Instead of sensory evaluation, we designed an evaluation method of the setting function of hairstyling products, based on an original theory focusing on changes in bending stress observed when a load with continuous bending is applied to human hair. Specifically, we developed a device to measure bending stress to quickly and objectively evaluate the condition of human hair, particularly its dynamic properties such as the setting function, following the application of hairstyling products. This device generates a load with continuous bending while applying a pendulum motion to a hair tress, one end of which is anchored. The setting function and holding power of resins of various molecular weight and ionic properties were evaluated using this device. The results demonstrated a close correlation with those obtained by experts' sensory evaluation. The evaluation results of bending stress and holding rate confirmed that the combined use of two different resins could improve the function of setting preparations. Evaluation using this device was able to substitute for sensory evaluation, and offers quick objective evaluation and detection of changes in the holding power of hairstyling products over time. We conclude that evaluation using this device is a promising new method.

Fructose-1, 6-diphosphate (FOP), a glycolytic metabolite is reported to ameliorate inflammation and inhibit the nitric oxide production in murine macrophages stimulated with endotoxin. It is also reported that FOP has cytoprotective effects against hypoxia or ischemia/reperfusion injury in brain and heart. In this study, we examined whether FDP has protective effects on UV-induced oxidative damage in skin cell culture system and human skin in vivo. FDP had a protective role in UVB-induced LDH release and ROS accumulation in HaCaT although it did not show direct radical scavenging effect in the experiment using 1, 1-diphenyl-2-picrylhydrazyl (DPPH). FDP also preserved cellular GSH content after UV irradiation in HaCaT and normal human fibroblast culture system. Cellular oxidative stress induces multiple downstream signaling pathways that regulate expression of multiple gene including MMP-1 and collagen, we examined the effects of FDP on UV-induced alteration of these protein expression in fibroblast culture and human skin in vivo. The increased MMP-1 expression in fibroblast and human skin by UV irradiation was significantly decreased by FDP. FDP also prevented the UV-induced decrease of collagen expression in fibroblast and human skin. Moreover, the decreasing the intracellular levels of reducing equivalents in human fibroblast by glutathione (GSH) depletion lowered the UVA dose threshold for reduction of procollagen expression, indicating that the differences of glutathione contents define the susceptibility of fibroblasts towards UV-induced reduction of procollagen expression. FDP also preserved cellular GSH content after UV irradiation, indicating that FDP has protective effects on UV-induced reduction of procollagen expression, which are possibly through maintaining intracellular reducing equivalent. Based on these premises, we examined the effect of daily use of a moisturizer containing FDP on facial wrinkle in comparison with vehicle moisturizer lacking FDP. In the clinical study, FDP significantly decreased facial wrinkle compared with vehicle alone after 6 months of use. Our results suggest that FDP has anti-aging effects in skin by increasing cellular antioxidant system and preventing oxidative signal and inflammatory reaction. Therefore FDP may be useful anti-aging agent for cosmetic purpose.

In this study, in an attempt to search for functional cosmetic ingredients from higher fungal, we have produced exopolysaccharides (GF-l, approximately carbohydrate 75％, protein 25％) and polysaccharide (GF-2) of mycelium extract, by submerged culture of Grifolafrondosa. For applications in anti-aging cosmetic field, we investigated the diverse biological activities. Antioxidant activity and inhibition of Matrixmetalloproteinases (MMPs) were investigated enzymatic assays by measuring the superoxide scavenging activity using xanthine-xanthine oxidase system and the proteolytic activity of MMPs using EnzChek Collagenase/Gelatinase kits, respectively. GF-l polysacchairde showed inhibition of superoxide radical by 90％ at a concentration of 0.2％ (w/v) and inhibition of collagenase by 45％ at 0.2％ (w/v). GF-2 polysaccharide of mycelium extract also exhibited good antioxidant activity. However, MMPs inhibition activity was relatively lower level compared to GF-l polysaccharides. The treatment of human dermal fibroblast (HDF) with GF-l and GF-2 polysaccharides increased the proliferation of fibroblast by approximately 23-25％ at a concentration of 0.5％ (w/v), also showed collagen synthesis increase in HDF by about 50％ at 0.5％ (w/v) compared to that of untreated control. We also report the moisturizing effects of polysaccharides in cosmetic products (O/W emulation) and its own ingredient, in vitro and in vivo. The GF-1 polysaccharide showed higher moisturizing ability than sodium hyaluronate, which is the most commonly used moisturizers ingredient. These results suggest the GF-l polysaccharide, protein-bound polysaccharide, may be used as an ingredient for new moisturizing and anti-aging cosmeceuticals.

An innovative biopolymer known as the Rhizobian gum has been developed in France, which shows some dramatic refreshing effect on the skin. The origin of this innovative project takes its source in the natural environment, and in particular the natural environment of the roots of sunflowers and wheat, where a symbiotic bacterium has been discovered. It is a Rhizobium bacterium, which is hosted by the roots, and which is able to synthesize a specific polymer showing a dramatic water binding capacity. This polymer is in particular synthesized in period of drought, and its biological role is to concentrate the small amount water present in the soil in order to take it available for the root, which becomes then able to absorb it. This vital mechanism allows the plant to survive despite a severe climatic environment. This basic research has been conducted in collaboration whit the French National centre of scientific Research (CNRS), and has lead to the isolation of the Rhizobium bacteria. Rhizobian gum is a branched biopolymer consisting in the repetition of a polysaccharide unit of 3 molecules of glucose, 3 molecules of galactose and 1 molecule of glucuronic acid, whit one pyruvate group an average 1.6 acetyl groups. The fresh effect of Rhizobian gum is a strong sensorial impact that 100 ％ of the consumers are able to perceive, and which is judged very pleasant by most of them. In addition to this, a large majority of consumers are perceived, and which is judge very pleasant by most of them. In addition to this, a large majority of consumers also feel a very pleasant relaxing sensation. Smoothness and softness are also felt by most consumers and qualified positively by most of them. These qualities guarantee a strong impact on today's consumers.

In cosmetic area, retinol is prominent ingredient for anti-wrinkle but unstable against light, heat, oxygen and so on. Therefore the stabilization of retinol is required. Here, we capsulated doubly retinol in the SLM(Skin Lipid Matrix) that makes three dimensional lamellar structure similar to skin, after formation of primary liposome (retinol-nanoemulsion). First, we make primary liposome from retinol / hydrogenated lecithin / polysorbate20 / caprylic ＆ capric triglyceride / ethanol / and so on, and the mean diameter to 70 nm, using microfluidizer passed three times at 800 Bar, repeatedly. Then we produce DC-liposome (doubly capsulated-liposome) that was encapsulated primary liposome with SLM made of hydrogenated phosphatidyl choline / caprylic ＆ capric triglyceride / 1, 3-butylene glycol / ceramide3 / cholesterol /etc. We measured for color stability against light and heat with chromameter. As a result of this experiment, we observed DC-liposome was more than from 1.5 to 3 times as stable as general liposome. Livability of retinol has improved from 2 to 6 times when we analyzed it by HPLC. Also, penetration effect of DC-liposome has improved.

Skin aging process on pre-menopausal women is a problem that needs to be prevented as early as possible. The decrease of oestrogen level which is one of the intrinsic factors of the skin aging process will affect the skin biological process, due to oestrogen receptors on the skin. A number of researches conducted on pre-menopausal women with the allocation of oestrogen hormone resulted in delaying the skin aging process. The administration of soy isoflavon, a phytoestrogen found in daily food, on pre-menopausal women is hoped to be able to prevent skin aging process, even clinically or molecular biologically. This research aims to explain the benefit of administering of soy isoflavon on skin aging process. The design of the research is randomised controlled trial (RCT). As many as 60 pre-menopausal women were collected with simple random sampling method. Soy isoflavon is an independent variable, while skin aging process is a dependent variable assessed from the hydration, sebum level, average roughness, depth of wrinkles, skin clarity, length of the telomere. Analysis was conducted using t and MANDVA tests and.the result showed a significance (F = 10,439; p = 0,001) over the allocation of soy isoflavon to the whole variable dependent, including the telomere length and the skin hydration, meant that allocation of soy isoflavon could delay skin aging process.

Normal human skin, constantly challenged by environmental micro-organisms, has an innate ability to fight invading microbes through antimicrobial peptides. These peptides, described in both plant and animal kingdoms are able to inactivate a broad spectrum of micro-organisms. Mammalian defensins constitute one of the most common antimicrobial peptide family. Among the three human beta-defensins hBD1, hBD2 and hBD3 produced in epithelia, only hBD2 and hBD3 are inducible and additionally have been described as expressed by differentiated keratinocytes at site of inflammation and infection. The aims of these studies were to define a cell culture model in which the basal production of hBD could be detected and up-regulated in order to enhance skin auto-protection against micro-organisms. A specific Polymerase Chain Reaction method have been developed for hBD2 and hBD3 mRNA detection in non-differentiated monolayer keratinocytes cell culture. We have been able to demonstrate that in vitro, hBD2 and hBD3 expression in normal human keratinocytes could be detected and enhanced by TNF-alpha and IFN-gamma, in hypercalcic culture conditions. This research opened the possibility of the development of cosmetic active compounds, able to induce the expression of skin natural antibiotic peptides responsible about microflora ecology of the skin.

