Methods utilizing compositions containing sacred lotus (methyltransferase) to treat aging skin

FIELD OF THE INVENTION

The present invention relates to the use of Sacred Lotus seed extract in a cosmetic, dermatological and/or dietary compositions for combating aging and methods for treatment and prevention of aging using such compositions. The present invention also concerns the use of components of the Sacred Lotus plant, such as methyltransferase and especially L-isoaspartyl methyltransferase, ascorbic acid, glutathione, etc.

BACKGROUND

Within the dermis are highly stable fibers of collagen and elastin. Collagen, the most abundant protein in the body, has a high tensile strength thus preventing skin from being torn by over stretching. Elastin, also a protein, allows movement. As skin ages elastic tissue increases but it loses the ability to stretch and recover. This loss of resiliency and elasticity is accompanied by increased stiffness, sagging and wrinkling. Changes in collagen solubility and cross-linking contribute to loss of elasticity.

On the cellular level, aspartyl and asparaginyl residues are prominent sites of age related damage in proteins. These damaged sites have been characterized in a variety of proteins, but are particularly common in the long-lived proteins. Enzymatic mechanisms for reversing damage to DNA are well established and have been shown to be essential for extended lifespan.

Experiments performed in vitro with recombinant and chemically modified polypeptides have shown that the presence of an L-isoaspartyl residue may alter both enzymatic activity and the binding of other molecules.

Limiting the accumulation of these residues within cells is currently believed to be important; all human cells examined thus far contain an L-isoaspartyl/D-aspartyl protein methyltransferase that has been proposed to serve this function. It is also believed that this methyltranseferase can recognize both D-aspartyl and L-isoaspartyl residues. In addition, it is thought that this enzyme may have the ability to reverse at least part of the damage to protein molecules.

Although the human isoaspartyl protein repair methyltranseferase has been purified from red blood cells and had its protein sequence determined, in addition to harvesting a variant in a bacterial system, the availability and use of methyltransferases has been limited.

On or around Nov. 14, 1995, however, it was reported that scientists germinated a 1,288 year old Sacred Lotus seed. The research reported in the November issue of the American Journal of Botany, began in 1982, when Jane Shen-Miller, a plant physiologist at the University of California at Los Angeles (UCLA), obtained seven brown, oval-shaped Sacred Lotus seeds from the Beijing Institute of Botany.

In 1983, Jane Shen-Miller filed through the hard shells of four of the ancient Sacred Lotus seeds and watched three of them sprout. She then dried and burned the seedlings so she could use radiocarbon dating to establish the ages, the oldest of which was 1,288 years old.

According to the November, 1995 report, one of those ancient Sacred Lotus seeds had been in the ground for over 1,200 years; it therefore has been postulated that the Sacred Lotus seeds act as embryos that must be kept going until such seeds are germinated. Up until this point, geneticists knew only about proteins that repaired damaged DNA. But findings have suggested that the L-isoaspartyl methyltransferase (MT) enzyme, found in the Sacred Lotus seeds and nearly all other organisms, may have the ability to repair other proteins--those that make up cells and tissues, thus slowing tissue decay.

In these ancient Sacred Lotus seeds, the MT enzyme was present at levels comparable to modern day Sacred Lotus seeds. Damaged proteins did not accumulate within the ancient Sacred Lotus seeds, suggesting that the MT enzyme, possibly along with other constituents, kept the ancient Sacred Lotus seeds alive for so many years.

Notwithstanding the above, it is unknown as to whether use of methyltransferase or extracts or components of the Sacred Lotus plant in topical or oral compositions would be effective in combatting aging, repairing damaged skin and/or restoring skin to a more youthful appearance. Moreover, there are no known acceptable products available which incorporate methyltransferase or extracts or components of the Sacred Lotus for combatting dermatological aging, repairing damaged skin and/or restoring skin to a more youthful appearance.

Consequently, there exists a need for acceptable delivery systems which incorporate methyltransferase or extracts or components of the Sacred Lotus plant for effectively treating and preventing aging, repairing damaged skin and restoring skin to a more youthful appearance.

SUMMARY OF THE INVENTION

In brief, the present invention alleviates and overcomes certain of the above-mentioned problems and shortcomings of the present state of the art through the discovery of novel acceptable delivery systems which embody methyltransferase or extracts or components of the Sacred Lotus plant for effectively treating and preventing aging, repairing damaged skin and restoring skin to a more youthful appearance and methods of using same.

Accordingly it is the object of the invention to provide a general method for prevention or alleviation of damage to the skin associated with aging through the topical and/or internal use of methyltransferase in combination with a suitable carrier or vehicle.

Another object is to restore the skin to a more youthful appearance.

Another object is to use the extract of the Sacred Lotus (Nelumbo nucifera) seed as a natural source of methyltransferase. It has now been observed, surprisingly and unexpectedly, that by using an extract of the Sacred Lotus seed by topical application as an active agent, signs of aging can be reduced, eliminated or even reversed.

