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187 Modulation of keratinocyte growth by culture filtrate from dermatophytes
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JSID Abstracts/ Joumal of Dermatological Science 10 (1995) 61-102
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MODULATION OF KERATINOCYTE GROWTH B,Y CULTURE FILTRATE FROM DERMATOPHYTES. X&.&n . R. Tsubpignp H. Department of Dermatology, Juntendo University School of Medicine, Tokyo. Dermatophytes cause superficial mycoses. and of which the clinical manifestations vary depending on the species, body site and immunologicaI conditicm of the host. We speculated that the hyperkeratotic or ecxmatous reaction of tie host WBSpartly due to the direct interaction between keratinocytes and the organism. In this study, we examined the effect of culture filtrates on keratinocyte growth. Two swains of Trichophyton rubrum and T. mentagrophytes were inoculated in keradnocyte baaI medium (KBM) for 6 days at mom temPsrature, and the culhm filtrate collected. Keradnocytes, precultivated in KGM for 48 h, were cultivated in KBM containing different concentrations of the culture filtrates for 4 days at 37 ‘C. Cell wtb was monitored by cell count sod a fluorometric assay with ahmar r he. The addition of T. dwum-culnUe filtrate from 2 to 10 96 stimulated kemtinocyte growth approximately twofold, while the addition of T. mentagrophytes-culture t&ate significantly suppressed cell growth in the same range of concentrations. These results contribute 10 the understanding of the difference in the clinical manifestations caused by T. rubrum and T. mentogrophytes.
A NEW APPROACH TO STAPHYLOCOCCUS AUREUS IN SKIN DISEASES ANALYSIS OF ANl.IBODY TO .SI.APHYLOKINASE. X. * Y Ooto. Dermatology,Saitama Medical Center, Saitma Medical School. Kawagce, Japan. A serum marker for Staphylococcus aureus-infection has not been commonly utilized yet in clinical dermatology, when comparea to AS0 and ASK in strcptowccal infection. We measured antibody level to staphylokinasc (STA), a protein product of Staphylcccccus aureus, in various skin diseases, in order to assess Practical and investigative significance of anti-STA. Level of anti-STA antibody was quantified with ELlSA tcclmique. In comparison to namal healthy subjeds, patients with celhditis and other skin infection showed significandy high anti-STA antibody level, and dds level was not correlated to AS0 or ASK, both of which am antibodies to stmptocoxal products. Patients with nummular eczema, erydmna nodomm and atopc dermatitis showd elevated antiWA. suwting modulating effects d staphyIomccal infection in t&e di-. However, petients with psoriasis did not show increase in the antibody level to STA. Western Motdng confirmed s immunoreaction bands to STA in the blood samples pedfic mm the patients with high anti-STA titex. We ccoclude that antibody to STA may be a useful indicator to investigate the effects d StaphyIccnccus aurws-infection which potentially modulates various skin di-.
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DETECTION OF BORRELIA DNA, NOT B. BURGDORFERI SENSU STRICTO, BUT B. GARINII OR VS461 IN MORPHEA AND LICHEN SCLEROSUS ET ATROPHICUS
ELECTRON MICROSCOPIC OBSERVATION, OF BACTERIAL I CELLS ON CHRONIC SKIN ULCERS. v Deoartment of Dermatoloev. Okavama Uivcrsitv Medical School, Okiyama, Japan. I’ I ‘I%exe have been no descriptions in the literature about ultmstructuraI observations of bacterial cells on the surfaces of chronic skim ulcers. The psistenee of bacteria on chmoic ulcers plays some role in delaying wound bealmg. The purpose of this sNdy is to examine how bacterial cells colonize or infcd the surface9 of skm ulcers. Samples were taken from 10 chronic skin ulcers (radiodermatitis, one ; necmtizing fasciitis, one ; decubitus ulcer, eight) for bacterial analysis and electron microscopic obscrvarion. The isolated organisms were S mtrel(s in four patients, P. aeruginoso + E. avium in one patien!, P. aeruginom + E. coli in one patient, and were not detected in four pabents. Bacterial microcolonies were observed in six patients and fibril-like structures around bacterial cells were seen in five patients. In one ulcer Ruthenium red (RR)-stained sample WBSinvcstigatcd using electron microscopy and abundant RR positive structurw were seen around bacterial cells. l&se tindings of bacterial microcolonies with fibril-lie structures around the bacterial 41s indicate the formation of biofilms. We consider that the bacterias may form biofilms on chronic skin ulcers and prolong wound healing. (This work was done in collaboration with Dr. M. Fukui, Kyoto University.)
