT






variant 2

GCAGGTAATACAGTATATTCAT






(LOC480344);

AAGAATATCAATCTTGGGGCTA






mRNA

AAATGCCTTGATTCTTTGCACC








TCTTTTACAAGTCCTTACGTTG








AATTACTAATTGATAAGCAGCA








GCTTCCTACATATAGTAGGAGA








CTGCCACGTTTTTGCTATCATG








ATTGGCTGGGCCTGCTGCTGTT








CCTAGTAAGGTAT


 


35
Diazepam
CfaAffx.14836.1.S1_s_at
<0.01
PREDICTED: Canis
100
AATGGTGCCATCTTACTGAGGG



binding


familiaris

ATTTTGTAGGCTGTTTTATAGA



inhibitor


similar to

TTTTCCTAAGCCTCTGGTTGCA






peroxisomal D3;

GTGATAAATGGTCCAGCCATAG






D2-enoyl-CoA

GAATCTCCGTCACCATTCTCGG






isomerase

GCTATTCGATCTTGTGTATGCT






isoform 1

TCCGACAGGGCAACATTTCACA






(LOC478706);

CTCCTTTTACTCACCTGGGCCA






mRNA

AAGTCCAGAAGGATGTTCCTCC








TATACTTTTCCCAAGATAATGG








GCCAAGCCAAGGCAGCAGAGAT








GCTCATGTTTGGAAAGAAGTTA








ACAGCTAGAGAAGCCTGTGCTC








AAGGACTTGTTACTGAAGTTTT








TCCCGATAGCACTTGTCAGAAA








GAAGTTTGGACCAGGCGGAAAG








CATATTCAAAACTCCCCCGAAA








TACCTTGCATATTTCCAAACAG








AGCATCAGAAATCTTGAGAAAG








AAAAGCTACATGCTGTTAACGC








AGAAGAAAACAGCGTCCTCCAG








GAAAGGTGGCTGTCAGACGAAT








GCATAAATGCAGTCATGAGCTT








CTTATCCCGGAAGGCCAA


 


36
Tumor
Cfa.1611.1.A1_s_at
<0.01
PREDICTED: Canis
97.90874
ATGATAGTTGCCATGCCAACCA



protein


familiaris

GCTCCAGAATTACCGCAATTAT



p53


similar to

TTGTTGCCTGCAGGGTACAGCC



binding


tumor protein

TTGAGGAGCAAAGAATTCTGGA



protein


p53 binding pro-

TTGGCAACCCCGTGAAAACCCT






tein; 1; tran-

TTCCACAATCTGAAGGTACTCT






script variant 4

TGGTGTCAGACCAACAGCAGAA






(LOC478274);

CTTCCTGGAGCTCTGGTCTGAG






mRNA

ATCCTCATGACCGGGGGGGCAG








CCTCTGTGAAGCAGCACCATTC








AAGTGCCCATAACAAAGATATT








GCTTTAGGGGTATTTGACGTGG








TGGTGACGGATCCCTCATGCCC








AGCCTCGGTGCTGAAGTGTGCT








GAAGCATTGCAGCTGCCTGTGG








TGTCACAAGAGTGGGTGATCCA








GTGCCTCATTGTTGGGGAGAGA








ATTGGATTCAAGCAGCATCCAA








AATACAAACATGATTATGTTTC








TCACTAATACTTGGTCTTAACT








GATTTTATTCCCTGCTGTTGTG








GAGATTGTGNTTNNNCCAGGTT








TTAAATGTGTCTTGTGTGTAAC








TGGATTCCTTGCATGGATCT


 


 4
Ubiquitin
CfaAffx.275.1.S1_s_at
<0.001
PREDICTED: Pan
97.19626
GATTTGGCCCGTGACCCTCCAG



conjugating


troglodytes

CACAATGTTCTGCAGGTCCTGT



enzyme E2D 3


LOC461941

TTGGGATGATATGTTTCATTGG






(LOC461941);

CAAGCCACAATTATAGGACCTA






mRNA

ATGACAGCCCATATCAAGG


 


 5
NEDD8
Cfa.12556.1.A1_s_at
<0.001
PREDICTED: Canis
99.12473
GGAATGGGCTACTCTACTCATG



ultimate


familiaris

CAGNCAAGCAGGNCCTGCATCA



buster-1


similar to NEDD8

GGCCAGTGGGAACCTGGACGAA






ultimate buster-

GCCCTGAAGATTCTTCTCAGCA






1 (NY-REN-18

ATCCTCAGATGTGGTGGTTAAA






antigen)

TGATTCAGATCCTGAAACGANC






(LOC475542);

AACCAGCAAGAAAGTCCTTCCC






mRNA

AGGAAAACATTGACCAACTGGT








GTACATGGGCTTCGACGCTGTG








GTGGCTGATGCTGCCTTGAGAG








TGTTCAGGGGAAACGTGCAGCT








GGCAGCTCAGNCCCTCGCCCAC








AACGGAGGAACTCTTCCTCCTG








ACCTTCAGCTCTTGGTGGAAGA








CTCTTCATCAACGCCATCCACG








TCCCCTTCCGACTCCGCAGGTA








CCTCTAGTGCCTCAACAGATGA








AGATATGGAAACCGAAGCTGTC








AATGAAATACTGGAAGATATTC








CAGAACATGAAGAAGATTATCT








TGACTCAACACTGGAAG


 


37
BCL2-
CfaAffx.6742.1.S1_s_at
<0.01
Canis familiaris
100
GGCCCACCAGCTCTGAGCAGAT



associated


BCL2-associated

CATGAAGACAGGGGCCCTTTTG



X protein


X protein (BAX);