Also until the present invention there were no acceptable vehicles utilizing methyltransferase from a natural source such as lotus seed and its other natural components in an elegant cosmetic and/or basic pharmaceutical composition and/or dietary formulation. Thus, another object of the present invention are formulations of Sacred Lotus seed in dermatologicals, such as lotions, creams, sunscreens, cleansers, and various skin care formulas to improve the youthful appearance of the skin, repair visible damage, and reduce the signs of aging including loss of elasticity, fine lines, wrinkles, blemishes, spots and discolorations.

Another object is the formulation of dietary supplements providing the anti-aging benefits of methyltransferase and antioxidants found in the Sacred Lotus seeds.

A suitable carrier or vehicle will include the formulation of creams, gels, lotions, powders, tablets, capsules, and liquid preparations for the skin to repair the damage from aging, reduce further damage and restore skin to a more youthful appearance.

These and other objects are achieved by the present invention which is directed to a topical or oral formulation for the protection of the skin against damage caused by aging and a method for preventing or alleviating such damage and restoring it to a more youthful appearance by employing such in a topical or oral formulation.

The formulation is a suitable cosmetic or dermatologically or orally acceptable non-toxic, non-allergenic carrier containing methyltransferase and other components of the seed or of the sacred lotus plant.

The above features and advantages of the present invention will be better understood with reference to the accompanying FIGS., detailed description and examples. It should also be understood that the particular methods and formulations illustrating the present invention are exemplary only and not to be regarded as limitations of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS

Reference is now made to the accompanying FIGS. in which is shown illustrative results of exemplary embodiments of the present invention from which its novel and unique features and advantages will be apparent.

FIG. 1 depicts before and after results on a neck and hands following administration of the Anti-Aging Moisture Concentrate, as set forth in Example 1.

FIG. 2 likewise depicts before and after results on a neck following administration of the Anti-Aging Moisture Concentrate, as set forth in Example 1.

FIG. 3 depicts before and after results on faces following administration of one of the topical compositions set forth in Examples 2-6.

FIG. 4 depicts before and after results on faces following administration of one of the topical compositions set forth in Examples 2-6.

FIG. 5 depicts before and after results on facial skin following administration of one of the topical compositions set forth in Examples 2-6.

FIG. 6 depicts before and after results on facial skin around the mouth following administration of one of the topical compositions set forth in Examples 2-6.

DETAILED DESCRIPTION

By way of illustrating and providing a more complete appreciation of the present invention and many of the attendant advantages thereof, the following detailed description and examples are given concerning the novel delivery systems which embody methyltransferase or extracts or components of the Sacred Lotus plant for effectively treating and preventing aging, repairing damaged skin and restoring skin to a more youthful appearance and methods of using same.

The present invention uses Sacred Lotus Seed extract in skin and dietary formulas to combat aging. It is believed that the use of Sacred Lotus seeds in cosmetics and dietary supplements accomplish anti-aging effects based on anti-aging factors (enzymes such as methyltransferase) which are present in the seeds.

Moreover, Sacred Lotus seeds contain certain anti-oxidants such as vitamin C and glutathione which may contribute to anti-aging effects on skin along with methyltransferase and may even contain other beneficial factors.

In accordance with the present invention, methyltransferase and/or the natural compounds found in an extract of Sacred Lotus seeds may be used in an effort to repair age related signs of the skin such as lines, spots, wrinkles, and/or loss of elasticity. Other sources of methyltransferase that may be used in the invention are components of Sacred Lotus plants, extracts of Yellow Lotus seeds or components of Yellow Lotus plants, wheat germ oil, liver, brain, testes and bio-fermentation or chemical synthesis for example.

An extract is prepared, for example, as follows. Maceration is the preferred process since no heat is used which may destroy or alter temperature sensitive components; however percolation, digestion, infusion and decoction are within the scope of the invention as more scientific information becomes available.

A twenty percent extract of NELUMBO NUCIFERA is generally used and prepared by placing 200 grams of finely milled untreated whole seeds, including husks and pithes, in a stoppered container with about 750 ml of a 50/50 wt/wt mixture of purified water and propylene glycol U.S.P. and allowed to stand for a period of at least 3 days in a warm place with frequent agitation, until soluble matter is dissolved. The mixture is filtered and, after most of the liquid has drained, the residue on the filter is washed with sufficient quantity of the solvent mixture; the filtrates are combined to produce 1000 ml.

It is also within the scope of the present invention to use different amounts of the seed, other parts of the plant as well as other species solvents and mixtures.

While it is believed that Sacred Lotus "Nelumbo nucifera" is a preferred species and the seed is a preferred part, other parts of the plant or other species, such as the Yellow Lotus "Nelumbo lutea," containing these "anti-aging" enzymes or constituents are believed to be suitable alternatives for accomplishing the objectives of the present invention.