Lever. Department of Dermatology, Niigata University, Niigata, Japan. Wayne State university, Detroit, USA, Bbxhard-KarlsUniversit% TUbingen. Germany, Otto-Guericke-Universitat Magdeburg. Magdeburg, Germ-my. The association of B. burgdorferi with morphea and lichen sclemsus et atrophicus has been suggested, but is still controversial. We extracted DNA from the biopsies of 19 morphea and 34 LSA cases of U. S., Japanese and German patients. ‘l%e samples were subjected to amplify for general bomlia sequence polymerase chain reaction, and type specific amplification for B. burgdorferi sensu stricto. B. garinii, and VS 461. Five morphea and two LSA specimens, all of them from Japan or Germany, yielded positive result for B. garinii or VS 461, but not for B. burgdorferi sensu sticto. The specificity was confirmed by restriction enzyme digestion. The involvement of European stmin bonelia in the development of morphea and LSA was suggested.
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NEGATIVE CHARGE DISTRIBUTION WITHIN THE EPIDERMIS AND ITS ASSOCIATION WITH CELL-CELL ADHESION AND THE ACTION OF EXFOLIATIVE TOXIN. E.\ Af& Department of Dermatology and $Cell Biology, Okayama University Medical School, Okayama. Japan. ImraepidermaJ cleft occurs within the granular layer of the epidermis by the action of exfoliative toxin (ET). In the gmnular layer of the epidermis, there are many anionic sites. We examined the correlation between the cleft formation caused by ET and anionic sites using mouse embryo&4 skin. ET induced cleft either within the suprabasal layer of the developing skin, and within tbe gramdar layer of the fully developed skin. Negative charge distribution also changed as stratification proceeds. Negative charge emerged within tbe 14-day-ol d embryonal skin, and was gathered in the granuiar layer as skin matures. Glycosaminoglycao (GAG) chains, which are highly negatively charged, were neutralized with cationic molecule, polyethyleneimine (PEI). PEI induced cleft formation within the supraba.wI layer of the skin of the 15day-old embryos, and within the granular layer of the neonataI skin. These findings indicate that GAG chains may function in cell-cell adhesion within the epidermis, and ET may cleave the epidermis as tbe result of the disruption of proteoglycan-mediated cell-cell adhesion.
ANTIBIOTIC SUSCEPTIBILITIES OF PROPIONtBACTElUUM ACNES ISOLATED FROM ACNE VULGARIS : A 25-YEAR RETROSPBCTIVB STUDY. I. Kurokawa’)*, S. Nishiiimaz), S. Kawabata*. “Department of Dermatology, Hyogo Prefectural Tsukaguchi Hospital, Amagasaki, Hyogo, Japan. “Department of Dermatology, Kori Branch Hospital, Kansai Medical University, Neyagawa, Osaka, Japan. “Division of Dennatologicals 8 Ophthabnologicals, Ako Research Institute, Otsuka Pharmaceutical Co. Ltd., Ako, Hyogo, Japan. We have evaluated antibiotic susceptibilities of Propionibacterium ~lcnes (P. owes) isolated from acne lesions during the last 25 years retrospectively. Clinical isolates of P. acnes were tested for antibiotic susceptibilities using an agar plate dilution method. Although highly resistant strains to erythromycio and clindamycin were observed in about 10% of all strains, the others were verv sensitive to these drums. No resistant strains were observed against minocycline and nadifloxacin. During the last 25 years, no obvious resistant tendency of P. ocnes was not found by evaluating antibiotic susceptibilities of P. acnes.
