SCUR ABSTRACTS Abstracts for the Society for Cutaneous Ultrastructure Research (SCUR) 23rd Annual Meeting Grand Hotel, Gardone Riviera, Brescia, Italy, April 11-13, 1996
~ Sessions Abstract Numbers
Session 1: Platform presentations 1-4 Keratinocytes Session 2: Platform presentations 5-8 Melanocytes, antigen-presenting celis, endothelial cells Session 3: Invited lecture 9 "Interaction between cytotoxic lymphocytes and target cells" Session 4: Platform presentations 10-13 The effects of UV radiation on epidermal cells Session 5: Platform presentations 14-19 Confocal laser scanning microscopy and immunoelectronmicroscopy in the study of skin biology Session 6: Invited lecture 20 "Recent developments in correlative morphology: ultra high resolution scanning electron microscopy used in molecular and cell biology" Session 7: Platform presentations 21 - 24 r Diagnostic electron microscopy Session 8: Invited lecture 25 "Cytoplasmic markers in histiocytic syndromes" Session 9: Short communications 26 - 35 SCUR Dermopathology Club • Session 10: Platform presentations 36 - 39 Bullous diseases Session 11: Invited lecture 40 "Lysosomes and programmed cell death: reflections on keratinization" Poster session 41-59
Scientific Program Committee Exhibitors at the Annual Meeting
Giuseppe De Panfilis, M.D. , Chairman Beiersdorf S.p.A. Jean Kanitakis, M .D. Bio Nike istituto Chi111ico Italiano S.a.S. G. Carlo Manara, M.D. Cady Paris Dieter Metze, M.D. Far111ila DermicaI S.r.I. Giorgio Pasolini, M.D. Istituto Ganassini Janssen-Cilag S.p.A. Laboratoires Pharmaceutiques de La Roche Posay Italia S.p.A. Laboratori Guieu S.p.A. Prodotti Formenti S.r.1. Local Organizing Committee Rydelle Laboratoires Business-Johnson Wax S.p.A. Schering S.p.A. Giuseppe De Panfilis, M.D. Schering Plough S.p.A. Antonietta Lonati, M.D. Azienda Ospedaliera Spedali Civili di Brescia G. Carlo Manara, M.D. Va letudo S.r.l., Divisione Biogena Giorgio Pasolini, M .D. Ya111anouchi Pharm3 S.p.A.
0022-202X/96/S10.50 Copyright © 1996 by The Society for Investigative Dermatology, Inc.
655 656 AUSTltACT S TH E JOUR NAL O F INVESTIGATIVE DEI1..M AT O LOGY
1 2 C IIANGES IN KERATINOCYTE CELL-CELL ADIIESION AND CYTOSKELETON THE EX PllESSION OF INTERM EDI AT E FILAMENTS OF HUM AN REARRANGEMENTS INDUCED DY I NIIIIlITORS OF SER INE-T IIR EONI NE KERATINOC YT ES IN VITRO DIO r IO o s ON A FEEDEll -LAYER. Fm llc"sca PROTEIN PI-IOSI' Jl ATASES. Ma «'k Haftck Mirc jllc Serres a nd Daniel Sc hmit!. Pri ll ll :l no. 1.0101 Domenici. Gi:lIIl1i Gerlini. Nicola Pimnincl li . Paolo Romall lloli. INSERM U.346/ CNR S, Dept. of Dennatology, E. Herriot Hos pilal, Lyon, France. I)e parllllc ni or Human An il lomy a nd Hi slology and In slitul c o f Dc rnmtology. Protein pho5phorylation steps arc involved in the regul ation of cell -cell and cell Uni versity of I=l ore nce. Ita ly. substr.1tc adhesio n, We treated cell cultures of nomlal human and HaCaT kcratinocytcs Norl11 al human kc ral inOt:y tcs can proliferate. diffe re ntiate and str.lIi fy in vitro on a with potcnt inhibitors of se rine- threonine protein phosphalascs. okadaic acid (500 oM) feede r laye r of IClhall y irradialed or lIl ilo rn ycilHreatcd 3T3 fi broblasts: un der these and calyc ulin A (50 nM), in order to study th eir innuence on the cell adhesion. cO TICl iti o ll s th e), e>..pre ss llIa ny dirfe reniiati onlllark e rs. incl ud ing inte rmcdi ate fil a mc nt Kcralinocylcs grown in nonnal calcium medium fonned monolaycrs joined together by pru te in s. I II o rder 10 assess th e role o f feede r c ells 0 11 th e pro liferatio n ~lIl d numerous villosilics and infrequent dcsmosomes. In places, the cultures showed diffe rl' llt ia ti o tl or kcratinocytes we a na lysed. by inHIlUilO lluoresccll ce a nd e lec tron stratifi cation and expressed numerou s dcsmosol11 cs in th e nattened suprnbasal layers. mi crosl·oPY. 1I 0 fll1:11 human keratinocYles grown on pl .. sti c in n !'ilandard m edium Afler 60 min. exposure to the drugs, the ce ll s lost their ce ll ·ccll c ontac ts hut remained ( OlJ l becco '~ modifi ed llIi nimlllll esse nl ia llllediulll . with 200 mg/L e nCl 2 ;! !ld 10% retal attac hed to the plasti c and showed no ultrastruc tural signs o f cell suffering. The I:a lr se rum ). Kc [';tliIl Ol:ytes were gro w lI fo r c ight consccuti ve passagcs and ana lyzed al supmbasallaycrs delached "cn bloc". As the basaJ cell s "ro unded up", their intennediate th e fi rsl. Ibird. fifth and sc vc llih passngc. Tilc cells reached cOllllue tl ce a t ;lJl passages . maments, which, initially, we re uniformly dislTibutcd in the cyto plasm and formed well with the e.'\ce pli o ll of thc first OtI C, but ncver exprcssed v ill1 cl1tin o r cytokcralins. as de fined bundles in association with desmosomes, re tmc ted under lhe inOucnce of drugs i llui c ~ lI e d by illlrnunorluo rcscel1 cc wilh ilrrinity i501<1l e 3 4 THE DEVELOPMENT OF A STRUCTURALLY COMPETENT EPIDERMAL RE PLI CATI ON TECHNtQUES FOR SCANN tNG· AND FRE EZ E FRACTU RE ELECT RON BARRIER iN AIR-EXPOSED KERATiNOC YTE CULTURES: A TIME COURSE MICROSCOPY TO VISUA LI SE TH E ULTRASTRUCTU RE OF HUMAN STRATUM STUDY. M. Farlasch M. Ponec M. Rosdy. Depl. of Derm atol ogy, Univ. of CORNEUM IN RELATI ON TO ITS CURVATURES . Fe r Soies Remle G V'HI cl er Molen Joost Erlangen, Germ any, Depl. of Dermalology, Univ. of Leiden, The Nelherlands, M. van .! Nonrdl'lIdc Ik llk K Koenen A. Micke Mot!l!l\aas. Department uf Electron Microscopy. Laboraloire SKINETHIC, Nice, France. Univcrsity of Ld llell , Tht: Netherl ands. Recentl y we could show that an air-exposed (AlE) keralinocyle cullure The li vill1; hlllllll ll s!ratulII COrucUIII consists of w rneucylc$ cmbedded ill lipi d hila)'e rs. and is reconstituted in chemically defined medium formed a slructurally and usu:tll y I.:u ntparetl witlt :I brid wali. Howcver tit is wa ll possesscs Ill 5 6 TilE lJS E OF X-RAY MICROANALYSIS TO DETEJ{ MINE M ELAN IN T YPES IN I·IUM AN C1 IARACTERI SA TI ON OF GI-PHASE IN NEVUS CELL NEVU S AN D SKIN MELANOSOMES. II K Ktw T! S: 1I N Smj' J vall \Iii[ MC\I!cll A M Mnrnmaas 'Int! S p ..... 1'! MALI GNANT MELANOMA I3 Y D UI3LE LABE LLI NG IN TISS UE SECTI ON La htifali lry lelr Elcc trcll l Mic mSl.: llPY ;llld DC P;lrtlllclU of Dcm l:unlogy. l..t:idcll Ulli vC l'sit)' Hospital . S teFli' Ilqxl('I'Ill'UUl , tyhnjna !3pcln(fJC' h. !)uhlc:;. Pe ter A hnlC'yn C linic for Lcitlcil. T he Nl.: ll u.!rlam..ls The hi gh ind dcm.:c nf skin cam:cr in p:uiclUs willi skill type I (reu hair :lIld palc skin, DCt'Ilmtnlogy oflhc Rllhr· Uni versit y nl Boehulll, Gennuny COrll[l,m.:ll \(l iudiv idual:-. with skill type VI (lI cgruitl) has ura wu the ,lIlCUli o u III the role ()f pi!l Ul CIlt Malignancy is cClIlnected with Ihe loss of controll ed cell pro life ration. Decrease of I llIclaui u) ill ti ll.: prolCC lioli againsl advc rsc crfcI.: ts of UV liVlu . III hUl1Ia ll mdanoc YIl.:s 11,1,'0 types of certain ce ll ph3ses leads to an increase of cleavage aCli vity at the levcl o f single cells. I1Idall il1 I.: an he rccllglliz I.: lI : pheomdallill. pn:lI ominamly prcselll in :i kiu ty pe 1 and cumdunin. The aim or th is study was \0 dctcllll inc lhe S·phase rraction a lllQunl in prolircmling mai nl y fou nd in type VI 1IIc1 : u. o~.:y U!s. Pheomdallill is h OWl! tll gC lLeratc free radicals under the cell po pulati ons lIsing Bromodeoxyurid ine / M II3 I (Ki67) double lubcll ing on lo r· illtl ueuI.:e III UV· Ii!jhl ;tlltl eUludaniu aCls unde r Ihe sounc I.:onditi Ull" as a free r;utical ~a ve ll gc r . Thc ult ra lo tnu.; lurc of mdallu:-ou mcs from Ihe twu skin types is differel\( too. McI;lIIosulU CS uf skin type I nmli n lixcd pal1l ffin sections. 