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Photo-Hemolysis Associated with Protoporphyrinemia
PHOTO-HEMOLYSIS ASSOCIATED WITH PROTOPORPHYRINEMIA PRELIMINARY REPOET*
LEONARD C. HARBER, M.D., ALAN FLEISCHER, B.S. AND RUDOLF L. BAER, M.D.
Since 1961 when Magnus, Jarrett, Prankard, ated at a target distance of 6.25 centimeters. The and Rimington (1) first described a new disease intensity of the incident radiation was approxientity, viz. erythropoietic protoporphyria, there mately 1100 microwatts per centimeter square. have been reports of similar cases by several inMeasurement of Hemolytis was carried out by vestigators (2, 3, 4, 5, 6). In all the reported two methods: 1) direct cell counts: aliquots of cases, there were markedly elevated erythrocyte the red blood cell suspensions were counted in a protoporphyrin levels associated with cutaneous No. 500 Levy Counting Chamber (hemocytomphotosensitivity. This form of photosensitivity eter), at 10 minute intervals. Hemolysis was may express itself in diverse cutaneous lesions in- evident in the reduction of the number of red cluding erythema, vesicles, wheals and eczema. cells counted; 2) spectrophotometrically: the conWe wish to report the preliminary results of stud- centration of hemoglobin in the red blood cell ies concerning the in vitro hemolysis of red blood supernatant was measured in a Coleman junior cells by ultraviolet light using cells from a pa- spectrophotometer. tient having erythropoietic protoporphyrinemia. The patient is a seven year old girl who has RESULTs vesicular and crusted skin lesions on the light exThe results of the erythrocyte protoporphyrin posed areas on and off since the age of 4 and who has been under our observation for the past 2 determinations listed in Table I indicate that the years. Erythrocyte protoporphyrin blood levels patient's protoporphyrin levels obtained on 4 (7) were assayed on 4 occasions and were found consecutive specimens examined over a two to be significantly elevated. month period were significantly elevated as comPreparation of Red Blood Cells for Hemolysis pared to the levels observed in the control speciStudies: In each test run, the effects of ultraviolet mens. light on the patient's red blood cells and on the Erythrocyte protoporphyrin levels were also red blood cells of a normal control subject were examined in the patient's parents and in 3 sibcompared. Prior to irradiation, the cells were lings. The values obtained were essentially norwashed three times and resuspended in a 1:400 ma except for one brother, age 10, who, although dilution of a 0.9% NaC1 phosphate buffer. asymptomatic, hadj a protoporphyLight Source: The red blood cells were irradi- completely rin blood level of 90 pg/100 ml red blood cells ated with light from a bank of 4 Westinghouse (Normal value <70 pg/100 ml red blood cells). F20 T12 BL "blacklight" fluorescent tubes. The The results of the red blood cell hemolysis emission spectrum of these tubes is mainly bestudies using the spectrophotometric technic are tween 3200A° and 4500A°. In order to minimize the infrared radiation and the minute amount of presented in Table II. Abnormal hemolysis was radiation below 3200A the light was directed noted in the patient's specimen on each of four through a Corning 1-69 filter and through a win- occasions. The results of the red blood cell hemoldow glass filter, 3 mm thick. The red blood cells ysis studies using the direct cell count method were enclosed in quartz receptacles, and irradi- can be seen in Table III. There appeared to be a direct correlation between the length of irradia* From the Department of Dermatology of New York University Schools of Medicine, New York, tion and the degree of hemolysis. It should be New York. noted that the hemolysis shown in Tables II and This study was supported by Grant Number III was due to radiation between 3200A and DA-49-193-MD-2275 from the U.S. Army Medical
Research and Development Command.
4500A.
ragne, B.S. Received for publication October 30, 1963.
t Radiation above 3200A was measured using an Eppley thermopile.
Technical Assistance given by Mary E. Mo-
483
484
TifE JOURNAL OF INVESTIGATIVE DERMATOLOGY
TABLE I
Wegner (8) appear to indicate that the protopor phyrin molecule is also capable of photosensitiz-
Eryghrocyge prooporphyrin levels (JLg/100 ml red blood cells)
ing mice.
Patient's Red Blood Control Subject's
6/ 5/63 6/13/63 7/12/63 7/30/63
Cells
Red Blood Cells
683 1780
20
428
28
333
63
35
TABLE II Red blood cell hemolysis; opica dnsjgy* of supernatan ag 41 OOA No Light
Light
Specimen
Patient Control Patient Control
A (6/ 5/63) B (6/12/63) C (7/12/63)
D (7/30/63)
0 0 0 0
0 0 0 0
0.17 0.06 0.13 0.15
0.02 0.01
0.00 0.01
* Coleman Junior Spectrophotometer.
