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235 Analysis of cell kinetics of the human skin by a novel method using bromodeoxyuridine
JSID Abstracts / Jownal of Dermatological Science 10 (1995) 61-102
238
235 ANALYSIS OF CELL KINETICS OF THE HUMAN SKIN BY A NOVEL MEl.HOD USING BROMODEOXYURIDINE ., & Nskano’* T. I&,,&, 80d F. Murata’. ‘Department of Dermatology, ‘I?epartment or Analomy. Facully ol Medicine. Kagasbima University. Kagoshima, Japan. Bromodeoxyuridine (BrdUrd) is a tbymidine analogue which is of S-phase cells. incorporated into DNA-synthesizing nuclei lmmunocytochemical detection of BrdIJrd has become an well established method for cell kinetic studies. We topically applied BrdUrd dissolved in isotonic sodium chloride solution an the human skin with occlusive dressrog method (ODM) alter cape-stripping (X30) of the skin. The skrn was excised and fined in 10% phosphate-buffered formalin and embedded in paraffin. Deparaffinized tissues were immunohistochemically examiod using a0 anti-BrdUrd monoclonal antibody. We found that BrdUrd well penetrated into the dermis through the epidermis by tape-stripping and ODM and that lymphocyte proliferarion by malignant Lymphoma and heavy lymphocytic
infiltration by inflammatory skin diseases were well differentiated with this method This method could be applied for cell kinetic studies in various
skin diseases
ACETYLATOR PHENOTYPING IN PATIENT WITH pPHENYLENEDlAMlNE (PPD) ALLERGY. Y.Kawakubo*. M. N-E.. M. Ohkide Department of Dermatology, Tokai University School of Medicine, Isehara, Japan and #Universitats-Hautklinik Freiburg FRG. Not all haptens can react directly with a carrier protein. Several of them needtote metabolically activated, mainly through oxidative process. Therefore, these substances are dasiited as prohapten. Conjugating systems for the xenobiotics are assumed to have detoxifying effect and we of them, namely N-acetylation, is proved to show interindividual difference in the capacity. The aim of the present study is to investigate the N-acetylating capacity among patients with PPD allergy and normal subjects. Aswereported previously, the N-acetylating capacity in the hair root is well correlated with that judged from urinary caffein metabolites, we em@oyed caffein-method for the study. Seventy-four patients with contact dematitis were investigated. From patch test results, seven patients were sensitized by PPD. Acceding to urinary caffein metabolites, we observed three groups, fast, intermediate and slow acetylators in the normal subjects and PPD-negative patients, respectively. However, we could not find no rapid acetylator in the PPD-alergic patient group. Therefore acetylator phenotype might be a gxdmarker for thesensitivity toPPD.
236
239
HlSTOCHEMlCAL STAINING OF NITRIC OXIDE SYNTHASE (NADPHDIAPHORASE) IN NORMAL HUhSAN SKIN C. Mod and Y. Sas& Dqmbnent of Dennatolcw, Kwume University School of Medicine, Kwwme, Japan. Nitric oxide (NO), a toxic gas with fire-radical pnpaties, serves as a neunmal messenger in the inhibitory mnadrenergic nonclmliic aenoua regulation of smooth muscle, and *t mediatar of cell pmlifeTation and i&mmation of vtiLnls tissues. NO is generated auymatically from L-argininc by a wnstib~live, cytcdic, Ca~/calmodulin-activated NO synthape (NOS): NADPH- and tetrahydmbiopterindepadent cytochrome P-450~type bemo~roieia. Dir& detection of NOS activity is diRic,dt. bowever the bistockmical d&e&ion of NADPH-diaobomse RJADPH-d) is widely ied to repmat NOS activity In the present shdy;NOS catalytic acti&
I-BP72
was detacd in normal human skin. Unfiied cryostat skin sections wm brie@ fixed with acetone,then incubated with PBS coataiaing NADPH and aitro blue tctrazolium (NBT). Water-insoluble NBT famazaa (NADPH-d) was ~rcseat ia the @amis, blood vessels, twreckx pilli mu& and eccriae glands, boww, pai~bual nerves were devoid of the ma&a poducts. ‘Ibe pnscnt vault showed that NOS is present in the epidermis, blood w&s and anwtor pilli muscles of normal human skin. Sinw NO is gmeratcd by NOS. NO may be releasedfrom UK epiti awl/or dermis, andmayactconMmitantlyonthe~.