'l1,e resulting quotient referred to IlS labell ing index. (l rc rdativel y cl et:l nlll lucent . tltey cUllsist frequently of fusclJ gra nult.:s :md iI spcl.: ific fille structure is al lows interprcwtion of tcmpomry shift ing of cell cyc le plmses. 'll,C labelli ng index ge nerall y uhse rvelJ . The melli lloSHIlI C,'; of skin type VI lire larger and mo re ekc lmlL dellsc. COIII W.IS dClcllllincd fo r ker8 li nocytes. ncvus cells and nml ignant mclulloma ccll s with plcxClI gr'l1I ule lo ami a fiue slnn;tu re. li ke ill type 1. arc oul y rarel y obse rved. Apan from uUlrphnln resulting quotients o f' 11.7 • 22.8% fo r nevus cei l nevi and 15.4 - 78.7% for liY. a difh:rellce in the I.: helllical ClIl11pusitiulI . i. e .• the sulphu r I.; oncentratiou. hc tweell the two malignant mclanOrtl<1. Additionally the melanoma cells were detected elcctro n IIId:-IIIu.,.ol11 e t y pe ~ C: III he esta hl ishelJ. It has beell dc.snibcd Oil hasis tJf X- ra y micman:llytical tX RMA) d;II:1 Ihill lIIe\;tll()S(U lles of skin type I havc a rd:uive high sulphur cuncentra tion, wherells microscopicall y by using Ihe ant ibody I-1MB 45. An expanded supcrlic ial spreading mel anot.:y tes fro l1l ski n type VI ctllilain low alll(lunt ... of sulphu r. U... ing the same h..-..: hnillue we could mal ignant melanoma wilh regression zone exhibited VCt) ' hi gh indices in the demonSlr;lIe II nUlUlce ill this onsc rv;uinll . TIle sulphur cO IH:el1lr,ui nn or mehwns() t1I es within lhc stune nodul ar as well as in the pcripheml unci in regression zones. Top values of labell ing imlividual appeared to he site depelldeli lo Melallocytes in Ihe skin of a type I illdi vidl lil l exhihit a 11Iw indi ces have been documented in melanoma metastas is. Compari son of all proli· sulphur t.:nm.:ell tnllio ll . while the mdanOSOlIlCS in the hai r hulb of th t: salllC indiv idual arc. o n aver· 'c rating cel ls by Ihei r ind ices showed more S·plmsc cell s in maligna nt pigmcnted :t!;c. rid I in sulphur. Similar site depe udellt variations we re nhserved ill skill type VI md:ulU suIIICS. ;:1 The result lo of thi lo study Slll lW that XR MA allows the dt:lfactt:ri7.1ttioll llf melanin tYI'e 0 11 ha... is uf tUIll OUrs. This indical es thut mel anomas displuy decreased G I-phase us n the ~ tl l phtl r c()m:emr:ltion in Ille n H.: 1:IIHI .~ u m e s. We will , therefore, usc XRM A data to delel'llli lle consequence of 311 (! xlrcmely short resting stale (GO·phase). Our resu\ts lead to thc elt;lII!;elo fro m pll el1 - In eUllI cianili atlll vice ve rsa in stud ies thai arc a imed (u manipulate the concl usion that Ihc 1:.L bclling index can be used as a reliable indicator 10 1' Ilmlignancy. mcl :UHlge ll esis ill vilrtl. VOL. 107, NO.4 OCTO I3 EI1. 1996 A BSTR.ACTS 657 7 8 EFFECTS OF CYCLOSPO RI N-A ON T HE MORPHOLOGY OF HUMAN E FFECTS OF TRANSFORMING GROWTH FACTOR fl3 ON C ELLS OF LANGER HANS CELLS IN C ULTURED EPIDERMAL S HEETS. ErilIl= VASCULf\R \VALL. Tonny Karlsrnark Takasj Kobayasi Dept. of Demlatology, Prig!lano. Glallnl Gerlini. Nicolft Pimninclli. Pnolo ROnli1l1l1oli. Be nvenuto Ginnnolli . Rigshospilal. Uni v o f Copcnhagen, Denmark. Ins~{Jt ut :- of Derm atology and Department of Human An atom y and Hi stology. UOIver:slty of "'l orence. haly. It has bcen supposcd that transforming growth factor rn (TGFJ33) has some cflects Epldennal sheets obtained by di spasc trc.11mcllt frolll dCmlalOIll CS of 1I 0nna i s kin arc on dermal fibroblasts In a previous mecting. the authors have demonstrated that TGFp a n i~po r1 a nt 1001. 1,0 in vesti ga te th e fine structure of Lll ngcrlmTl s cell s in varioll s I promotcd pcricytes transformalion to myofibroblasls in the vascul ar wall This study ex~ nm en t a l conditions. Since th e number, dcndrilicity and expression of charac teri sti c was intend ed to estimate the effects of TGFj33 on th c transformat io n and to compare a ntigens (H0-Dr and D In) of L1n gc rh 9 10 INfERAcrlON BETWEEN CYTOTOXIC LYMPHOCYTES AND TARGET CELLS . UV-INDUCED ULTRASTRUCTURAL MODIFICATIONS OF Q. Arancja. A Slrjnl'uro • .e...... J:J:a. C Ra monj, C M Au sjell o. A Cassone. KERATINOCYTES IN HUMAN EPIDERMIS: PROTECTION ISOlU to S upcriorc di Sani t:'t. Viale Regin a E1 cna 299. 00 16 1 Romc. Italy. BY VITAMIN E. P.U. Giacom oni ., I.F. Nadaud ., E. Straface. The mechani sms of the cytotoxic acti on of natura l kille r (N K) cells and ly01phokine-acti vntcd kille r (LAK) cell s against tumor cell targets arc relati G. Donelli. W. Malomi & M. Heencn # - • Dept de Rccllcrdre de L'Orial vely we ll dcfin ed. Ultr:lstructural studies de mons trated that the extent of umcomc, C/troilly-unle, Fran ce - # Service dc Dcrmat%gic, Hopita/ Erasrtlc, cell-to-cell contact in target-crfector conjugatcs is c rit ical for tri ggcring th e Bruxcl/cs, Be/gillm and Dept oJ U1 trastructllres, Is titllto SlI pcriort: di Sarrita, Roma. activation path ways o f cytotoxic crfectors. The fornlillion of thc so-call ed clo Italy. sed c h amber bctwecn effector and la rget cells fa vors inte rcellul ar s ignal We have explon:!d the morphological modifications induced by UV ling and all ows lyti c fa c tors to reach th e target. With regurd 10 the intera radialion fro m It solar simulator on the epidermis of informed volonleers (2 to clion of the cytotox ic lymphocytes wi Lh mi c roo rganisms. onl y a ve ry limited 3 minimal erythemDl doses) and biopsies were taken befon:! and 2, 4 and 7 amount o f infonll ati on is available. Thercfore. we studicd the inte rac li on i" hours aft er the irradiation. Biopsies wen:! fixed and prepared for o pLical and vitro between hum an lymph ocytes (NK und LAK ) and th e opportunis lic electron microscopy analys ts. pathogen Ca/ldida a l bi cal/.f. Ou r observati ons del1l onstr:lle th ll l hum :l11 NK and 2 and ... hours aft er the irradiation. nudear chromatin condenses and L..AK cells differ in lheir ability to bind to th c differen t form s of growth o f C. keratinocytes undergo spongiosis, i.e. cell· to-cell contacts arc lost and the cell albicafl s and Ih at the 1I1 0st cx tcnsive and ti ght cst contacts occur betwecn LAK surfaces become irreeular. In the