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Gene analysis in patients with tyrosinase activity-negative albinism
115 LEUKOTRIENE PSEUDOMONAS
64 ELEVATES RESISTANCE INFECTION IN MICE.
T. Demitsu, H. Katayama and H. Yaoita, Jichi Medical School, Tochigi-ken.
TO SALMONELLA
Department
AND
GENE ANALYSIS ALBINISM
Ti%%m$“S 9.2%(co"tt-ol).
AN ADHESIVE AND SPREADING FACTOR PRODUCTION OF ONE CANCER CELL LINE OF HUNAN HAIR FOLLICLE ORIGIN. H. Katasama, Department of Dermatolasr, Tochiri
Cell adhesion to substratum is B pivotal step for growth of anchorage dependent cells such LS keratinocytes. Recently several proteins that mehiate cell adhesion and spreading hare been found and characterized. In this communication, I present an evidence that one cancer cell line of human hair follicle origin, malignant trichilemmal cyst. produces such a kind of adhesive and spreading factor. This cell adheres to substratum tightly in the absence of serum and the adhesion is not released without the use of trypsin. Preooating of culture well with serum-free culture supernatant of this cell induced good adhesion and spreadins as compared with unprecoating OT that with bovine serum albumin. Furthermore this cell can grow The growth curve showed a more rapid continuously in the absence of serum. growth in the absence of serum than that i6 the pmsence of serum. SDS-PACE of this serum-free culture supernatant resealed numerous protein bands. Autocrine secretion theory (cancer cells produce and resuond to their own growth factor)is emerging-as .aunifying cbncept in the search for the molecular and cellular basis of malignant transfor&ion. This cell may be growing in such a fashion. The key Factor might be an adhesive factor.
ALTERED EXPRESSION OF A THIRD ACTIN ACCOMPANYING MALIGNANT TRANSFORMATION AND/OR PROGRESSION IN MOUSE 616 MELANOMA CELLS AND HUMAN PIGMENT TISSUES
, M.
S. Taniguchi' Okamoto', H. Sadano', J. Nakayama' T.Kakuna9a3, T. Babal, and Y. Hori2. Medical Inst. of Bioregulat/onl, and Dept. of Dermatology, Faculty of Medici&, Kyushu Univ., Fukuoka 812; Research Inst. of Microbial Diseases, Osaka Univ.3, Suita 565. Of the many phenotypes in neoplastic cells , we directed OUP attention to changes in expression of actin, a protein involving in control of cell morphology and motility. 1) In mouse 816 melanoma cell lines, we found a new type of actin (Ax), the expression of which altered with metastatic potential. We cloned and sequenced full length cDNA encoding AX clarifying that AX is similar to 8actin in the coding region but different in the untranslated region. 2) A third actin, similar to smooth muscle a-actin, is co-expressed with B-and y-actin in human benign pigment tissues (blue news and novus pigmentosus) and early passage cells from those tissues, while the third actin was not evident in human malignant melanoma cells in culture or the tissues. Knowledge of the depression of the third actin in human malignant melanoma is important for diagnosis as a transformation sensitive marker for human pigment cells.
,
pi--l*
IN VIVO AND IN VITRO LICHEN
OF DIMETHYL
SULPHOXIDE
AMYLOIDOSUS
W. L Lo and C. K. Wong. Department and Yang-Ming
EFFECTS
Medical
of Dermatology,
College. Taipei,
Taiwan,
Veterans General Hospital
Republic
of China
Significant clinical results have been observed in lichen amyloidosus patients with either topical or oral administration of dimerhyl sulphoxide (DMSO). Skin explants from lichen amyloidosus patients were cultured in medium containing 0.5% DMSO. After incubation of 4, 6, 8, and 12 weeks, explants were processed for electron microscopical examination. In addition to clinical investigation, this paper will report the ultrastructural changes of amyloid in the presence of DMSO.
ACTIVITY-NEGATIVE
We have succeeded to isolate cDNA of mouse tyrosinase (Tase) (Nucleic Acids Res. 14, 2413, 1986) with the aid of our monoclonal antibody to mouse Tase (J. Invest. Dermatol. 85, 426, 1985). By using the cDNA as a probe, we have tried to determine DNA polymorphism in a respective patient with Tase activity-negative albinism and with Hermansky-Pudlak syndrome. They expected to show DNA polymorphism different from that of a healthy person. Though various kinds of restriction enzymes such as Barn HI, Bgl II, Eco RI, Hae III, Hind III, Hinf I, Kpn I, Pvu II, Sal I, Sma I, Xba I. and Xho I were used in our experimental determination, apparent DNA polymorphism was not detected among them. From these results. we can onlv conclude that tyrosinase activity-negative albino, at ieast in our two cases, is not caused by deletion of entjre tyrosinase gene, but may be caused by small change(s) such as a point mutation.