CTTCAGGGTTTCATCCAAGATC



(BAX)


mRNA

GAGCAGGGCGAATGGGGGGAGA








GACACCTGAGCTGCCCTTGGAG








CAGGTGCCCCAGGATGCATCCA








CCAAGAAGCTGAGCGAATGTCT








CAAGCGCATCGGAGATGAACTG








GACAGTAACATGGAGTTGCAGA








GGATGATCGCAGCTGTGGACAC








AGACTCTCCCCGTGAGGTCTTC








TTCCGAGTGGCAGCTGAGATGT








TTTCTGATGGCAACTTCAACTG








GGGCCGGGTTGTTGCCCTCTTC








TACTTTGCCAGCAAACTGGTGC








TCA
















TABLE 12







Summary of genes important for rebound potentiation


and DNA integrity










Gene




Expression




Compared



Gene
to Control
Role





Gamma-aminobutyric acid
↑
Involved in single channel


(GABA) A receptor,

conductance (Cl-channel)


gamma 2




Calmodulin
↓
Influx of calcium results in




calcium/calmodulin




complex which activates




CaMKII and calcineurin


Calcinuerin
↓
Involved in the pathway for




RP suppression


Calcium/calmodulin-
↓
Involved in induction and


dependent protein kinase II

suppression of RP


Adenylate cyclase-
↓
Adenlyl cyclase is involved


associated protein 1

in suppression of RP


Protein Phosphatase I
↓
Dephosphorylates




components in stress-activated 




pathways. Active PP-1 




results in CaMKII inhibition 




and RP suppression


Diazepam binding inhibitor
↓
Displaces benzodiazepine




Down regulates the effects




of GABA


Tumor protein p53 binding
↓
Keep the cell from


protein

progressing through the cell




cycle if there is damage to




DNA present.


Ubiquitin conjugating
↓
The regulated proteolysis of


enzyme E2D 3

proteins by proteasomes


(and NEDD8 ultimate

removes denatured,


buster-1)

damaged or improperly




translated proteins from




cells and regulates the level




of proteins like cyclins or




some transcription factors


BCL2-associated X protein
↓
Accelerates programmed




cell death by binding to, and




antagonizing the apoptosis




repressor BCL2









Effect of Nutrition on Genes Involved with Glucose Metabolism

Twenty four genes associated with glucose metabolism are down regulated in animals fed the super senior diet which would suggest that these animals are utilizing fat (fat oxidation) instead of glucose as a fuel source. The compositions disclosed herein may be part of a therapeutic regime in diabetic animals and/or for obesity prevention or treatment in an animal. These down regulated genes are identified and their putative role in glucose metabolism described in detail below in Tables 13 and 14.







TABLE 13







Genes involved in Glucose Metabolism

















Best
% match







current
of probe



Sequence


P-
BLAST
sequence to



ID No.
Gene
Probe
value
annotation
BLAST hit
Probe Target Seq.
















38
Phosphorylase
Cfa.10856.1.S1_at
<0.01
PREDICTED: Canis
99.3392
GAAAGTTCACCACTGCATGTTT



kinase


familiaris

TATGATCAGATAACTCATTGAA






similar to

ATGAGTCTTTGCTCTTTAGACT






phosphorylase

AAATTCCCACCTAGTACTGCCA






kinase beta;

TTAAAATGAATTTGCCAGCTGG






transcript

TGTGCATACTGGAAATGAAAAG






variant 2

ATACTGAAAGAATGGAACGAAT






(LOC478139);

GGTGAGCTTAACTCAGTGGCAC






mRNA

TGTCATACTGGAAAAATACAGT








AAAATCATAAAAACAGATCTGC








CAGCTGATGTTTTTATTCTCAG








AAACAGCATTGTTGATAATATT








TTAGTATACAGAGCTACTGTAC








AATTTTTACCTTGNAAACATGA








CTGTGGTTTTGTATTTGTGTTG








ACTTTAGGGGTTGGGATAAAAT








NCAGTATAATATATACCTTATC








AAACNTTTTCTTTGAGCTCTTA








CTAAAAATATGGCATGCATAAG








ATTGTTCAGAAGAGTAGACTGT








TAACCTAGTTTGTA


 


39
Phosphorylase
Cfa.10412.1.A1_s_at
<0.01
PREDICTED: Canis
99.36306
CTTCCAGAGCTGAAGCTGGCCA






familiaris

TTGATCNAAATTGACAATGGCT






phosphorylase;

TCTTCTCTCCCAAGCAGCCTGN






glycogen; liver;

CCTCTTCAAAGATTTAATCAAT






transcript

ATGCTATTTTATCATGACAGGT






variant 1

TTAAAGTCTTCGCAGACTATGA






(PYGL); mRNA

AGCCTATGTCAAGTGTCAAGAA








AAAGTCAGCCAGCTGTACATGA








ATCCAAAGGCCTGGAACACAAT








GGTACTCAAAAACATAGCTGCC








GCAGGGAAGTTCTCTAGTGACC








GAACAATTAAGGAATATGCCAG








GGACATCTGGAACATGGAACCT








TCAGATCTCAAGATTTCCCTAT








CCAATG


 


40
Glycogen
Cfa.913.1.A1_s_at
<0.01
PREDICTED: Canis
99.49622
GACTCCACCGGAGGCAATTGCA



synthase


familiaris

CTGTGTAGCCGTCTGCTGGAGT



kinase 3


similar to

ATACACCAACTGCCCGATTGAC






Glycogen

ACCACTGGAAGCTTGTGCACAT






synthase kinase-

TCATTTTTTGATGAATTAAGGG






3 beta (GSK-3

ACCCAAATGTCAAACTACCAAA






beta); tran-

TGGGCGAGACACACCTGCACTC






script variant 1

TTCAACTTCACCACTCAAGAAC






(LOC478575);