intercellu,lar lilmina, round, electro n dense ce lls a nd fun gal germ tubes. Nonetheless. th e hi ghl y in te rac tive LAK cell s bodies are observed, which in some instances a,n:! connected to one of the cell. are unable to kil l or inhibit thc growth of C. a lbi ea l/ ,\'. as scarcely interacti ve This suggests that these bodies are ceUular blebs, cut by section planes which NK c ells. evcn when the largel is the ge rm tube. In addition. recent in some instances cross the cell-to·bleb junction, and in several other instances observatio ns point o ul th (ll C. a lbi cclIIs. IIflcr th c ime fll clio ll with e rfec tor do nol. cells. is cap abl e of modulatin g Ih e cx pression of some surface compone nt s 7 hours after the irradiation, keratinocytes do no longer display which can e nter cyto tox ic lymph ocytes inducing evid ent mo rpho logical morphological modifica Lions in keeping with the expected repair. damage . Fi nally. th e interac ti on betwee n C. albiealls and NK / LAK cell s Pn:!tn:!ating the epidermis with topical applica lion o f a cream containing 1 :t:. induces a rapid and elc·.. alcd IFN-'Y and GM-CSF producti on by th e cytotox ic vitamin E dramatically protects epidermis against Lhe cellular nnd nuclear cells. These observations indieill c a potentiall y import,lIIt rol e. ahhough nO Il alteralions observed 2 hours after UV. directl y inhibitory. of thcse inlmunoeffeetors in umi-emldida defe nse whi ch can occ ur by aCliva ting phagocyti c cell s or influe nc in g regul :ll o ry T lym phocy tes. 11 12 UV-INDUCED BLEBBING OF NORMAL HUMAN I> ERMAL EL tMINATION Of APOPTOTIC AND VITAL MELANOCYTES AFTER UV A I KERATINOCYTES IN CULTURE IS HINDERED BY IRRADIAT ION. Manina n harach- Buhles. "nlomas Kromcr, Stcfa l! el :111m!!!.!. ~ a-TOCOPHEROL. P.U. Giacomoni"', E. Straface, G. Donelli & Altmeyer. Dermatological Clini c of th e Ruhr-University. Bochulll. Gennany W. Malorni - .. Laborat o;rcs de Reclll:rc}1e de L'Odal-umcome, Clleuilly-Larue, Physiologically in adults Illelan ocytes arc located in th e cpidcmlis, but 1I 0t in th e Frana and DqJt o/Ultrastnlctllrcs, Istitllto SlIpcriorc di Sa"ita, Roma, Italy. dennis. In this study we invcstigated the di stribution and uitrastlllclUral alt cl'a tiolls o r Upon exposure to UV radiation, normal human keratinocytes in melall ocylCs lin er repeliled high dose irTllciiation with UV A I (340-400 11111 ). culture lose cell-to-cell and cell-to-subs trate contacts, round up, In 14 probands th e volar sid e of the forearm was irradint ed with 20 J I cm'! 10 x undergo zeiosis (which is cha racte rized by the appearance of many \\~t hin 14 days and bioJl sicd imlllcdiatcly aficr Lhc last irradiati on. surface ble bs) and eventually detach and floa t in the medium. Lightmicroseopic:IUy melanin was detectcd by mcn us of Fontana-Musson stailling. Twenty four h ours after UV irradiation with a dose roughly equivalent Scrics of cpou-el1lb edded, scmithill scctions were processcd for compu!er- suppon ed to one human MED (minimal erythemal d ose) the fraction of Lhree-dimensional reconstruct ions of basal Il1 clauocytcs. Ultrathin sections were damaged-cells (cells with blebs) is 70 % for both UV A and UVB. cxalilin ed for Illclanocytcs in and bclow thc cpidennis. About 60 % of d e tached cells dis play segmented nuclei (apoptotic Afier irradiati on with UV A I multiple cells below th e dennal- epidenllal basclllelll cells) after 200.000 ;, m' UVA. This number is 90 % after 1 200 Jim' membrane stain ed posit ive by the Fontana Masson-stain ing. Ult ra stnlcturnlly these UVB. cells could be identified as mclall ocytes in thc denni s. Somc of these Illelanocytcs Treating cells with 30J.1M a-tocopherol before UV irradiation reduces ex hibited signs of degencration with intraccllular Iysosollles, whercas others werc the fraction of blebbing cells to 4 % after UV A or to 20 % after UVB and absolutely inlncl. nle int act IIlclanocytes were cncircled by an eX1rl!cellular shcath the fraction of apoptotk cells drops to 30 % or to 60 % afte r UVA or composed of granular components, looking li ke the basal lamina o r the dcnno UV8, resp ectively. cpidcnll:ll juncti oll . 'Ille demlO-epidcnnal basement mcmbrane was fo ca lly Treating cells with vitamin E a ft er UV radiation reduces the fraction of int errupted or lIlultiplicatcd. Focally aggrcgates of a fin e fi lamentary material blebbing cells to 25 % (UV A) or to 20 % (UVB) and the fra ction of exhibiting I11 clanosomes could be dctectcd in the upper dermis. apoptotic cells among detached ones remain practically cons ta nt after High dosc irradiation with UV A I cnuscs an in creased apoPlosis bUI also II dennal UVA (70 %) as well as after UVB (60 %). elimination of vilal IIlcllluoeytes. "nle obsclv ation of vit nl Illclanocy1es ill til e dCl1l1i s corrclates with thc ultrastructural observations ill acquired dcnnnlmcinnocytosis. 658 AUSTRACTS THEjOUI,NAL OF INVEST IGAT IVE IJER.MATOLOGY 13 14 MICIWSCO I'I CA L AI.T EI< ATIONS 01- MOUSE I.AN(;ElmANS CELLS UI'ON CONFOCAL LASER SCANNING MICROSCOPY - A USEFUL TOOL FOR UI.T RAV IO LET Ii RADI ATI ON AT LOW DOSI-:..1.3.uill ROI11:-t "llo li . Slef;mo Hac.:ci. INVESTIGATION OF IMMUNOFLUORESCENCE STAININGS. l ~ ra n l'l:"'c Ol " ri<'II;IIII), ~!.Y.!!£ Slrci lci u. Dl.! pHrt lllc nl o f I-IUtll 15 16 DESMOSOMAL AND ADHERENS JUNCTION-ASSOCIATED ULTRASTRUCTURAL LOCALIZATION OF PROTEINS IN DARIER DISEASE AND HAILEY-HAILEY DISEASE_ CORNIFICA TION-ASSOCIATED PROTEINS IN HUMAN C. Kowalewski'. A. Winkielman', D.Schmitt#, M.Blaszczyk', S. Jablonska', EPIDERM IS. (+) and M. Haftek#. Dept. or Dermalology. Warsaw School or Medicine, Polan d Masashi Ak iya mal.2, Hiroshi Shimi zul • Taku ji Masuna ga l . Ak ira (1/) U 346. INSERM, Dept. or Dermalology. Hospilal E.Hcrriol, Lyo n, France Ishi kol. Kozo Yoneda), Soo-Yo ul Kim". Tnke ii Nishi kawa). The aim or th e study was to in vestigat e th e expression and ultrastructural loca lization I Department oj' Dermatology, Keio Un ive rsity School o j' Med icine: of desmosomal and ad hcrcns junction (AJ)-associalcd protei ns in normal and lesional 2Division o r Derm atology. Kilasato In stitute Hospi lnl; )Dcparlllleltl oj' epidermis of Darier disease (DO) and Hailey-Hailey disease (I·IHO). In additi on to immunofluorescence (IF) and post-embedding imll1unogold electron microscopy (JEM) Dermatology. Kyoto Un ive rsil Y Facult y or Medi cine, Japan: 4Pacilic th c laser scan ning confocal microscopy was uscd to dem onstrate chan ges in distribution Corp . R & D Center, Korca. of cytoskelctal protcins IF studies have shown th e presence of dcsmosomal and AJ Ullrastruclural locali zati on or keratin s. ril aggrin /profilaggrin , associated proleins in th e areas or cell-cell contact (CCe) in unin vo lved epidermis. In lori cri n, small proline-rich prolein s (S PR) I and 2. in volu crin , and aca ntholytic keratinocytes in both diseases, IF pattern