A composition according to the present invention for topical application contains an effective amount of the anti-aging agent Sacred Lotus seed extract from 0.05% to 100% by weight of the extract relative to the total weight of the composition, and preferably from 0.10% to 10% concentration by weight. It should be understood that the topical compositions may be used at any appropriate daily intervals, depending of course upon the particular type of composition formulated. For instance, the night serum, as set forth in Example 3 hereinafter, is to be administered once-a-day at night after proper cleansing, whereas the wrinkle serum of Example 2 may be administered twice-a-day in the morning and evening. The spot-lightening formulation of Example 4, on the other hand, may be applied, after cleansing, initially on a daily basis for about 6 to 8 weeks, at which time fading will gradually occur. Administration then may be reduced to only 1 to 2 times per week, after cleansing, to prevent reappearance of spots. The wrinkle alpha hydroxy formulation set forth in Example 6 may be applied once or twice daily following cleansing; however, if dryness occurs, use of this product may be reduced to 1 to 3 times per week.

A composition according to the present invention for oral administration contains an effective amount of the anti-aging agent Sacred Lotus seed (with husk, shell and plumele) from 0.01% to 100% by weight of the ground seed or 0.05% to 100% by weight of the extract relative to the total weight of the composition, and preferably 0.03% to 0.05% of the Sacred Lotus seed concentration by weight (300 mcg to 500 mcg per 840 mg tablet or capsule). More particularly, and by way of example, an oral composition containing ground Sacred Lotus seed can be prepared as follows. The whole Sacred Lotus seed with husk, shell and plumele (pith or embryo), supplied by W.T.H., 130 Wing Lok St., 8/F, Hong Kong, is first ground through a coffee mill having a fine screen (#24 Mesh or 700 microns particle size). The ground seeds are than treated with ionizing radiation with an absorbed dose from about 7.19 kGy minimum to 7.69 kGy maximum for sterilization purposes. The irradiated ground seeds may then be powder blended with other selected components to formulate oral delivery systems in accordance with the present invention, as discussed in greater detail hereinafter.

A composition according to the present invention may be in any of the cosmetic or pharmaceutical forms which are generally used for topical application such as liquids (both aqueous and non-aqueous solutions), creams (both oil-in-water and water-in-oil, O/W & W/O emulsions), gels (both aqueous and non-aqueous), lotions, serums, ointments, paste, powders, liposomes, laminates, microspheres, tablets and powders.

An oral composition according to the present invention may be in any of the dosage forms which are generally used for dietary supplements such as liquids, gels, powders, tablets, caplets, capsules and soft gels. While it is generally preferable to formulate the oral dosages to each contain, for example, about 350 mcg of Sacred Lotus seed for administration twice daily, it should be understood that any effective dosage strength may be formulated for administration at any effective daily interval, so long as the objectives of the instant invention are not defeated.

Compositions of the present invention may also contain additives, such as water, alcohols, oils (mineral, vegetable, animal and synthetics), glycols, colorants, preservatives, emulsifiers, gelling agents, gums, esters, hormones, anti-oxidants, silicones, polymers, fragrances, flavors, sunscreens, other active ingredients, antioxidants, acids, bases, buffers, vitamins, minerals, salts, polyols, proteins and their derivatives, essential oils, other enzymes, co-enzymes and extracts, surfactants, detergents, soaps, anionics, non-ionics, ionics, waxes, lipids, UV filters, stabilizers, fillers, celluloses, glycans, amines, solubilizers, thickeners, sugars and sugar derivatives, ceramides, sweeteners and the like, so long as such additives do not defeat the objectives of the present invention.

A composition according to the present invention as an effective anti-aging agent may be employed alone, that is without the use of additional actives, or the Sacred Lotus seed may be used to enhance other ingredients. Moreover, it should be understood by those versed in this art that the present invention contemplates treating the skin with either the topical compositions or the oral compositions of the instant invention or concommitantly with both types of compositions. It is believed that when both the oral and topical compostions of the present invention are administered, such combination therapy will promote both good health and radiant skin, especially when the oral compostions include micronutrients, such as in the oral composition set forth in Example 7 hereinafter.

One preferred composition combines the Sacred Lotus seed with glycocyamine, hyaluronic acid, and alpha hydroxy acids for their role in glycosaminoglycan (a protein sugar complex essential for normal hydration in the skin) synthesis plus antioxidants, such as vitamins A, C and E, extracts of pine bark, grape seed and green tea, catalase, superoxide dismutase and coenzyme Q-10 to protect the skin from free radicals and alleviate further cellular damage.

Other items such as lecithin, squalene, panthenol, vitamin D3, jojoba oil, olive oil, glycerin and other moisturizers also appear to benefit with the addition of Sacred Lotus seed in providing moisture into the skin enhancing its repair process, promoting elasticity and making the skin appear younger.

Hydroquinone, sulfur and salicylic acid also appear to exhibit synergism with Sacred Lotus Seed extract.

The following examples are given for illustrative purposes only to delineate some of the features of the invention and are not intended to be limiting. As to exemplary formulations set forth below, the quantities are given in percent by approximate weight (% wt) or approximate units (IU) unless otherwise noted based on the total weight of the composition. The term qs means to use a sufficient quantity by weight to bring the entire composition to 100%. Whenever possible International Nomenclature Cosmetic Ingredient (INCI) names are used.

