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Amount of Ascorbic Acid in Normal Epidermis1
PRELIMINARY AND SHORT REPORT AMOUNT OF ASCORBIC ACID IN NORMAL EPIDERMIS* JOSEPH 0. PRIE5TLEY, M.D.t AND RoscoE C. FOSTER, B.S.t
In undertaking investigations of the function of
grams of skin (wet weight). At that time no satisfactory method was available for separating epidermis from corium without some chemical alteration (1).
ascorbic acid in the skin no reference could be found in the literature concerning the amount of ascorbic acid in epidermis. Jensen and Poulsen in 1942 made an extensive study of the concentration of ascorbic acid in 181 specimens of skin obtained by punch biopsy from 72 persons. They extracted the tissue with 8% mctaphosphoric acid solution
Since then Van Scott has demonstrated that epidermis can be stripped from tbe corium after the skin has been stretched (2). METHOD
TABLE I
Using the same methods described above, ascorbic acid determinations were done in 10 specimens Ascorbic Acid, mg.
Area
Cause of Amputatioo
of normal skin obtained from surgical amputations. The skin had been prepared for surgery by scrubbing with soap and water and then washing
per 100gm. wet weight
Epi- Co- whole dermis rium Skio
Breast Breast Leg Leg
1
2 3 4
Breast Leg
5
6 7 8 9* 10
Breast Breast Breast Breast
with zcphiran chloride and alcohol. In cases where
immediate determinations could not be done the skin was stored at —20° C. All determinations were done within twenty-four hours after surgery.
Carcinoma 8.35 3.48 Carcinoma 8.26 1.00 Sarcoma of bone 4.94 0.18 Arterial throm- 6.64 3.37 bosis Carcinoma 6.63 4.98 6.89 3.08 Diabetic gangrene Carcinoma 12.80 6.40 Carcinoma 5.41 3.23 Carcinoma 11.31 4.25 7.30 Carcinoma 13.40 7.00 9.90
Subcutaneous tissue was removed from the corium
by dissection, and the epidermis was separated from the corium by the Van Scott stretch method.
Each specimen of tissue was extracted with 8% metaphosphoric acid solution, the liquid portion decanted and centrifuged, and the supernatant titrated with a standardized solution of 2, 6-dichlorophenol indophcnol.
The amount of ascorbic acid found in the epi-
dermis and corium specimens are listed in Table I.
* Negro
5UMMART
The amount of ascorbic acid in the epidermis and corium of ten specimens of normal skin was determined. The amount of ascorbic acid found in
and titrated this extract with a solution of 2,6-
dichlorophcnol indophcnol which had been standardized with an aqueous solution of ascorbic acid of known concentration. In 95% of the specimens of normal skin they found concentrations of
the 10 specimens of normal epidermis varied from 5.41 to 13.40 mg. per 100 gm. wet weight, and all
specimens consistently showed much more ascorbic acid in the epidermis than in the corium.
ascorbic acid varying from 0 to 10 mg. per 100 * From the Section of Dermatology, Depart-
REFERENCES
ment of Medicine, University of Chicago, Chicago,
Illinois. (Chief of Service: Stephen Rothman,
1. JENSEN, T. AND POUL5EN, E.: On the ascorbic
acid content of normal and pathological skin. Acta Dcrmatovenereologica. Vol. xxii 241.
M.D.) Present address: 1403—1407 Exchange Building, Memphis 3, Tenn. t Present address: Howard University Dental School, Washington, D. C. Received for publication August 25, 1958.
1942.
2. VAN SCOTT, E. J.: Mechanical separation of the
epidermis from the corium. J. Invest. Dermat., 18: 377, 1952.
3
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
94
linolenic acid extract. Arch. This pdf is a scanned copy UV of irradiated a printed document.