of desmoplakin was difll,se Iransglut amin ase I (TG I) we re studied by posl-emmbedd in g whcreas plakoglobin and dcsmoglein I prcsent ed a dott pattern and clumps. IEM showed immunoelectron mi croscopy using a cryofi xali on-c ryosubstitulion an accumulat ion or desmoglein in the cytoplasm. however no internalization or meth od in normal human epiderm is. A nti body-binding sites were desmosomes wa s observed . The AJ-associ ated proteins were distributed al ong the demonstrated by imlllullugoid and sil ve r enh an cclll clll . Filaggrin and kera tinocyte plas ma membrane with accumulation in microvilli. Laser sca nning confoca l kerat ins I and 10 were loL:a lil.cd in kerat ohyalin granu les ill th e granul ar microscopy showed th aI Ih e co nn ecti on between partially dissociated cells was always cell laye r and lil aggrin was di spersed cl ilTusely in Ih e cytoplasm or Ih e mediated by both keratin and F-actin . Normal ex pression or desmoso mal and AJ first cornilled ce ll s. Loricrin , SPR 1 and 2. and involu crin were located associated proleins in uninvolved cpidermis and the similarities in Ihe distribution of to th e cell membrane or th e corniri ed ce ll s and to the cornified ce ll cYlOskeleta l and CCC-proteins in aca nth olytic keratinocylcs of both DO and HHD could suggest that the loss of CCC in the se two different diseases is due to the run ctiona l envelope. TG 1 was associated wit h th e cell membrane of Ih e granul ar changes in thc deslllosomal protein s. followed by mechanical disruption of AJ. and nol to anei spin ous cell s. These rindings support th e conlrihulion of TG I 10 Ih e a primary defect in th e sy nthesis of proteins fOflnin g ece formati on or cornified ce ll envelope ;11 vivo. 17 18 SURFACE MOLECULES INVOLVED IN TH E PHY SICAL CONTACT EVIDENCE FOR VESICLE-ASSOCIATED IN VIVO EXPRESSION OF CANDIDA ALBICANS BETWEE N LANG ERHAN S CELLS AND T LYMPHOCYTES. M. SECRETORY ASPARTIC PROTEINASE IN OROPHARYNGEAL CANDIDOSIS: RESULTS Concha l. .I. Peguet-Navarro' , MA . Vidal' , I. Caorsi , D. Sehm iW OF AN IMMUNOELECTRONMICROSCOPIC STUDY. Martin Schalferl, H a n s~ . Korting1. I[ nstl.tuto de I-hstologta y Patolo!;ta, Qnl verSi'ilii07\i'l'stra) de t illie, Margarete Borg-von Zepelin2, Markus Ollert3. , Department of D erma t o lO~Y, Ludwig - Va ldI via, Chtle: ' INSERM U346, Hopltal Ed. HetTIot , Lyon . I·rance. Ma ximillans· University . Munich, 2lnstilute of Hygiene, Univ ersity of Gollingen, Department II IS wel l kn own thai tn order 10 aCliva te T cell s Langerhans cell s (LC) of Biochemistry and Molecular Biology, University of Hamburo. Gennany. expose. nlllnunogenl c peplide/MH C class . II f~a gm e nt s a nd deliver approprtale acces.sory SIgnals. Probably , durtng tillS tnt eractlon, complex Isoforms of secretory aspartic proteinase of Candida albicans have been claimed to be adheSIon tn echamstn s arc required to keep both cell s in li gh t conlacJ. In a major virulence factor of th e opportunistic yeast both in in vitro and in animal studies. prevIOus work \~C have characteri zed ill vivo and then ill vitro two Although its importance for inva sion in general Is beyond doubt deta iled informations on the difTercn ll ~ p es or tnl ereellul ar junctions involved in the phys ical interaction pathogenetiC role of secretory aspartic proteinase are sti ll lacking. The same applies to the between LC and T cells, i.e . glycocalyx-glycocalyx contacls and question If the enzyme definitely Is expressed In vivo. and nol only In vitro or ex vivo. For in intercellular bridges. In this sludy, we analyzcd 01C conlribulion of several vivo Investigations parts of the lesional oral epithelium were collecled from a HIV-infected molecules known to medIate T ce ll aClivallon in these adhesion r!roccsses. patient with oropharyngea l candidosis. Using the antJ-secrelory aspa rtic proleinase murine Ea rl y kmetl c expenmenls showed that both Ihe length of cellular monoclonal antibody FX- 10 immunoelectronmicroscopy was perform ed (pre-embedding gold inlcracti on and LC/T lYmp hocyte ralio regulaled cluster ronllalion. Then il labelling). Thus it was possible to directly demonstrate expression of secretory aspartic was apparent th at LC-T cell cluslerin g triggered upregulation or CD I a proteInase for the first lime in vivo and in man. Besides, th e role of the enzyme during the ICA M-l , B7- 1 and B7-2 on LC. Consistently monoclonal antibodies Interaction of yeast and mucosal ce ll cou td be elucidated. Secretoy aspartic proteinase is against Ihe latt er molecul es excel'l B7- 1, and anti-LFA-3 efficien tly actively secreted by the Candida ce ll in association with membran e bound vesicles showing a brocked LC-T ce ll cluslering by 70-80%. AI immunoelectron mi croscop~ , high affinity to the epithelial surfaces. In particular secretory aspartic proteina se can be whereas LC's discrete plasma membrane zones in close contact with T delected at the site of close interaction betwee n Ca ndida and epithelial celfs. thus suggesting a cell s re vealed CDla, 87-1 , B7-2, CD40, LFA-3 and ICAM-I labell ing, ra te of the enzyme in host-fungal ad hesiv e processes. The findIngs further support the theory inl ercellular bridges were Immunoreacti ve to ICAM-I only. These data of a decisIve role of Candida aspartic proteinase in host cell invasion by Candida albican s. This show that CD la, B7-2, LFA-3 and ICA M-I are potent molecules th at Is of interest for both pathogeneSis and therapy as Inhibition of secretory aspartic proteinase dri ve th e LC self pept ide presentation to T ce ll s and also provide a mig ht prove an important alternative in the prevention and treatment of candidosis. rati onale ror th e dominant role played by ICA M-I on LC-T cell antigen specific clustering. VOL. 107, NO.4 OCTOI3ER 1996 ABSTRACTS 659 19 20 NORMAL HUMAN EPIDERMIS HARBOURS OCCASIONAL MACROPHAGE-LIKE CELLS. AN RECENT DEVELOPMENTS IN CORRELATIVE MORPHOLOGY: .. IN SITU" IHMUNOELECTRONMICROSCOPY STUDY .. lL..E.4Q.c.l..i.nL(.l..l... ..--Al:i-.l:1Omm.a.a..a. .L2 3) A L~--..AJ.Ql1at i 11) 1\ Ml.l.l..dcr I J) H Mal:1:_e.l..l..1-UJ--.B.tl. UL TR.AJUGH RESOLUTION SCANNING ELECTRON ~rmeer I " G Bowden ~nd G On penOIls tIl , (1) Dept MlCROSCOPY USED IN MOLECULAR AND CELL BIOLOGY. J Dermatology, BreSCia, Italy; (2) Dept Dermatology and (3) Electron Microscopy. Leiden, Tho Netherlands; (4) zooprophylactic loot. , Robert P. Apkarian. Emory University, Atlanta, Georgia USA Brescia , Italy; (5) Dopt Pathology , Halifax, NS, Canada . Modem high resolution secondary electron and scanning transmit Although Langerhane celle (LeI are well eotablished as antigen presenting cells (APC) constitutively harboured in normal human ted electron microscopy (HRSEM and STEM) techniques provide epidermis , the conatltutive preoeDeD in normal human o pidermis of cell and molecular biologists unprecedented topographic imaging other APe, ouch ae macrophageo (Mph). 