EXAMPLE 1

______________________________________ Anti-Aging Moisture Concentrate ______________________________________ Part I: Water 78.10 Diazolidinyl Urea 0.30 Methylparaben 0.20 Part II: Hydroxyethylcellulose 0.30 Part III: Stearamidopropyl Dimethylamine Lactate 1.50 Glycerin 2.00 Cetyl Alcohol 4.00 Glycol Stearate 3.50 Propylparaben 0.10 Part IV: Sacred Lotus seed extract 10.00 ______________________________________

Procedure:

Part I is mixed and heated to 80 C. Part II is slowly added with mixing; then covered and continued mixing for 30 minutes. Part III is mixed and heated also to 80 C. Part III is then added to Part I with mixing. With continued mixing the heat is removed and cooling started. At 40 C Part IV is added. At 35 C mixing is stopped.

A thick luscious fat and oil free lotion was obtained to moisturize aged and dried skin.

EXAMPLE 2

______________________________________ Anti-Aging Wrinkle Serum ______________________________________ Part I: Water 75.36 Glycerin 1.50 Tetrasodium EDTA 0.10 Panthenol 0.30 Diazolidinyl Urea 0.30 Methylparaben 0.25 Sodium PCA 0.50 Part II: Magnesium Aluminum Silicate 0.90 Part: III: Glyceryl Dilaurate 0.50 Sunflower Oil 10.00 Olive Oil 2.50 Cetyl Alcohol 2.70 Glyceryl Stearate PEG-100 3.00 Propylparaben 0.10 Part IV: Tocopheryl Acetate 0.50 Retinyl Palmitate 0.05 Coenzyme Q-10 0.01 Part V: Corn Oil, Glyceryl Oleate, Propylene Glycol, 0.05 BHA, BHT, Propyl Gallate, and Citric Acid Magnesium Ascorbyl Phosphate 0.05 Catalase 0.25 Superoxide Dismutase 0.25 Beta Carotene 0.01 Green Tea Extract 0.10 Grape Seed Extract 0.10 Pine Bark Extract 0.10 Sacred Lotus seed extract 0.20 25% Sodium Hydroxide qs to pH 7.00 0.02 Fragrance 0.30 ______________________________________

Procedure:

Part I is mixed and heated to 80 C. Part II is slowly added with mixing; then covered and continued mixing for 30 minutes. Part III is mixed and heated also to 80 C. Part III is then added to Part I with mixing. With continued mixing the heat is removed and cooling started. At 50 C Part IV is added. At 40 C Part V is added followed by fragrance. At 35 C mixing is stopped. Ph is measured and adjusted if necessary.

An enriched serum that readily dispenses from a pump to alleviate wrinkles.

EXAMPLE 3

______________________________________ Anti-Aging Night Serum For Dry Skin ______________________________________ Part I: Water 51.62 Glycerin 2.00 Tetrasodium EDTA 0.05 Panthenol 1.50 Diazolidinyl Urea 0.20 Methylparaben 0.20 Part II: Hydroxyethylcellulose 0.10 Part III: Propylene Glycol 3.00 Cetearyl Phosphate 1.00 Cetearyl Alcohol 1.00 DEA Cetyl Phosphate 2.20 Cetyl Alcohol 2.00 Glycol Stearate 1.00 Soybean Oil 3.00 Propylparaben 0.10 Squalene 3.00 Polysorbate 20 0.40 Jojoba Oil 2.00 Cyclomethicone 2.00 Phenyl Dimethicone 3.00 Titanium Dioxide 1.00 Part IV: Tocopheryl Acetate 1.50 Coenzyme Q-10 0.01 Retinyl Palmitate 0.20 Part V: Oat Beta Glucan 1.00 Magnesium Ascorbyl Phosphate 0.30 Catalase 1.00 Superoxide Dismutase 2.00 Beta Carotene 0.05 Green Tea Extract 1.00 Grape Seed Extract 1.00 Pine Bark Extract 1.00 Sacred Lotus seed extract 4.00 N-Acetyl Glycocyamine 2.00 Saccharide Isomerate 2.00 Sodium Hyaluronate 2.00 Histidine HCL 0.50 25% Sodium Hydroxide qs to pH 7.00 0.02 Fragrance 0.25 ______________________________________

Procedure:

Part I is mixed and heated to 80 C. Part II is slowly added with mixing; then covered and continued mixing for 30 minutes. Part III is mixed and heated also to 80 C. Part III is then added to Part I with mixing. With continued mixing the heat is removed and cooling started. At 50 C Part IV is added. At 40 C Part V is added followed by fragrance. At 35 C mixing is stopped. Ph is measured and adjusted if necessary.

A serum that is used at night to restore moisture to dry skin.

EXAMPLE 4

______________________________________ Anti-Aging Spot Lightener ______________________________________ Part I: Water 91.79 Hydroquinone 2.00 Ascorbic Acid 1.00 Sodium Metabisulfite 0.15 Diazolidinyl Urea 0.30 Methylparaben 0.20 N-Acetyl Glycocyamine 1.00 Citric Acid 0.10 Part II: Polysorbate 20 2.00 Fragrance 0.20 25% Sodium Hydroxide 0.26 Sacred Lotus seed extract 1.00 ______________________________________

Procedure:

Part I is mixed until completely clear. Adjust pH of Part I to 5.50 +/-0.10 with 25% Sodium Hydroxide solution. Premix Part II and add slowly to Part I with constant mixing. When clear the Sacred Lotus Seed Extract is added and stirred until completely clear. A thin solution that is easily used from a roll-on applicator to target age spots, freckles, uneven skin tone and damage caused by the sun.