TGTCAAGTAATCCACCTCTAGC






mRNA

TACCATCCTTATTCCTCCTCAT








GCTCGGATTCAAGCAGCTGCTT








CAACCCCTACAAATGCCACAGC








AGCCTCAGATGCTAATGCCGGA








GACCGTGGACAGACGAACAATG








CCNCTTCTGCATCAGCTTCTAA








CTCCACCTGAACAGTCCCGAGC








AGCCAGCTGCACAGGAAGAACC








ACCAGTTACTTGAGTGTCACTC








A


 


22
Calmodulin
Cfa.4168.1.S1_at
<0.01
PREDICTED: Canis
52.54237
CCACCCATGGTGACGATGACAC






familiaris

ACATCCTGGTGGCATGCGTGTG






similar to

TTGGTTTAGCGTTGTCTGCGTT






calmodulin 1;

GTACTAGAGCGAAAATGGGTGT






transcript

CAGGCTTGTCACCATTCACACA






variant 3

GAAATTTAAAAAAAAAAAAAAA






(LOC480416);

ANNNNGANAAAAAACCTTTACC






mRNA

AAGGGAGCATCTTTGGACTCTC








TGTTTTTAAAACCTCCTGAACC








ATGACTTGGAGCCAGCAGATTA








GGCTGTGGCTGTGGACTTCAGC








ACAACCATCAACATTGCTGATC








AAGAAATTACAATATACGTCCA








TTCCAAGTT


 


29
Protein
CfaAffx.408.1.S1_s_at
<0.01
PREDICTED: Canis
99.64664
TTCAGTTCCTGTCTCATGGCCG



Kinase C


familiaris

CTCCCGGGACCATGCCATCGCC






similar to

GCCACTGCCTTCTCCTGCATCG






myeloid-associ-

CTTGTGTGGCTTATGCCACCGA






ated differenti-

AGTGGCCTGGACCCGGGCCCGT






ation marker

CCCGGAGAGATCACCGGCTACA






(LOC611521);

TGGCCANTGTGCCGGGCCTGCT






mRNA

CAAGGTGCTGGAGACCTTTGTG








GCCTGCATCATCTTCGCCTTCA








TCAGCAACCCCTCCCTGTACCA








GCACCAGCCGGCCCTGGAGTGG








TGTGTGGCCGTCTACTCCATCT








GTTTCATCCTGGCGGCTGTGGC








CATCCTACTGAACCTGGGGGAC








TGCACCAACATGCTGCCCATCT








CCTTCCCCAGTTTCCTGTCGGG








CCTGGCCCTGCTCTCCGTCCTG








CTGTATGCCACGGCTCTGGNTC








TCTGGCCGCTCTACCAGTTCAA








CGAGAAGTATGGTGGCCAGCCC








CGTCGGTCGAGGGATGTTAGCT








GCGCCGACAGGCACACCTACTA








CGTGTGTACCTGGGACCGCCGC








CTGGCTGTGGCCATCCTGACAG








CCATCAACCTGCTGGCTTACGT








GGCTGACCTGGTGTAC


 


30
Protein
Cfa.15485.1.A1_s_at
<0.01
PREDICTED: Canis
100
GGAGCAGTCAGAACTAAGACAT



Kinase C


familiaris

GGTCCGTTTTACTATATGAAGC



Binding


similar to

AGCCACTCACCACAGACCCTGT



Protein


protein kinase C

TGATGTTGTACCGCAGGATGGA






binding protein

CGGAA






1 isoform b;








transcript








variant 11








(LOC477252);








mRNA




 


41
Hexokinase
Cfa.19125.2.S1_at
<0.01
Macaca fasci-
76.70683
TAATGACTGCCAACTCACTGTT



3


cularis testis

TGTTGGAGTTATATGCAGAAAT






cDNA; clone;

AAAGNCCAAGTCTTCAGAAACA






QtsA-14856;

GGCTTCAGGATGCCCTCACCAG






similar to human

GGATGGAAGAGGCAGGCTGCAG






receptor associ-

CAAAGAGATGCAGAGTTCCCTT






ated protein 80

GCACATCTCGACTTAAATGAGT






(RAP80); mRNA;

CTCCCATCAAGTCTTTTGTTTC






RefSeq:

CATTTCAGAAGCCACAGATTGC






NM_016290.3

TTAGTGGACTTTAAAAAGCAAC








TTAACGTTCGGCAAGGTAGTCG








GACACGGACCAAAGCAGGCAGA








GGAAGAAGGAGAAAACCCTGAA








TTTCTAGGGTCCAGACACCCGA








CAAAACCATTAGCAATAGGGGT








GGGCCGTGTCATTAAGTCTTAG








TGGCTTCTGTTTCATTGTTGAA








CAAGTTTTTTGGCCCNGCAGTT








TTCACCACCAGCACCAACTCAG








CATTCTTGTTTTGATGTTTTCT








ATAAGCTATACAGACAATTGTG








TATAGTATTCTGTTTTATAACA








GTCTGGATTCACTT


 


42
Fructose
CfaAffx.26135.1.S1_s_at
<0.01
PREDICTED: Canis
100
AGTGGCGCTGTGTGCTGAAAAT



1,6 bisphos-


familiaris

TGGGGAACACACTCCCTCAGCC



phatase


aldolase A;

CTTGCGATCATGGAAAATGCCA






transcript

ACGTTCTGGCCCGTTAT






variant 1








(LOC479787);








mRNA




 