TABLE III Red blood cell hemolysis; direct cell flg5* Irradiation Time
EEC Patient
EEC Control Subject
46 42 32 24 25 22
66
minutes
0 5 15 25 35 45 55 65
9.8 0
The data presented in Table III appear to follow the Reciprocity Law and indicate that the longer the light exposure the greater the degree of hemolysis (9). Previous investigations by Cook
(10), Blum (9) and Meyerstein (11) utilizing "blacklight" have shown that hemolysis occurs in rwrmal human red blood cells after extended periods of time, ranging up to 10-42 hours. They attributed this to the photosensitizing effect produced by the small amounts of protoporphyrin which are normally present in all red blood cells. It should therefore be stressed that hemolysis in the cells from our patient due to the markedly increased concentration of the photosensitizing substance occurred within 10 minutes after the start of irradiation. In view of the very fa.st and pronounced in vitro hemolysis, it is noteworthy from the clinical viewpoint thM our patient manifested vesicular and crusted lesions and not purpuric ones. The most likely explanation is that the protoporphyrin molecule which is known to be present in the serum (5) is also present in the epidermis and cutis. Consequently, the photosensitizing action could well produce damage to these cutaneous cells and lead to changes compatible with the diverse clinical features which have been reported. Since no cases of purpura, anemia or icterus have
been reported, it may be assumed that the amount of light passing through the blood vessel
walls is insufficient to initiate a clinically significant degree of photochemical reaction in the intravascular red blood cells. StJM1&ARY
64
In vitro studies were done on photochemically mediated hemolysis of red blood cells from a 7 year old girl with erythrocytic protoporphyria. Markedly elevated red blood cell hemolysis wa DISCUSSION observed spectrophotometrically and in direct In all reported cases of erythropoietic proto- cell counts. The hemolytic reaction is attributed porphyria the abnormal photosensitivity has been to the abnormally high concentration of proto. associated with elevated erythrocytic protopor- porphyrin in the red blood cells. phyrin levels. Our findings of red blood cell heREFERENCES molysis reinforce the concept that the protopor* No. 500 Levy Counting Chamber.
phyrin molecule is a strong photosensitizer capable of damaging or destroying human cells in which a sufficient concentration of light and protoporphyrin are brought together. Studies by
1. MAGNUS, I. A., JARRTT, A., PRANEAED, T. A. J. AND RIMINGTON, C.: Erythropoietic
protoporphyria. A new porphyria syndrome
with solar urticaria due to protoporphyrinanemia. Lancet, 2: 448, 1961.
2. LANGH0F, II., MULLER, II. AND RnT8CHL, L.:
PHOTO-HEMOLYSIS ASSOCIATED WITH PROTOPORPHYRINEMIA
Untersuchungen zur Familiaren, Protoporphyrinamiachen Lichturticaria. Arch. Kiln. Exp. Derm., 212: 506, 1961. 3. HASGER-ARONSON, B.: Erythropoietic proto-
485
trophotometric niicromethod for the quantitative determination of the free erythrocyte protoporphyrin. J. Biol. Chem., 172: 459, 1948.
porphyria. Lancet, 1: 1073, 1962.
8. WEGNER J.: Untersuchungen uber die Licht-
protoporphyria. Amer. J. Dis. Child., 104:
phyrin. Z. Ges. Inn. Med., 3: 123, 1948. 9. BLUM, H. F.: Photodynamic Action and Dis-
4. PORTER, F. S. AND LOWE, B.: Erythropoietic 537, 1962.
5. bun, G., MAGNUS, I. A. AND RIMINUT0N, C.:
Erythropoietic protoporphyria in sisters. Brit. J. Derm., 75: 225, 1963.
6. REDEKER, A. G., BERKE, M. AND LEVAN, N.:
Erythropoietic protoporphyria with eczema
solare. Arch. Derm., 86: 569, 1962.
7. GRINSTEIN, M. AND WINTROBE, M. M.: Spec-
sensibñisierung weiber Mause durch Por-
eases Caused by Light. New York, N.Y.,
Reinhold Publishing Company, 1941. 10. CooK, J. S. AND BLUM, H. F.: Dose relationships and oxygen dependence in ultraviolet and photodynamic hemolysis. J. Cell. Comp. Physiol., 53 (1): 41, 1959. 11. MEYERSTZIN, W.: Photographic phenomena with blood films. J. Phot. Sci., 2: 174, 1954.