Chemists must take into consideration more factors to formulate with inorganic sunscreens than many other active ingredients including organic sunscreens. Because the UV radiation attenuation grade particles of Titanium Dioxide and Zinc Oxide are in the nanometer range, the state of their dispersion in the product film on the skin governs their efficacy and aesthetics.(omitted)

Lip chapping is a serious cosmetics problem, though remedies other than moisturizing have not been proposed. We investigated changes in the surface configurations of lip corneocytes and increased CD activity and improved chapping severity. Our results suggest that lip chapping can be activities of desquamation-regulating proteinases associated with lip chapping. Using scanning electron microscopy, villus-like projections were observed on the inner surfaces of most corneocytes from normal lips, whereas those with flatter surface were predominant in chapped lips. Further, cell surface area increased with the severity of lip chapping. Cathepsin D (CD)-like and chymotrypsin-like proteinase, which are also present in skin as desquamation-regulating proteinases, were detected in lip corneocytes, though only CD activity was found to decrease in severely chapped lips. Hydration was also lower in areas of lip chapping. Sequential topical application of apricot extract essence characterized as similar to senile xerosis rather than dry skin such as winter xerosis, as it shows a delayed transition of corneocytes through the stratum corneum, and the reduced CD activity may be one of the mechanisms that is further decreased by low hydration. We propose that an enhancement of both CD activity and lip moisture may be effective to improve lip chapping.

Most consumers have noted hair damage following coloring treatments. Proper evaluation of hair bleaching products must be performed using quantitative assessments of hair damage, though they are difficult, because of the slight fluctuations in hair composition. In the present study, we utilized a sensitive evaluation method for hair damage and found that the amount of soluble protein fraction extracted from hair under a reducing condition, termed labile protein, dramatically increased after bleaching.(omitted)

The cutaneous network of unmyelinated nerve fibers is extremely dense, and closely interacts with the many cell types present in dermis and epidermis, including keratinocytes, fibroblasts, Langerhans cells, and melanocytes. Cell communication involves various neuroendocrine factors, with cell differentiating and proliferative activities, or inflammatory properties. Thus, nervous cells in the skin not only create a sensory system connected to the central nervous system, but also mediate many of the biological activities of the skin.(omitted)

It has been known that the color of skin reflects the blood flow within. In lips, the capillaries close to the skin surface are numerous; hence lips are redder than the rest of face. However, dermatological research on lips is not as advanced as research on facial or body skin, and little was known about the relationship between relatively dull-colored lips and skin blood flow. The physiological differences between colorful and dull-colored lips were studied by a two-dimensional laser Doppler blood flow analyzer, a spectrometer for the measurement of the degree of oxygen saturation, and a confocal microscope for observing inside lips non-invasively. Dull-colored lips and the corner of lips (dull-colored compared to the center) showed relatively poor blood flow and lower oxygenated hemoglobin. It was found that colorful lips (generally the young) had a blood flow that tended to run straight in parallel with the skin surface. This unique blood-capillary structure can express clear red blood. Those with dull-colored lips had lost this unique structure. Their blood ran perpendicularly from the deep of the skin and down back again into the deep part as like the blood circulation patterns of facial skin. Therefore, the lips of the latter group had fewer blood capillaries near skin surface in the lips than that of the colorful-lips group. A lipstick containing a-glucosyl-hesperidin, which is derived from certain citrus fruits and can enhance blood circulation, was applied for evaluating its effects. Blood flow was increased 30 min after the application. After two weeks of daily application, the lips' condition became noticeably less dull. These findings suggest that the decrease of blood flow in dull-colored lips is caused by the loss of the unique capillary structure and the use of the lipstick to increase blood flow can give a vivid color to lips.

OMC is essentialiy necessary compound in sun goods as organic UV protecting products. But the skin-trouble problem is raising because of skin penetration of OMC. In this study, non-capsulated pure OMC was compared with Organic-Inorganic-Nano-hybrid OMC for skin penetration force and SPF degree. Organic- Inorganic Nano-Hybrid OMC is OMC trapped in the pore of the mesoporous silica synthesized by the sol-gel method after OMC is nanoemulsified in the system of the hydrogenated Lecithin/ Ethanol/caprylic/capric triglyceride/OMC/water. OMC- nano- emulsion was obtained by a microfluidizing process at 1000bar and then micelle size in the nanoemulsion solution is 100-200nm range. Mesoporous silica nano-hybrid OMC was prepared by the process; surfactant was added in dissolved OMC-Nanoemulsion, then the rod Micelle was formed. OMC-nanoemulsion was capsulated in this rod Micelle and then silica precursor was added in the OMC-nanoemulsion solution. Through the hydrolysis reaction of the silica precursor, mesoporous silica concluding OMC-Nanocapsulation was obtained. The nano-hybrid surface of this OMC-Nanoemulsion-Inorganic system was treated with polyalkyl-silane compound. OMC-Mesoporous silica Nano-hybrids coated with polyalkyl-silane compound show the higher sun protecting factor (SPF Analyzer: INDEX 10-15) than pure OMC and could reduce a skin penetration of OMC. The physico-chemical properties of these nano-hybrids measured on the SPF index, partical size, strcture, specific surface area, pore size, morphology, UV absorption, rate of the OMC dissolution using SPF Analyzer, Laser light scattering system, XRD, BET, SEM, chroma Meter, HPLC, Image analyzer, microfluidizer, UV/VIS. spectrometer.

The wrinkles correspond to the most obvious expression of skin ageing and are manifested by changes on the organization and dermal structure. In the extracellular matrix, decreased quantities of collagens and glycosaminoglycans as well as a deterioration of the fibrillary network is noted, result in a reduction of dermal thickness. In addition, the activity of the collagenases increases in contrast to the synthesis of collagen fibers. Nor are cells spared during the aging process. We thus studied and compared the contractile capacity as well as the synthesis capacity of normal human fibroblasts and human fibroblasts obtained from biopsies of forehead wrinkles. The capacity of the fibroblasts to be adhered to the collagen network and to maintain a three-dimensional structure of dermis was studied on a model of equivalent dermis. The metabolic activity was studied by evaluating the capacities of synthesis of collagen I, main component of dermis. Human fibroblasts resulting from the forehead wrinkle contract less the gel of collagen than the normal human fibroblasts and present an activity of biosynthesis of collagen I less important than normal human fibroblasts. These results show that fibroblasts with aging present a deceleration of their metabolic activity and lose their capacity of adhesion to collagen fibers thus limiting the possibility of organizing the dermal tissue. We investigated the potential of an active ingredient able to compensate for the reduction of the metabolic activity and to restore the contractile capacity of fibroblasts obtained from forehead wrinkles. This effect was compared with a reference molecule: the vitamin C.

In recent years, a miniature spectrometer has been extensively developed due to the combination of fiber optics and semiconductor detector arrays. This type of miniature spectrometer has advantages of low price and robustness because of the capability of mass production and no moving parts are required such as lenses, mirrors and scanning monochromator. In this study, for skin diagnostics, a portable near infrared (NIR) system has been developed using a LlGA microspectrometer, which is photo-diode arrays-type.(omitted)

To search and development a new material with superior melanogenesis inhibitory activity, the bioactivities (obs. pl$_{50}$) of alkyl-3,4-dihydroxybenzoyl esters and N-alkyl-3,4-dihydroxybenz-oyl amides as substrate molecules were measured in mouse melanoma cells. And also, we have studied that 3-D QSARs (3 dimensional Quantitative Structure-Activity Relationships) between molecular interaction field of substrates and the bioactivities were analyzed using CoMFA (Comparative Molecular Field Analyses) method. When cross-validation value (q$^2$) is 0.68 at 3 components, the Pearson correlation coefficient ($r^2$) is 0.900. From the basis on the findings, the model was appeared by the contour map such as steric field and electrostatic field relationships between quantitative structure and the bioactivity of the various substrate derivatives. Measured bioactivities (obs. pl$_{50}$) of unknown compounds are very similar to predicted activity (pred. pl$_{50}$) according to the CoMFA model. As the results of prediction, we could conclude that the bioactivities were increased by creation of R$_1$ substitution of 5,5-dime-thylhexoxy, 6,6-dimethylheptyl, 1-amino-6,6-dimethylheptyl group etc and R$_2$ substitution of hydroxy, methyl, methoxy group etc.p etc.

It has been shown that transparent microemulsion gels of flexibly adjustable viscosity containing less than 10％ w/w surfactants can be obtained just by cooling down through a new 1-step preparation procedure (via 4 different in-situ colloidal systems) using PIT-technology with a range of emulsifiers, co-emulsifiers and lipids. They can be used for a wide scope of applications. Specific examples have been given with antiperspirant formulas that proof the stability of these thickened microemulsions even with high content of electrolytes and low pH. We propose that the structure of these microemulsion gels can be described by cross-linking of nano-fine oil droplets via an ABA-triblock copolymer. A new generation of transparent antiperspirant roll-ons with significantly improved skin tolerance can now be formulated as an attractive alternative to alcohol-based products.