1e still matter of debate. We most recently s howed, however, t hat the CD36 moleculo and tho "in modes. Appropriately prepared biological specimens viewed by s itu" immunoelectronmicroAcoPY (IEM) techn iquo can be conoidered a HRSEM provide vistas of macromolecular structures within the v ery suitable oystem to addreoo thio question , oinee Mph, other t han LC-l.ike cells of dend r itic lineage, expreas CD36 on the cell ourface. context of complex compartmentalization such as organelles, cells, The a.Lm of t he present study was, therefo re, to investigate the CD36- tissues, and organs. Isolated preparations of vesicles, viruses, expressing cells withi n normal human epidermio "in vivo", by means of .. in situ" IEM performod atter ultracryomicro tomy of normal human skin proteins, nucleic acids, and many other biologically significant b i opsies. Analys is of colloidal-gold- immuno labelled epidermal molecul es previously imaged only by TEM are currently being ultracryosections , carried out scrutinizing , cell by cell, a total epidermal area corres ponding to 1000 basal cells, revealed that , reexamined with enhanced topographic contrasts available from field whereas keratinocyts s , LC and lymphocytes did not show any gold emission in-lens electron optics. The latest specimen preparation partic~es on the c e ll ourface , four EC showing morphological characters of Mph-like cello, located in the spinous layer, were trends involving cryo-immobilization, chromium coating, and cryo consistently gold-dec orated on their plasma-membrane. CD36-positive imaging in conjunction with HRSEM and STEM promise more Mph- like cells might t rigger in normal human epidermio "down regulatory" Signals, a s already demonstrated for CD36-pooitivQ Mph accurate molecular imaging than was possible with chemical fixation. pOpulating the epiderl'llls of UV-irradiated and 1esional skin . 21 22 ELEcrRON MICROSCOPIC AND GENETIC STUDY OF ICHTHYOSIS THE BEHAVIOUR OF SARCOPTES SCABIEI IN THE SKIN. Mil;hrll; Eimiani. CONGEN1TA TYPES I-IV. KiWi-Maria Niemi Department of Dermatology, .GarIl! MaualI:nIa. .GarIl! Alessandrini'. Qig!\tiQ ~ I.lIW Anl1Iw.I.i. Helsinki University Central Hospital, Finland. Department of Dermatology and ' In stitute of Histology of Siena University. Italy The behaviour of Sarcoptes Scabiei var. Hominis in its natural habitat is poorly By investigating the ultrasructural features of skin specimens of 48 patients understood mainly because of the lack of an in vitro or in vivo propagation system. with a clinkal picture of ichthyosis congenita recessiva the following groups To obtain more information on its life-cycle we performed an ultrastructural study were distinguished: Type I 23 patients from "15 families, Type n 10 patients of 35 burrows obtained from 12 patients affected by common scabies. Attractive from 9 families, Type III 5 patients from 5 famili es, Type IV 1 patient. For the images of the tunnel, parasite body and egg architecture were obtained by scanning typing Heidelberg criteria were used. In addition, 9 patients from 7 families electron microscopy. With this tcchnique we a1so documented the presence of holes had uJ trastructural features such as dispersed keratohyalin or vesicular in the tunnel roof probably representing aeration structures. By transmission structures which did not alone fit to any of the above mentioned groups. electron microscopy we showed that keratinocytes around burrowing mites (also Molecular genetic studies on recessive congenjtal ichthyoses in Finnish ahead of the mite capitulum) are heavily damaged andlor necrotic. ]n the posterior population progress in collaboration with Elina Laiho, Jaakko Ignatius, Juha mid-gut of the mite we documented the presence of fecal pellets containing keratinocyte micro-organelles such as melanosomes and mitochondria. For the first Kere, Ulpu Saarialh o-Kere and Aamo Palotie. Two previously undescribed time we disclosed the steps leading to the adhesion of mite eggs to the burrow point mutations in the transglutaminase I gene (TOM1) have bee n detected so floor. We demonstrated that the typical finger-like projections of the outer layer of far in e lectron microscopic types I a nd II ; one of them was present in almost a ll the egg shell gradually disappear where the eggs are in contact with the tunnel patients with IC Type II in heterozygous form. Linkage study of these 48 floor. As a consequence the inner layer of the egg shell fuse with the amorphous patients and those of previous genetic studies indicate genetic he terogeneity in material lining the tunnel floor thus securing the eggs to the skin. Our data confirm recessive ichthyosis congenita. that Sarcoptes Scabiei feeds on keratinocytes and strongly suggest that it produces as substance with proteolytic activity that has a key role in its life cycle allowing burrowing, fceding and cggs~burrow adhesion. 23 24 RAREf'ACfION OF VESSELS IN CIRCUMSCR ID ED AND SYSTEM IC SCLEROSIS. IUIlUe!'AC'I'ION t)\o" V \o; SS I '; I A~ IN LICllEN SCLl';l{OS US le'l' Barbara Pan?. Ma ina Ba ch r ch· l3uhles StCf.1 U el GUlllmal Peter Altmc cr A' I' H.OP III CUS. MllI"i on 1\1I' {0 1" SI.(> uhnn ... 1 (;ammal M al'llnn Demmlological Clinic of th e Rllhr·Universit y. Bochul11, Germany Bllchllrnc : h·nuhll'~ »01(' 1" All IlI I'VC'I" . Dormaloiog'ical clinil: or t ho III circumscribed (CS) and systemic sclerodenna (SSe) II rarefaction of blood· l{ullr·Univor!'it.y l3ochum, Cormany leading capillaries can be observed parallel to :I n ill crease of collagen. In thi s Li chen !'<:lol'ol-; lI!' 0 1. at.rophicu!i i:. dill it.:lIl1y chan.u:I.('''; zC' (( hy investigation we reconstructed th e Architecture of the vessels of the superficial and porcelullI.whilC' plaQlI o:-l wil. h hUIlI Ol"I' hllge:;;., hi ~ 1. o l o J;i (; .. lI y by I"l.ll" orll cl.ion of bl ood vc!'scls. In lhrc{' Jlal.i (' nl~ W(l jlPrrornwd deep vascular plexus in CS aud SSe and exaillined lh e uhrnstnlcturc of the observed compllt.c ,-,:.; upporlcd t.hrcC'· dilllC'll sional rcconstl'lll:IIO Jl !' uf 1.11<' blind end of vessels. Oiopsies of th e sclerotic plaque of CS and thc sclcrotic arca of supe rficia l vcs:-l{' I!'< l.I :. illl; scriHI 7 ~Im 11&1:: s( 'c li o ll ~ . Tlm'{' hi ~ l o I O t;i c; " the involved forcann of palients suffering from SSc were taken aud cmbedded in s t ll gc~ wnro diITercntiat.cd. In) II P inril trative s lagP I.ho 11 1f1 amllllllory Epan 812. Series of semithin sect ions sClvcd for eO lUpu( cr~s upp0l1 cd three· mononuclear infilt.rate rCl-l ch('s up 1.0 t.i1 £' O llid (' rnli ~. The C' dC' nl a I.Oll s dimensional recoll stm ction. Dctectcd blind endin gs of vessels were processed for sl.ngC' if!; dlllrllctoriw d by thr{'{' zones: a natlCllI(' 25 26 CYTOPLASMIC MARKE:RS IN HIS TIOCYTIC SYNDROMES . RU9gcro Caputo . MULTIP LE MOLL'S GLAND CYST S OF TH E EYE LI DS. LlGHT- AND Institute of Dermatologic Sciences , University of Milan , ELECTRON- M ICROSCO PIC STUDY. 