EXAMPLE 5

______________________________________ Anti-Aging Acne Blemish Treatment ______________________________________ Part I: SD Alcohol 40-B 67.85 Salicylic Acid 2.00 Sacred Lotus seed extract 1.00 Witch Hazel Distillate 10.00 Glycerin 10.00 Part II: Retinyl Palmitate 0.05 Fragrance 0.10 Polysorbate 80 10.00 ______________________________________

Procedure:

Mix Part I until completely clear keeping covered to avoid any alcohol loss. Mix Part II until uniform. Add Part II to Part I and mix until clear.

A simple topical that is effective especially on oily skin without excessive irritation.

EXAMPLE 6

______________________________________ Anti-Aging Alpha Hydroxy Acid Wrinkle Treatment ______________________________________ Part I: Water 76.61 N-Acetyl Glycocyamine 1.00 Aloe (10X conc.) 0.20 Magnesium Acorbyl Phosphate 0.50 Diazolidinyl Urea 0.20 Methylparaben 0.10 Part II: Glycolic Acid 70% 7.14 Blackcurrent Extract, Bilberry Extract, 1.00 Citric Acid, Glycolic Acid, Lactic Acid and Malic Acid Sugar Cane Extract, Citric Acid, Glycolic 1.00 Acid, Lactic Acid and Malic Acid Part III: Polysorbate 20 1.50 Retinyl Palmitate 0.05 Fragrance 0.20 Tocopheryl Acetate 0.50 25% Sodium Hydroxide 8.00 Sacred Lotus seed extract 2.00 ______________________________________

Procedure:

Part I is mixed until completely clear. Part II is also mixed until completely clear. Part II is added to Part I and mixed until clear. Adjust pH of Parts I & II to 5.00 with 25% Sodium Hydroxide solution. Premixed Part III is added and mixed until clear followed by Sacred Lotus Seed Extract addition.

A liquid combination of various alpha hydroxy acids with Sacred Lotus Seed Extract shown to be effective in reducing the appearance of lines and wrinkles. Also minimized pore size and improved the texture of the skin imparting a more youthful and radiant appearance.

EXAMPLE 7

______________________________________ Anti-Aging Dietary Supplement ______________________________________ Vitamin A Palmitate 1750 IU Natural Beta Carotene 750 IU Lycopene 0.375 mg Lutein 0.375 mg Vitamin E (d-alpha tocopheryl acetate and 50 IU mixed tocopherols) Cholecalciferol 200 IU Vitamin C 150 mg Thiamine HCL 4 mg Riboflavin 5 mg Niacinamide 20 mg Pyridoxine HCL 6 mg Folic Acid 200 mcg Vitamin B-12 5 mcg Pantothenic Acid 7.5 mg Biotin 75 mcg Calcium (Carbonate & Citrate) 12.5 mg Magnesium Oxide 18.75 mg Iron (Fumarate) 7.5 mg Zinc (Sulfate & Gluconate) 7.5 mg Manganese Gluconate 2 mg Selenium (L-Selenomethionine & Citrate) 35 mcg Chromium Nicotinate 37.5 mcg Copper Gluconate 0.5 mg Green Tea Extract 7.5 mg Grape Seed Extract 7.5 mg N-Acetyl Glycocyamine 15 mg Citrus Bioflavanoids 37.5 mg N-Acetyl Cysteine 5 mg Sacred Lotus seed 350 mcg Magnesium Stearate (Lubricant) 8.4 mg Stearic Acid (Binder) 42 mg Microcrystalline Cellulose (Tablet Aide) qs 840 mg ______________________________________

Procedure:

All of the above was mixed in a powder blender until completely homogenous. The mixed powder was fed into a tablet press and compressed into tablets.

A unique blend of protective antioxidants and essential nutrients with the entire Sacred Lotus Seed (in food preparations the pithe (embryo) or plumule is removed due to bitter taste) used to promote good health and radiant skin. This product maybe taken, for example twice per day. i.e., take one tablet with breakfast and one tablet with lunch or dinner.

EXAMPLE 8

During the course of a Dermatologist's clinical study it was discovered that a lotion containing Sacred Lotus Seed extract alone resulted in a decrease in the appearance of aging sun-damaged skin on the hands. The lotion used in this Example 8 was the Anti-Aging Moisture Concentrate set forth in Example 1 above. In two participants, the lotus seed extract alone was used in a 10% concentration with either a placebo or active formulated lotion on the hands.

Side by side comparison of the treated vs. untreated hand of one participant showed obvious differences. The skin of the treated hand was more porcelain in appearance, there was reduction in fine wrinkling, dryness and the hand appeared less aged and less sun-damaged. In another participant, one hand was treated, and before and after photographs showed similar results. See FIG. 1.

