kDa 3.0 I 21.5- 14.4- 6.0- 3.5- EFFECT OF PERMING/BLEACHING ON CUTICLES 3.5 .,, .,, I pl 4.0 I 3.93 ,. ,. ,. ,. ,, 4.5 I / ., ., 1------- .,, .,, .,, .,, ,. 10 o t I I I I I I ' I I I I I 20 40 60 80 Contents of isoelectric variants of S1 00A3 (%) I ' I ' ' I 225 5.0 I 100 Figure 4. Proportion of various isoelectric variants of S l00A3 in bleached hair upon 2D PAGE analysis. The upper panel shows the shift of S 1 00A3 protein oxidized by 1 % hydrogen peroxide to the lower pl. Each spot is at indicated pl. The lower panel shows the acidic pl shift of S 100A3 spots in an H 2 02 concentration­ dependent manner. the range of the concentration of hydrogen peroxide needed for cuticle delamination and its fragmentation into small peptides. We postulate that this molecular transformation represents whole cuticle oxidation, as most cuticle proteins, including S100A3, contain a higher portion of cysteines (12). Although most oxidized proteins were retained in cuticles even if some of the disulfide bridges were converted into cysteic acid, we postulate that the oxidized cuticles become more fragile due to the cleavage of disulfide bridges between cuticle proteins. A decrease in intermolecular crosslinks by bleaching treatment may result in the fragmentation of the cuticles into smaller peptides. Our advanced model now suggests that the individual cuticles exhibit distinct physical properties, namely cuticle durability against mechanical stress, depending on the expe­ rienced hair treatment chemicals. This will enable us to improve our hair-care products suitable for each hair condition.

226 JOURNAL OF COSMETIC SCIENCE Perm I' Washing Bleach� L Figure 5. Advanced model encompasses hair damage mechanisms induced by both perming and bleaching treatments. This model starts from the "actual cuticle" chipped away at its edge in the second stage of our previous hair damage model (5 ). REFERENCES (1) I. Marenholz, C. W. Heizmann, and G. Fritz, Sl00 proteins in mouse and man: From evolution to function and pathology (including an update of the nomenclature), Biochem. Biophys. Res. Commun., 322, 1111-1122 (2004). (2) K. Kizawa, H. Uchiwa, and U. Murakami, Highly expressed SlO0A3, a calcium-binding protein, in human hair cuticle, Biochim. Biophys. Acta, 1312, 94-98 (1996). (3) K. Kizawa, H. Troxler, P. Kleinert, T. Inoue, M. Toyoda, M. Morohashi, and C. W. Heizmann, Characterization of the cysteine-rich calcium-binding S 100A3 protein from human hair cuticles, Biochem. Biophys. Res. Commun., 299, 857-862 (2002). (4) T. Takizawa, T. Takizawa, S. Arai, K. Kizawa, H. Uchiwa, I, Sasaki, and T. Inoue, Ultrastructural localization of SlO0A3, a cysteine-rich, calcium binding protein, in human hair shafts revealed by rapid-freezing immunocytochemistry,J. Histochem. Cytochem., 47, 525-532 (1999). (5) T. Inoue, I. Sasaki, M. Yamaguchi, and K. Kizawa, Elution of Sl00A3 from hair fiber: New model for hair damage emphasizing the loss of S100A3 from cuticle, J. Cosmet. Sci., 51, 15-25 (2000). (6) J. A. Swift and B. Bews, The chemistry of human hair cuticle. I. A new method for the physical isolation of cuticle,]. Soc. Cosmet. Chem., 25, 13-12 (1974). (7) S. S. Sandhu and C. R. Robbins, A simple and sensitive technique, based on protein loss measurement, to assess surface damage to human hair,]. Soc. Cosmet. Chem., 44, 163-175 (1993). (8) M. M. Bradford, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding, Anal. Biochem., 72, 248-254 (1976). (9) U. G. Fohr, C. W. Heizmann, D. Engelkamp, B. W. Schafer, and J. A. Cox, Purification and cation binding properties of the recombinant human S 100 calcium-binding protein A3, an EF-hand motif protein with high affinity for zinc,]. Biol. Chem., 270, 21056-21061 (1995). (10) G. Fritz, C. W. Heizmann, and P. M. Kroneck, Probing the structure of the human Ca2 + - and Zn 2 + -binding protein S 1 00A3: Spectroscopic investigations of its transition metal ion complexes, and three-dimensional structural model, Biochim. Biophys. Acta, 1448, 264-276, (1998). (11) H.J. Ahn and W. S. Lee, An ultrastructural study of hair fiber damage and restoration following treatment with permanent hair dye, Int.]. Dermatol., 41, 88-92 (2002). (12) M.A. Rogers, L. Langbein, H. Winter, I. Beckman, S. Praetzel, and J. Schweizer, Hair keratin associated proteins: Characterization of a second high sulfur KAP gene domain on human chromosome 21,J. Invest. Dermatol., 122, 147-158 (2003).