Recently inside nano liposome particles or nano-emulsions which contain tough-melting physiology activity materials or the coefficient of low organism utilization promote the coefficient of organism utilization, so this part has been studied a lot because they can absorb selectly cosmetics, specially physiology activity materials, into the skin. Also, in particle size, cells interstitial lipid interval are 30~50nm, so nano-emulsions that the size is similar to 30~50 nm are made to study for absorbing quickly into the skin. And transparent skin which contains oils in common skin lotion dosage form has become the center of public interest. The used nano-emulsions in this study were unsaturated lecithin/co-surfactant! ethanol/ oil / water. And polysorbate 20/ polysorbate 80/ Dicetyl phosphate/hydrogenated .caster oil/ isoceteth-20/SLS were used in co-surfactant. The used oils were cyclomethicone and caprylic/capric triglyceride. The manufacturing process was that microfluidizer was fixed in 1000bar and transit times were changed from 1 to 10 times. From transparency and particle size, the transparency sequence was SLS> polysorbate 20= polysorbate 80> isoceteth-20> dicetyl phosphate >hydrogenated caster oil and the particle size was small. Specially cyclomethicone nano-emulsions, when we made unsaturated lecithin /SLS /ethanol/water/cyclomethicone, cyclomethicone 5％ was good for transparency. And 20％ of this was used for making transparent skin toner in common skin dosage form.

Nor-aporphine derivatives have been discovered which interfere with the flux of Calcium into and out of the cell interior. It has been shown that adrenergic antagonists that block the Calcium exchange lead to an inhibition of Protein kinase C activity, thus blocking tyrosinase activation. Di-acetyl-dimethoxy-methyl-nor-aporphine is a semi-synthetic molecule of natural origin with very high potency. On B16 melanocytes as well as in normal human melanocytes the decrease in melanin synthesis reaches -50％ at a level of 40 ppm in the culture medium. On a molar concentration basis, this is 50 to 70 times stronger than Kojic acid inhibition. Yet, the cell viability is not affected. Reversibility studies show that after washing out of the active compound, melanogenesis returns to normal levels. Possible mechanisms of the activity are discussed. Tests carried out on SkinEthic(R) three-dimensional models of the epidermis and in vivo clinical studies on Asian population confirm the strong inhibition of melanogenesis. Safety evaluation of these molecules, on the other hand, demonstrates good skin tolerance and absence of toxicity.

Living cells are continuously subject to all sorts of stress such as ultraviolet rays on skin cells. Tests made in various laboratories show that when young fibroblasts (Le. at the beginning of their proliferate life) were repeatedly put under stress at subletal doses, they acquired a phenotype similar to Senescence Induced Prematurely by Stress (SIPS). The work presented hereafter was made on a new model of senescence induced prematurely by stress from ultraviolet Brays (UVB). The human fibroblast model was put under repeated UVB stress, causing SIPS. Several ageing biomarkers were used in order to characterise the cells that underwent stress:. an increase in the proportion of positive cells with senescence associated $\beta$-galactosidase activity (SA $\beta$-gal) measured by a specific coloration,. the proportion in the different morphological stages that fibroblasts undergo during culture visualised by microscopic observation,. the expression of genes known for overexpressing during senescence, particularly fibronectin and apolipoprotein J, measured by Real Time-PCR,. the common deletion of 4,977 bp in mitochondrial DNA, evaluated by nested PCR. Studying the variation of these 4 biomarkers, we have evaluated the protective effect of a Laminaria digitata extract (LDE) that can be used as a natural active ingredient for anti-ageing cosmetics.

Reactive oxygen species are capable of damaging biomolecules such as lipids, proteins, and DNA, which can not only lead to various diseases, but also oxidative damage resulting aging. In our previous study, Cercis chinensis (Leguminosae) showed a potent antioxidant activity. Nineteen compounds were isolated through antioxidant activity-guided fractionation. The C. chinensis extract and some of the constituents exhibited a potent antioxidant activity on the free radicals and lipid peroxidation and a notable protective effect on the t-BuOOH induced oxidative damage. In vivo test of skin damage induced by UVB irradiation, the extract of C. chinensis and a constituent, piceatannol, exhibited a significant protective effect. The life-span of the HEK-N/F cells were extended by 1.21-2.12 fold as a result of the continuous administration of 3 $\mu\textrm{g}$/ml of the C. chinensis extract and the active constituents compared to that of the control. These observations were attributed to the inhibitory effect of the C. chinensis extract and its constituents on the age-dependent shortening of the telomere. Thus, C. chinensis was demonstrated to protect the skin cells against oxidative stress and inhibit thereby the cellular aging, followed by expectation as anti-aging cosmetic ingredient.

It is well known that ascorbic acid (VC) is an important factor for several physiological reactions. In the skin, VC works as an anti-aging agent due to removing of oxidative stress generated by UV irradiation and stimulation of collagen synthesis. Thus, developing more effective VC derivatives is an important issue in creating anti-aging skin care products. Our study succeeded to develop a novel ascorbic acid derivative, ascorbic acid tetra-2-hexyldecanoate (VC-IP), which is a lipid-soluble pro-VC. The purpose of this study was to indicate the effects of VC-IP as pro-VC and anti-aging agent. First, it was examined whether VC-IP is converted to VC in physiological conditions. Since VC was detected from the cell extracts treated with VC-IP, it was indicated that VC-IP is a pro- VC.(omiited)

Recently several studies have indicated that ubiquinone-l0 (CoQ-10) decrease with aging in various organs of the human body. The decrease of ubiquinone-10 in the aging process suggests the reduction of both energy production in mitochondria and anti-oxidation with aging. Based on these findings, Ubiquinone-10 is being focused at as an anti-aging agent in the cosmetic market.(omitted)

In recent years, the chemical modification of polymers has been a driving force in many industrial sectors. There was evidence that this might also be the case of perfluoropolyethers (PFPEs) in the personal care industry. To investigate this potential, a systematic study has been conducted at Solvay Solexis SpA (formerly Ausimont SpA), through the analysis of the organic chemistry suitable to modify PFPE structures, and through the examination of the properties of PFPE derivatives thus produced from the viewpoint of a cosmetic formulator.(omitted)

UV radiation is the most dangerous stress factor among permanent environmental impacts on human skin. Consequences of UV exposure are aberrant tissue architecture, alterations in skin cells including functional changes. Nowadays new kinds of outdoor leisure-time activities and changing environmental conditions make the question of sun protection more important than ever. It is necessary to recognize that self-confident consumers do not consider to change their way of life, they demand modern solutions on the basis of new scientific developments. In the past one fundamental principle of cosmetics was the use of physical and organic filter systems against damaging UV-rays. Today new research results demonstrate that natural protecting cell mechanisms can be activated. Suitable biological actives strongly support the protection function not from the surface but from the inside of the cell. A soy seed preparation (SSP) was proven to stimulate natural skin protective functions. The major functions are an increased energy level and the prevention of DNA damage. These functions can I be defined as biological UV protection. The tumor suppressor protein p53 plays a key role in the regulation of DNA repair. p53 must be transferred into the phosphorylated form to work as transcription factor for genes which are regulating the cell cycle or organizing DNA repair. A pretreatment with SSP increases the phosphorylation rate of p53 of chronically UV-irradiated human keratinocytes significantly. According to the same test procedure SSP induces a dramatic increase in the expression of the tumor suppressor protein p14$^{ARF}$ that is supporting the p53 activity by blocking the antagonist of p53, the oncoprotein Mdm2. Mdm2, a ubiquitin E3-ligase, downregulates p53 and at the same time it prevents phosphorylation of p53. The positive influence of the tumor suppressor proteins explains the stimulation of DNA repair and prevention of sunburn cell formation by SSP, which was proven in cell culture experiments. In vivo the increased skin tolerance against UV irradiation by SSP could be confirmed too. We have assumed, that an increased repair potential provides full cell functionality.y.