JSID Abstracts/ Joumal of Dermatological Science 10 (1995) 61-102
187
190
MODULATION OF KERATINOCYTE GROWTH B,Y CULTURE FILTRATE FROM DERMATOPHYTES. X&.&n . R. Tsubpignp H. Department of Dermatology, Juntendo University School of Medicine, Tokyo. Dermatophytes cause superficial mycoses. and of which the clinical manifestations vary depending on the species, body site and immunologicaI conditicm of the host. We speculated that the hyperkeratotic or ecxmatous reaction of tie host WBSpartly due to the direct interaction between keratinocytes and the organism. In this study, we examined the effect of culture filtrates on keratinocyte growth. Two swains of Trichophyton rubrum and T. mentagrophytes were inoculated in keradnocyte baaI medium (KBM) for 6 days at mom temPsrature, and the culhm filtrate collected. Keradnocytes, precultivated in KGM for 48 h, were cultivated in KBM containing different concentrations of the culture filtrates for 4 days at 37 ‘C. Cell wtb was monitored by cell count sod a fluorometric assay with ahmar r he. The addition of T. dwum-culnUe filtrate from 2 to 10 96 stimulated kemtinocyte growth approximately twofold, while the addition of T. mentagrophytes-culture t&ate significantly suppressed cell growth in the same range of concentrations. These results contribute 10 the understanding of the difference in the clinical manifestations caused by T. rubrum and T. mentogrophytes.
A NEW APPROACH TO STAPHYLOCOCCUS AUREUS IN SKIN DISEASES ANALYSIS OF ANl.IBODY TO .SI.APHYLOKINASE. X. * Y Ooto. Dermatology,Saitama Medical Center, Saitma Medical School. Kawagce, Japan. A serum marker for Staphylococcus aureus-infection has not been commonly utilized yet in clinical dermatology, when comparea to AS0 and ASK in strcptowccal infection. We measured antibody level to staphylokinasc (STA), a protein product of Staphylcccccus aureus, in various skin diseases, in order to assess Practical and investigative significance of anti-STA. Level of anti-STA antibody was quantified with ELlSA tcclmique. In comparison to namal healthy subjeds, patients with celhditis and other skin infection showed significandy high anti-STA antibody level, and dds level was not correlated to AS0 or ASK, both of which am antibodies to stmptocoxal products. Patients with nummular eczema, erydmna nodomm and atopc dermatitis showd elevated antiWA. suwting modulating effects d staphyIomccal infection in t&e di-. However, petients with psoriasis did not show increase in the antibody level to STA. Western Motdng confirmed s immunoreaction bands to STA in the blood samples pedfic mm the patients with high anti-STA titex. We ccoclude that antibody to STA may be a useful indicator to investigate the effects d StaphyIccnccus aurws-infection which potentially modulates various skin di-.
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DETECTION OF BORRELIA DNA, NOT B. BURGDORFERI SENSU STRICTO, BUT B. GARINII OR VS461 IN MORPHEA AND LICHEN SCLEROSUS ET ATROPHICUS
ELECTRON MICROSCOPIC OBSERVATION, OF BACTERIAL I CELLS ON CHRONIC SKIN ULCERS. v Deoartment of Dermatoloev. Okavama Uivcrsitv Medical School, Okiyama, Japan. I’ I ‘I%exe have been no descriptions in the literature about ultmstructuraI observations of bacterial cells on the surfaces of chronic skim ulcers. The psistenee of bacteria on chmoic ulcers plays some role in delaying wound bealmg. The purpose of this sNdy is to examine how bacterial cells colonize or infcd the surface9 of skm ulcers. Samples were taken from 10 chronic skin ulcers (radiodermatitis, one ; necmtizing fasciitis, one ; decubitus ulcer, eight) for bacterial analysis and electron microscopic obscrvarion. The isolated organisms were S mtrel(s in four patients, P. aeruginoso + E. avium in one patien!, P. aeruginom + E. coli in one patient, and were not detected in four pabents. Bacterial microcolonies were observed in six patients and fibril-like structures around bacterial cells were seen in five patients. In one ulcer Ruthenium red (RR)-stained sample WBSinvcstigatcd using electron microscopy and abundant RR positive structurw were seen around bacterial cells. l&se tindings of bacterial microcolonies with fibril-lie structures around the bacterial 41s indicate the formation of biofilms. We consider that the bacterias may form biofilms on chronic skin ulcers and prolong wound healing. (This work was done in collaboration with Dr. M. Fukui, Kyoto University.)
