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Studies on the Eczematous Sensitization1
STUDIES ON THE ECZEMATOUS SENSITIZATION* II. THE EFFECT OF THE ENGENDERING OF AN ECZEMATOUS SENSITIZATION ON THE THRESHOLD OF REACTION TO PRIMARY IRRITANTS
J. B. HAEBERLIN, JR., M.D., R. M. OLIVER, M.D. AND A. ROSTENBERG, JR., M.D.
A well-known clinical observation is that an existing dermatitis, regardless of type or etiology, may be aggravated or kept activated by contact with irritants, such as soap and water, cleaning solutions, solvents, topical medications, rough
or woolen clothing, and the like. Various theories have been advanced to explain this phenomenon, such as polyvalence of sensitization or lowering the threshold of reaction to primary irritants. The interrelationship between primary irritation and eczematous sensitization is poorly understood, but that primary irritation affects the development of an eczematous sensitization has been definitely established (1, 2). It should be borne in mind that many cc-
zematous sensitizers are also primary irritants. To our knowledge, the influence of the development of an eczematous sensitization on the threshold of reaction to primary irritants has not been studied. The purpose of this study was then to determine the influence, if any, of the genesis of an eczematous sensi-
tization on an individual's threshold of reactivity to primary irritants. METHOD
To answer this question we (1) determined in a series of individuals the threshold irritant dose to various substances by patch tests with graded dilutions of these materials, (2) then in one half of the subjects endeavored to engender an eczematous sensitization, and (3) after a suitable interval both groups were "repatched" with the original substances to determine whether there was any change in the threshold of irritation. The substances chosen as primary irritants were (1) hydrochloric acid, (2) tincture of green soap, and (3) croton oil. Dilutions were made up of the first two in water and of the last in olive oil. After preliminary trials on other individuals, the dilutions were selected so that the most concentrated gave a weakly positive reaction in the majority of individuals tested, whereas, the least concentrated gave a negative in the majority of the individuals tested. This provided a range of dilutions which should make observable a shift in the threshold, should this occur as a result of this experiment. The actual dilutions used are given in table 1. Because of the variability of reactions to testing with primary irritants considerable care was required to make the test applications as uniform as possible. * From the Department of Dermatology and the Allergy Unit of the University of Illinois Colleges of Medicine and Pharmacy. The expenses of this investigation were in part defrayed by the Asthmatic Children's Aid.
Received for publication August 28, 1947. 27
28
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
To this end the chamber patch as described by Rokstad (2) was used with uniform pieces of blotting paper and approximately uniform amounts of test solu-
tion dropped from dropping bottles. It was found that maximum clarity of reading was obtained by removing the patches after approximately 24 hours, and allowing several hours to elapse before reading the tests. In reading the tests, the papule formation as described by Rokstad was ignored. The reactions were of three morphologic types and were recorded according to the code: 1 = erythema corresponding to the circle of the blotting paper, 2 = erythema spreading from the site in a more diffuse fashion, and 3 = follicular pustules over the area. In each type stronger reactions were recorded as "a" and milder reactions as
For the purpose of sensitization 2,4 dinitrochlorobenzene was employed. This chemical was chosen because it is known to be a potent sensitizer and because it is a substance not likely to be encountered by our subjects in ordinary life. The technic employed for sensitizing was the dropping on of approximately 0.03 cc. of a 30 per cent acetone solution on the volar surface of the arm. At TABLE 1 1
2
3
4
5
per cent per cent per cent per cent per cent
HC1
Tincture of green soap Croton oil
1 10
2.5
2.5
5
10
20
30
5
10
40 20
20 50 40
In water In water In olive oil
the same time, approximately 0.03 cc. of a 0.1 per cent acetone solution was applied to the volar surface of the other arm in order to be certain that none of the subjects were already sensitive to this material. Because of the necessity of observing our patients over a reasonably long period of time, and because of the fact that we wished to do as many as possible of them at the same time, we did not believe that ordinary clinic patients would be suitable, and accordingly, inmates of a tuberculosis sanatorium were used.1 RESULTS
Table 2 gives the readings to graded dilutions of primary irritants in eight cases before and after sensitization, and a like number of controls in whom no sensitization was engendered. 1 The fact that the subjects were tuberculous may have affected our results. We had a rather low yield of sensitizations and also the development of the sensitivity seemed slower than our previous experience in the sensitization of non-tuberculous individuals had led
us to expect. We are aware that there is a considerable literature on the interference of a tuberculous infection with the development of other allergic phenomena, but we thought that because of the other reasons mentioned these subjects would nevertheless be most suitable.