Rhamnose-rich (RROP-s) and fucose-rich (FROP-s) oligo-and polysaccharides were prepared and extensively characterised by physical and chemical procedures [1,2] and compared to L-fucose. Their biological properties were then studied on human skin fibroblast cell cultures, human skin explant cultures and on hairless rat skin, using a variety of cell-biological, biochemical and computerised morphometrical procedures. Among the most important properties we could establish, the following are of particular interest for the tretment and prevention of age-dependent modifications of human skin (loss of skin-tissue, cells and matrix, wrinkle formation and others) : stimulation of cell proliferation (by $^3$[H]-thymidine incorporation and the MTT test), scavenging of reactive oxygen species (ROS) using several different procedures, and protease (MMP-2 and MMP-9) down-regulation. A topical preparation, using RROP-s and FROP-s, and/or L-fucose, was shown to increase cell proliferation, dermal matrix synthesis, efficient scavenging of ROS-s and to increase also the thickness of dermal tissue when applied for 4 weeks on hairless rat skin, accompanied by the densification of collagen bundles as well as by an increase of elastin synthesis. Using fluorescent labeled FROPs, it could be shown that these oligosaccharides react with cell-membrane receptors and especially with the elastin-laminin-receptor and the fucose-mannose receptor, but they penetrate also in the cell nucleus, suggesting the possibility of a direct action on the regulation of gene expression. When applied to the human skin of a team of voluntary women encompassing all age-groups, the efficiency of FROP-containing preparation could be confirmed using indentometry and computerised evaluation of skin micro-relief, as well as evaluation of periorbital wrinkles. It appears therefore that these preparations correspond to all the requirements of active anti-aging principles.

Two ways of making clear emulsions\ulcornerMicroemulsion Visible light λ = 400-700 nm Microemulsion, d < 50 nmㆍRefractive Index Matching(omitted)

Influence of gelatinase on basement membrane (BM) structure was investigated by using a skin equivalent (SE) model. The results showed that (1) gelatinase produced by cells degraded the BM and (2) the addition of matrix metalloproteinase-specific inhibitor to the SE medium accelerated the formation of BM structure, indicating that gelatinase is involved in BM impairment. The activity of gelatinase was also studied in healthy human facial skin tissues. The result of in situ zymography revealed gelatinase activity around the basal layer of the epidermis, where BM integrity was severely compromised. Therefore, this enzyme was suggested to be associated with BM decomposition in human facial skin. To assess the behavior of gelatinase in stratum corneum (SC) non-invasively, an immunological study was performed. Since positive immunostaining of pro-gelatinase B was observed in SC stripped from sun-exposed skin, whereas no positive staining detected in SC of non-irradiated skin, gelatinase in the epidermis could be non-invasively detected by measuring gelatinase in SC. Gelatinase in SC of healthy female volunteers was monitored using a special film that sensitively and conveniently detects gelatinase. Ninetr percent of SC from facial skin (l00 women, 40's-50's) was gelatinase-positive. On the other hand, SC from non-irradiated skin was negative. These results strongly suggest that (1) gelatinase is constantly produced in the facial epidermis of most middle-aged woman during their daily life, and (2) the enzyme might be involved in the aging-related degeneration of both BM and the matrix fibers of the upper layer of the dermis, acting as a very important aging factor. Strong inhibitory activity against gelatinase was found in turmeric extract and identified curcumin as the major ingredient. Topical application of cream containing turmeric extract significantly decreased the number of gelatinase-positive SC clusters in human facial skins. These results indicated that turmeric is an effective ingredient to prevent skin from photo aging by suppressing chlonically upregulated gelatinase activity by UV and to improve skin condition.

Hair fragility is a major concern to most people of African decent due to frequent chemical treatments, thermal appliance use and environmental exposure. It is extremely important to both cosmetic scientists and African-American consumers to understand the internal and external hair damage caused by these factors.(omitted)

A new method was developed to prepare microcapsules involving hydrophobic components. A totally new "silicone-resin-polypeptide" was used as the wall materials. The polypeptide was made by hydrolysis of collagen and silk protein and so on, and that was combined with silicone. This microcapsule was easily prepared from silicone-resin-polypeptide in water solution. The ratio of encapsulation in the microcapsule was not only high level as 90％, which had never been reached, but also the particle size could be controlled to obtain very small size (average particle size: 2${\mu}{\textrm}{m}$). Moreover, these microcapsules were resistant to high shearing forces and were stable over a long time period. This stable microcapsule was not crushed in pressure with finger spreading, so the core materials hardly touch the skin directly. Application in cosmetics by using microcapsule involving UV absorbents (2-ethylhexy1-4-methoxycinnamate (OMC) and 4-tert-butyl-4' -methoxydibenzoyl-methane (BMDBM)) was examined. It was possible to apply organic UV absorbents in water-rich formulations without any surfactant by using this microcapsule. This formulation demonstrated a good moisturizing and soft skin feel. Therefore, the microcapsule was applied to hair care products. As a result, the sunscreen hair lotion with microcapsule was able to prevent from damaging and decoloring of hair color by UV rays. As just, it was suggested that this microcapsules were be widely applied in cosmetics.cosmetics.

The problem of detecting individual differences of chemically similar liquids remains to be unsolved for various areas of the natural sciences, such as medicine, biology, nutrition, and cosmetics 1. This problem is of crucial importance for the cosmetic sciences, including perfumery and aromatherapy. For example, some certain subtle differences of smell and taste between chemically similar liquids are very difficult to detect by using conventional methods of analysis, such as gas chromatography (GC) and the like (except under unique advanced techniques that are expensive, time consuming, and are not readily available). However, a trained human nose and! or mouth of a perfumer and/ or taster can detect differences of that kind. There are two conventional ways to make a fragrance.(omitted)

Up until today, the key to contouring has been resumed in these two alternatives, either limiting the adipocyte storing capacity by modulating lipogenesis, or by stimulating lipolysis to eliminate adipocyte lipid content. Another interesting way could be the regulation of adipocyte differentiation. In this work, we have evaluated the effect of a brown algal extract of Sphacelaria scoparia (SSE) on the differentiation of pre-adipocytes into adipocytes. A pre-adipocyte line (3T3-L 1) was used. The differentiation was evaluated by the measure of produced lipids thanks to red oil coloration and spectrophotometry, and also by the expression of adipocyte differentiation markers: enzymes such as fatty acid synthase (FAS) and stearoyl CoA desaturase (SCD), or membrane proteins such as glucose transporters (GLUT -4) and fatty acid transporters (FAT) expressed on the surface of human adipocytes. These genes are under control of two transcription factors: CAAT-enhancer binding protein (c/EBP alpha) and sterol response element binding protein (SREBP1). All these markers were analysed at different stages of differentiation by RT -PCR. Sphacelaria extract (SSE) inhibits pre-adipocytes differentiating into adipocytes following a dose-dependant relation, using a kinetics similar to retinoic acid. It decreases the expression of mRNA specific to FAS, FAT, GLUT -4, SCD1, c/EBP alpha and SREBP1. Moreover, SSE regulated on collagen 1 and collagen 4 expression. A stimulation of collagen 1 was also measured in human skin fibroblasts. Thus, SSE performs as a genuine differentiation inhibitor and not only as a lipogenesis inhibitor, and could be used in slimming products.

The prime objective of many facial wrinkle-reduction treatments is to achieve visible improvement. Thus the visibility of before/after treatment differences is often part of an efficacy assessment. This paper investigates whether the background knowledge of the people acting as observers in such assessments is of impact on the results: e.g. the subjects themselves are familiar with their faces, skin professionals have much experience in judging skin quality, and thus both might be more sensitive to small changes. In a clinical study 44 Female subjects were regularly treated during a period of 12 weeks with one of three wrinkle-reduction methods: K, Land M (placebo). Photographs were taken before treatment and at 6 and 12 weeks. The photographs were judged by 3 types of observers:ㆍ24 Lay observers were given the 0＆6-week and the 0＆12-week photo pairs of all subjects to indicate the one with the least wrinkles in a two-alternative forced-choice (TAFC) procedure.ㆍThe subjects themselves were given the 0＆6-weel and the 0＆12-week pair of their own photos (8 replications) to indicate the photo with the least wrinkles (TAFC).ㆍA trained panel of skin professionals (N=3) each gave 9-point Fitzpatrick wrinkle-severity scores for all individual 0-week and 12-week photos. We found that the lay observers perceived the same differences as the subjects themselves: significant improvements after 12 weeks for treatment K (p<0.0005 and p=0.005, respectively), no visible effects for treatments Land M, and, most importantly, a significant difference between treatments K and M/placebo (p=0.02 and p=0.04, respectively). Also the trained panel found this difference between K and M (p=0.013), but here it was due to a significant deterioration over time of the 'placebo-treated' wrinkles (M, p=0.03). Thus in conclusion we have found no indications that extra knowledge - in the form of familiarity with the own face or in the form of professional training - results in the identification of more treatments that show significantly visible wrinkle-reduction.

Active ingredients have been around in cosmetics for a long time but have they really resulted in active cosmetic products\ulcorner In order to achieve this, the right active needs to be delivered to the right location at the right concentration for the correct period of time. And the extent (and therefore the concentration) of this delivery depends on the formulation. From a rather theoretical approach based on the polarity of the active ingredient, the stratum corneum and the oil phase, the Relative Polarity Index was established that enables the selection of a suitable emollient for ensuring skin penetration of the active ingredient. Practical examples subsequently show the validity of this approach that demonstrates that one can regulate the delivery of an active molecule (and therefore the efficacy of a cosmetic formulation) by selection and control of the emollient system. Cosmetic formulations are generally quite complex mixtures and subsequent experiments using different emulsifier systems indicated that this component of a cosmetic formulation could also have an impact on steering the active ingredient to the right layer of the skin, although it is too early to be able to derive general rules from this.