43
Glyceral-
AFFX-Cf_Gapdh_3_at
<0.01
Canis familiaris
100
AGCTCACTGGCATGGCCTTCCG



dehyde


glyceraldehyde-

TGTCCCCACCCCCAATGTATCA



3-phosphate


3-phosphate

GTTGTGGATCTGACCTGCCGCC



dehydrogenase


dehydrogenase

TGGAGAAAGCTGCCAAATATGA






(GAPDH); mRNA

CGACATCAAGAAGGTAGTGAAG








CAGGCATCGGAGGGACCCCTCA








AAGGCATCCTGGGCTACACTGA








GGACCAGGTGGTCTCCTGTGAC








TTCAACAGTGACACCCACTCTT








CCACCTTCGACGCCGGGGCTGG








CATTGCCCTCAATGACCACTTT








GTCAAGCTCATTTCCTGGTATG








ACAATGAATTTGGCTACAGCAA








CCGGGTGGTGGACCTCATGGTC








TACATGG


 


44
Glucose 6-
Cfa.19351.1.S1_at
<0.01
Homo sapiens
15.11194
GAATGTGTTGGCAGACTGAGGC



phosphate


cDNA FLJ30869

CCCCCATGTTTTTAATGCGCAC



dehydrogenase


fis; clone

TGGGGACAACCATCTAAGGTCT






FEBRA2004224

AGAAACTTTTGGACCATAGGAA








AGATAGGTTTATGGTCCTCTTC








CAGATGCAGCCCTAGGAGAGCA








TTCCCATGGGGTCTCTGGATCC








CTTTCNTTGCTCTGTGAGGCTC








TGTGACCACCTTTTGNNNTGNN








GGGGGCAGGGGGNCTTCCTCAG








CTCCGCCTCCAGTGCCCCCAGG








TCCCCCACGGCTCACAGTCCNT








GAAAATTCAGAGCTGCCCTGTA








AGGATTTTGTCCACTGGGCAAT








TCAGATATACTTCGATATCCCT








GAGAAAGAAGAGGCAGCAGCAA








ACACTCCCNAGGGCATCTGTCT








CAGNANTCTCTCNTTGNATGAG








ACAGAAGCCTACTTTTCAGAAA








NCTTATCANGGNTACTTTATAA








GAAACTTTTTTTTTTTTNCTAA








AATCAGACAAAAGGTGGCTTNT








GCATATTCTTNATTAATAACTG








TGTCTTTGTCTCCTCTGCTTAA








CTTTAGGA


 


45
Enolase
CfaAffx.30133.1.S1_s_at
<0.01
PREDICTED: Canis
97.72257
GGTACATCACGCCTGATCAGCT






familiaris

GGCTGACCTCTACAAGTCCTTC






similar to

ATCAGGGACTACCCAGTGGTGT






T21B10.2b;

CTATCGAAGACCCCTTCGACCA






transcript

GGATGACTGGGAAGCTTGGCAG






variant 1

AAATTCACTGCCAGCGCTGGAA






(LOC479597);

TCCAGGTGGNGGGGGANGATCT






mRNA

CACCGTGACCAACCCAAAGCGG








ATTTCCAAGGCTGTGGGCGAGA








AATNGTGCAACTGCCTCCTGCT








TAAAGTGAACCAGATTGGCTCT








GTGACCGAGTCTCTTCAGGCGT








GCAAGCTGGCCCAGTCCAATGG








GTGGGGCGTCATGGTGTCGCAT








CGCTCCGGGGAGACCGAAGATA








CCTTCATCGCTGACCTGGTGGT








GGGANTCTGCACTGGGCAGATC








AAGACGGGTGCACCATGCAGAT








CTGAGCGCTTGGCCAAGTACAA








CCAGATCCTCAGAATTGAAGAG








GAACTGGGTAGCAAGGCCAAGT








TCGCCGGCAGAAGCTTCAGAA


 


46
Lactate
Cfa.300.1.S1_at
<0.01
PREDICTED: Canis
97.99427
ATCTGACCTGTTACTCAAGTCG



dehydrogenase


familiaris

TAATATTAAAATGGCCTAAGAA






similar to L-

AAAAACATCAGTTTCCTAAAGT






lactate dehydro-

TACACATAGGAATGGTTCACAA






genase A chain

AACCCTGCAGCTATGTCCTGAT






(LDH-A) (LDH

GCTGGATGAGACCTGTCTTGTG






muscle subunit)

TAGTCCTAAATTGGTTAACGTA






(LDH-M) (Prolif-

ATATCGGAGGCACCACTGCCAA






eration-inducing

TGTCATATATGCTGCAGCTACT






gene 19 pro-

CCTTAAACCAGATGTGTATTTA






tein); tran-

CTGTGTTTTGTAACTTCTGATT






script variant 1

CCTTCATCCCAACATCCAACAT






(LOC476882);

GCCTAGGCCATCTTTTCTTCTT






mRNA

CAGTCACATCCTGGGATCCAAT








GTATAAATTCAATATTGCATGT








ATTGTGCATAACTCTTCTA


 


47
Citrate
Cfa.10361.2.S1_at
<0.01
PREDICTED: Canis
98.49624
AGTATGCCAGATCGGAACCTTT



lyase


familiaris

TTCCCATTTACAGTTCATGTTA






similar to

ATCCAATTTTTTTTATTATCTC






citrate lyase

ACTGGCCAGTTATTCCTTTAAA






beta like

AATGAACTTCCTTCTTTTTGAT






(LOC476974);