Lever. Department of Dermatology, Niigata University, Niigata, Japan. Wayne State university, Detroit, USA, Bbxhard-KarlsUniversit% TUbingen. Germany, Otto-Guericke-Universitat Magdeburg. Magdeburg, Germ-my. The association of B. burgdorferi with morphea and lichen sclemsus et atrophicus has been suggested, but is still controversial. We extracted DNA from the biopsies of 19 morphea and 34 LSA cases of U. S., Japanese and German patients. ‘l%e samples were subjected to amplify for general bomlia sequence polymerase chain reaction, and type specific amplification for B. burgdorferi sensu stricto. B. garinii, and VS 461. Five morphea and two LSA specimens, all of them from Japan or Germany, yielded positive result for B. garinii or VS 461, but not for B. burgdorferi sensu sticto. The specificity was confirmed by restriction enzyme digestion. The involvement of European stmin bonelia in the development of morphea and LSA was suggested.
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192
NEGATIVE CHARGE DISTRIBUTION WITHIN THE EPIDERMIS AND ITS ASSOCIATION WITH CELL-CELL ADHESION AND THE ACTION OF EXFOLIATIVE TOXIN. E.\ Af& Department of Dermatology and $Cell Biology, Okayama University Medical School, Okayama. Japan. ImraepidermaJ cleft occurs within the granular layer of the epidermis by the action of exfoliative toxin (ET). In the gmnular layer of the epidermis, there are many anionic sites. We examined the correlation between the cleft formation caused by ET and anionic sites using mouse embryo&4 skin. ET induced cleft either within the suprabasal layer of the developing skin, and within tbe gramdar layer of the fully developed skin. Negative charge distribution also changed as stratification proceeds. Negative charge emerged within tbe 14-day-ol d embryonal skin, and was gathered in the granuiar layer as skin matures. Glycosaminoglycao (GAG) chains, which are highly negatively charged, were neutralized with cationic molecule, polyethyleneimine (PEI). PEI induced cleft formation within the supraba.wI layer of the skin of the 15day-old embryos, and within the granular layer of the neonataI skin. These findings indicate that GAG chains may function in cell-cell adhesion within the epidermis, and ET may cleave the epidermis as tbe result of the disruption of proteoglycan-mediated cell-cell adhesion.
ANTIBIOTIC SUSCEPTIBILITIES OF PROPIONtBACTElUUM ACNES ISOLATED FROM ACNE VULGARIS : A 25-YEAR RETROSPBCTIVB STUDY. I. Kurokawa’)*, S. Nishiiimaz), S. Kawabata*. “Department of Dermatology, Hyogo Prefectural Tsukaguchi Hospital, Amagasaki, Hyogo, Japan. “Department of Dermatology, Kori Branch Hospital, Kansai Medical University, Neyagawa, Osaka, Japan. “Division of Dennatologicals 8 Ophthabnologicals, Ako Research Institute, Otsuka Pharmaceutical Co. Ltd., Ako, Hyogo, Japan. We have evaluated antibiotic susceptibilities of Propionibacterium ~lcnes (P. owes) isolated from acne lesions during the last 25 years retrospectively. Clinical isolates of P. acnes were tested for antibiotic susceptibilities using an agar plate dilution method. Although highly resistant strains to erythromycio and clindamycin were observed in about 10% of all strains, the others were verv sensitive to these drums. No resistant strains were observed against minocycline and nadifloxacin. During the last 25 years, no obvious resistant tendency of P. ocnes was not found by evaluating antibiotic susceptibilities of P. acnes.