BATS
INDUCTION IN SYSTEMIC ORGANS OF AFTER SEVERE BURN INJURY M. Hatoko*,
S. Hirai, H. Tada. T. Mummatsu, and T. Shimi. Department of Dermatology, Nara Medical University. Kashiham, Japan. III order to know Ihe stress response of the systemic organs to severe bum injury, we have investigated the induction of 72-kD heat shock protein(HSP72) in various organs (brain, hypophysis, lung, h~,liver,pancreas.spleen,kidney,adrenal gland,andskeleial muscle) of the severely burned rats. Full thickness bum was developed on the rats’ skin by immersing the rats into the hot water (9o’C ) for 3 sec. At 0.24, and 48 hr after bum injury. the HSP72 expression of various organs was examined by Western blot analysis. At 24hr after bum injury, the level of HSPn increased in hypophysis, lung, heart, and kidney. In all organs examined, the expression of HSF72 increased 48hr after bum inJury. The level of the HSPZ? was the highest in hypophysls and the lowest in brain and adrenal gland 48hr after injury. These results indicate that severe bum inJuy actually causes the stress response in systemic organs.
240 SEASONAL VARIATION OF L’ VALUE OF PIGMWIBD ARRAON FEMALE FACE MEASURED BY REMO’IE COLOR SRNSUW SYSTEM. M.Miauaki*. Slbksda. M.Nl Sbiaeido butimte of Beauty 8chcea. Tokyo, “Sbiido Rewcb Ceater, lbkobama, Japan It is known ttmt tin color of pi~moated area 011female face cbaogc” by seaxms. In order to eoafirm tbia phenomenon, w attempted to menwe akio color of pigmented area and non-pigmented area on female face every month for a year. Vohmteem comprises 20 femalea from 34 to 49 yun old, who had pigmentation on their fieea. Tbe &in color M amewxl by meaaariog L*.a*.b* values of the face image takeo by ITV camem. Daring tbh pxiod, the amount of W mm measured with WAB Radiometer (Euro LT. Co&d). The L* values of both pigmentsa area and ooa-pigmented area were dk?m?asingfmmAprutoAuguatwithtlw blcreueoftbeamoaatofw.Bat the degree of the obange of pigaented area b more than that of mmpigmentedtru.Attersumma,L*vrluarwtasd~tothcvrlwmolsurrd in F&wry. lbere reaolb aaggeated that L* vabm of p&pawed ama ebaqjed easily sccordii to the amomlt of w.
238
235 ANALYSIS OF CELL KINETICS OF THE HUMAN SKIN BY A NOVEL MEl.HOD USING BROMODEOXYURIDINE ., & Nskano’* T. I&,,&, 80d F. Murata’. ‘Department of Dermatology, ‘I?epartment or Analomy. Facully ol Medicine. Kagasbima University. Kagoshima, Japan. Bromodeoxyuridine (BrdUrd) is a tbymidine analogue which is of S-phase cells. incorporated into DNA-synthesizing nuclei lmmunocytochemical detection of BrdIJrd has become an well established method for cell kinetic studies. We topically applied BrdUrd dissolved in isotonic sodium chloride solution an the human skin with occlusive dressrog method (ODM) alter cape-stripping (X30) of the skin. The skrn was excised and fined in 10% phosphate-buffered formalin and embedded in paraffin. Deparaffinized tissues were immunohistochemically examiod using a0 anti-BrdUrd monoclonal antibody. We found that BrdUrd well penetrated into the dermis through the epidermis by tape-stripping and ODM and that lymphocyte proliferarion by malignant Lymphoma and heavy lymphocytic
infiltration by inflammatory skin diseases were well differentiated with this method This method could be applied for cell kinetic studies in various
skin diseases
ACETYLATOR PHENOTYPING IN PATIENT WITH pPHENYLENEDlAMlNE (PPD) ALLERGY. Y.Kawakubo*. M. N-E.. M. Ohkide Department of Dermatology, Tokai University School of Medicine, Isehara, Japan and #Universitats-Hautklinik Freiburg FRG. Not all haptens can react directly with a carrier protein. Several of them needtote metabolically activated, mainly through oxidative process. Therefore, these substances are dasiited as prohapten. Conjugating systems for the xenobiotics are assumed to have detoxifying effect and we of them, namely N-acetylation, is proved to show interindividual difference in the capacity. The aim of the present study is to investigate the N-acetylating capacity among patients with PPD allergy and normal subjects. Aswereported previously, the N-acetylating capacity in the hair root is well correlated with that judged from urinary caffein metabolites, we em@oyed caffein-method for the study. Seventy-four patients with contact dematitis were investigated. From patch test results, seven patients were sensitized by PPD. Acceding to urinary caffein metabolites, we observed three groups, fast, intermediate and slow acetylators in the normal subjects and PPD-negative patients, respectively. However, we could not find no rapid acetylator in the PPD-alergic patient group. Therefore acetylator phenotype might be a gxdmarker for thesensitivity toPPD.