TCCAAGCTTATGATTTTACTGC






mRNA

TCATTAATGTGTTACAAATATG








CACTTAATGATTTCACAGGGAG








ATAAAATAGTGAAGAGAGATGG








GCTGAGGGGCTGTTAGGACTTT








AATGAAACAGATCTTTCCCGAA








TATTTCTCCCTTCACATTTCTC








ACATTAGATGTTTCCCACATTG








TTCTACTCCACACTATAAATAA








TTTTAAGGCCAATCTTAAAAAA








TGGTAGTTAAGTGAAGGGGTTG








TGTTTATTTCACTAGAAATCTG








ATAAAACGAGAGATGACATAGA








AAAAGTTATCATTTTTGTTCAT








ACAGATGGCTTCTAAAAATAAA








TCTTCAAAACTGATTACTTTTA








ACCTCCACCTCCCAAAATGAAA








CATCCCTACATTTGAACTGCTA








GGTGAGAACTCTGAAAGCCCTC








ATCC


 


48
Glycerol
CfaAffx.21204.1 S1_s_at
<0.01
PREDICTED: Canis
100
GGGTACATCCTATGGCTGCTAT



kinase


familiaris

TTCGTCCCCGCGTTTTCAGGGT






similar to

TATATGCACCTTACTGGGAGCC






glycerol kinase

CAGTGCAAGAGGGATCATCTGT






isoform 2; tran-

GGGCTCACTCAATTCACCAATA






script variant 8

AATGCCATATTGCTTTTGCTGC






(LOC480872);

ATTAGAAGCTGTTTGTTTCCAA






mRNA

ACCCGGGAGATTTTGGATGCCA








TGAACCGAGACTGCGGAATTCC








ACTCAGTCATTTGCAGGTAGAT








GGAGGAATGACCAACAACAAAA








TTCTTATGCAACTACAAGCAGA








CATTCTATATATCCCAGTAGTG








AAGCCCTCGATGCCAGAAACAA








CTGCCCTGGGAGCTGCCATGGC








AGCCGGGGCTGCGGAGGGAGTT








GGTGTTTGGAGTCTTGAACCCG








AGGATCTGTCAGCAGTCACGAT








GGAGCGATTTGAACCCCAGATC








AATGCTGAGGAAAGTGAAATTC








GTTACTCTACATGGAAGAAGGC








TGTGATGAAGTCAGTGGGCTGG








GTTACAACTCA


 


49
Transketolase
CfaAffx.13684.1.S1_s_at
<0.01
Homo sapiens
86.53846
GAAGATCTGGCCATGTTTCGGT






transketolase

CCATCCCCACTGCTACGATCTT






(Wernicke-

TTACCCAAGTGACGGGGTGTCA






Korsakoff

ACAGAGAAGGCGGTGGAATTAG






syndrome); mRNA

CAGCCAATACAAAGGGCATCTG






(cDNa clone MGC:

CTTCATCCGGACCAGCCGCCCA






15349 IMAGE:

GAAAACGCCATCATCTATAACA






4310396);

ACAATGAGGATTTCCAAATCAA






complete cds

ACAAGCCAAGGTGGTCCTGAAG








AGCAAGGATGACCAGGTGACTG








TGATTGGGGCCGGAGTGACCCT








ACATGAGGCCTTGGCTGCTGCT








GAACTGCTGAAGAAAGAGAAGA








TCAACATTCGTGTGTTGGACCC








CTTCACCATCAAGCCCCTGGAC








AGAAATCTCATTCTCGAAAGCG








CCCGTGCGACCAAGGGCAGGAT








CGTCACCGTGGAGGACCATTAC








TATGAAGGTGGCATAGGTGAGG








CAGTGTCCTCTGCCTTGGTGGG








TGAGCCTGGCATCACCGTCTCC








CGCCTTGCAGTTGGTGAGGTAC








CAAGAAGCGGGAAGCCAGCTGA








GCTGCTGAAGATGTTTGGCATT








GACAGGGACGCCATCGCACAAG








CTGTGAGGGACCTTGTCGCCAA


 


50
Ribulose
Cfa.13084.1.A1_s_at
<0.01
Homo sapiens
57.79468
CCCCAAGGAGATGAGGAGCGAT



phosphate


SLIT-ROBO Rho

GACCCCAGCAACAGGAANAACA



3-epimerase


GTPase acti-

GCCCACTGAAGGGCTGGTGTGT






vating protein 2

GTGTNCTTCACGTGCCAGAAGA






(SRGAP2); mRNA

GAAGTTTAGATCCTCCCAGGGG








AATCGCAATGTTGTGGCGTCCT








GACTTGTATGTCACGTTTTGTG








TAAAAATGGTATATTCTTTAAA








ATAGTGTTGATAACTGGAATAT








TGTATGTATGCTTGGAGATGCT








TTGTGTGAACCTAAGACTGTCA








CTCAACAGATGTTGGATTGGG


 


51
Ribose 5-
Cfa.335.2.S1_at
<0.01
PREDICTED: Canis
100
AGCCTTTCTACTGACCCTGCAA



phosphate


familiaris

GAGTGGAGCGTGTTCACCTTGA



isomerase


similar to

ACCCCCAGCGTGCAGCTGAGGT






ribose 5-phos-

AGACATGCCTCTCCAGGAGCCT






phate isomerase

TTGCCTTAATGCATCTGTGCCA






A (ribose 5-

GACAGACGGCTGG






phosphate








epimerase)








(LOC475755);








partial mRNA




 


52
Cytochrome
CfaAffx.4942.1.S1_s_at
<0.01
PREDICTED: Canis
100
GGCAGTTTGAAAATAAAGTTCC



c oxidase


familiaris

AGAGAAACAAAAGCTATTTCAG



polypeptide


similar to

GAGGATAATGGAATTCCAGTGC



VIIa-liver/


cytochrome c

ATCTAAAGGGTGGAGTAGCTGA



heart,


oxidase;