A novel branched wax has been developed for the control of the hardness of oil-wax gels. Using this wax, glossier application and smoother texture but tough lipstick can be obtained. Oil-wax gels are oily solids composed of liquid and crystalline solid oils (waxes). They are widely used in various cosmetic products, especially lipsticks. The control of gel hardness is one of the most important techniques in improvement of the lipstick quality. Addition of small amounts of commercial branched paraffin wax (e.g. microcrystalline wax, b-PW) to n-paraffin wax (n-PW) has been commonly used to increase gel hardness. However, gel hardness is very sensitive to the quantity of b-PW and the gel obtained is not always hard enough for practical use. In this study we examined the relationship between the gel hardness and the properties of the wax crystal in the gel. We have found that, when b-PW is added to n-PW, the wax crystal size becomes smaller (hardening the gels) and its crystallinity is decreased (softening the gels) simultaneously. Considering this result, we have developed a novel branched wax, Bis(polyethylenyl)- tetramethyldisiloxane (named ESE). ESE molecules are composed of a central tetramethyldisiloxane unit (branch unit) with polyethylene units at both ends. The central unit may suppress crystal growth while the ends are expected to prevent a decrease in wax crystallinity during crystallization. When ESE is added to n-PW, the wax crystal obtained becomes smaller without decreasing in crystallinity; consequently, the gel hardness is dramatically increased. By using ESE, the total amount of wax in a lipstick can be decreased by 30％ without spoiling the stick toughness, thereby achieving glossy application and smooth texture.

An innovative biopolymer known as the Rhizobian gum has been developed in France, which shows some dramatic refreshing effect on the skin. The origin of this innovative project takes its source in the natural environment, and in particular the natural environment of the roots of sunflowers and wheat, where a symbiotic bacterium has been discovered. It is a Rhizobium bacterium, which is hosted by the roots, and which is able to synthesize a specific polymer showing a dramatic water binding capacity. This polymer is in particular synthesized in period of drought, and its biological role is to concentrate the small amount water present in the soil in order to take it available for the root, which becomes then able to absorb it. This vital mechanism allows the plant to survive despite a severe climatic environment. This basic research has been conducted in collaboration whit the French National centre of scientific Research (CNRS), and has lead to the isolation of the Rhizobium bacteria. Rhizobian gum is a branched biopolymer consisting in the repetition of a polysaccharide unit of 3 molecules of glucose, 3 molecules of galactose and 1 molecule of glucuronic acid, whit one pyruvate group an average 1.6 acetyl groups. The fresh effect of Rhizobian gum is a strong sensorial impact that 100 ％ of the consumers are able to perceive, and which is judged very pleasant by most of them. In addition to this, a large majority of consumers are perceived, and which is judge very pleasant by most of them. In addition to this, a large majority of consumers also feel a very pleasant relaxing sensation. Smoothness and softness are also felt by most consumers and qualified positively by most of them. These qualities guarantee a strong impact on today's consumers.

The use of both an Alpha Hydroxy Acid, Citric Acid, and a Skin Moisturizer, Glycereth-26, formulated into a hand dishwashing detergent have been shown to be synergistic in their effects on certain skin health parameters. Each ingredient was evaluated alone and together in a hand dishwashing detergent via a 9-week use test. Panelists washed dishes using the sponge method commonly used in Asian markets and a 1:7 dilution of the concentrated dish detergent. Panelist's hands were evaluated initially and at 3-week intervals for nine weeks. After nine weeks panelist's hands showed significant improvements in Moisture Absorption and Transepidermal Water Loss (TEWL). A synergistic effect on TEWL was found between the AHA and the Moisturizer. These effects, showing an improvement in the condition of the panelist's skin are impressive, particularly since they were observed from a dishwashing product that is highly diluted and at near neutral pH during the washing process.

Sphingolipids are important structural components of the stratum corneum lipids and serve the epidermal permeability barrier function. Recent investigations on biological activities of sphingolipids have revealed that they have a number of important biological functions in the cell such as cell proliferation and differentiation, anti-inflammation, mediation of signal transduction and many more.(omitted)

To investigate the potency of some natural extracts as skin whitening agents, in this study, 25 natural plants were prepared from natural sources including medicinal plants, such as Angelica dahurica using methylene dichloride, ethyl acetate, n-butyl alcohol, and water as the extraction and/or the partitioning solvents. These natural extracts were subsequently subjected to in-vitro DOPA auto-oxidation test in the media containing human or mushroom tyrosinase as the oxidation promoting enzymes. Most of the extracts showed relatively higher enzyme inhibition(omitted)

Silk Sericin(SS) is a natural protein extracted from cocoon of bombix mori and shows moisturizing effect to the skin due to a number of hydroxyl groups in the structure. But its application to cosmetics is limited due to its poor solubility in water. In order to solve this drawback and expand its application to cosmetics, polyethyleneglycol(PEG) was conjugated with sericin by reacting activated polyethyleneglycol(ActPEG). Reaction site of sericin is tyrosine residue, which was determined by using $^1$H-NMR. Random coil structure of sericin was transformed to beta-sheet structure by conjugating polyethyleneglycol. It was confirmed that melting point of sericin-PEG conjugate was lowered compared to that of each sericin and PEG due to the interaction between sericin and PEG in crystalline structure. Self-assembled sericin-PEG nanoparticle was obtained by dialyzing with alcohol solution of sericin-PEG conjugate against water. The particle is spherical and has 200-400nm of size. The moisturizing ability of sericin-PEG nanoparticle was much higher than that of sericin itself. Incorporation of vitamin A into sericin-PEG nanoparticle was carried out by diafiltration method. The content of incorporated Vitamin A in sericin-PEG nanoparticle was 8.9 wt％. Releasing behaviour of vitamin A incorporated into nanoparticle was tested in phosphate buffer, pH 7.4 at 37$^{\circ}C$. and half-life of Vitamin A release was 43hrs. Sericin-PEG nanoparticle exhibited higher moisturing effect than sericin itself and distilled water, respectively. No toxicity and irritation were observed in animal tests. It can be expected that the self-assembled sericin-PEG nanoparticle can be developed for cosmetics.

By using sequentially efficacy tests based on tyrosinase, the key enzyme of melanogenesis, then a cell line of melanocytes cultured in vitro, we have been able to detect the whitening potential of a plant extract and then to develop a new whitening Active Ingredient whose the whitening potential was confirmed on cultured melanocytes. Through a phytochemical approach, it seems that the whitening potential could be due to the "tannin" fraction of plant extract. A complementary work is planned to explain more precisely which fractions are responsible for the whitening potential and a clinical test is in progress on 30 Asian skin type volunteers to show the whitening efficacy on human volunteers.

Metabolic disease such as diabetes, which is caused by stress or imbalanced diet, has been increasing. A diabetic tend to suffer from a delay or difficulty of wound healing. The extract of SHIKON (SK), that is the root of Lithospermun erythrorhison, has been reported to have an effect on healing for normal wound, but has never studies for intractable wound so far. Therefore we examined the effect of SK extract on wound healing with healing impaired mouse model. Full-thickness round wounds were created on the backs of db/db mice and applied SK, and we observed neovascularization and collagen synthesis, distribution of apoptotic cells, and vascular endothelial growth factor (VEGF)- positive cells in granulation tissue. After two weeks, a number of capillary vessel and collagen synthesis were increased in SK-treated wounds. Infiltration of VEGF-positive neutrophils was also seen in the wound, besides apoptotic fibroblasts and endothelial cells were appeared in the granulation tissue. After three weeks, the wound closed completely with SK-treated but not in control. These results suggest that SK enhanced neovascularization by VEGF and this kind of apoptosis process makes the scar smooth. In this study, it is obvious that SK also accelerates healing of intractable wound.

Functional cosmetic ingredients such as L-ascorbic acid, retinoic acid, indole-3-acetic acid, salicylic acid, acidic dye(indigo carmine) are intercalatively encapsulated by skin-friendly metal hydroxides and oxides matrices. Such functional organic-inorganic nanohybrids are realized via chemical coprecipitation and surface coating reactions. The hetero-structural nature of these nanohybrids, their particle morphology and textural characterizations are mainly discussed on the basis of powder X-ray diffraction, electron microscopies, and high performance liquid chromatographic analyses. The cosmetic ingredients encapsulated in inorganics show greatly improved storage stability, sustained releasing property as well as higher transdermal transfer efficiency.

The importance of cornified envelope (CE)-bound lipids to epidermal barrier function is increasingly being recognized. In the present study, we intentionally damaged the cornified layer of hairless mice by ultraviolet irradiation and sodium dodecyl sulfate (SDS) treatment, and assessed the changes in epidermal barrier function by measuring Trans Epidermal Water Loss (TEWL). We also measured changes in the amount of CE-bound lipids using thin layer chromatography (TLC). The results showed that both treatments increased TEWL and decreased CE-bound lipids (omega-hydroxy cerami de and omega-hydroxy acid). In addition, investigation of the chronological changes in TEWL revealed an inverse relationship between TEWL and CE-bound lipids, and a correlation between CE-bound lipids and epidermal barrier function. We then measured the amount of CE-bound lipids in the cheek and the medial side of the upper arm in humans. The results showed that because the cheek receives external stimulation on a daily basis, the amount of CE-bound lipids was significantly lower, while the level of TEWL was higher. These observations, together with those from the animal study, indicate that CE-bound lipids are related to epidermal barrier function.