29
STUDIES ON ECZEMATOUS SENSITIZATION
TABLE 2 HC1
GREEN SOAP
INITIAL
CROTON OIL
CASE
NO.
TO
12345 i2j345 l2I345
SENSITIZED TO
2,4
RETEST TO
IT
SEESTANTS
Group A. Sensitized to 2,4 dinitro chlorobenzene 0
lb
lb lb is is
0
?
lb lb lb
0 0
0 0
0 0
0 0
is is lb Is la
0 0
0
lb is Is 0 lb lb lb 0
0
0
0
0
0 lb 2b 2b
4/
0
0 0
0
lb lb is
0 0
0 0
0 0
0 0
0 0
0 0
4/14/47
0
0 0
lb
0
Before
0
0
?
lb lb la 0
0
0
lb
0
0
0
0
3b 4/21/47 5/27/47 6/17/47 6/17/47
0
lb lb is 0 lb lb
0
After
?
?
0
0
0
lb
lb
Before
0
0
lb lb
lb
0
lb
3s
0
?
0
is is
2b 2b 3b 3a
0
is lb
is
After
lb
lb
Before After
0 0
0
0 0
Before After
0 0
0
Before After
0
0
After
0 0
0 0
0 0
Before After
0 0
0
lb
Before
After
Before
1
2 3
4
5
6 7
8
0
0
lb 0
0 0
0
7
la
0
is is lb lb
0 0
0
0
0 0
lb 0
0 0
4/ 7/47
Sb 2b 2b
lb lb is 0 lb lb lb
0
Sb
2b 2b
0
0
0
lb
0 lb 0 0 lb lb lb is
0
0
lb lb
lb lb lb 0 0 lb lb 0 2b 0
0 0
Sa
2s 3/31/47
Sb
Sb
0
0
0
0 0
0 0
0 P
0
0
6/ 9/47
6/17/47 6/17/47
5/27/47 6/17/47 6/17/47
4/28/47 5/27/47 6/
0
0
5/27/47 8/ 9/47
7/47 5/27/47
lb 0
lb lb is lb lb Is is is 0 lb -lb 0 0 lb lb lb lb lb is 0 lb lb
3b 2b
9/47
6/
9/47
4/14/47 5/27/47 7/1/47 7/1/47
3b 4/21/47 5/27/47 7/1/47 7/1/47
4/28/47 5/27/47 7/1/47 7/1/47
Group B. Control 9
10
ii 12
13
14
15
16
Initial Final
0 0
0 7
0
?
lb lb 0 lb 1' 0
0
Initisi
0
0
0
0
0
0
0 0
Sb 0
0
lb 0 lb is 0
lb lb
Final
0
0
0
Initial Finsi
0 0
0 0
lb is la 0 0 0 lb 0
0 0
P
is is 0 lb lb 0
Initial
0
0
lb
0
0
0
0
Final
0
0
0
lb is 0
0
Initial Final
0 0
0 0
0
0 0
Initial Final
0 0
0 0
0
Initial Final
0 0
0 0
0
lb
ib 0
Initial Final
0 0
0 0
lb
P
is
0
0
The numbers 1 through
Is
?
0
0 is lb lb lb
is 0
lb is is
P
la 0
0
0
0
lb lb Ia 0 lb is 0
0 0
lb ls is P
lb lb
lb la
7 0
lb
lb lb
0 0
lb lb lb lb lb lb
0,
P
0 0
Ia
0
Sat the tops
0
0
6/9/47
3/31/47
6/ 9/47
Sb
Sb 4/ 7/47
0
6/ 9/47
0
lb lb 0
0 0
lb
is
is
is
0
0
0
0
0 0
0 0
0 0
lb is
4/7/47
6/9/47
4/7/47
6/9/47
4/ 7/47
6/17/47
0 0
ib lb lb 4/ 7/47 0
Sb 2b
7/1/47
0
0
0
Sb Sb 4/14/47
0
0
0
0
0
lb lb lb 0
Sb Sb
6/ 9/47
0
of the columns refer to the corresponding dilution strengths given in table 1.
SUMMARY AND CONCLUSION
With the technic employed the development of an eczematous Sensitization did not have any effect on the threshold of irritation to the primary irritants
30
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
used. We do not believe that it is possible to generalize from our observations that an eczematous sensitization may never affect cutaneous irritability towards primary irritants. REFERENCES 1. LANDSTEINEE, K., AND DISOMMA, A. A.: Studies on the sensitization of animal8 with
simple chemical compounds. VIII. Sensitization to picric acid; subsidiary agents and mode of sensitization. J. Exper. Med., 72: 361, 1940. 2. ROKSTAD, I.: Skin reactions caused by fractions of oil of turpentine and hexanitrodiphenylamine, Acta Dermat.-venereol., Vol. 26, supplement 15, 1946.