TGCCCTCCTGTATAGAGCCACT



mitochondrial


subunit 7a 3

ATGATGCTTACAGTTGGTGGAA



precursor


(LOC611134);

CAGCATATGCCATGTATCAGCT






mRNA

AGCTGTGGCTTCTTTTCCCAAG








AAGCA


 


53
Cytochrome
Cfa.15065.1.S1_at
<0.01
PREDICTED: Canis
99.75961
GGTCCGCAGTCGTTCTGTGCGG



c oxidase


familiaris

TCATGTCTGTGCTGGTGCCGCA



subunit


similar to

GCTGCTGAGGGGCCTAACAGGC



VIII liver


Cytochrome c

CTCACCCGGCGGCTCCCGGTGC



form


oxidase polypep-

ATCGTGCCCAGATCCATTCCAA






tide VIII-liver;

GCCGCCGCGGGAGCAGCTCGGG






mitochondrial

ACCATGGATGTTGCCGTTGGGC






precursor (Cyto-

TCACCTNCTGCTTCCTGTGTTT






chrome c oxidase

CCTCCTGCCATCGGGCTGGGTC






subunit 8-2)

CTGTCACACCTGGAGAGCTACA






(LOC476040);

AGAAGCGGGAGTGAAGGGGGCT






mRNA

GTCCTGTCCCTCACCCTGTGAC








CTGACCACCCCTGGCCTGTCCT








GATCATGTCTGCTGCATTCCTG








GCCGGCCTTCCATGGATCATGT








CCTTCAATTACAGTGACCTCTT








CTACAGTCATGACCTCTTGATT








TCTCCATGGTGACATCCTGGGA








CCAAACATATTGGTTTATAA


 


54
Ubiquinol-
Cfa.1425.2.A1_at
<0.01
PREDICTED: Canis
27.18053
CTTATGCATTCCTTCCAAAATT



ùcytochrome


familiaris

GGATCATTTAGGTCAAATTATT



c reductase


similar to

TGATGTTAAATCATAAGTTTTC






Ubiquinol-

ATTTGCTTACATTTACGATATC






cytochrom-c

AGCGTCAGCTACGGAATCAATC






reductase com-

TGCTGAAGGACCGTGGCTGGCG






plex core pro-

GCGTGTACGATCCAGCAACCAG






tein 2; mito-

CGCCTGGGACCCGACTTCATCC






chondrial pre-

AGGAACCCCTCAGAAGACTCCA






cursor (Complex

CTGACATTAGGAAGACTCATAA






III subunit II);

GAACCTTACAAGAAAAAGTATC






transcript

AACCCCATCAAAACGGCAGAAA






variant 1

AGAAACATATCTTGTTATTAGT






(LOC479815);

AGCTGAAATTCCATTTTCTACA






mRNA

TGTTGCCATACCTTATAAAAAC








TACACTAAGCTACGCTTAAGGA








AATACATTTTCTTAAATAAATT








AGAATTGAAACCAATTTTTAAG








TAAATCTAGGGNTTCAATTTAT








TCTCATTGNGTNTTGTTTCTGG








TGCAATCATGAANAACAGCATN








CTATTAACCAACCTTGGTCCCA








TGTACATAA


 


55
ATP synthase
CfaAffx.3186.1.S1_s_at
<0.01
PREDICTED: Canis
98.57651
AATTGGGACTGTGTTTGGGAGC






familiaris

CTCATCATTGGTTATNCCAGGA






similar to ATP

ATCCCTCTCTGAAGCAACAGCT






synthase; H +

CTTCTCCTACGCCATTCTGGGC






transporting;

TTTGCCCTCNCGGAGGCCATGG






mitochondrial FO

GGCTTTTTTGCCTGATNGTGGC






complex; subunit

CTTTCTCATCCTCTTNGCCATG






c isoform 2a

TGAAGGAGTCGTCTCCACCTCC






precursor

CATAGGTCTTTCTCCCATGTCT






(LOC477595);

TGTCTGCCCTGTATGCCCTGTA






mRNA

TGTTCCTTTTCCTATACCTCCC








CAGGCAGCCTGGGGAAAGTGGT








TGGCTCAGGGTTTGACA


 


56
NADH-
Cfa.4415.1.S1_at
<0.01
PREDICTED: Canis
98.20789
GGTGACTTTGGACGTCCGTTCC



ubiquinone


familiaris

TGCTCTGTGGAGGCNNTGCTTC



oxido-


similar to NADH-

GTTCCGGGCCTTGCGGCAACTC



reductase


ubiquinone

GGTNTTTCCTTCCCCTGCGCGG






oxidoreductase

GAGACCTCTGCCACAACCATGT






MLRQ subunit

TACGCCAGATCATCGGTCAGGC






(Complex I-MLRQ)

CAAGAAGCATCCGAGCTTGATC






(CI-MLRQ)

CCCCTCTTCATATTTATTGGGG






(LOC477682);

CAGGAGGTACTGGAGCAGCGCT






mRNA

GTATGTATTGCGCTTGGCATTG








TTCAATCCAGATGTTAGTTGGG








ATAGGAAGAATAACCCAGAACC








TTGGAACAAACTGGGTCCCAAT








GATCAATACAAGTTCTACTCAG








TGAATGTAGATTACAGCAAACT








GAAGAAAGAAGGTCCAGACTTC








TAAATGAAATGTTTCACTATAA








AGCTGCTTAGAATGAAGGTCTT








CCAGAAGCCATCCGCACAATTT








TCCACTTATCCAGGAAATATTT








CCCCTCTAAATGCACGAAATCA








TGTTGGTGTATTGTGTTGGGGT








TTACACTNNANNANTAAATATC








TGAAACTTGANANGTGTCACTA








TTTAATGCTGAAAATTTGCTCT








GAACTTTA


 