EXAMPLE 9

In another study using the Anti-Aging Moisture Concentrate set forth in Example 1 above, the neck of one patient showed dramatic results, according to side by side comparisons of treated vs. untreated skin of the neck. The patient then went on to treat the untreated side, which was the left side, and this also showed the dramatic improvement seen on the previously treated right side. See FIG. 2.

There was also unexplained improvement in the elasticity in a right to left comparison. See FIG. 2 which are photographs that were taken of a pinch test of the skin. On the treated right side of the neck, pinching of the skin revealed very little and difficulty in pinching up any loose skin, and the untreated left side of the neck, skin was easily pinched up to a much greater degree and the skin fold was larger.

EXAMPLE 10

In order to test the effectiveness of the Sacred Lotus Seed Extract when combined with other ingredients another study was done. Two different groups were given the various formulations containing Sacred Lotus seed extract, as set forth in Examples 2-6 set forth above, to use for twelve weeks. The first panel consisted of eight professional models. The second used fourteen women employed in other occupations. The third is a combination of the two. See Tables 1, 2 and 3, respectively. Tables 4, 5, 6 and 7 concern Leaf Width of Germinated Seeds, L-Isoaspartyl Methyl transferase, Steroconfigutation of Aspartyl Residues and Age Determinations, respectively. More particularly, Table 4 illustrates leaf growth of the 332 year old Sacred Lotus seed compared to a 10 year old plant. Table 5 reports the results of methyltransferase assay on a ca 416, 95 and 10 year old Sacred Lotus seeds and fresh Yellow Lotus seeds. Table 6 shows measures of aspartyl residues in L & D configurations of the seeds used in Table 5. Table 7 lists ages of the old Sacred Lotus seeds that can still germinate and reports the methods used.

The Dermatologist noted dramatic improvements in the patients using the formulas containing Sacred Lotus Seed Extract. The formulas increased elasticity, reduced visible pore size, promoted a more refined skin texture, reduced the appearance of crow's feet, lines and wrinkles, reduces blemishes, lightened dark spots promoted firmer, younger looking skin. See FIGS. 3, 4, 5 and 6.

The left column lists the various categories sought to be improved, for example brown spots, blemishes, smaller pores, etc. The next columns give the results of the panel members. The last column is the average percent improvement of the responses given in each category. The top row identifies the participant by number. The bottom row gives overall result of the improvement. Some items are blank because no improvement was needed in that particular category.

TABLE 1 __________________________________________________________________________ PANEL SURVEY MODELS IMPROVEMENT 1 2 3 4 5 6 7 8 AVERAGE % __________________________________________________________________________ BROWN SPOTS 80 90 100 90 80 100 70 87.10% LESS BLEMISHES 80 90 70 90 90 100 100 88.60% SMALLER PORES 100 90 90 80 80 90 100 100 91.30% MOUTH WRINKLES 80 90 100 90 80 100 80 88.60% CROWS FEET 80 100 100 80 80 80 100 90 88.60% LINES FOREHEAD 80 80 90 90 80 80 100 90 86.30% TEXTURE 90 90 100 100 100 100 100 97.10% MOISTURE 90 90 100 70 90 100 80 88.60% SMOOTHER SOFTER 90 100 90 100 100 100 100 100 97.50% LESS ROUGHNESS 80 100 90 100 100 80 100 100 93.80% RADIANCE 90 100 100 100 100 100 100 90 97.50% YOUNGER LOOKING 80 100 100 100 100 100 100 90 96.30% DIFFERENCES 100 90 90 100 100 90 100 100 96.30% __________________________________________________________________________

TABLE 2 __________________________________________________________________________ MDR PANEL SURVEY IMPROVEMENT 1 2 3 4 5 6 7 8 9 10 11 12 13 14 AVERAGE __________________________________________________________________________ % BROWN SPOTS 80 100 80 90 80 90 100 90 70 70 90 90 70 84.60% LESS BLEMISHES 80 100 70 70 90 90 100 80 90 70 70 80 100 83.80% SMALLER PORES 80 100 80 90 90 90 100 90 90 90 90 100 100 91.50% MOUTH WRINKLES 90 100 80 90 90 70 70 90 100 90 90 87.30% CROWS FEET 100 80 90 90 70 70 90 90 90 100 87.00% LINES FOREHEAD 80 100 80 90 90 90 80 70 90 90 90 90 86.70% TEXTURE 90 100 90 100 100 90 100 90 90 90 80 100 100 93.80% MOISTURE 100 100 90 90 90 90 90 100 90 100 80 100 100 93.80% SMOOTHER SOFTER 100 100 90 90 90 90 100 100 100 100 90 80 100 100 95.00% LESS ROUGHNESS 100 100 80 90 90 90 100 90 100 90 80 100 100 92.30% RADIANCE 90 100 80 100 100 90 100 100 90 100 100 90 100 100 95.70% YOUNGER LOOKING 80 100 90 90 90 90 100 90 100 100 90 100 100 93.80% OVERALL DIFFERENCES 90 100 90 100 100 90 100 100 90 90 100 90 100 100 95.70% __________________________________________________________________________

