400 JOURNAL OF THE SOCIETY OF COSMETIC CHEMISTS Biotechnology and the cosmetics industry: impact of new production technologies James w. Bracke, Ph.D., and Kipling Thacker, Diagnostic, Inc., 1955 W. County Road B-2, St. Paul, MN 55113 Applications for certain biotechnology-derived raw materials in cosmetics and other product areas are necessitating the development of novel manufac- turing methods. Many of the newly developed compounds being produced are technically com- plex to prepare and handle, labile, and required in large quantities of high purity. Some recent devel- opments in production technology will be con- sidered, concentrating on the manufacture of chemical specialties of biological origin. The devel- opment of a new process for producing hyaluronic acid will be used as an example. The advantages and disadvantages involved in using manufacturing techniques which employ aerobic and anaerobic cell-culture production systems with genetically modified components will be discussed. Particular attention will be given to the current capabilities, limitations, and future development needs of such processes. Some significant developments in down- stream processing which have particular relevance to biological materials will also be discussed with respect to their anticipated impact on the develop- ment of biological processes. SCIENTIFIC SESSION II Photographic standardization of dry skin Ronald L. Rizer, Ph.D., John C. Seitz, and Thomas S. Spencer, Ph.D., S.C. Johnson& Son, 1525 Howe Street, Racine, WI 53403 Photographic standards were developed to provide visual definition for a range of scaling, cracking, and erythema observed with typical dry skin. All photographs were obtained with a Nikon f-3 cam- era and 105 mm macro lens under standardized conditions. Photographic standards were selected by viewing over 1,000 photographs taken during a study to evaluate the effect of lotion treatment on dry skin. Normal and most severe cases of dry skin were assigned 0 and 9, respectively. Intermediate grades of untreated dry skin were ranked in order of dry skin severity by an iterative procedure where all photographs were reviewed independently by several qualified readers, and repeated until all readers agreed with each other's selections. When clinical photographs taken under standardized con- ditions in a comparative study were evaluated using the photographic standards, significant improve- ments were shown in scaling, cracking, and ery- thema with lotion treatments. Photographic stan- dards of dry skin provided visual definition and documentation of skin condition. Also, they lend themselves to blind, independent evaluations of dry skin by multiple observers. Enhancement of the sensitivity of the Buehler method by use of the Hill Top Chamber D. L. Conine, Ph.D., J. L. Lichtin, Ph.D., and M. Polikandritou, Hill Top Research, P.O. Box 42501, Cincinnati, OH 45242 Tests of delayed contact hypersensitivity were con- ducted on guinea pigs using Buehler methodology to compare the sensitivity of the method using the Hill Top Chamber (HTC), a new plastic occlusive chamber and patch system, and the Webril patch (WP), Skin reactions were induced at significantly lower concentrations for samples tested with the HTC than for those tested with the WP, Four compounds, 1-chloro-2, 4-dinitrobenzene (DNCB), neomycin sulfate (NS), p-phenylenediamine (PPDA), and potassium dichromate (PD) were used as allergens. For each compound, the following steps were carried out: (1) three 0.4 ml induction applications, one per week, on the same site on the same animal (2) two week period of relaxation (3) a challenge application on a naive site and (4) rechallenge applications. For PPDA, EC50 were significantly lowered from 0.04% to 0,02% in ani- mals induced with the WP and HTC, respectively. For PD, EC•0 were significantly lowered from a calculated value of 11.0% to. 0.6% in animals induced with the WP and HTC, respectively. Con- centrations of 20% and 10% NS in petrolatum produced significantly higher percentages of re- sponders in animals sensitized via the HTC than in those with the WP. Similar results were found in comparisons involving DNCB. Effect of the chemical irritants anthralin and benzoyl peroxide on mouse skin epithelial cell protein production ChristopherJ. Molloy, Jeffrey D. Laskin, Ph.D., and Michael A. Gallo, Ph.D., UMDNJ-Rutgers Medi- cal School, Piscataway, NJ 08854 The widely used topical agents anthralin (An) and benzoyl peroxide (BP) are potent skin irritants. In the mouse skin model these compounds are hyper- plasiogenic and effective skin tumor promoters. Using the animal model, we compared the effect of these compounds with the tumor promoter 12- O-tetradecanoylphorbol-13-acetate (TPA) on pro- tein production in epidermal cells. The compounds were applied topically to the shaved dorsal skin of