The fuzzy rat that expresses hypersecretion of sebum and hyperplastic sebaceous glands is a genetic mutant for the study of many pharmacological aspects especially human acne. Through this model, we examined the effects of several phospholipids on the secretion of sebum after topical application. The phospholipid derivatives were phosphatidylcholine (PC), hydrogenated phosphatidylcholine (HPC), phosphati dylserine (PS) and hydrogenated phosphatidylserine(HPS). All agents were dissolved into the vehicle (1, 3-Butanediol, ethanol and water) at 0.5％ weight volume and applied on the dorsal area of the fuzzy rat. To observe histological changes, the skin biopsies were stained with Oil Red O and the size and morphology of sebaceous gland was observed under microscope. Topical treatment with PC and/or HPC showed a marked decrease in sebum excretion. Especially hydrogenated PC (HPC) appeared to have more predominant sebosuppressive function than any other treatment. The other agents such as PS and HPS showed a marginal effect on sebum secretion. With the sebosuppressive activity, HPC and PC seem to have a good potential application on acne treatment. In order to obtain more insights into possible mechanisms behind the above observations, effects of each phospholipid on the expression of peroxisome proliferator-activated receptor (PPAR) genes were investigated. Recently, it has been demonstrated that expression and activation of PPAR subtypes appear to modulate the accumulation of cytoplasmic fat droplets that characterizes the sebocyte differentiation(1). It was also previously suggested that PPAR${\gamma}$ antagonist would seem possible to interfere sebum production without side effects (2). In this study we examined the diverse effects of the tested phospholipids on the expression of several PPAR genes based on reverse transcription-polymerase chain reaction (RT-PCR) from the topically treated skin of fuzzy rats. The results and possible implications are discussed.

The matrix metalloproteinases (MMPs) are a family of enzymes responsible for degrading connective tissue. MMPs catalyze the breakdown of collagen from the extracellular matrix, leading to wrinkle formation and accelerated skin aging. Furthermore, ultraviolet irradiation causes increased expression of certain MMPs. In the extracellular matrix turnover, MMPs are interacting with endogenous regulators named tissue inhibitors of metalloproteinases (TIMPs). Using peptide substrate assays, it has been demonstrated that TIMP-MMP complexes interact highly specifically with $K_{i}$ values of 10$^{-9}$ -10$^{-16}$ M. Therefore applications for TIMP as inhibitor of collagen degradation are suggested for cosmetic anti-aging products to prevent wrinkle formation and loss of elasticity. To date four TIMP proteins (TIMP-1, TIMP-2, TIMP-3 and TIMP-4) have been identified which show a high degree in sequence similarity. The production of human TIMP-2, a 194-residue nonglycosylated protein, was performed by fed-batch culture of Escherichia coli. TIMP-2 accumulated in the bacterial cells in an insoluble form as inclusion bodies. The inclusion bodies were solubilized and the protein refolded to yield the native TIMP-2 in the active form. The integrity of the protein was confirmed by mass analysis, Edman sequencing and gel shift experiments with authentic samples. The inhibitory activity of the refolded and purified TIMP-2 was demonstrated with MMP-1 and MMP-2 assays using synthetic fluorogenic peptide substrates.s.

Biotin is a water-soluble vitamin used as a skin conditioning agent and promotes the formation of intercellular lipid layers through increased lipid synthesis, which improves the skin's natural barrier function. The anti-inflammatory effects of biotin have been investigated using in vitro assay models, such as MTT assay, measurements of concentrations of nitric oxide(NO), prostaglandin E2(PGE$_2$), and inhibition rate of 5-lipoxygenase(5-LOX). In comparison with biotin, other plant extracts were tested at the same time which were kudzu vine extract, sage extract, paeonia extract, and dipotassium glycyrrhetinate. Nitric oxide is a signal molecule with functions such as neurotransmission, local vascular relaxation, and anti-inflammation in many physiological and pathological processes. NO can cause apoptosis and necrosis of target cells such as keratinocytes and is generated from L-arginine by nitric oxide synthase (NOS). Prostanoids, including prostaglandins and thromboxanes, are generated by the phospholipase $A_2$/cyclooxygenase(COX) pathway, and leukotrienes are generated by the 5-lipoxygenase pathway from arachidonic acid. Prostaglandin E2 recently have been shown to be beneficial in the resolution of tissue injury and inflammation, also has been implicated as an immunosuppressive agent and plasma levels of PGE$_2$ are elevated in patients sustaining thermal injury. Lipoxygenase metabolites from arachidonic acid have been implicated in inflammation, anti-inflammatory activity of the raw materials was evaluated in vitro by the offered inhibition of lipoxygenase.

Korean ginseng(Panax Ginseng C.A. Meyer) known as a oriental miracle drug is an important medicinal plant. Ginseng has been used for geriatric, tonic, stomachic, and aphrodisiac treatments for thousands years. Also, it is an antibiotic and has therapeutic properties against stress and cancer. Ginseng is widely distributed all over the world. Among them, Korean mountain ginseng has the most valuable effect on pharmaceuticals. The roots of mountain ginseng contained several kinds of ginsenosides that have many active functions for the human body. However, the study of mountain ginseng has a limit because the mountain ginseng is very expensive and rare. So, we artificially cultured mountain ginseng adventitious roots using the bioreactor culture system. We induced callus from original mountain ginseng, directly dug up in mountain and aged about one hundred ten years. Separated adventitious roots were precultured in 500ml conical flasks and then, transferred in 20L bioreactors. The adventitious roots of mountain ginseng were harvested after culturing for 40days, dried and then, extracted with several solvents. In this study, we investigated the whitening effect, anti-wrinkle effect and the safety of tissue cultured adventitious roots extract of mountain ginseng in order to identify the merit as a cosmetic ingredient. Particularly, extract of mountain ginseng adventitious roots showed whitening and anti-wrinkle effects. The inhibitory effect of this extract on the melanogenesis was examined using B-16 melanoma cell. When B-16 melanoma cells were cultured with adventitious root extract, there was a dramatically decrease in melanin contents of 8-16 melanoma cell. And we identified this extract inhibited Dopa auto-oxidation significantly. Also, when transformed mouse fibroblast L929 cells were treated with this extract, there was a significant increase in collagen synthesis. The results show significant inhibited melanization and wrinkle without inhibiting cell viability.

A recent development in cosmetics has been the pursuit of wrinkling in the skin. The cosmetics composed of anti-wrinkle agent stand out from the point of view of environmental contamination and pollution. Among them, Indole-3-acetic acid (IAA), studied with wrinkling pigmentation and swelling conditions in the area of the eye, showed clinically significant reduction in depth of lines during one month trial using skin treatment. But, IAA has shown some problems when used in cosmetic formulations, such as stability, permeability and toxicity. The results of the clinical examination were shown that its permeability and toxicity didn't matter. To increase the stability of IAA, antioxidants such as Licorice, ubiquinone, tocopherol, Baicalin, ferulic acid, BHT, ascorbic acid, sodium metabisulfite, and so on were employed in cosmetic formulations. Our main purpose is the study for the stability efficiency and effect of each other of cream formulations containing optimal dosage antioxidants (o/w type emulsion), This study evaluated wrinkle reduction effect of IAA, which is used in cosmetics.

A human Cu, Zn-superoxide dismutase (Cu, Zn-SOD) was fused with a Tat PTD of HIV-1 to produce a novel anti-aging ingredient, Tat-SOD for cosmeceuticals. Test of stability and evaluation of transduction efficacy and enzymatic activity suggest Tat-SOD is an effective active ingredient for anti-aging treatment.

We evaluated activities of various plant leaf extracts and found the availability against skin aging in the leaf extract of star fruit (Averrhoa carambola L), and developed Star Fruit Leaf Extract BG30 as an ingredient of cosmetics. Star Fruit Leaf Extract BG30 was found to show scavenging activities of reactive oxygen species and an inhibitory effect on the activity of matrix metalloproteinase-1. It showed increasing activity of type I collagen and recovery effect from damage of UV-B irradiation in human fibroblast. We performed the separation of the active principal from Star Fruit Leaf Extract BG30 to give isofurcatin 2"-Ο-$\alpha$-L-rhamnopyranoside, which showed increasing activity of type I collagen. To examine the anti-wrinkle effect of Star Fruit Leaf Extract BG30, seven volunteers applied a Star Fruit Leaf Extract BG30 1 ％ cream in double blind manner to one-side of the corner of their eye and the placebo cream to the opposite side. Clinical evaluation of wrinkling was performed every week for 5 weeks using a silicone rubber replica. A statistically significant improvement of Star Fruit Leaf Extract BG30-treated site was seen in decreased wrinkles. Star Fruit Leaf Extract BG30 results in clinically visible improvement in wrinkling when used topically for 5 weeks.