24. Wynn, C. H. and Iqbal, M.: Isolation of rat
skin lysosomes and a comparison with liver Path., 80: 91, 1965. and spleen lysosomes. Biochem. J., 98: lOP, 37. Nicolaides, N.: Lipids, membranes, and the 1966.
human epidermis, p. 511, The Epidermis
Eds., Montagna, W. and Lobitz, W. C. Acascopic localization of acid phosphatase in demic Press, New York. human epidermis. J. Invest. Derm., 46: 431, 38. Wills, E. D. and Wilkinson, A. E.: Release of 1966. enzymes from lysosomes by irradiation and 26. Rowden, C.: Ultrastructural studies of kerathe relation of lipid peroxide formation to tinized epithelia of the mouse. I. Combined enzyme release. Biochem. J., 99: 657, 1966. electron microscope and cytochemical study 39. Lane, N. I. and Novikoff, A. B.: Effects of of lysosomes in mouse epidermis and esoarginine deprivation, ultraviolet radiation and X-radiation on cultured KB cells. J. phageal epithelium. J. Invest. Derm., 49: 181, 25. Olson, R. L. and Nordquist, R. E.: Ultramicro-
No warranty is given about the accuracy of the copy.
Users should refer to the original published dermal cells. Nature, 216: 1031, 1967. version of1965. the material. vest. Derm., 45: 448, 28. Hall, J. H., Smith, J. G., Jr. and Burnett, S. 41. Daniels, F., Jr. and Johnson, B. E.: In prepa1967.
Cell Biol., 27: 603, 1965.
27. Prose, P. H., Sedlis, E. and Bigelow, M.: The 40. Fukuyama, K., Epstein, W. L. and Epstein, demonstration of lysosomes in the diseased J. H.: Effect of ultraviolet light on RNA skin of infants with infantile eczema. J. Inand protein synthesis in differentiated epi-
C.: The lysosome in contact dermatitis: A ration. histochemical study. J. Invest. Derm., 49: 42. Ito, M.: Histochemical investigations of Unna's oxygen and reduction areas by means of 590, 1967. 29. Pearse, A. C. E.: p. 882, Histochemistry Theoultraviolet irradiation, Studies on Melanin, retical and Applied, 2nd ed., Churchill, London, 1960.
30. Pearse, A. C. E.: p. 910, Histacheini.stry Thearetscal and Applied, 2nd ed., Churchill, London, 1960.
31. Daniels, F., Jr., Brophy, D. and Lobitz, W. C.: Histochemical responses of human skin fol-
lowing ultraviolet irradiation. J. Invest. Derm.,37: 351, 1961.
32. Bitensky, L.: The demonstration of lysosomes by the controlled temperature freezing section method. Quart. J. Micr. Sci., 103: 205, 1952.
33. Diengdoh, J. V.: The demonstration of lysosomes in mouse skin. Quart. J. Micr. Sci., 105: 73, 1964.
34. Jarret, A., Spearman, R. I. C. and Hardy, J. A.:
Tohoku, J. Exp. Med., 65: Supplement V, 10, 1957.
43. Bitcnsky, L.: Lysosomes in normal and pathological cells, pp. 362—375, Lysasames Eds., de Reuck, A. V. S. and Cameron, M. Churchill, London, 1953.
44. Janoff, A. and Zweifach, B. W.: Production of inflammatory changes in the microcirculation by cationic proteins extracted from lysosomes. J. Exp. Med., 120: 747, 1964.
45. Herion, J. C., Spitznagel, J. K., Walker, R. I. and Zeya, H. I.: Pyrogenicity of granulocyte lysosomes. Amer. J. Physiol., 211: 693, 1966.
46. Baden, H. P. and Pearlman, C.: The effect of ultraviolet light on protein and nucleic acid synthesis in the epidermis. J. Invest. Derm.,
Histochemistry of keratinization. Brit. J. 43: 71, 1964. Derm., 71: 277, 1959. 35. De Duve, C. and Wattiaux, R.: Functions of 47. Bullough, W. S. and Laurence, E. B.: Mitotic control by internal secretion: the role of lysosomes. Ann. Rev. Physiol., 28: 435, 1966. the chalone-adrenalin complex. Exp. Cell. 36. Waravdekar, V. S., Saclaw, L. D., Jones, W. A. and Kuhns, J. C.: Skin changes induced by
Res., 33: 176, 1964.
