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Paraphenylenediamine Sensitivity
PARAPHENYLENEDIAMINE SENSITIVITY COMPARATIVE PATCH TEST REACTIVENESS OF THE ORTHO, META AND PARA ISOMERS
L. EDWARD GAUL, M.D.
In 1944 Rostenberg and Kanof (1) investi- Case 1 showed a patch test reaction to PPD, gated the extent and direction the 2:4 dinitro- characterized by a button-like induration and chlorobenzene molecule could be changed and flaring edema. It required about 4 weeks to heal. still produce reactions in already sensitized per- Case 2 and 3 displayed the button-like indurasons. They found that the number of reactors tion, but no flaring edema. The healing time was followed closely the resemblance of the isomers to the primary sensitizing structure. Of course,
the 2:4 gave the greatest number of reactions, then the 2:5 and the 3:4. The isomer which deviated the most and gave the least number of reactions was 3:5. Rothman, Orland and Flesch (2) in 1945 showed that ortho and meta-procaine were non-reactive in the presence of sensitivity
to para-procaine. In 1952 Rajke (3) noted that ortho-compounds did not react in the presence of
sensitivity to the para-group, and the failure of nupercaine to react was attributed to the fact
less than 2 weeks. The isomers were patch tested
as 1% concentrations in petrolatum. Care was taken to avoid contamination. Before presenting the results, the dermatologic background of the subjects will be reviewed since the level of sensi-
tization had an important bearing on the outcome of the tests.
Case 1, female aged 28, had experienced 9 attacks of dermatitis. The onset occurred in 1949 with a dermatitis from a hair dye. Sulfadiazine induced a drug reaction in 1950, procaine penicillin
that it was not a derivative of para-amino-
in 1950, and dental procaine 2 reactions in 1958. Positive patch tests had been recorded to procaine,
benzoate. Reactions from meta-phenylenediamine were considered due to contamination by the para-isomer. The isomers of 2:4 dinitrochlorobenzene re-
condensation (4). Case 2, female aged 38, was seen for a generalized, patchy, erytheinatous dermatitis which ap-
tained some reactiveness; whereas those of para-
phenylenediamine (PPD) were non-reactive.
2:4 toluene diamine and butyraldehyde aniline
peared first over the breasts. A week before the onset, phthalylsulfathiazol had been administered. She took a total of 6 grams in a 48 hour period. At the age of 19, there was a soft tissue
These findings suggested that certain information might be obtained from patch testing the isomers laceration which required suturing. A sulfonamide of known sensitizers. If all the isomers were re- might have been applied topically. At this time active, then the particular compound might have also a generalized urticaria developed from tetaa broader spectrum of sensitivity; and if only one nus immunization. There had been no previous
sulfonamide administration to her knowledge. There had been one injection of penicillin seven years ago with no reaction. Dental procaine was used 14 years ago with no reaction. There was a reaction to 2:4 toluene diamine but not to phthasensitizing structure of a compound and knowing lylsulfathiazol by patch test. this, more precise information should be available Case 3, female aged 50, had a hand dermatitis for immunochemical research. Finally, a better for 6 months. For many years, spectacle bows and understanding of the reactivity of isomers might wrist watches had caused itching and "bumps" to of the isomers was reactive, then the spectrum might be narrower, perhaps limited to the derivatives of the particular isomer. Patch testing isomers should also disclose the limits of the
be valuable for interpreting the direction of cross-
sensitization. Paraphenylenediamine lent itself well to this undertaking because of the availability of its isomers and the prevalence of this type of sensitivity. Three subjects comprised the clinical material.
appear at the sites of contact. In 1955 a parenteral procaine penicillin caused local redness with pain and tenderness at the injection site. There was an
associated sensitivity to nickel and 2:4 toluene diamine. Procaine had been tolerated. She did not recall taking sulfonamides. The hand dermatitis was traced to a nickel plated cigarette lighter and case.