A-l,
A-IS
WITH TYROSINASE
Y. Tomita, S. Shibahara* and H. Tagami. Department of Dermatology and *Department of Applied Physiology, Tohoku University School of Medicine. Sendai.
of Dermatology,
The effects of exogenous leukotriene Bq(LTB4) on the resistance of nose peritoneal macrophages against Salmonella and Pseudomxas (P.) infections were studied. In vitro, LTB4 added to macrophage monolayers at final concentrations of lo-12-lo-8M, enhanced their phagocytosis of S. typhimurium to 2.3 times the control level and that of P. aeruginosato 1.8 times the control level. The intracellular kilTin rates were also elevated bv the addition of LTBd: for S. t himirium 83 3%(LTB4) vs 59.l%(;ontrol) and for P. adruginora, In viva, intraperito?ieally injected LTB4(5 nq) enhanced the clearance at 24 h of intraperitoneally injected-s. typhimurium from the mouse peritoneal cavity and ipleen, but this Fffect disappeared by 48 h. Activation of macrophages by exogenous LTB4 seemed to have contributed to such an augwnted resistance of macrophages to bacterial infection. This study suggested a possible use of LTB4 for bacterial infectious diseases whereby phagocytes are able to play a key role in host defense. (Co-investigators: T. Saito-Taki & M. Nakano. Department of Microbiology, Jichi Medical School)
0
IN PATIENTS
ON
SELECTIVE THERMAL NEUTRON CAPTURE THERAPY OF MALIGNANT MELANOMA USING MELANOMA-SEEKING lDB-COMPOUNDS. l.RADIDBIDLDGICAL BASIS
M. Ichihashi! A. Sasase! T. Hiramoto! H. Fukuda? K. Yoshino3 and Y. Mishima! Dept. of Derm., Special Institute of Cancer NCT, Kobe Univ. Sch. Med.,Kobel, National Institute of Radiological Sciences, Chiba2 and Department of Chemistry, Faculty of Science, Shinshu University, Matsumoto3 Aims: EvaluatipB of the lethal effect of thermal neutron capture Bl-paraboronophenylalanine therapy using r~~~l,~~A,~:'lo~b:~~~ured melanoma cells and determination of RBE of basic conditions for the treatment of human melanoma in viva. Results: lDBl-BPA-NCT exhibited enhanced lethal effect on both p melanotlc and amelanotic melanoma cells, but not on non-melanoma RBE of lDBl-BPA-NCT (106: 2.5pg/ml) at Do was 4.2, about 2 cells. times greater than that of fast neutron. Conclusions: These results indicate that ~OB~-BPA actively and selectively accumulated in both melanotic and amelanotic melanoma cells, and further lOBl-BPA-NCT is shown to kill melanoma cells more efficiently than fast neutron radiation.
64 ELEVATES RESISTANCE INFECTION IN MICE.
T. Demitsu, H. Katayama and H. Yaoita, Jichi Medical School, Tochigi-ken.
TO SALMONELLA
Department
AND
GENE ANALYSIS ALBINISM
Ti%%m$“S 9.2%(co"tt-ol).
AN ADHESIVE AND SPREADING FACTOR PRODUCTION OF ONE CANCER CELL LINE OF HUNAN HAIR FOLLICLE ORIGIN. H. Katasama, Department of Dermatolasr, Tochiri
Cell adhesion to substratum is B pivotal step for growth of anchorage dependent cells such LS keratinocytes. Recently several proteins that mehiate cell adhesion and spreading hare been found and characterized. In this communication, I present an evidence that one cancer cell line of human hair follicle origin, malignant trichilemmal cyst. produces such a kind of adhesive and spreading factor. This cell adheres to substratum tightly in the absence of serum and the adhesion is not released without the use of trypsin. Preooating of culture well with serum-free culture supernatant of this cell induced good adhesion and spreadins as compared with unprecoating OT that with bovine serum albumin. Furthermore this cell can grow The growth curve showed a more rapid continuously in the absence of serum. growth in the absence of serum than that i6 the pmsence of serum. SDS-PACE of this serum-free culture supernatant resealed numerous protein bands. Autocrine secretion theory (cancer cells produce and resuond to their own growth factor)is emerging-as .aunifying cbncept in the search for the molecular and cellular basis of malignant transfor&ion. This cell may be growing in such a fashion. The key Factor might be an adhesive factor.
ALTERED EXPRESSION OF A THIRD ACTIN ACCOMPANYING MALIGNANT TRANSFORMATION AND/OR PROGRESSION IN MOUSE 616 MELANOMA CELLS AND HUMAN PIGMENT TISSUES
, M.
S. Taniguchi' Okamoto', H. Sadano', J. Nakayama' T.Kakuna9a3, T. Babal, and Y. Hori2. Medical Inst. of Bioregulat/onl, and Dept. of Dermatology, Faculty of Medici&, Kyushu Univ., Fukuoka 812; Research Inst. of Microbial Diseases, Osaka Univ.3, Suita 565. Of the many phenotypes in neoplastic cells , we directed OUP attention to changes in expression of actin, a protein involving in control of cell morphology and motility. 1) In mouse 816 melanoma cell lines, we found a new type of actin (Ax), the expression of which altered with metastatic potential. We cloned and sequenced full length cDNA encoding AX clarifying that AX is similar to 8actin in the coding region but different in the untranslated region. 2) A third actin, similar to smooth muscle a-actin, is co-expressed with B-and y-actin in human benign pigment tissues (blue news and novus pigmentosus) and early passage cells from those tissues, while the third actin was not evident in human malignant melanoma cells in culture or the tissues. Knowledge of the depression of the third actin in human malignant melanoma is important for diagnosis as a transformation sensitive marker for human pigment cells.