ABSTRACTS 401 mice. 24 and 48 hr. later skin fragments were pulse labelled with 35S-methionine to assay total protein production. After labelling, proteins were extracted and separated using 2-dimensional gel electropho- resis with isoelectric focussing in the pH range 5-7 in the first dimension, and 10% polyacrylamide gels in the second dimension. Using this technique we were able to detect over 100 individual proteins under control, TPA, An, and BP treated animals. TPA and An treatment were found to alter the production of at least 10 distinct proteins, 9 of which appear to be keratins. BP treated epidermis closely resembled control skin, suggesting that it may act by a mechanism differing from that of TPA and An. Further studies using pulse labelled skin fragments should facilitate the characterization of proteins involved in the skin irritation process and the mechanism by which BP and An act. An alternative to direct panelist-judge interac- tion in evaluating underarm deodorants S. Mark Henry, Ph.D., G. Jacobs, and V. F. Cotty, Bristol-Myers Products, 225 Long Avenue, Hillside, NJ 07207 This report describes a novel method which elimi- nates the unpleasant and problematical close con- tact between deodorant panelists and judges. Instead of direct sniffing of the axilla, samples of axillary secretions are obtained using the bottom end of disposable glass tests tubes rotated briefly in the axillary vault. The tubes are removed, then handed to judges positioned out of sight of the subjects. The judges then score each pair of tubes and record their data. Results correlate well with tests conducted in the conventional manner. In addition to eliminating the potential problems of subject-judge contact and recognition, this technique isolates the axillary odor from smoke, fragrances, and other odors which may be present on the subject's body or clothing. The axillary odor remains on the tube for several hours if covered and refrigerated. The effects of dl-alpha-tocopheryl acetate on ultraviolet light-induced epidermal ornithine decarboxylase activity Peter T. Pugliese, M.D., Gery B. Saylor, David C. Salter, D. Phil., and Carl Nocka, Ph.D., Xienta, Inc., P.O. Box 98, Bernville, PA 19506 Ornithine decarboxylase (DOC) activity increases in response to ultraviolet (UV) light irradiation. Applying Vitamin E (dl-alpha-tocopheryl acetate) prior to UV irradiation produces a 92% reduction in induced-ODC activity. Our mathematical model predicts that this reduction is due to both a physical blocking of the UV light and a biochemical effect within the epidermis. Vitamin E can be an effective blocking agent if incorporated into a sunscreen, and may also prevent damage caused by the UV light which reaches the stratum comeurn. Finding a preservative system suitable for aloe vera gel Max Ceballos, Aloe Laboratories of Texas, Inc., 2809 E. Grimes, Harlingen, TX 78550 The '*ideal" preservative system for aloe vera gel consists of two components: physical heat and chemicals. The effect of different temperatures with time was examined for aloe vera gel in terms of pH, color, enzyme activity, and bacterial counts. The most lethal temperature was 90øC for 15 minutes, provided a stabilizing agent such as citric acid has been added. The "contact" power and long-range effects of twenty-five different preserva- tives on "raw" aloe vera gel and pasteurized gel were evaluated over a 7 day period. The results indicate these preservatives are geared to gram negative organisms, since they are very ineffective against total counts and, in particular, molds and yeast. Since most of these preservatives are pur- ported to be broad spectrum, these results are surprising. The preservative with the best general performance was diazolidinyl urea + methyl - and propyl - paraben and propylene glycol (Ger- maben II). Release of zinc pyrithione from shampoo bases and the effect of various additives Almas Babar, Ph.D., Christina Kawilarong, and Fotios M. Plakogiannis, Ph.D., Arnold & Marie Schwartz College of Pharmacy and Health Sciences/L.I.U., 75 DeKalb Avenue, Brooklyn, NY 11201 Based on previous work on the use of ZNPT in shampoos, it was demonstrated that its antidan- druff activity greatly depends upon the release rate of the ZNPT from the shampoo base. Therefore a series of ZNPT shampoos were prepared contain- ing several additives: alcohol (95%) 5, 10, and 15% DMSO 5, 10, and 15% PEG-400 5, 10, and 15% and urea 5, 10, and 15%. The release rates from these shampoos were determined at time intervals of 15, 30, and 45 min., and compared with a control shampoo and two commercial products. It was found that the release rate decreased in the follow- ing order: alcohol (95%) propylene glycol DMSO, uvea PEG-400 control commercial products. The data also demonstrated that the release rate was influenced by the percentage of the additive present in the formulation and the time of