57
Facilitated
Cfa.1370.1.A1_at
<0.01
Homo sapiens
23.95833
TTGGAAGGATGGATGCTTGCCC



glucose


cDNA FLJ44038

CAGGTCATGGACACCTCCACAA



transporter/


fis; clone

ATCATCTAGTTTCCCAGTATTT



Glucose


TESTI4028880;

TTATAAATGGAGATTGGGCTCC



transporter-


highly similar

ATGACACTTTACTTGGTCTTCC



like


to Glucose

TTCTTACATAGGTTTTTTGATT



protein III


transporter

ACCCTTTCTCTCCTTGGTGCTT



(GLUT3)


type 3; brain

ATATACTTAAGACCCTTTAGCC








AAACCCTTGCCAATGACAGTAT








TTCAGTCACTAGTTCTCACTGT








TTCCTCTGATCATTGAGCCTTT








GGAAAAAAAATCTCACAGAGCT








TATATGTAATGGGGCTTGGTTG








AACAGATGACTTCCTGTAACTG








CACCTCTACTTTTGGCTTCTCA








AAAACAGTGGGTTGGCAGTAAT








GCAGCGTGGAAGTTTTCCCATT








TCTCAGTGAC
















TABLE 14







Summary of Genes involved in Glucose Metabolism










Gene




Expression




Compared



Gene
to Control
Role





Phosphorylase kinase
↓
Necessary for activation of




glycogen synthase which




stores glucose as glycogen


Phosphorylase
↓
Necessary for glycogen




conversion to glucose 1-




phosphate which feeds into




glycolysis


Glycogen synthase kinase 3
↓
Necessary for activation of




glycogen synthase which




stores glucose as glycogen


Calmodulin
↓
Necessary for activation of




glycogen synthase which




stores glucose as glycogen


Protein Kinase C
↓
Necessary for activation of




glycogen synthase which




stores glucose as glycogen


Protein Kinase C Binding
↓
Necessary for activation of


Protein

glycogen synthase which




stores glucose as glycogen


Hexokinase 3
↓
Necessary for glucose




conversion to pyruvate to




enter the TCA cycle


Fructose 1,6 bisphosphatase
↓
Necessary for glucose




conversion to pyruvate to




enter the TCA cycle


Glyceraldehyde 3-
↓
Necessary for glucose


phosphate dehydrogenase

conversion to pyruvate to




enter the TCA cycle


Glucose 6-phosphate
↓
Involved in pentose


dehydrogenase

phosphate pathway


Enolase
↓
Necessary for glucose




conversion to pyruvate to




enter the TCA cycle


Lactate dehydrogenase
↓
Involved in converting




private to lactate


Citrate lyase
↓
Necessary for citrate




conversion to oxaloacetate




which feeds acetyl-CoA




into the fatty acid synthesis




pathway


Glycerol kinase
↓
Necessary for changing




glycerol into DHAP which




feeds into glycolysis


Transketolase
↓
Involved in pentose




phosphate pathway


Ribulose phosphate 3-
↓
Involved in pentose


epimerase

phosphate pathway


Ribose 5-phosphate
↓
Involved in pentose


isomerase

phosphate pathway


Cytochrome c oxidase
↓
Associated with the


polypeptide VIIa-

production of ATP (energy


liver/heart, mitochondrial

source) in the electron


precursor

transport chain which is




associated with the TCA




cycle


Cytochrome c oxidase
↓
Associated with the


subunit VIII liver form

production of ATP (energy




source) in the electron




transport chain which is




associated with the TCA




cycle


Ubiquinol-cytochrome c
↓
Associated with the


reductase

production of ATP (energy




source) in the electron




transport chain which is




associated with the TCA




cycle


ATP synthase
↓
Associated with the




production of ATP (energy




source) in the electron




transport chain which is




associated with the TCA




cycle


NADH-ubiquinone
↓
Associated with the


oxidoreductase

production of ATP (energy




source) in the electron




transport chain which is




associated with the TCA




cycle


Facilitated glucose
↓
Involved in glucose uptake


transporter/Glucose




transporter-like protein-III




(GLUT3)

## Claims

1. A method for improving the quality of life of a senior or super senior animal comprising feeding the animal a composition comprising:
at least about 9% by weight protein;
at least about 5% by weight fat; and
at least about 0.05% by weight of at least one omega-3 polyunsaturated fatty acid.

2. The method of claim 1 wherein the method comprises feeding the animal the composition in an amount effective to improve the animal's quality of life, wherein improved quality of life is evidenced by an improvement in one or more characteristics selected from the group consisting of alertness, vitality, cartilage protection, maintenance of muscle mass, and skin and pelage quality.

3. The method of claim 1 wherein the method comprises feeding the animal the composition in an amount effective to improve alertness.

4. The method of claim 1 wherein the method comprises feeding the animal the composition in an amount effective to improve vitality.

5. The method of claim 1 wherein the method comprises feeding the animal the composition in an amount effective to protect cartilage.

6. The method of claim 1 wherein the method comprises feeding the animal the composition in an amount effective to maintain muscle mass.

7. The method of claim 1 wherein the method comprises feeding the animal the composition in an amount effective to improve skin and pelage quality.

8. The method of claim 1 wherein the animal is selected from the group consisting of a cat, a dog, and a horse.

9. A method for improving the quality of life of a senior or super senior animal comprising feeding the animal a composition comprising:
at least one omega-3 polyunsaturated fatty acid selected from the group consisting of docosahexaenoic acid and eicosapentaenoic acid;
at least one antioxidant; and at least one nutrient selected from the group consisting of choline, manganese, methionine, cysteine, L-carnitine, lysine, and mixtures thereof.