Received for publication February 25, 1960. 231
232
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
* FILTER PAPER SQI1Ag CoNrAer
FIG. 1
ortho was non-reactive. The healing time of the reactions revealed still another difference. It was The patch tests were removed in approximately 20 days in case 1, 3 days in case 2 and less than 24 hours. Case 1 was tested with para, meta and one day in case 3. ortho isomers. Cases 2 and 3 were tested with only the meta and ortho. The reactiveness of the COLOR CHANGES OF THE ISOMERS isomers is graphed in Figure I. The severity of The presence of a PPD test on the Elastoplast the para reaction in ease 1 is shown. Itching was intense. A day later, the meta reaction appeared, patch discolors the skin. This is a valuable marker very mild compared to the para. There was no for delayed reactions and to identify other tests itching. Some 30 hours later, the ortho reaction for comparing degree and healing time. For some developed, even less intense than the meta. Two time in PPD sensitivity, the degree of blackness days later, all the reactions had crested. Now has been watched. It was thought that the skin another difference was evident. A flaring, non- contained a substance, perhaps an enzyme, that inflammatory edema featured the para. Its selectively affected and changed its color. The broadest spread was 5 centimeters. As the flaring color changes of the isomers of PPD are correlated stabilized, itching lessened and became nil. The with their patch test results in Table I. The tested meta showed only barest flaring edema and the patch contained the usual paper squares with a ortho none at all. Case 2 developed an erythro- dab of test material in the center. The untested edema to the meta isomer and no reaction to the patch was prepared in the same way and left exortho. Case 3 showed a plus-minus reaction to the posed in the air. The pam patch test was jet black. The test in meta and this vanished the following day. The PATCH TEST RESULTS WITH ISOMERS
233
PARAPHENYLENEDIAMINE SENSITIVITY
TABLE
I
Color changes in isomers of phenylenediamine
Color Changes
Tested patch Para* light gray Meta*
black
black
black
black
black
yellow
yellow
yellow
yellow
yellow
yellow
yellow
yellow
yellow
yellow
light tan Ortho*
light tan Test patch in air Para Meta Ortho
Days
(faint black specks)
blue
blue
blue
blue (still darker)
blue
yellow yellow
(darker) yellow yellow
yellow yellow
brown yellow
brown yellow
1
5
10
15
20
* Color in petrolatum.
air remained blue. The skin effected a rapid and complete color change in the para isomer. The meta and ortho isomers showed essentially the same color changes. The skin was without influence. Their color changes were not associated with antigenicity. The presence of black dots on the meta test paper and a few black specks on the ortho test paper, and the absence of these color changes in the air test, points to contamination of the meta and ortho isomers by the para. Fieser
(4) notes that ortho-quinones of the benzene series have oxidation-reduction potentials higher
ness detected contamination from the para isomer
and indirectly served as a most critical test for the lack of reactiveness of the meta and ortho isomers. The dye chemist might find a patch test
highly specific for contamination of isomers. Cases 2 and 3 with much lower levels of skin reactiveness substantiated the conclusions in case 1. The ortho isomer was non-reactive in cases 2
and 3. SUMMARY
The ortho and meta isomers of phenylenedliaquinones, which means that ortho-phenylenedia- mine were found to be nonreactive in the presmine is less easily oxidized than para-phenylene- ence of para-phenylenediamine sensitivity. Antidiamine, and thus is less likely to form colored genicity rests in the para-isomer. Color changes, ortho - quinone oxidation products. Meta- or the pathways of oxidation and reduction taken phenylenediamine does not normally form qui- by the isomers of phenylenediamine, help to exnone structures. Mayer (5) has emphasized many plain the presence or absence of antigenicity. times that in para-phenylenediamine sensitivity, Information on the patch test reactiveness of
by 85—95 millivolts than the isomeric para
antigenicity rests in the quinone structure. The color changes help to explain the patch test results in Figure I. Recall that case 1 had had multiple drug and dermatitic reactions, also "caine" sensitivity, which had produced a high level of skin reactiveness. The para isomer pro-
voked an exaggerated reaction, one unduly severe. The meta and ortho isomers contained sufficient para to induce a delayed and much less intense response. The high level of skin reactive-
chemical isomers could lead to a better understanding of immunochemistry and cross-sensitization.
REFERENCES 1. Rosrsnnmu, A. AND KANOF, N. M.: Studies on
eczematous sensitization. J. Invest. Dermat., 6: 201, 1945.
2. ROTRMAN, S., ORLAND, F. J. AND FLESCR, P.:
Group specificity of epidermal allergy to procaine. J. Invest. Dermat., 6: 191, 1945.
234
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
3. RAJKA, G.: On hypersensitivity to the "paragroup". Acta Allergologica, 5: 11, 1952. 4. FIESER, L. F. AND FrasER, Mary: "Organic Chemistry," 3rd ed., p. 712. Reinhold, N. Y., 1956.
5. MAYER, R. L.: Discussion. Fisher, A. A., Pelzig,
A. and Kanof, N. B.: Persistence of allergic eczematous sensitivity and the cross-sensi-
tivity pattern to paraphenylenediamine. J. Invest. Dermat., 30: 9, 1958.