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.
J. B. HAEBERLIN, JR., M.D., R. M. OLIVER, M.D. AND A. ROSTENBERG, JR., M.D.
A well-known clinical observation is that an existing dermatitis, regardless of type or etiology, may be aggravated or kept activated by contact with irritants, such as soap and water, cleaning solutions, solvents, topical medications, rough
or woolen clothing, and the like. Various theories have been advanced to explain this phenomenon, such as polyvalence of sensitization or lowering the threshold of reaction to primary irritants. The interrelationship between primary irritation and eczematous sensitization is poorly understood, but that primary irritation affects the development of an eczematous sensitization has been definitely established (1, 2). It should be borne in mind that many cc-
zematous sensitizers are also primary irritants. To our knowledge, the influence of the development of an eczematous sensitization on the threshold of reaction to primary irritants has not been studied. The purpose of this study was then to determine the influence, if any, of the genesis of an eczematous sensi-
tization on an individual's threshold of reactivity to primary irritants. METHOD
To answer this question we (1) determined in a series of individuals the threshold irritant dose to various substances by patch tests with graded dilutions of these materials, (2) then in one half of the subjects endeavored to engender an eczematous sensitization, and (3) after a suitable interval both groups were "repatched" with the original substances to determine whether there was any change in the threshold of irritation. The substances chosen as primary irritants were (1) hydrochloric acid, (2) tincture of green soap, and (3) croton oil. Dilutions were made up of the first two in water and of the last in olive oil. After preliminary trials on other individuals, the dilutions were selected so that the most concentrated gave a weakly positive reaction in the majority of individuals tested, whereas, the least concentrated gave a negative in the majority of the individuals tested. This provided a range of dilutions which should make observable a shift in the threshold, should this occur as a result of this experiment. The actual dilutions used are given in table 1. Because of the variability of reactions to testing with primary irritants considerable care was required to make the test applications as uniform as possible. * From the Department of Dermatology and the Allergy Unit of the University of Illinois Colleges of Medicine and Pharmacy. The expenses of this investigation were in part defrayed by the Asthmatic Children's Aid.
Received for publication August 28, 1947. 27
28
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
To this end the chamber patch as described by Rokstad (2) was used with uniform pieces of blotting paper and approximately uniform amounts of test solu-
tion dropped from dropping bottles. It was found that maximum clarity of reading was obtained by removing the patches after approximately 24 hours, and allowing several hours to elapse before reading the tests. In reading the tests, the papule formation as described by Rokstad was ignored. The reactions were of three morphologic types and were recorded according to the code: 1 = erythema corresponding to the circle of the blotting paper, 2 = erythema spreading from the site in a more diffuse fashion, and 3 = follicular pustules over the area. In each type stronger reactions were recorded as "a" and milder reactions as
For the purpose of sensitization 2,4 dinitrochlorobenzene was employed. This chemical was chosen because it is known to be a potent sensitizer and because it is a substance not likely to be encountered by our subjects in ordinary life. The technic employed for sensitizing was the dropping on of approximately 0.03 cc. of a 30 per cent acetone solution on the volar surface of the arm. At TABLE 1 1
2
3
4
5
per cent per cent per cent per cent per cent
HC1
Tincture of green soap Croton oil
1 10
2.5
2.5
5
10
20
30
5
10
40 20
20 50 40
In water In water In olive oil
the same time, approximately 0.03 cc. of a 0.1 per cent acetone solution was applied to the volar surface of the other arm in order to be certain that none of the subjects were already sensitive to this material. Because of the necessity of observing our patients over a reasonably long period of time, and because of the fact that we wished to do as many as possible of them at the same time, we did not believe that ordinary clinic patients would be suitable, and accordingly, inmates of a tuberculosis sanatorium were used.1 RESULTS
Table 2 gives the readings to graded dilutions of primary irritants in eight cases before and after sensitization, and a like number of controls in whom no sensitization was engendered. 1 The fact that the subjects were tuberculous may have affected our results. We had a rather low yield of sensitizations and also the development of the sensitivity seemed slower than our previous experience in the sensitization of non-tuberculous individuals had led
us to expect. We are aware that there is a considerable literature on the interference of a tuberculous infection with the development of other allergic phenomena, but we thought that because of the other reasons mentioned these subjects would nevertheless be most suitable.
29
STUDIES ON ECZEMATOUS SENSITIZATION
TABLE 2 HC1
GREEN SOAP
INITIAL
CROTON OIL
CASE
NO.