10. The method of claim 9 wherein the omega-3 polyunsaturated fatty acid in the composition is DHA and wherein the composition comprises at least about 0.02% by weight DHA as measured on a dry matter basis.

11. The method of claim 9 wherein the omega-3 polyunsaturated fatty acid in the composition is DHA and wherein the composition comprises from about 0.02% to about 0.40% by weight DHA as measured on a dry matter basis.

12. The method of claim 9 wherein the omega-3 polyunsaturated fatty acid in the composition comprises EPA and wherein the composition comprises at least about 0.1% by weight EPA as measured on a dry matter basis.

13. The method of claim 9 wherein the omega-3 polyunsaturated fatty acid in the composition comprises EPA, and wherein the composition comprises from about 0.1% by weight to about 1% by weight EPA as measured on a dry matter basis.

14. The method of claim 9 wherein the omega-3 polyunsaturated fatty acid in the composition comprises a mixture of DHA and EPA, and wherein the composition comprises at least about 0.02% by weight DHA and at least about 0.1% by weight EPA on a dry matter basis.

15. The method of claim 9 wherein the composition comprises one or more antioxidants selected from the group consisting of vitamin E, vitamin C, taurine, beta-carotene, carnitine, lipoic acid, and cystine.

16. The method of claim 9 wherein the composition comprises at least about 500 IU/kg vitamin E, at least about 50 ppm vitamin C and at least about 600 ppm taurine.

17. The method of claim 9 wherein the composition further comprises at least about 1000 ppm choline.

18. The method of claim 9 wherein the composition fed to the animal is an animal treat or an animal toy.

19. The method of claim 9 wherein the composition fed to the animal is a nutritional supplement.

20. A method for improving the quality of life of a senior or super senior small or regular breed canine comprising feeding the animal a composition comprising:
from about 60% to about 70% by weight carbohydrate;
from about 15% to about 25% by weight protein selected from the group consisting of animal protein and vegetable protein;
from about 5% to about 7% by weight fat selected from the group consisting of animal fat and vegetable fat;
from about 2.5% to about 4% by weight of at least one omega-3 polyunsaturated fatty acid;
from about 1% to about 2% by weight fiber;
from about 1% to about 2% by weight minerals; and
from about 0.5 to about 1.5% by weight vitamins.

21. A method for improving the quality of life of a senior or super senior large breed dog, wherein the method comprises feeding the animal a composition comprising:
from about 60% to about 70% by weight carbohydrate;
from about 15% to about 25% by weight protein selected from the group consisting of animal protein and vegetable protein;
from about 5% to about 7% by weight fat selected from the group consisting of animal fat and vegetable fat;
from about 3% to about 5% by weight of at least one omega-3 polyunsaturated fatty acid;
from about 1% to about 1.5% by weight fiber;
from about 0.5% to about 1% by weight minerals; and from about 0.75 to about 1.25% by weight vitamins.

22. A method for improving the quality of life of a senior or super senior cat, wherein the method comprises feeding the animal a composition comprising:
from about 30% to about 35% by weight carbohydrate;
from about 40% to about 50% by weight protein selected from the group consisting of animal protein and vegetable protein;
from about 12% to about 15% by weight fat selected from the group consisting of animal fat and vegetable fat;
from about 1% to about 2% by weight of at least one omega-3 polyunsaturated fatty acid;
from about 3% to about 5% by weight fiber;
from about 1% to about 2% by weight minerals; and
from about 1% to about 2% by weight vitamins.

23. The method of claim 1 wherein the method comprises feeding the animal the composition in an amount effective to improve the animal's quality of life, wherein improved quality of life is evidenced by improvement in one or more biological pathways selected from the group consisting of blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport.

24. The method of claim 1 wherein the method comprises feeding the animal the composition in an amount effective to improve the animal's quality of life, wherein improved quality of life is evidenced by a change in expression of one or more genes which encode proteins associated with or related to biological pathways selected from the group consisting of blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport.

25. A method to treat an animal suffering from a disorder or disease associated with or related to a biological pathway selected from the group consisting of blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport comprising administering to said animal a super senior pet food composition.

26. The method of claim 25 wherein said super senior pet food composition comprises at least about 9% by weight protein, at least about 5% by weight fat, and at least about 0.05% by weight of at least one omega-3 polyunsaturated fatty acid.

27. The method of claim 25 wherein said super senior pet food composition further comprises at least one omega-3 polyunsaturated fatty acid selected from the group consisting of docosahexaenoic acid (“DHA”) and eicosapentaenoic acid (“EPA”).

28. The method of claim 25 wherein said super senior pet food composition further comprises at least one antioxidant and at least one nutrient selected from the group consisting of choline, manganese, methionine, cysteine, L-carnitine, lysine, and mixtures thereof.

29. The method of claim 25 wherein said super senior pet food composition comprises the components disclosed in Table 1 or Table 1A.

30. A method to treat an animal suffering from a disorder or disease associated with or related to a biological pathway selected from the group consisting of blood clotting and platelet activation and aggregation, bone and muscle integrity, inflammatory responses, cartilage degradation and pain response, DNA damage and repair pathways, neural function, glycogen synthesis and degradation, glycolysis, gluconeogenesis, the pentose phosphate pathway and electron transport comprising modulating the expression level of one or more genes listed on Tables 5-14 in said animal in order to mimic the pattern of expression seen in vivo after administration of a super senior pet food composition.

31. The method of claim 30 wherein said super senior pet food composition comprises the components disclosed in Table 1 or Table 1A.

32. The method of claim 23 wherein said animal is selected from a group consisting of a senior or super senior large breed canine, regular breed canine, small breed canine or feline.