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Cholesterol Sulfate Inhibits Proteases that are Involved in Desquamation of Stratum Corneum  Junko Sato, Mitsuhiro Denda, Jotaro Nakanishi, Junko Nomura,

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Presentation on theme: "Cholesterol Sulfate Inhibits Proteases that are Involved in Desquamation of Stratum Corneum  Junko Sato, Mitsuhiro Denda, Jotaro Nakanishi, Junko Nomura,"— Presentation transcript:

1 Cholesterol Sulfate Inhibits Proteases that are Involved in Desquamation of Stratum Corneum 
Junko Sato, Mitsuhiro Denda, Jotaro Nakanishi, Junko Nomura, Junichi Koyama  Journal of Investigative Dermatology  Volume 111, Issue 2, Pages (August 1998) DOI: /j x Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 Increasing the abnormal scales on the backs of mice after topical application of CS. Skin surface appearance of a normal vehicle-treated control mouse (a) versus a cholesterol sulfate-treated mouse (b). Animals were treated with 80 μl daily for 3 d. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 Increased thickness of the stratum corneum layer from mice treated with CS. (a) The stratum corneum of vehicle-treated control. (b) The stratum corneum from animals treated with 80 μl cholesterol sulfate (10 mM) daily for 3 d. Scale bars: 5 μm. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 Application of cholesterol sulfate resulted in an increased number of stratum corneum layers as compared with vehicle treatment (n = 6, mean ± SD). Cholesterol sulfate versus control, p < 0.01. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 The level of anti-DG I-reactive protein in cholesterol sulfate-treated mice was higher than that of vehicle-treated mice (n = 3, mean ± SD). Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

6 Figure 5 CS inhibited cell dissociation from the stratum cornuem sheet. The stratum corneum sheet (1 mg) was incubated in a detegent mixture for 24 h at 37°C with cholsterol sulfate (n = 3, mean ± SD). Significant differences: 100 μM cholesterol sulfate versus control, p < 0.05; 1 mM cholesterol sulfate versus control, p < 0.05. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

7 Figure 6 CS inhibited trypsin in the enzyme assay systemin vitro. Two micromoles of CS, 2 μM of phosphatidylcholine, 2 μM of palmitic acid, 2 μM of cholesterol, and 2 μM of tauroric acid were examined for inhibitory activity using porcin pancreatic trypsin (n = 3, mean ± SEM). Significant difference: cholesterol sulfate versus control, p < 0.05. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

8 Figure 7 Cholesterol sulfate is a competetive inhibitor of trypsin. (a) Lineweaver–Burk plot of the inhibition of trypsin by cholesterol sulfate (▵, 3 × 10–7 M;□, 2 × 10–7 M);○, no cholesterol sulfate. (b) Dixon plot of trypsin by cholesterol sulfate (○, 2.0 × 10–5 M 3107-V;▵, 1.0 × 10–5 M 3107-V;□, 5.0 × 10–6 M 3107-V; +, 2.5 × 10–6 M 3107-V). Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

9 Figure 8 Cholesterol sulfate is a competitive inhibitor of chymotrypsin. (a) Lineweaver–Burk plot of the inhibition of chymotrypsin by cholesterol sulfate (▵, 2.98× 10–6 M;□, 2.23 × 10–6 M);○, no cholesterol sulfate. (b) Dixon plot of chymotrypsin by cholesterol sulfate (○, 1.21 × 10–4 M 3120-V;▵, 6.06 × 10–5 M 3120-V;□, 3.03 × 10–5 M 3120-V). Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

10 Figure 9 Increasing the abnormal scales on the backs of mice after topical application of chymostatin and leupeptin. Skin surface appearance of a normal vehicle-treated control mouse (a-1) versus a chymostatin and leupeptin treated mouse (a-2) and its binary images of a normal vehicle-treated control mouse (b-1) versus a chymostatin and leupeptin treated mouse (b-2). Scales were assessed by measurement of its area using a binary image and were quantitated by NIH image. Increased scales on mice by topical application of chymostatin and leupeptin in comparison with that from vehicle-treated mice (c) (n = 4, mean ± SD). Cholesterol sulfate versus control, p < 0.05. Journal of Investigative Dermatology  , DOI: ( /j x) Copyright © 1998 The Society for Investigative Dermatology, Inc Terms and Conditions

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