TO
12345 i2j345 l2I345
SENSITIZED TO
2,4
RETEST TO
IT
SEESTANTS
Group A. Sensitized to 2,4 dinitro chlorobenzene 0
lb
lb lb is is
0
?
lb lb lb
0 0
0 0
0 0
0 0
is is lb Is la
0 0
0
lb is Is 0 lb lb lb 0
0
0
0
0
0 lb 2b 2b
4/
0
0 0
0
lb lb is
0 0
0 0
0 0
0 0
0 0
0 0
4/14/47
0
0 0
lb
0
Before
0
0
?
lb lb la 0
0
0
lb
0
0
0
0
3b 4/21/47 5/27/47 6/17/47 6/17/47
0
lb lb is 0 lb lb
0
After
?
?
0
0
0
lb
lb
Before
0
0
lb lb
lb
0
lb
3s
0
?
0
is is
2b 2b 3b 3a
0
is lb
is
After
lb
lb
Before After
0 0
0
0 0
Before After
0 0
0
Before After
0
0
After
0 0
0 0
0 0
Before After
0 0
0
lb
Before
After
Before
1
2 3
4
5
6 7
8
0
0
lb 0
0 0
0
7
la
0
is is lb lb
0 0
0
0
0 0
lb 0
0 0
4/ 7/47
Sb 2b 2b
lb lb is 0 lb lb lb
0
Sb
2b 2b
0
0
0
lb
0 lb 0 0 lb lb lb is
0
0
lb lb
lb lb lb 0 0 lb lb 0 2b 0
0 0
Sa
2s 3/31/47
Sb
Sb
0
0
0
0 0
0 0
0 P
0
0
6/ 9/47
6/17/47 6/17/47
5/27/47 6/17/47 6/17/47
4/28/47 5/27/47 6/
0
0
5/27/47 8/ 9/47
7/47 5/27/47
lb 0
lb lb is lb lb Is is is 0 lb -lb 0 0 lb lb lb lb lb is 0 lb lb
3b 2b
9/47
6/
9/47
4/14/47 5/27/47 7/1/47 7/1/47
3b 4/21/47 5/27/47 7/1/47 7/1/47
4/28/47 5/27/47 7/1/47 7/1/47
Group B. Control 9
10
ii 12
13
14
15
16
Initial Final
0 0
0 7
0
?
lb lb 0 lb 1' 0
0
Initisi
0
0
0
0
0
0
0 0
Sb 0
0
lb 0 lb is 0
lb lb
Final
0
0
0
Initial Finsi
0 0
0 0
lb is la 0 0 0 lb 0
0 0
P
is is 0 lb lb 0
Initial
0
0
lb
0
0
0
0
Final
0
0
0
lb is 0
0
Initial Final
0 0
0 0
0
0 0
Initial Final
0 0
0 0
0
Initial Final
0 0
0 0
0
lb
ib 0
Initial Final
0 0
0 0
lb
P
is
0
0
The numbers 1 through
Is
?
0
0 is lb lb lb
is 0
lb is is
P
la 0
0
0
0
lb lb Ia 0 lb is 0
0 0
lb ls is P
lb lb
lb la
7 0
lb
lb lb
0 0
lb lb lb lb lb lb
0,
P
0 0
Ia
0
Sat the tops
0
0
6/9/47
3/31/47
6/ 9/47
Sb
Sb 4/ 7/47
0
6/ 9/47
0
lb lb 0
0 0
lb
is
is
is
0
0
0
0
0 0
0 0
0 0
lb is
4/7/47
6/9/47
4/7/47
6/9/47
4/ 7/47
6/17/47
0 0
ib lb lb 4/ 7/47 0
Sb 2b
7/1/47
0
0
0
Sb Sb 4/14/47
0
0
0
0
0
lb lb lb 0
Sb Sb
6/ 9/47
0
of the columns refer to the corresponding dilution strengths given in table 1.
SUMMARY AND CONCLUSION
With the technic employed the development of an eczematous Sensitization did not have any effect on the threshold of irritation to the primary irritants
30
THE JOURNAL OF INVESTIGATIVE DERMATOLOGY
used. We do not believe that it is possible to generalize from our observations that an eczematous sensitization may never affect cutaneous irritability towards primary irritants. REFERENCES 1. LANDSTEINEE, K., AND DISOMMA, A. A.: Studies on the sensitization of animal8 with
simple chemical compounds. VIII. Sensitization to picric acid; subsidiary agents and mode of sensitization. J. Exper. Med., 72: 361, 1940. 2. ROKSTAD, I.: Skin reactions caused by fractions of oil of turpentine and hexanitrodiphenylamine, Acta Dermat.-venereol., Vol. 26, supplement 15, 1946.
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.