1'. Co mbemale l !. Ka nitakiS£..Ji. I . R. C .C . S . Ospedal e Maggiore , Milan , Italy l 3 3 The aim of this lecture is to illustrate the most important Dupln C. Parraud M. Guigon . Dept. of Dermato logy (1). and cytopl asmic markers present in different histi ocytoses a nd to Ophthal mology (3). "Desgenelles " Hospital; (2) Dept. of Dermatology, suggest a quantitative evaluation of these structures . Eight Ed. Herrio t Hospital. Lyon , foron ce. different c ytoplasmic markers have been stu died in eight A GO-year-old-woman t reated with bromocriptin for a prolac tln his tiocytic syndromes , namely histiocytosis X ( HX) , juvenile secreting pituitary adenoma developed o n the free m argin of bOlh xanthog ranuloma (JXG) . generalized eruptive histiocytoma eyel ids multiple cyst ic tumours conta ining a ser o u s Iluid . No (GElI) I benign cephalic histiocytosis (BCH) , papular xanthoma (PX) , xanthoma disseminatum (XD) , multicentric reticular ec toderma l m a l fo rm ations were prese nt a nd fami ly history was histiocytosis (MRH) and sinusal histiocytosis (SH) . Dense a nd unnoti ea ble. Hi sto logy showed mult i p le ro und cystic cavities multivesicular bodies , myeloid bodies a nd coated vesicles are containing a granular PAS- positive material and occasio nally lamellar present in all the studied histiocytoses , although scored , keratin. Their wall was lined by 3-4 layers or cuboidal cells expressing when quan itatively evaluated, in different numbers in each epithelial membrane antigen. Ul trastructurally the ce lls of the syndrome . Langerhans granules are present only in HX , a nd pleo morphic cytoplasmic inclusions only in MRH . Comma - shaped epithelial wall disclosed numer o us round cytoplasmic i nclusions with bodies , mainly observed i n BCH (+++), are prese n t also in HX a granula r -a mo rpho us material and occasio na lly lipi dic and/ or (+1 , JXG (++) , GE H (+) and SH (++) , whilst regular laminated myel inoid inclusion s. The ce lls of the innermost r ow prese nted b odies are present in GEI-I (+++) and IIX (+/++). f'inally , fatty numerous surface microvilli. Som e clear ce lls were observed. devoid of dro plets are found i n IIX (+) , JXG (+++) , PX (+++) , XD (+++) Birbec k grJ.nules. melanosomes and des mosomes. Mo ll's gland cyst s ilnd fJl RH (+) , but not in GEH, BeH and SH. In our opin ion, o n ly the Langerhans gra nules and probably the pleomorphic (or apocrine hidrocystom as o f the eyelids) are rare, usually solitary c yto plasmic inclusions are s pecific market;"s useful to classify lesions; t hey m ay be ll1ultiple in the case of a very rare hereditary the his tiocytic syndro mes . ectod erm a l dysplas i a syndrome (S chOpf-Sc hal tz-Passarge) . We spec ulate that hy per prola tinac mia induced hyperproliferation of the ells lining the excretory duct of MolI's glands. leading to its o bstructio n and formatio n of retention cyst s. 27 28 I'A PU 1. 0 ERYT II I(Q DI'RMA (O FUJ I). I.I G IIT - AND EI.ECTRON AN ULTRA STRU CTURAL STUDY ON THE LONG-TERM SU RVIVAL M ICROSCOP IC STUDY 0 1' A NEW CASE. I. Kan itaki s I.. Mise rv M. CASE OF HA RLEQU IN ICHTII YOSIS. VQshjkj Thn jrt llc ilj Kmujko NQlUura I·.l ure A. ClauLl\". Dep t. of De rmatology. Ell. Ilerriot lIos pital. 69437 Ejl'Q Ym nadn M asnYllki ShjmiZl! SCls uko Wn shjo HjroShi KII\Vnhilfil Dept. of I.\'on ex 03. Fra nce . Dcnnnto logy, Mi c uni versity School of Mccl icill c and Pediatri cs unit, Yamamoto . P:Jpuloery tilfoderlll:J or Ofuj i (PO) i :-. a cond itio n d esc ribed in 1984; so j';jr abo ut 50 r ases havc bee n repon ed in the lite rature. most o f I lospitai. JIlJ};'lI1 . wllil'h (35) concerned japan c~c patients. Our patient was a 73-year Harlcquin ichthyosis is n severe nnd usua ll y fatn l hercditary skin disorder. (J ld Ill an or so uth - ca~ l asian o rigin that presented with a pruritic Onl y fe w affected infants have survived Ihe fi rst year o f life, <1 lthoug h th ere arc eruptio n m ad' o f diffuse d ~ lrk red polygonal papules predo lllin;:t ting several CtiSC reports of this di sease. Since we have treated n bnby o f harlequin on the trunk Cl nd limbs but sparing the body fo ld s. Thi, co ndition icl ll hyosis for lllorC th an a year, wc rcport the r ~lfC casc and the uilrastmcturc of 11 ..1<..1 been presenl for three years and had been lreated with local th c skin as compared with fa la!. A female infant was delivered vagi nally [I t 34 ~tl' ro iL1~. UV H. PU VA 29 30 FAM ILIAL DYSK ERATOTIC COMEDONES - A IIISTOLOGICAL, LOCALIZED CONTINUOUS PEELING SKIN SYNDROME. A. Bru ,ascQ IIISTOCIlEM ICAL, AND ULTRA STRUCTU RAL ST UDY. Son' a Reimann mo S. Veraldi G Tadini R Caputo Institute of Dermatological Sciences, University of Rutten· Di cier Metze Dept Dcrmato l. , Uni versity MOnster. Dcrmatopalhol Lab., Mil an . I.nly. fricdrichshafcn, Germany We report a 26-year-old fema le patient affecled by thin scaling lesions with Familial dyskerat otic comedones is a rare autosoma l dominant dermatosis We underlying erythema, exclusively locali zed to the palms and dorsal side of the fingers report on two sisters who prcscnlcd wi th iso lated papul es wit h a kcralolic plug on the The patient stalCd that the dermatitis developed at the age of 7, and it was extremities and. to a lesser extent, on the lnmk. The persi stent. occasionally inn amcd asymptomatic. The parent s were consanguineous Histopathological examination and pruritic lesions li rsl appeared in childhood and gradua ll y increased in number with sho wed hyperkeratosis with a separation of the stratum corneum just above the stratu m lime Bi opsies were examined hi stologicall y, ult rastructurally and granulosum. All these features allowed to exclude palmo-plantar keratodermas. imlTIunohi slochcmically A deep invagination of the epidermis was fi lled with laminated keratolysis exfo liativa (duc to fungal infection or pompholyx). epidermolysis bullosa onho- and parakeratoti c material. The epid ermi s showed dyskeratosis and incompl ete simplex superficiali,. and ,uggeSled .he diagnosis of" peeling skin syndrome" (PSS) acanth olys is o f the kcratinocylcs in the upper stratum spinosum. Immunolabeling for By electron microscopy. th e corneocytcs o f the first 5-6 layers showed lipid vacuoles fil aID;rin showed an irregular di stribution of keratohyalin , The expression of involucrin and nuclear debris. In the upper layers. an int racellul ar cleavage was demonstrated, with and th e overall distribulion of various keratins was preserved. The mitoti c rate appeared fragmentation of the kerat in fi lament s. The cell -to-cell ad hesion was firmly maintained increased Confoca l la ser scanning mi croscopy of nu orcscence-stainings for by desmosoma l disks. No other significant ultrastructural abnorm alities were notcd. deslllogiein , E-cadherin. keratin and