In this study, we have investigated the potent anti-aging effect of DA-3711, a cosmetic ingredient derived from artificial skin culture. The artificial skin was originally developed as a skin replacement for the treatment of chronic skin wounds. To produce DA-3711, neonatal human fibroblasts were seeded into biocompatible collagen/chitosan/glycosaminoglycan (GAG) scaffolds and cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with fetal bovine serum and nonessential amino acids. Analysis of the culture broth (DA-3711) showed that growth factors such as VEGF, TGF-$\beta$, KGF were present at significantly higher levels that in the culture broth of fibroblasts cultured in monolayer. The biological activity of DA-3711 was assessed by measuring in vitro cell proliferation and collagen synthesis of normal human fibroblasts. Fibroblasts treated with 10％ DA-3711 showed a 2-fold higher proliferation and 2 to 4-fold higher collagen synthesis than untreated cells. DA-3711 also exhibited anti-oxidative effects, since cells under peroxide-induced oxidative stress showed a 30％ higher viability in DA-3711-containing medium than in medium without DA-3711 addition. The results suggest that DA-3711 may have anti-aging effects by stimulating skin regeneration and protecting against oxidative stress.

A chemical investigation of Peucedanum praeruptorum has resulted in the isolation of 3 khellactone derivatives, which have inhibitory effects on melanogenesis in B16 mouse melanoma cell lines. The khellactone derivatives were isolated from the crude extract of the roots of Pecedanum praeruptorum by a combination of adsorption chromatography and HPLC. The structures of isolated compounds were identified as 3', 4'-diangeloyl-cis-khellactone, 3'-angeloyl-4'-senecioyl-cis-khellactone and, 3', 4'-disenecioyl-cis-khellactone by $^1$H NMR, $^{13}$ C NMR and mass spectral studies and by comparisons of spectral data with reported literatures. These khellactone derivatives can be a good candidate for new skin whitening agent due to its strong inhibitory activity and safety.

Yew (Taxus cuspidata Sieb.) chose that grow as medicine, food, decorative plant in Korea's Kyong-Gi province surroundings. Extracts of yew extracted leaf of 250 g and stems of 300 g with 1,3-butylene glycol (1,3-BG), propylene glycol (PG) and water. As results, external appearance of leaf extract of yew was slightly brown clear extract. The pH was 5.3$\pm$0.5, and specific gravity was 1.012$\pm$0.05, and refractive index was 1.375$\pm$0.05. Also, appearance of stem's extract was slightly brown clear extract, and the pH was 5.4$\pm$0.5, and specific gravity was 1.016$\pm$0.05, and refractive index was 1.358$\pm$0.05. Oil of yew separated from seeds, and extracted polysaccharide high purity from fruits. As a result, specific gravity of oil was 0.987, and obtained 40％ of yield. Total polyphenols amount of yew extract is detected 0.563％ in leaves, 0.325％ in stems, whereas total tannins amount contained 0.054％ and 0.037％ each in leaves and stems. As effect in cosmetics, the anti-oxidative effect by DPPH method is 75.0％ in leaves, and stems was 64.0％. Collagen synthesis rate was shown high activity by 54.16％ in stem's extract, 33.18％ in leaves' extract. Also, PPE-inhibitory activities were 13.7％ and 23.5％ each in leaves and stems. Anti-inflammatory effect of yew seed oil displayed superior effect of 41 ％ than control. Polysaccharide's molecular weight that is gotten from fruits was 5$\times$10$^4$~ 3$\times$10$^{5}$ dalton, and got 20.0$\pm$5％ of yield.

A new colloidal silver (Ag) system is present in which a fine colloidal Ag is in situ deposited onto functionalized porous poly (ethylene glycol dimethacrylate) (poly (EGDMA)) microspheres. The effectiveness of Ag deposition was investigated considering the surface characteristics of poly (EGDMA) microspheres. The result reported in this study illustrates that the control of surface area and surface functionality (in this study, a hydroxyl group) of poly (EGDMA) microspheres is an important factor that determines practically the degree of deposition of colloidal Ag. The x-ray analysis showed that Ag nanoparticles were dispersed evenly inner and outer surfaces and had a face center cubic (fee) phase. In the preservative efficacy test, the Ag-containing poly (EGDMA) microspheres had a powerful anti-bacterial activity, showing a high potential for a new preservative in cosmetic industry.

DA-3711 is a novel, anti wrinkle agent containing growth factors derived from human skin culture. In this study, effect of topical application of DA-3711 on chronic ultraviolet B (UVB)-induced skin damage was evaluated in hairless mice model. Exposure to UVB for 8 weeks induced apparent wrinkles on the back skin of the mice. The dorsal surfaces were exposed to UVB for a further 8 weeks, during which the surfaces where treated daily by topically application of a lotion containing either 35％ or 70％ of DA-3711. For comparison, lotion base, lotion base containing Cylasphere Retinol(equation omitted) (2500 I.U., Coletica) and NouriCe $l^{(R)}$ (Biozhem) were applied topically once a day for 8 weeks.(omitted))

Fructan, a polysaccharide existing in plants or produced by microorganisms, is a sugar polymer of fructose with $\beta$-2,6 linkages. In this study, we investigated some cosmeceutical properties of Fructan such as moisturizing effect, cell proliferation effect, anti-inflammation effect and cell cytotoxicity. Zymomonas mobilis, a microorganism producing Fructan, was cultured in a medium containing 10％ sucrose and 2％ yeast extract as main components for 24 hours at 37$^{\circ}C$ and pH 7. Fructan was obtained by precipitation from the cultured medium by adding alcohol (alcohol ratio of 1:3) after removing the enzyme by centrifuging. Fructan exhibited almost same moisturizing effect as hyaluronic acid and cell proliferation effect on human fibroblast and keratinocyte as well. Moreover, on cell proliferation test on bio-artificial skin constructed by 3-dimensional(3-D) culture after inducing primary skin inflammation with 0.5％ sodium lauryl sulfate (SLS), the 3-D artificial skin treated with 0.01 mg/ml, 0.05mg/ml of Fructan exhibited higher cell proliferation than the 3-D artificial skin treated with SLS only. On anti-inflammation test on 3-D artificial skin evaluated by measuring secreted quantity of interleukin-1$\alpha$ (IL-1$\alpha$) which is a pre-inflammatory mediator induced by SLS, the quantity of IL-1$\alpha$on the 3-D artificial skin treated with 0.01 mg/ml, 0.05mg/ml of Fructan was less than the one on the 3-D artificial skin treated with SLS only. As a result of these studies, Fructan has anti-inflammation effect against inflammatory reaction by a skin irritant as well as cell proliferation effect in bio-artificial skin. Fructan was also evaluated as a safe material without any toxicity in safety tests using fibroblasts and animals.

A kojic acid derivative, kojic acid 7-O-$\beta$-D-tetraacetylglucopyranoside(KTG) was synthesized. Regio-and stereo-selective glycosylation at 7-postion in kojic acid with $\beta$-D-pentaacetylglucose was achieved with high yield(80％) by the use of Lewis acid and organic base in nonpolar solvent. KTG was hydrolyzed in methanol by the aid of sodium methoxide to give kojic acid 7-O-$\beta$-D-glucopyranoside(KGP). KGP is freely soluble in water and soluble in methanol and ethanol. Its structure was comfirmed by $^1$H-NMR and $^{13}$ C-NMR. Tyrosinase activity inhibition of KGP was measured with mushroom tyrosinase compared with ascorbic acid, kojic acid and arbutin. KGP showed higher tyrosinase inhibition activity($IC_{50}$/=33.3 $\mu\textrm{g}$/ml) than ascorbic acid(63.2 $\mu\textrm{g}$/ml) and arbutin(91.8 $\mu\textrm{g}$/ml) but lower inhibition activity than kojic acid(8.3 $\mu\textrm{g}$/ml). To test free-radical scavenging activity, we used 1, 1-diphenyl-2-picrylhydrazyl(DPPH) as a free-radical source. Free-radical scavenging activity of KGP was very low($SC_{50}$/>1000 $\mu\textrm{g}$/ml) compared with ascorbic acid($SC_{50}$/=2.68 $\mu\textrm{g}$/ml) and arbutin($SC_{50}$/=180$\mu\textrm{g}$/ml). Melanin formation inhibition of KGP was measured in B16 melanoma, compared with kojic acid, arbutin and Vitamin C. Inhibition activity of KGP for melanin formation was not found within test concentrations.