In undertaking investigations of the function of
grams of skin (wet weight). At that time no satisfactory method was available for separating epidermis from corium without some chemical alteration (1).
ascorbic acid in the skin no reference could be found in the literature concerning the amount of ascorbic acid in epidermis. Jensen and Poulsen in 1942 made an extensive study of the concentration of ascorbic acid in 181 specimens of skin obtained by punch biopsy from 72 persons. They extracted the tissue with 8% mctaphosphoric acid solution
Since then Van Scott has demonstrated that epidermis can be stripped from tbe corium after the skin has been stretched (2). METHOD
TABLE I
Using the same methods described above, ascorbic acid determinations were done in 10 specimens Ascorbic Acid, mg.
Area
Cause of Amputatioo
of normal skin obtained from surgical amputations. The skin had been prepared for surgery by scrubbing with soap and water and then washing
per 100gm. wet weight
Epi- Co- whole dermis rium Skio
Breast Breast Leg Leg
1
2 3 4
Breast Leg
5
6 7 8 9* 10
Breast Breast Breast Breast
with zcphiran chloride and alcohol. In cases where
immediate determinations could not be done the skin was stored at —20° C. All determinations were done within twenty-four hours after surgery.
Carcinoma 8.35 3.48 Carcinoma 8.26 1.00 Sarcoma of bone 4.94 0.18 Arterial throm- 6.64 3.37 bosis Carcinoma 6.63 4.98 6.89 3.08 Diabetic gangrene Carcinoma 12.80 6.40 Carcinoma 5.41 3.23 Carcinoma 11.31 4.25 7.30 Carcinoma 13.40 7.00 9.90
Subcutaneous tissue was removed from the corium
by dissection, and the epidermis was separated from the corium by the Van Scott stretch method.
Each specimen of tissue was extracted with 8% metaphosphoric acid solution, the liquid portion decanted and centrifuged, and the supernatant titrated with a standardized solution of 2, 6-dichlorophenol indophcnol.
The amount of ascorbic acid found in the epi-
dermis and corium specimens are listed in Table I.
* Negro
5UMMART
The amount of ascorbic acid in the epidermis and corium of ten specimens of normal skin was determined. The amount of ascorbic acid found in
and titrated this extract with a solution of 2,6-
dichlorophcnol indophcnol which had been standardized with an aqueous solution of ascorbic acid of known concentration. In 95% of the specimens of normal skin they found concentrations of
the 10 specimens of normal epidermis varied from 5.41 to 13.40 mg. per 100 gm. wet weight, and all
specimens consistently showed much more ascorbic acid in the epidermis than in the corium.
ascorbic acid varying from 0 to 10 mg. per 100 * From the Section of Dermatology, Depart-
REFERENCES
ment of Medicine, University of Chicago, Chicago,
Illinois. (Chief of Service: Stephen Rothman,
1. JENSEN, T. AND POUL5EN, E.: On the ascorbic
acid content of normal and pathological skin. Acta Dcrmatovenereologica. Vol. xxii 241.
M.D.) Present address: 1403—1407 Exchange Building, Memphis 3, Tenn. t Present address: Howard University Dental School, Washington, D. C. Received for publication August 25, 1958.
1942.
2. VAN SCOTT, E. J.: Mechanical separation of the
epidermis from the corium. J. Invest. Dermat., 18: 377, 1952.
3
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
94
linolenic acid extract. Arch. This pdf is a scanned copy UV of irradiated a printed document.