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.
L. EDWARD GAUL, M.D.
In 1944 Rostenberg and Kanof (1) investi- Case 1 showed a patch test reaction to PPD, gated the extent and direction the 2:4 dinitro- characterized by a button-like induration and chlorobenzene molecule could be changed and flaring edema. It required about 4 weeks to heal. still produce reactions in already sensitized per- Case 2 and 3 displayed the button-like indurasons. They found that the number of reactors tion, but no flaring edema. The healing time was followed closely the resemblance of the isomers to the primary sensitizing structure. Of course,
the 2:4 gave the greatest number of reactions, then the 2:5 and the 3:4. The isomer which deviated the most and gave the least number of reactions was 3:5. Rothman, Orland and Flesch (2) in 1945 showed that ortho and meta-procaine were non-reactive in the presence of sensitivity
to para-procaine. In 1952 Rajke (3) noted that ortho-compounds did not react in the presence of
sensitivity to the para-group, and the failure of nupercaine to react was attributed to the fact
less than 2 weeks. The isomers were patch tested
as 1% concentrations in petrolatum. Care was taken to avoid contamination. Before presenting the results, the dermatologic background of the subjects will be reviewed since the level of sensi-
tization had an important bearing on the outcome of the tests.
Case 1, female aged 28, had experienced 9 attacks of dermatitis. The onset occurred in 1949 with a dermatitis from a hair dye. Sulfadiazine induced a drug reaction in 1950, procaine penicillin
that it was not a derivative of para-amino-
in 1950, and dental procaine 2 reactions in 1958. Positive patch tests had been recorded to procaine,
benzoate. Reactions from meta-phenylenediamine were considered due to contamination by the para-isomer. The isomers of 2:4 dinitrochlorobenzene re-
condensation (4). Case 2, female aged 38, was seen for a generalized, patchy, erytheinatous dermatitis which ap-
tained some reactiveness; whereas those of para-
phenylenediamine (PPD) were non-reactive.
2:4 toluene diamine and butyraldehyde aniline
peared first over the breasts. A week before the onset, phthalylsulfathiazol had been administered. She took a total of 6 grams in a 48 hour period. At the age of 19, there was a soft tissue
These findings suggested that certain information might be obtained from patch testing the isomers laceration which required suturing. A sulfonamide of known sensitizers. If all the isomers were re- might have been applied topically. At this time active, then the particular compound might have also a generalized urticaria developed from tetaa broader spectrum of sensitivity; and if only one nus immunization. There had been no previous
sulfonamide administration to her knowledge. There had been one injection of penicillin seven years ago with no reaction. Dental procaine was used 14 years ago with no reaction. There was a reaction to 2:4 toluene diamine but not to phthasensitizing structure of a compound and knowing lylsulfathiazol by patch test. this, more precise information should be available Case 3, female aged 50, had a hand dermatitis for immunochemical research. Finally, a better for 6 months. For many years, spectacle bows and understanding of the reactivity of isomers might wrist watches had caused itching and "bumps" to of the isomers was reactive, then the spectrum might be narrower, perhaps limited to the derivatives of the particular isomer. Patch testing isomers should also disclose the limits of the
be valuable for interpreting the direction of cross-
sensitization. Paraphenylenediamine lent itself well to this undertaking because of the availability of its isomers and the prevalence of this type of sensitivity. Three subjects comprised the clinical material.
appear at the sites of contact. In 1955 a parenteral procaine penicillin caused local redness with pain and tenderness at the injection site. There was an
associated sensitivity to nickel and 2:4 toluene diamine. Procaine had been tolerated. She did not recall taking sulfonamides. The hand dermatitis was traced to a nickel plated cigarette lighter and case.