,
pi--l*
IN VIVO AND IN VITRO LICHEN
OF DIMETHYL
SULPHOXIDE
AMYLOIDOSUS
W. L Lo and C. K. Wong. Department and Yang-Ming
EFFECTS
Medical
of Dermatology,
College. Taipei,
Taiwan,
Veterans General Hospital
Republic
of China
Significant clinical results have been observed in lichen amyloidosus patients with either topical or oral administration of dimerhyl sulphoxide (DMSO). Skin explants from lichen amyloidosus patients were cultured in medium containing 0.5% DMSO. After incubation of 4, 6, 8, and 12 weeks, explants were processed for electron microscopical examination. In addition to clinical investigation, this paper will report the ultrastructural changes of amyloid in the presence of DMSO.
ACTIVITY-NEGATIVE
We have succeeded to isolate cDNA of mouse tyrosinase (Tase) (Nucleic Acids Res. 14, 2413, 1986) with the aid of our monoclonal antibody to mouse Tase (J. Invest. Dermatol. 85, 426, 1985). By using the cDNA as a probe, we have tried to determine DNA polymorphism in a respective patient with Tase activity-negative albinism and with Hermansky-Pudlak syndrome. They expected to show DNA polymorphism different from that of a healthy person. Though various kinds of restriction enzymes such as Barn HI, Bgl II, Eco RI, Hae III, Hind III, Hinf I, Kpn I, Pvu II, Sal I, Sma I, Xba I. and Xho I were used in our experimental determination, apparent DNA polymorphism was not detected among them. From these results. we can onlv conclude that tyrosinase activity-negative albino, at ieast in our two cases, is not caused by deletion of entjre tyrosinase gene, but may be caused by small change(s) such as a point mutation.
A-l,
A-IS
WITH TYROSINASE
Y. Tomita, S. Shibahara* and H. Tagami. Department of Dermatology and *Department of Applied Physiology, Tohoku University School of Medicine. Sendai.
of Dermatology,
The effects of exogenous leukotriene Bq(LTB4) on the resistance of nose peritoneal macrophages against Salmonella and Pseudomxas (P.) infections were studied. In vitro, LTB4 added to macrophage monolayers at final concentrations of lo-12-lo-8M, enhanced their phagocytosis of S. typhimurium to 2.3 times the control level and that of P. aeruginosato 1.8 times the control level. The intracellular kilTin rates were also elevated bv the addition of LTBd: for S. t himirium 83 3%(LTB4) vs 59.l%(;ontrol) and for P. adruginora, In viva, intraperito?ieally injected LTB4(5 nq) enhanced the clearance at 24 h of intraperitoneally injected-s. typhimurium from the mouse peritoneal cavity and ipleen, but this Fffect disappeared by 48 h. Activation of macrophages by exogenous LTB4 seemed to have contributed to such an augwnted resistance of macrophages to bacterial infection. This study suggested a possible use of LTB4 for bacterial infectious diseases whereby phagocytes are able to play a key role in host defense. (Co-investigators: T. Saito-Taki & M. Nakano. Department of Microbiology, Jichi Medical School)
0
IN PATIENTS
ON
SELECTIVE THERMAL NEUTRON CAPTURE THERAPY OF MALIGNANT MELANOMA USING MELANOMA-SEEKING lDB-COMPOUNDS. l.RADIDBIDLDGICAL BASIS
M. Ichihashi! A. Sasase! T. Hiramoto! H. Fukuda? K. Yoshino3 and Y. Mishima! Dept. of Derm., Special Institute of Cancer NCT, Kobe Univ. Sch. Med.,Kobel, National Institute of Radiological Sciences, Chiba2 and Department of Chemistry, Faculty of Science, Shinshu University, Matsumoto3 Aims: EvaluatipB of the lethal effect of thermal neutron capture Bl-paraboronophenylalanine therapy using r~~~l,~~A,~:'lo~b:~~~ured melanoma cells and determination of RBE of basic conditions for the treatment of human melanoma in viva. Results: lDBl-BPA-NCT exhibited enhanced lethal effect on both p melanotlc and amelanotic melanoma cells, but not on non-melanoma RBE of lDBl-BPA-NCT (106: 2.5pg/ml) at Do was 4.2, about 2 cells. times greater than that of fast neutron. Conclusions: These results indicate that ~OB~-BPA actively and selectively accumulated in both melanotic and amelanotic melanoma cells, and further lOBl-BPA-NCT is shown to kill melanoma cells more efficiently than fast neutron radiation.