filamentous actin suggested a di sarrangement o f These ult rastructural feat ures support the diagnosis of PSS. However, our case seems the adherens structures and cytofil arnent s Ult ra struclu ra l examination revealed focal to be peculi ar because PSS is defined as a generali zed disease that frequently spares the di ssolution o f deslllosollles associated with a perinuclear aggregation of keratin palms and sales. We therefore suggest to call this entity as "locali zed continuous PSS ". filam enl s whereas ad herens junctions remained intact Dyskeratotic kcratinocytcs were characlcrized by clumped keraLO hyal in granul es In conclusion, our findings confirm that fami lial dyskerat otic comedones represent s a unique entity different from nevus comcdoni cus. keratosis pil ari s, M. Kyrl c, M. Durier and other fo cal acalltholyti c dyskcraLO tic di sorders VOL. 107, No.4 OCTOBER 1996 ABSTRACT S 661 31 32 U LTRASTRUCTURAL ABERRATIONS OF DERMAL ELASTIC ABNORMAL SPERM TAIL IN MICE LACKING THE INTERLEUKIN-1 GENE. FlBERS AND COLLAGEN FIBRILS IN LINEAR FOCAL ELASTOSI S H -J Schulze R Birchmeier" T Krieg, Dept. of Dermatology, Univ. of Kaln , fN AN ASIAN CHILD. Friedrich Breier Franz Trautinger Klaus Kasercr and and " Max-Planck-Institute of Geneti CS, Kaln, Germany. In vitro sludies have suggested that interleukin-1 (IL-1) exerts a number of Wol fgang lurecka. Departmenl for Dcm13tology and Inst itute for Clinica l important biological functions, e. g. stimulates growth and differentiation of T Pathology, Uni versity of Vienn a, Medical School. Austri a. and B cells, induces ICAM-1 expression, induces and synergizes with other A 13-year-old gi rl is presented with the clinical diagnosis of linear foca l elastosis (LFE) cytokines. To define the function of IL-1 in vivo, mice lacking IL-1 were on the limbs. Biopsy speci mens were obtained from lesional skin and embedded for generated following gene targeting in embryonic stem cells. These mice histology, immunohistochemistry for collagen types I and III and electron microscopy. were apparently healthy and fertile suggesling that IL-1 aclivity is dispensable during development. In the second generation, male mice By using a video analys ing system light microscopy in elastica stained sections revealed homozygous for the IL-1 defect but not heterozygous mutant or wild type an increase of elasti c ti ssue in the center of the lesion. Th e increase of number of clast ic mice were infertile, whereas in vilro insemination by means of fibers (EF) was statistica ll y significa nt and rcached focall y 100% EF were para ll ely intracytoplasmic sperm injection revealed normal fertilization. Semen arranged, thinned. elonga ted and rocally shaped like a paint brush nt their end s. analysis showed progressive backward-motility of the spermatozoa as Immunohistochemi stry ror co llagen type I and III revealed collagen bu ndles thinned, handicap for penetralion due to a uniform snapp-off of the tail in all sperms. adjacent to numerous EF. Uhrastnlctural investigation showed elonga ted or fragmented Histological analysis of the testical revealed identical malformations in late EF with electron dense material next to collagen fibers. In the matrix Ilumerous fin e stages of the spermatogenesis. Ultrastructurally, segments of the fibrous shealh in the main part of the flagellum were stringy and inconlinuous reticul ar, or granular electron dcnsc material interminglcd with elast ic fibcr mi crofibrils whereas the outer dense fibers and the structural conformation of th e were observcd. In LFE. rarely reported in young females, androgcllctic-. skin aging axonema appeared to be uninvolved. The data suggest that Ihe assembly of and mechanical ractors are discussed as pathogenetic causes or this cntity. sperm cytoskeleton may be sensitive to IL-1 expression. 33 34 BACILLARY EPITHELIOID ANGIOMATOSIS: TWO CASE REPORTS OSSIFYING FASCllnS OFTHE NOSE P Inoocenz! - M CarJeslmo - C Bosman - S Caty!er! P Innocenzj - S Gjustin! - C Bosman - S Catyied C llnlca Dermatologlca Universit3. "La Sapienza" Roma - ttalia Clinica Dermatologlca UniversJta -La Sapienza- Roma - ltalia We report the cllnicopalhological and ullraslructural study of two cases of A 24 year-old-woman presented wiih a seven monihs histDlY of a firm, bacillary epiihelioid angiomatosis in HIV positive patients. The first patient painless. nonulcerated, rapidly enlarging tumor of the nose. PhYSical was a 51-year- old, HIV pos itive , caucasian, homosexual male who examination and roulinary blood tests were normal. No nasal bone developped scattered papulo-nodular angiomatous lesions. Some lesions involvement was revealed by skull X- ray. His topathological examination were crusled and some had collarettes of scale, closely resembling a showed numerous spindle and dendritic cells with scattered os leoblasts . pyogeniC granuloma. The number of cuteneous lesions progressed daily. Osteoclasts were occasionally observed mainly in the lower portion of the The second patient was a 36-year- old transexual male, intravenous drug nodule. A markedly oedematous stroma and multiple ectatic vessels were abuser for len years , affected by chronic active hepatitis for many y~ ars . also evident. The presence of microtubular cytoplasmic structures observed Cutaneous examination revealed multiple angiomatous, crusted. done under electronmicroscopy examination revealed the myofibroblastic nature of shaped , brownish nodules involving face and legs. I-iistophatological the spindle ceUs, thus allowing the diagnOSis of ossifying fasciitis. findings were similar In both cases and showed lobular pattsrn of vasculal The patient was treated with conservative surgical therapy and no relapse proliferation involving the superficial dermis with an overlying flattened has occurred after 5 years of follow-up. epiderm is and lateral epidermal collarettes. Bacilli were found using Warthin -Starry silver stain. The ultraslruclural study demoslrated cluslers of bacilli located extracel1ularly within the connettive tissue surrounding blood vessels_ The bacteria had a trilamlnal wall with two electron- dense layers separated by an electron - lucent layer. The cytoplasm was coarsely granular and showed electron - dense areas sometimes separated from the wall by a uniform electron-lucent zone. 35 36 SPINDLE CELL HEMANGIOENDOTHEUOMA: TWO CASE REPORTS DIRECT IMMUNOELECTRON MICROS COPY ON THE CONJUNCTIVA IN D. Innocenzl - R, Boldrlnl U Bottonl C Bosman - S. Calvie ri OCULAR CICATRI CI AL PEMPHIGOID VERSUS SUBEPIDERMAL AUTOIMMUNE BULLOUS DERMATOSES WITH OCULAR INVOLVEMENT. Cllnlca Dermalologica Unlversit" "La Sapienza" Roma - Iialia REPORT OF 10 CASES. Cathcrinc Prost HervC Robin M ichel Heller FredCric Calix Two cases of spindle cell hemangioendothelioma of the right flank are Thanh HOil ng-Xuun Centre d'Etudcs et de Diagnostic des Malad ies 13ulleuscs. H osp it ~ll