To find a new substance with superior melanogenesis inhibitory activity, the bioactivities of alkyl-3,4-dihydroxy-5-substituted benzoate (A) and N-alkyl-3,4-dihydroxy-5-substituted benzamide (B) derivatives as substrate of tyrosinase were measured in mouse melanoma cells. And the bioactivities analyzed using comparative molecular similarity indices analysis (CoMSIA). From the CoMSIA model, when cross-validation value (q$^2$) is 0.713 at four components, the pearson correlation coefficient ($r^2$) is 0.900. Unknown compounds were predicted, using QSAR analyzed results from the CoMSIA methods. Excellent agreement was obtained between the measured and the predicted bioactivities of unknown compounds. As the results of prediction from CoMSIA, we could conclude that the bioactivities were increased from pl$_{50}$=3.18-4.80 to above 5.17 by creation of 6-methylheptyl, n-pentylphenyl and 2-hydroxypentylphenyl group etc,.,.

Ginsenosides, the major active ingredients of ginseng, show a variety of biomedical efficacies such as anti-aging, anti oxidation and anti-inflammatory activities. To understand the effects of compound K (20-O-D-glucopyranosyl-20 (S)-protopanaxadiol), one of the major metabolite of ginsenosides on the skin, we assessed the expression level of ∼ 100 transcripts in compound K-treated HaCaT cells using cDNA microarray analysis. Compound K treatment induced differential expression of 21 genes, which have been reported to be involved in the organization of ECM structure as well as defense responses in human skin cells. One of the most interesting findings is 2-fold increase in hyaluronan synthase2 (HAS2) gene expression by compound K. We found that change in expression of HAS2 gene represents a specific response of HaCaT cells to compound K because hyaluronan synthase 1, 3 was not changed by treatment with compound K. We also demonstrated that the compound K effectively induced hyaluronan synthesis in human skin cells and hairless mouse skin. The human clinical study indicates that topical application of compound K-containing oil-in-water emulsion showed improvement of xerosis, wrinkle and fine lines in the aged skin. We concluded that compound K has anti-aging effects by the induction of HAS2 gene expression and following hyaluronan synthase.

As melanin is a key material for skin pigmentation, inhibitors of melanin formation have been used to cosmetics and drugs to prevent hyperpigmentation. Therefore, search for effective inhibitor$ from various plants were attempted. For this purpose, I examined in vitro tyrosinase assay system. Tyrosinase showed a maximal activity at 4 units concentration of tyrosinase, 10 minutes, 42$^{\circ}C$ and pH 6.5.

Physiological lipid mixtures comprised of cholesterol, ceramide and free fatty acid better maintain epidermal homeostasis and have been recently used for dermatoses induced by skin barrier damage, for example for atopic dermatitis and xerotic skin. Itching and dry atopic dermatitis of the skin may be related to altered skin barrier function. In a previous study, the use of multi-lamellar emulsion (MLE), which is a lipid mixtures containing cholesterol, pseudoceramide and free fatty acid, has been shown to accelerate the recovery of the epidermal permeability barrier. In this study, we assessed the efficacy of MLE compared with a currently used anti-itch moisturizer (AIM), the active ingredients of which are menthol and camphor, on barrier recovery after barrier disruption. To clarify the effect of MLE and AIM after acute barrier perturbation, we measured the relation between transepidermal water loss (TEWL) and the barrier recovery rate at 3, 6, 24, and 48 hours after tape stripping hairless mice and then observed changes in the stratum corneum (SC), including the intercellular lipid structure and secretion of lamellar bodies, by electron microscopy. MLE treated skin recover skin barrier function more rapidly, and AIM treated skin delayed barrier repair. Morphological changes in the epidermis, of MLE treated skin revealed well-conserved lipid multi-lamellar structures at 24 h after tape stripping, whereas AIM treated skin showed altered lamellar bilayers within the SC interstices at 48 h. In addition, MLE treated skin showed an increase in the number of LBs and in their secretions and a decrease in the number of SC layers versus AIM treated skin. These results suggest that MLE may accelerate the production of an epidermal permeability barrier in hairless mice by increasing the number and secretion of LB and improve the dryness and itch associated with an altered epidermal permeability barrier.

Exposure of skin cells, particularly keratinocytes to various nuclear factor-kappaB ($\textrm{NF}_{-{\kappa}}\textrm{B}$) activators [e.g. tumor necrosis factor-$\alpha$, interleukin-1, lipopolysaccharides, and ultraviolet light] leads to phosphorylation and degradation of the inhibitory protein, $\textrm{I}_{{\kappa}}\textrm{B}$. Liberated $\textrm{NF}_{-{\kappa}}\textrm{B}$ is translocated into the nucleus where it can change or alter expression of target genes, resulting in the secretion of extracellular signaling molecules including melanotrophic factors affecting melanocyte. In order to demonstrate the possible role of $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation on the synthesis of melanotrophic factors from the keratinocytes, the activities of $\textrm{NF}_{-{\kappa}}\textrm{B}$ induced by melanogenic inhibitors (MIs) were determined in human HaCaT keratinocytes transfected with $\textrm{pNF}_{-{\kappa}}\textrm{B}$-SEAP-NPT plasmid. Transfectant cells released the secretory alkaline phosphatase (SEAP) as a transcription reporter in response to the $\textrm{NF}_{-{\kappa}}\textrm{B}$ activity and contain the neomycin phosphotransferase (NPT) gene for the dominant selection marker for geneticin resistance. MIs such as niacinamide, kojic acid, hydroquinone, resorcinol, arbutin, and glycolic acid were preincubated with transfectant HaCaT cells for 3 h and then ultraviolet B (UVB) was irradiated. $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation was measured with the SEAP reporter gene assay using a fluorescence detection method. Of the Mis tested, kojic acid ($IC_{50}$/ = 60 $\mu$M) was found to be the most potent inhibitor of UVB-upregulating $\textrm{NF}_{-{\kappa}}\textrm{B}$ activation in transfectant HaCaT cells, which is followed by niacinamide ($IC_{50}$/= 540 $\mu$M). Pretreatment of the transfectant HaCaT cells with the Mis, especially kojic acid and niacinamide, effectively lowered $\textrm{NF}_{-{\kappa}}\textrm{B}$ binding measured by electrophoretic mobility shift assay. Furthermore, these two inhibitors remarkably reduced the secretion level of IL-6, one of melanotrophic factors, triggered by UV-radiation of the HaCaT cells. These observations suggest that Mis working at the in vivo level might act partially through the modulation of the synthesis of melanotrophic factors in keratinocyte.

It has been known that adenine is a very important material in living cells. Because, adenine is a member of nucleotide base, so it takes part in DNA, RNA and ATP synthesis. There are many reports that adenine participated in ingredients, especially DNA, RNA, NADH and ATP, affect on the cell. As well adenosine, conjugated adenine to glycoside, was known to anti-wrinkle compound. But there is no report whether adenine shows a good effect on the skin, especially anti-wrinkle. So, in this study, we tested whether adenine affects cell proliferation, collagen synthesis, collagenase synthesis inhibition in human dermal fibroblasts. In addition, we performed clinical study with adenine cream. Cell proliferation effect was tested by MTT assay. Collagen and collagenase synthesis were measured by Immunoassay with ELISA kit. Clinical study was performed by IECK according to KFDA Functional Cosmetic method. The results of cell proliferation show that 10$^{-6}$ ~10$^{-8}$ ％ of adenine increases cell proliferation about 50 ％ compare with non-treated control. At 10$^{-7}$ ~10$^{-10}$ ％, adenine increases type I collagen synthesis about 50％, decreases type I collagenase about 22％ compare with non-treated control. The results of clinical study show that 0.05％ adenine treated group reduces wrinkle significantly compare with placebo treated group. Therefore adenine may be a new anti-wrinkle candidate, through increases cell proliferation and collagen synthesis dramatically. And it decreases collagenase synthesis. So adenine could be used as a new anti-wrinkle agent.

One of the important function of skin is the protection from harmful environments. Many studies were carried out to keep the skin from wrinkling and pigmenting. Skin wrinkle and pigmentation could be caused' by the unusual disruption of connective tissue, the formation of free radicals and the radiation of ultraviolet. In this study, the extracts obtained from 23 different kinds of medicinal plants were examined whether they have inhibitory effect on tyrosinase and elastase as well as free radical scavenging effect. All the extracts examined were obtained by using 70% (v/v) ethanol at $60^{\circ}C$. It has been found that there were two medicinal plants which have positive effects matching with the purpose of this study. The extract of Ephedra sinica stapf shows an inhibitory effect on tyrosinase ($IC_{50}$ = 83.7~$\mu\textrm{g}$/ml), on elastase ($IC_{50}$ = $690{\mu}\textrm{g}$/ml), and also shows free radical scavenging effect ($IC_{50}$ = 29.6$\mu\textrm{g}$/ml). The extract of Betula platyphylla Var. shows an inhibitory effect on elastase ($IC_{50}$ = 498.1$\mu\textrm{g}$/ml), and free radical scavenging effect ($IC_{50}$ = 9$\mu\textrm{g}$/ ml). The aim of the study is to find the natural compounds which have function in skin-lightening, anti-wrinkling and anti-oxidation, with safe and active ingredients in cosmetic.