Received for publication February 25, 1960. 231
232
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
* FILTER PAPER SQI1Ag CoNrAer
FIG. 1
ortho was non-reactive. The healing time of the reactions revealed still another difference. It was The patch tests were removed in approximately 20 days in case 1, 3 days in case 2 and less than 24 hours. Case 1 was tested with para, meta and one day in case 3. ortho isomers. Cases 2 and 3 were tested with only the meta and ortho. The reactiveness of the COLOR CHANGES OF THE ISOMERS isomers is graphed in Figure I. The severity of The presence of a PPD test on the Elastoplast the para reaction in ease 1 is shown. Itching was intense. A day later, the meta reaction appeared, patch discolors the skin. This is a valuable marker very mild compared to the para. There was no for delayed reactions and to identify other tests itching. Some 30 hours later, the ortho reaction for comparing degree and healing time. For some developed, even less intense than the meta. Two time in PPD sensitivity, the degree of blackness days later, all the reactions had crested. Now has been watched. It was thought that the skin another difference was evident. A flaring, non- contained a substance, perhaps an enzyme, that inflammatory edema featured the para. Its selectively affected and changed its color. The broadest spread was 5 centimeters. As the flaring color changes of the isomers of PPD are correlated stabilized, itching lessened and became nil. The with their patch test results in Table I. The tested meta showed only barest flaring edema and the patch contained the usual paper squares with a ortho none at all. Case 2 developed an erythro- dab of test material in the center. The untested edema to the meta isomer and no reaction to the patch was prepared in the same way and left exortho. Case 3 showed a plus-minus reaction to the posed in the air. The pam patch test was jet black. The test in meta and this vanished the following day. The PATCH TEST RESULTS WITH ISOMERS
233
PARAPHENYLENEDIAMINE SENSITIVITY
TABLE
I
Color changes in isomers of phenylenediamine
Color Changes
Tested patch Para* light gray Meta*
black
black
black
black
black
yellow
yellow
yellow
yellow
yellow
yellow
yellow
yellow
yellow
yellow
light tan Ortho*
light tan Test patch in air Para Meta Ortho
Days
(faint black specks)
blue
blue
blue
blue (still darker)
blue
yellow yellow
(darker) yellow yellow
yellow yellow
brown yellow
brown yellow
1
5
10
15
20
* Color in petrolatum.
air remained blue. The skin effected a rapid and complete color change in the para isomer. The meta and ortho isomers showed essentially the same color changes. The skin was without influence. Their color changes were not associated with antigenicity. The presence of black dots on the meta test paper and a few black specks on the ortho test paper, and the absence of these color changes in the air test, points to contamination of the meta and ortho isomers by the para. Fieser
(4) notes that ortho-quinones of the benzene series have oxidation-reduction potentials higher
ness detected contamination from the para isomer
and indirectly served as a most critical test for the lack of reactiveness of the meta and ortho isomers. The dye chemist might find a patch test
highly specific for contamination of isomers. Cases 2 and 3 with much lower levels of skin reactiveness substantiated the conclusions in case 1. The ortho isomer was non-reactive in cases 2
and 3. SUMMARY
The ortho and meta isomers of phenylenedliaquinones, which means that ortho-phenylenedia- mine were found to be nonreactive in the presmine is less easily oxidized than para-phenylene- ence of para-phenylenediamine sensitivity. Antidiamine, and thus is less likely to form colored genicity rests in the para-isomer. Color changes, ortho - quinone oxidation products. Meta- or the pathways of oxidation and reduction taken phenylenediamine does not normally form qui- by the isomers of phenylenediamine, help to exnone structures. Mayer (5) has emphasized many plain the presence or absence of antigenicity. times that in para-phenylenediamine sensitivity, Information on the patch test reactiveness of
by 85—95 millivolts than the isomeric para
antigenicity rests in the quinone structure. The color changes help to explain the patch test results in Figure I. Recall that case 1 had had multiple drug and dermatitic reactions, also "caine" sensitivity, which had produced a high level of skin reactiveness. The para isomer pro-
voked an exaggerated reaction, one unduly severe. The meta and ortho isomers contained sufficient para to induce a delayed and much less intense response. The high level of skin reactive-
chemical isomers could lead to a better understanding of immunochemistry and cross-sensitization.
REFERENCES 1. Rosrsnnmu, A. AND KANOF, N. M.: Studies on
eczematous sensitization. J. Invest. Dermat., 6: 201, 1945.
2. ROTRMAN, S., ORLAND, F. J. AND FLESCR, P.:
Group specificity of epidermal allergy to procaine. J. Invest. Dermat., 6: 191, 1945.
234
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
3. RAJKA, G.: On hypersensitivity to the "paragroup". Acta Allergologica, 5: 11, 1952. 4. FIESER, L. F. AND FrasER, Mary: "Organic Chemistry," 3rd ed., p. 712. Reinhold, N. Y., 1956.
5. MAYER, R. L.: Discussion. Fisher, A. A., Pelzig,
A. and Kanof, N. B.: Persistence of allergic eczematous sensitivity and the cross-sensi-
tivity pattern to paraphenylenediamine. J. Invest. Dermat., 30: 9, 1958.
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.