TABLE 3 __________________________________________________________________________ PANEL SURVEY TOTAL __________________________________________________________________________ IMPROVEMENT 1 2 3 4 5 6 7 8 9 10 11 12 __________________________________________________________________________ BROWN SPOTS 80 100 80 90 80 90 100 90 70 70 90 90 LESS BLEMISHES 80 100 70 70 90 90 100 80 90 70 70 80 SMALLER PORES 80 100 80 90 90 90 100 90 90 90 90 100 MOUTH WRINKLES 90 100 80 90 90 70 70 90 100 90 CROWS FEET 100 80 90 90 70 70 90 90 90 LINES FOREHEAD 80 100 80 90 90 90 80 70 90 90 90 TEXTURE 90 100 90 100 100 90 100 90 90 90 80 MOISTURE 100 100 90 90 90 90 90 100 90 100 80 SMOOTHER SOFTER 100 100 90 90 90 90 100 100 100 100 90 80 LESS ROUGHNESS 100 100 80 90 90 90 100 90 100 90 80 RADIANCE 90 100 80 100 100 90 100 100 90 100 100 90 YOUNGER LOOKING 80 100 90 90 90 90 100 90 100 100 90 OVERALL DIFFERENCES 90 100 90 100 100 90 100 100 90 90 100 90 __________________________________________________________________________ IMPROVEMENT 13 14 15 16 17 18 19 20 21 22 AVERAGE % __________________________________________________________________________ BROWN SPOTS 70 80 90 100 90 80 100 70 85.50% LESS BLEMISHES 100 80 90 70 90 90 100 100 85.50% SMALLER PORES 100 100 90 90 80 80 90 100 100 91.40% MOUTH WRINKLES 90 80 90 100 90 80 100 80 87.80% CROWS FEET 100 80 100 100 80 80 80 100 90 87.80% LINES FOREHEAD 90 80 80 90 90 80 80 100 90 86.50% TEXTURE 100 100 90 90 100 100 100 100 100 95.00% MOISTURE 100 100 90 90 100 70 90 100 80 92.00% SMOOTHER SOFTER 100 100 90 100 90 100 100 100 100 100 95.90% LESS ROUGHNESS 100 100 80 100 90 100 100 80 100 100 92.90% RADIANCE 100 100 90 100 100 100 100 100 100 90 96.40% YOUNGER LOOKING 100 100 80 100 100 100 100 100 100 90 94.80% OVERALL DIFFERENCES 100 100 100 90 90 100 100 90 100 100 95.90% __________________________________________________________________________

TABLE 4 __________________________________________________________________________ Leaf Width* of Sacred Lotus, SL (Nelumbo nucifera). 5th 6th 7th 8th 9th 10th 11th 3rd 4th N1 N2 N1a Pb N3 N4 N5 FRUIT DAY (mm) __________________________________________________________________________ .about.332 Yr-Old (oSL) oSL5 16 E 20 F 23 14.0 E 36 44.0 53.2 E 43 44.0 57.0 67.0 E 50 44.0 57.5 87.0 64.0 55 43.0 58.0 86.0 99.0 -- -- E 10 Yr-Old (dSL) dSL8 50 83.0 95.8 117 E E E 56 85.0 96.0 120 130 F F E 68 85.0 95.0 120 162 96.0 47.5 165 E E 73 86.0 95.0 121 163 100 48.0 183 F F 81 85.0 95.0 122 164 103 46.0 186 99.0 126 dSL 10** 50 61.0 65.0 85 F 56 61.0 64.0 82 130 -- -- E 68 63.0 66.0 131 127 81.5 -- F E 73 54.0 56.0 132 143 84.0 -- F F 81 dead 56.0 131 148 84.0 -- F F 78.0 __________________________________________________________________________ *Largest width of leaf ("tipto-tip" measurement); N node; N1a 2nd leaf of node 1; Pb leaf at plumule base; E emerging leaf; F folded leaf. **Cotyledon tips removed (48 h after imbibition) for methyltransferase assay (see Table 5).

TABLE 5 __________________________________________________________________________ L-isoaspartyl Methyltransferase in Cotyledons and Embryonic Axes of Sacred Lotus (SL, Nelumbo nucifera) and Yellow Lotus (YL, N. lutea). METHYLTRANSFERASE (pmol/min/mg protein) COTYLEDONS FRUIT IMBIBITION (hr) TIP HALF BASE HALF EMBRYONIC AXES __________________________________________________________________________ OLD SL (oSL) 6 (.about.416 yr) 22 (d) 0.19 0.28 0.44 7 (.about.95 yr) no 0.12 0.39 0.73 AVE + .sigma. 0.16 + 0.04 0.34 + 0.06 0.59 + 0.15 10 Yr-OLD SL (dSL)* 10 5 0.16 NA NA 11 48 0.02 NA NA I 12 5 0.26 NA NA 13 48 0.05 NA NA 14** no 0.59 NA 1.43 15 no 0.40 NA 0.72 16 no 0.16 0.43 0.62 17 no 0.14 0.37 0.76 II 18 5 0.10 0.30 0.59 19 5 0.20 0.26 0.74 20 48 0.15 0.31 0.70 21 48 0.12 0.21 0.67 AVE (II) + .sigma. 0.14 + 0.01 0.31 + 0.03 0.68 + 0.03 FRESH YL (fYL) 5 no 0.31 0.21 0.48 6 no 0.13 0.21 0.52 7 5 0.28 0.17 0.40 8 5 0.16 0.24 0.57 9 48 0.27 0.26 0.55 10 48 0.20 0.17 0.72 AVE + .sigma. 0.22 + 0.03 0.21 + 0.01 0.54 + 0.04 __________________________________________________________________________ *Two experiments (I, II). **Unhealthy cotyledons and plumules, and probably damaged pericarp. NA: not analyzed.