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.
LEONARD C. HARBER, M.D., ALAN FLEISCHER, B.S. AND RUDOLF L. BAER, M.D.
Since 1961 when Magnus, Jarrett, Prankard, ated at a target distance of 6.25 centimeters. The and Rimington (1) first described a new disease intensity of the incident radiation was approxientity, viz. erythropoietic protoporphyria, there mately 1100 microwatts per centimeter square. have been reports of similar cases by several inMeasurement of Hemolytis was carried out by vestigators (2, 3, 4, 5, 6). In all the reported two methods: 1) direct cell counts: aliquots of cases, there were markedly elevated erythrocyte the red blood cell suspensions were counted in a protoporphyrin levels associated with cutaneous No. 500 Levy Counting Chamber (hemocytomphotosensitivity. This form of photosensitivity eter), at 10 minute intervals. Hemolysis was may express itself in diverse cutaneous lesions in- evident in the reduction of the number of red cluding erythema, vesicles, wheals and eczema. cells counted; 2) spectrophotometrically: the conWe wish to report the preliminary results of stud- centration of hemoglobin in the red blood cell ies concerning the in vitro hemolysis of red blood supernatant was measured in a Coleman junior cells by ultraviolet light using cells from a pa- spectrophotometer. tient having erythropoietic protoporphyrinemia. The patient is a seven year old girl who has RESULTs vesicular and crusted skin lesions on the light exThe results of the erythrocyte protoporphyrin posed areas on and off since the age of 4 and who has been under our observation for the past 2 determinations listed in Table I indicate that the years. Erythrocyte protoporphyrin blood levels patient's protoporphyrin levels obtained on 4 (7) were assayed on 4 occasions and were found consecutive specimens examined over a two to be significantly elevated. month period were significantly elevated as comPreparation of Red Blood Cells for Hemolysis pared to the levels observed in the control speciStudies: In each test run, the effects of ultraviolet mens. light on the patient's red blood cells and on the Erythrocyte protoporphyrin levels were also red blood cells of a normal control subject were examined in the patient's parents and in 3 sibcompared. Prior to irradiation, the cells were lings. The values obtained were essentially norwashed three times and resuspended in a 1:400 ma except for one brother, age 10, who, although dilution of a 0.9% NaC1 phosphate buffer. asymptomatic, hadj a protoporphyLight Source: The red blood cells were irradi- completely rin blood level of 90 pg/100 ml red blood cells ated with light from a bank of 4 Westinghouse (Normal value <70 pg/100 ml red blood cells). F20 T12 BL "blacklight" fluorescent tubes. The The results of the red blood cell hemolysis emission spectrum of these tubes is mainly bestudies using the spectrophotometric technic are tween 3200A° and 4500A°. In order to minimize the infrared radiation and the minute amount of presented in Table II. Abnormal hemolysis was radiation below 3200A the light was directed noted in the patient's specimen on each of four through a Corning 1-69 filter and through a win- occasions. The results of the red blood cell hemoldow glass filter, 3 mm thick. The red blood cells ysis studies using the direct cell count method were enclosed in quartz receptacles, and irradi- can be seen in Table III. There appeared to be a direct correlation between the length of irradia* From the Department of Dermatology of New York University Schools of Medicine, New York, tion and the degree of hemolysis. It should be New York. noted that the hemolysis shown in Tables II and This study was supported by Grant Number III was due to radiation between 3200A and DA-49-193-MD-2275 from the U.S. Army Medical
Research and Development Command.
4500A.
ragne, B.S. Received for publication October 30, 1963.
t Radiation above 3200A was measured using an Eppley thermopile.
Technical Assistance given by Mary E. Mo-
483
484
TifE JOURNAL OF INVESTIGATIVE DERMATOLOGY
TABLE I
Wegner (8) appear to indicate that the protopor phyrin molecule is also capable of photosensitiz-
Eryghrocyge prooporphyrin levels (JLg/100 ml red blood cells)
ing mice.
Patient's Red Blood Control Subject's
6/ 5/63 6/13/63 7/12/63 7/30/63
Cells
Red Blood Cells
683 1780
20
428
28
333
63
35
TABLE II Red blood cell hemolysis; opica dnsjgy* of supernatan ag 41 OOA No Light
Light
Specimen
Patient Control Patient Control
A (6/ 5/63) B (6/12/63) C (7/12/63)
D (7/30/63)
0 0 0 0
0 0 0 0
0.17 0.06 0.13 0.15
0.02 0.01
0.00 0.01
* Coleman Junior Spectrophotometer.