236
239
HlSTOCHEMlCAL STAINING OF NITRIC OXIDE SYNTHASE (NADPHDIAPHORASE) IN NORMAL HUhSAN SKIN C. Mod and Y. Sas& Dqmbnent of Dennatolcw, Kwume University School of Medicine, Kwwme, Japan. Nitric oxide (NO), a toxic gas with fire-radical pnpaties, serves as a neunmal messenger in the inhibitory mnadrenergic nonclmliic aenoua regulation of smooth muscle, and *t mediatar of cell pmlifeTation and i&mmation of vtiLnls tissues. NO is generated auymatically from L-argininc by a wnstib~live, cytcdic, Ca~/calmodulin-activated NO synthape (NOS): NADPH- and tetrahydmbiopterindepadent cytochrome P-450~type bemo~roieia. Dir& detection of NOS activity is diRic,dt. bowever the bistockmical d&e&ion of NADPH-diaobomse RJADPH-d) is widely ied to repmat NOS activity In the present shdy;NOS catalytic acti&
I-BP72
was detacd in normal human skin. Unfiied cryostat skin sections wm brie@ fixed with acetone,then incubated with PBS coataiaing NADPH and aitro blue tctrazolium (NBT). Water-insoluble NBT famazaa (NADPH-d) was ~rcseat ia the @amis, blood vessels, twreckx pilli mu& and eccriae glands, boww, pai~bual nerves were devoid of the ma&a poducts. ‘Ibe pnscnt vault showed that NOS is present in the epidermis, blood w&s and anwtor pilli muscles of normal human skin. Sinw NO is gmeratcd by NOS. NO may be releasedfrom UK epiti awl/or dermis, andmayactconMmitantlyonthe~.
BATS
INDUCTION IN SYSTEMIC ORGANS OF AFTER SEVERE BURN INJURY M. Hatoko*,
S. Hirai, H. Tada. T. Mummatsu, and T. Shimi. Department of Dermatology, Nara Medical University. Kashiham, Japan. III order to know Ihe stress response of the systemic organs to severe bum injury, we have investigated the induction of 72-kD heat shock protein(HSP72) in various organs (brain, hypophysis, lung, h~,liver,pancreas.spleen,kidney,adrenal gland,andskeleial muscle) of the severely burned rats. Full thickness bum was developed on the rats’ skin by immersing the rats into the hot water (9o’C ) for 3 sec. At 0.24, and 48 hr after bum injury. the HSP72 expression of various organs was examined by Western blot analysis. At 24hr after bum injury, the level of HSPn increased in hypophysis, lung, heart, and kidney. In all organs examined, the expression of HSF72 increased 48hr after bum inJury. The level of the HSPZ? was the highest in hypophysls and the lowest in brain and adrenal gland 48hr after injury. These results indicate that severe bum inJuy actually causes the stress response in systemic organs.
240 SEASONAL VARIATION OF L’ VALUE OF PIGMWIBD ARRAON FEMALE FACE MEASURED BY REMO’IE COLOR SRNSUW SYSTEM. M.Miauaki*. Slbksda. M.Nl Sbiaeido butimte of Beauty 8chcea. Tokyo, “Sbiido Rewcb Ceater, lbkobama, Japan It is known ttmt tin color of pi~moated area 011female face cbaogc” by seaxms. In order to eoafirm tbia phenomenon, w attempted to menwe akio color of pigmented area and non-pigmented area on female face every month for a year. Vohmteem comprises 20 femalea from 34 to 49 yun old, who had pigmentation on their fieea. Tbe &in color M amewxl by meaaariog L*.a*.b* values of the face image takeo by ITV camem. Daring tbh pxiod, the amount of W mm measured with WAB Radiometer (Euro LT. Co&d). The L* values of both pigmentsa area and ooa-pigmented area were dk?m?asingfmmAprutoAuguatwithtlw blcreueoftbeamoaatofw.Bat the degree of the obange of pigaented area b more than that of mmpigmentedtru.Attersumma,L*vrluarwtasd~tothcvrlwmolsurrd in F&wry. lbere reaolb aaggeated that L* vabm of p&pawed ama ebaqjed easily sccordii to the amomlt of w.