TABLE 6 __________________________________________________________________________ Stereoconfiguration of Aspartyl Residues in Hydrolysates of Proteins of Seeds of Sacred Lotus (SL, Nelumbo nucifera) and Yellow Lotus (YL, N. lutea). D-ASP L-ASP COTYLEDON* EMBRYONIC COTYLEDON* EMBRYONIC FRUIT (Mol %) __________________________________________________________________________ OLD SL (oSL) oSL6 1.67 1.26 98.3 98.7 oSL7 1.76 1.50 98.2 98.5 AVE + .sigma. 1.72 + 0.04 1.38 + 0.12 98.2 + 0.05 98.6 + 0.1 10 Yr-OLD SL (dSL) dSL14 2.13 2.04 97.9 98.0 dSL16 1.66 1.52 98.3 98.5 AVE + .sigma. 1.90 + 0.23 1.78 + 0.26 98.1 + 0.2 98.2 + 0.2 FRESH YL (fYL) fYL5 1.69 1.52 98.3 98.5 fYL9 2.43 2.24 97.6 97.8 AVE + .sigma. 2.06 + 0.37 1.88 + 0.36 98.0 + 0.4 98.2 + 0.4 __________________________________________________________________________ *Cotyledon tissues from the base half

TABLE 7 __________________________________________________________________________ Age Determinations* Reported by Various Laboratories of Sacred Lotus (Nelumbo nucifera) from the Dry Lake Bed at Pulantien on Liaotung Peninsula, Northeastern China. RADIOCARBON CALENDAR MEAN TISSUE USED IN AGE RANGE PHYSICAL RADIOCARBON DATING (yr BP) A.D. AGE (yr) (DRY OR GERMINATION TESTED) REFERENCE __________________________________________________________________________ Ohga Collection.sup..diamond-solid. (1923) 100 .+-. 60** 1682-1807 256 .+-. 62 Charred residue Godwin and Willis, 1964 1040 .+-. 210** 770-1218 1,000 .+-. 224 Fruit (dry) Libby, 1955 Chinese Collection (1952) 40 .+-. 60 1887-1911 95 .+-. 12 Pericarp (dry fruit) Shen-Miller et al., 1994, this paper 270 .+-. 60 1527-1797 332 .+-. 135 Pericarp (germinated seed) Shen-Miller et al., 1994, this paper 300 .+-. 60 1495-1660 416 .+-. 82 Pericarp (ungerminated seed) Shen-Miller et al., 1994, this paper 340 .+-. 80 1447-1653 444 .+-. 103 Fruit (ungerminated) Priestly and Posthumus, 1982 430 .+-. 100 1414-1631 471 .+-. 108 Fruit (germinated) Priestly and Posthumus, 1982 705 .+-. 145 1212-1409 684 .+-. 99 Fruit (germinated) Shen-Miller et al., 1994, this paper 740 .+-. 185 1069-1409 755 .+-. 170 Fruit (ungerminated) Shen-Miller et al., 1994, this paper 915 .+-. 80 1023-1217 874 .+-. 97 Fruit (dry) Chang, 1978 1350 .+-. 220 434-977 1,288 .+-. 271 Fruit (germinated) Shen-Miller et al., 1994, this __________________________________________________________________________ paper *Radiocarbon age .+-. .sigma.(yr BP) based on the determined .sup.14 C content of the lotus fruit or pericarp and the .sup.14 C halflife of 5568 .+-. 30 yr (Libby, 1955), and following the conventions of Stuvier and Polach (1977); yr BP: years before present (before 1950); calendar range obtained from the A.D. range extrapolated from the .sup.14 C decadal calibration curve of Stuvier and Becker (1993), discarding thos ranges that fall after the lotus collection dates: mean physical age base on the mean age deduced from the calendar A.D. range. **Godwin and Willis redated the charred residue of Libby's fruit, originally dated as .about.1,040 yr BP. .sup..diamond-solid. Ohga, based on available geologic and historical records, estimated his Pulantien collection of the lotus fruits as .about.400 yr in age (Wester, 1973).?

The present invention may, of course, be carried out in other specific ways than those herein set forth without departing from the spirit and essential characteristics of the present invention. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive and all changes coming within the meaning and equivalency range of the appended claims are intended to be embraced herein.