TABLE III Red blood cell hemolysis; direct cell flg5* Irradiation Time
EEC Patient
EEC Control Subject
46 42 32 24 25 22
66
minutes
0 5 15 25 35 45 55 65
9.8 0
The data presented in Table III appear to follow the Reciprocity Law and indicate that the longer the light exposure the greater the degree of hemolysis (9). Previous investigations by Cook
(10), Blum (9) and Meyerstein (11) utilizing "blacklight" have shown that hemolysis occurs in rwrmal human red blood cells after extended periods of time, ranging up to 10-42 hours. They attributed this to the photosensitizing effect produced by the small amounts of protoporphyrin which are normally present in all red blood cells. It should therefore be stressed that hemolysis in the cells from our patient due to the markedly increased concentration of the photosensitizing substance occurred within 10 minutes after the start of irradiation. In view of the very fa.st and pronounced in vitro hemolysis, it is noteworthy from the clinical viewpoint thM our patient manifested vesicular and crusted lesions and not purpuric ones. The most likely explanation is that the protoporphyrin molecule which is known to be present in the serum (5) is also present in the epidermis and cutis. Consequently, the photosensitizing action could well produce damage to these cutaneous cells and lead to changes compatible with the diverse clinical features which have been reported. Since no cases of purpura, anemia or icterus have
been reported, it may be assumed that the amount of light passing through the blood vessel
walls is insufficient to initiate a clinically significant degree of photochemical reaction in the intravascular red blood cells. StJM1&ARY
64
In vitro studies were done on photochemically mediated hemolysis of red blood cells from a 7 year old girl with erythrocytic protoporphyria. Markedly elevated red blood cell hemolysis wa DISCUSSION observed spectrophotometrically and in direct In all reported cases of erythropoietic proto- cell counts. The hemolytic reaction is attributed porphyria the abnormal photosensitivity has been to the abnormally high concentration of proto. associated with elevated erythrocytic protopor- porphyrin in the red blood cells. phyrin levels. Our findings of red blood cell heREFERENCES molysis reinforce the concept that the protopor* No. 500 Levy Counting Chamber.
phyrin molecule is a strong photosensitizer capable of damaging or destroying human cells in which a sufficient concentration of light and protoporphyrin are brought together. Studies by
1. MAGNUS, I. A., JARRTT, A., PRANEAED, T. A. J. AND RIMINGTON, C.: Erythropoietic
protoporphyria. A new porphyria syndrome
with solar urticaria due to protoporphyrinanemia. Lancet, 2: 448, 1961.
2. LANGH0F, II., MULLER, II. AND RnT8CHL, L.:
PHOTO-HEMOLYSIS ASSOCIATED WITH PROTOPORPHYRINEMIA
Untersuchungen zur Familiaren, Protoporphyrinamiachen Lichturticaria. Arch. Kiln. Exp. Derm., 212: 506, 1961. 3. HASGER-ARONSON, B.: Erythropoietic proto-
485
trophotometric niicromethod for the quantitative determination of the free erythrocyte protoporphyrin. J. Biol. Chem., 172: 459, 1948.
porphyria. Lancet, 1: 1073, 1962.
8. WEGNER J.: Untersuchungen uber die Licht-
protoporphyria. Amer. J. Dis. Child., 104:
phyrin. Z. Ges. Inn. Med., 3: 123, 1948. 9. BLUM, H. F.: Photodynamic Action and Dis-
4. PORTER, F. S. AND LOWE, B.: Erythropoietic 537, 1962.
5. bun, G., MAGNUS, I. A. AND RIMINUT0N, C.:
Erythropoietic protoporphyria in sisters. Brit. J. Derm., 75: 225, 1963.
6. REDEKER, A. G., BERKE, M. AND LEVAN, N.:
Erythropoietic protoporphyria with eczema
solare. Arch. Derm., 86: 569, 1962.
7. GRINSTEIN, M. AND WINTROBE, M. M.: Spec-
sensibñisierung weiber Mause durch Por-
eases Caused by Light. New York, N.Y.,
Reinhold Publishing Company, 1941. 10. CooK, J. S. AND BLUM, H. F.: Dose relationships and oxygen dependence in ultraviolet and photodynamic hemolysis. J. Cell. Comp. Physiol., 53 (1): 41, 1959. 11. MEYERSTZIN, W.: Photographic phenomena with blood films. J. Phot. Sci., 2: 174, 1954.
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.