24. Wynn, C. H. and Iqbal, M.: Isolation of rat
skin lysosomes and a comparison with liver Path., 80: 91, 1965. and spleen lysosomes. Biochem. J., 98: lOP, 37. Nicolaides, N.: Lipids, membranes, and the 1966.
human epidermis, p. 511, The Epidermis
Eds., Montagna, W. and Lobitz, W. C. Acascopic localization of acid phosphatase in demic Press, New York. human epidermis. J. Invest. Derm., 46: 431, 38. Wills, E. D. and Wilkinson, A. E.: Release of 1966. enzymes from lysosomes by irradiation and 26. Rowden, C.: Ultrastructural studies of kerathe relation of lipid peroxide formation to tinized epithelia of the mouse. I. Combined enzyme release. Biochem. J., 99: 657, 1966. electron microscope and cytochemical study 39. Lane, N. I. and Novikoff, A. B.: Effects of of lysosomes in mouse epidermis and esoarginine deprivation, ultraviolet radiation and X-radiation on cultured KB cells. J. phageal epithelium. J. Invest. Derm., 49: 181, 25. Olson, R. L. and Nordquist, R. E.: Ultramicro-
No warranty is given about the accuracy of the copy.
Users should refer to the original published dermal cells. Nature, 216: 1031, 1967. version of1965. the material. vest. Derm., 45: 448, 28. Hall, J. H., Smith, J. G., Jr. and Burnett, S. 41. Daniels, F., Jr. and Johnson, B. E.: In prepa1967.
Cell Biol., 27: 603, 1965.
27. Prose, P. H., Sedlis, E. and Bigelow, M.: The 40. Fukuyama, K., Epstein, W. L. and Epstein, demonstration of lysosomes in the diseased J. H.: Effect of ultraviolet light on RNA skin of infants with infantile eczema. J. Inand protein synthesis in differentiated epi-
C.: The lysosome in contact dermatitis: A ration. histochemical study. J. Invest. Derm., 49: 42. Ito, M.: Histochemical investigations of Unna's oxygen and reduction areas by means of 590, 1967. 29. Pearse, A. C. E.: p. 882, Histochemistry Theoultraviolet irradiation, Studies on Melanin, retical and Applied, 2nd ed., Churchill, London, 1960.
30. Pearse, A. C. E.: p. 910, Histacheini.stry Thearetscal and Applied, 2nd ed., Churchill, London, 1960.
31. Daniels, F., Jr., Brophy, D. and Lobitz, W. C.: Histochemical responses of human skin fol-
lowing ultraviolet irradiation. J. Invest. Derm.,37: 351, 1961.
32. Bitensky, L.: The demonstration of lysosomes by the controlled temperature freezing section method. Quart. J. Micr. Sci., 103: 205, 1952.
33. Diengdoh, J. V.: The demonstration of lysosomes in mouse skin. Quart. J. Micr. Sci., 105: 73, 1964.
34. Jarret, A., Spearman, R. I. C. and Hardy, J. A.:
Tohoku, J. Exp. Med., 65: Supplement V, 10, 1957.
43. Bitcnsky, L.: Lysosomes in normal and pathological cells, pp. 362—375, Lysasames Eds., de Reuck, A. V. S. and Cameron, M. Churchill, London, 1953.
44. Janoff, A. and Zweifach, B. W.: Production of inflammatory changes in the microcirculation by cationic proteins extracted from lysosomes. J. Exp. Med., 120: 747, 1964.
45. Herion, J. C., Spitznagel, J. K., Walker, R. I. and Zeya, H. I.: Pyrogenicity of granulocyte lysosomes. Amer. J. Physiol., 211: 693, 1966.
46. Baden, H. P. and Pearlman, C.: The effect of ultraviolet light on protein and nucleic acid synthesis in the epidermis. J. Invest. Derm.,
Histochemistry of keratinization. Brit. J. 43: 71, 1964. Derm., 71: 277, 1959. 35. De Duve, C. and Wattiaux, R.: Functions of 47. Bullough, W. S. and Laurence, E. B.: Mitotic control by internal secretion: the role of lysosomes. Ann. Rev. Physiol., 28: 435, 1966. the chalone-adrenalin complex. Exp. Cell. 36. Waravdekar, V. S., Saclaw, L. D., Jones, W. A. and Kuhns, J. C.: Skin changes induced by
Res., 33: 176, 1964.