Saint Loui s 37 38 DIRE CT IMMUNOFLUORESCENCE ST UDIES OF SODIUM CHLORIDE ORAL OAP SON E THERAP Y NOR MALI ZE S THE DEFECTIVE ULTRASTRUCT URE SEPARATED SKIN IN SU B-E PIDERMAL AUTOIMMUNE BULLOUS DISEASES. AN D IM MUNO REACTI VI TY OF HE MI DES MO SOM ES AN D I MP RO VES SKI N C 8¢daoc PM Dung MJ. LehoPlet F. Labro usse JM Boonctbljmc P Bernard STABI LITY OF J UN CTIO NA L EB MI TI S TYPE PATIENTS. Ingrun An Deparunent of Ocrmatology and Pathology. Hopital Dupuyucn, Limoges, France. ton - lamprecht , Ulrike Ebschner . lnst. f . Ultrastruk turforsch . Direct immunoclcctronmicroscopy Cl EM ) allows a precise c lussification of der Haut , Universitats - Hautklinik , Heidelberg , Ger many . subepidermal auto-immune bu ll ous discases by fine localization of immune deposits The various clinical f orms of junctional E8 (JE8) are alon g the dcnno-cpidcnnal junc tion. Since thi s tec hn ique is time consumin g and charac t erized by hypoplasia of hemidesmos omes of different expensive, di rect immunonorescencc on sodiu m chloride separated skin (DIF Nne l) degree , l ack of subbasal dense p la tes , and jun ctional bli has been suggested as an alternative fo r rout ine diagnosis (Gammon JAAD 1990). Thc sters . Immunologically 1 9 - 0EJ - 1 and anti - kalinin antibodies aim of this study wa s 10 evalullte th e reliabi lity of D fF Nae l in comparison w ith direct fail binding or s h ow reduced reactivity , Lethal (Herlltz) IEM in 35 patients wit h subepidermal autoimm un e bull ous diseases. Add it ionall y, and benign for ms (mitis , inversa , localisata) are known . c irculating mHoantibodi es were detected by indi rect IF on NaCI separated skin and by No specific therapy directly i nfluencing the respect i ve western blol. Among the 26 pa tien ts with bull ous pemphigoid or pemphi goid gestati on is basic abnormality is available for any type of E8 . and deposit s in th e lamina lue ida (LL) by IEM. deposit s were found on the epid enn al We have treated a series of JE8 patients wi th Dapsone side of th e NaCl treated ski n in 20 cases, on the dennal side in 2 cases and on both sides and controlled its efficacy on clinical expression , s kin in 4 cases giving a re li ability of 77 %. 6 patients wi th cic 39 40 IDENTI FICATION OF TWO SPLI CING MUTATIONS IN COllAGEN TYPE VII L YSOSOMES AND PROGRAMMED CELL DEATH: REFLECTIONS ON GEN E (COl 7A l ) IN AN ITALI AN PATIENT AFFECTED BY RECESSIVE KERAT INIZATION. Geoffrey Rowden. Pathology Dcpt. Dalh ousie University & DYSTRO PHIC EPIDERM OLYS IS BUllOSA LOCAL/SA TA. .s. e.u:JiIlll, fl. Queen Elizabeth II Health Sciences Centre, Hali fax, Nova Scot in, Cl.lnadn. ~1, l.. ~ 1 , l'!. Z2lllli1, Q. Miuini2, M. QQlQmbj1. 1) Division 01 Biology and at Genelics, Department of Biomedical Sciences and Biotechnology and 2) InSlilule of Clinical The dramatic morphologic and fun ctional changes that occur the gn1l1ular/comified Dermalology, Unlversily 01 Brescia, 25123 Brescia, It aly. layer interface in the cpidcnn is are evidencc ofa tight ly controlled normal remodelling Colla gen type VII gene (COl 7A1) has been de monstrated to be altered in severa l process. Kera tinocYlcs trnnsfoml within hours fro m viable nucleated cell s to fi bre-filled variants of dystrophic epidermolysis bullosa (D EB), either with recessive or dominant squamcs. Our understanding of how thi s dcmoli tion proccss is achieved lags far behind mode of inhe rita nce, We observed a reduced expression of COl VI! mRNA and a our awe tit its effi ciency. In the late 1960s Iysosollles we re suggested as Ihe possible reduced level of COlVII , at the dermal-epidermal ju nction of the patient , if compared with his pare nts and a cont rol donor. The ultrastructural analysis of the de rma l source of the variolls digestive hydrolases. Attempts to resolve this questi on usi ng e piderma l junction of this fa mily is under study. We have id entified two mutations in a cytochemist ry and quantitation at the TEM level, however, demonstrated the scarcity pa tte nt a ff ected by a localisata variant of recessive dystrophic epide rmolysis bull osa of such organell es in kera linocytes, A review of th e known roles of Jysosomes in (l -RDEB). which is cha raclerized by th e less severe phenotype of the syndrome. kcratinocytes will be give n and an altemative proposal suggesting activation of non Th ese mutations are the lirst spli cing muta ti ons so lar described for COl7A 1 in DEB. Iysosomul located hydrolascs will be made, is -2 One mutation a paterna ll y in herited A-G tra nsition a t position of the donor Recent interest in the role of cell deat h and remodell ing vin the programmed or splicing s ll e of Intren 3, which re s ults in three abe rra nt mANAs depending on the s ki pping of a xon 3, on th e usage of a cryptic donor s ite in side exon 3, or on the Hpoptotic route may have fi nall y veri fi ed thi s suggestion. In situ ni ck end labelli ng of mai nt e nance of int ren 3. The second mutatio n is a ma te rn ally inhe rited G-A cl eaved nucleic acids exposed in the process ofapoptosis shows the regul ar occurrence tra ns ition a t pos ition -1 01 th e donor splicing s ite of intron 95. which causes th e of tile phenomenon in the gran ul ar laye r. PrcslI mubly the ell vi ronmental milieu of the acti va ti on of a cryptic donor site 7 nucl eotides upstrea m the normal site and results in uppcr epidcnnis ntVours the ac ti vation not only of the l1 ucl cases so typi cal of thi s fonn a de leted mAN A. All a be rra nt mR NAs contai n a shift of th e reading fra me giving rise of cell delction, but also the other hydrolascs necessary for the complete cell re to premature te rmin ation codons (PTes ). All elic specific analysis of the transcripts modelling and metamorphosis into rull y corni fi ed cell s. has shown that the patient synthesizes a n aliquot of norma l tra nscri pt, deriving from th e ma te rnal a lle le. These fi ndings a re in agre ement with the mi ld clinical phenotype observed in the iocalisata fo rm of RO EB shown by the patient. 41 42 UVA AND uv n I N DUCED ZEIOSIS IN A431 CELLS I N CULTURE : A MODIFIED CULTURE SYSTEM FOR ItuMA N Ef'UlERMAL CELLS FO R GRAF T1-I E EFFEcr OF M EDIUM COMPON ENTS. P.U. Gia cD mDni " E. straraee', T ING P URI'OSES: /\ N IN VIVO ST UDY IN IlIGS. AII I1 Ctl!r GM van Dum M aD' CH Vc rhllevell Tjllekc vall tl cr Nat A Mieke MQl1 l1 l1l1i1 S HIi Uk K KOI: rtc!l ails! Mariil POIXC A . Rougier ., W , Malomi .. - 4 Laboratoircs de L'Oreal - 94$50 C/u!ViUy-Larue - Fra" ce - l"'lh nrat( ,ry iilr Den ll awlogy. Ex perilllcllIll l Surgery, Electrt1 ll11li crnscopy, Biolll 43 44 USE OF KERATINOCYTE_ AND FIDROBL-AST-I'OI'UL-A TE D BI ODEGRADABLE TRANSFORMATION OF DESMOSOMES IS IMPAIRED IN THE IN COPOLYMER IN THE REGENERATION OF TUE SKIN: AN IN VITRO STUDY. VITRO RECONSTRUCTED HUMAN EPIDERMIS. I YiCanQY~S A M Annette OM van Porn Mary CH verhos:vcII Clt:IlJ!:!1S A vall IlIjl!s:rswjik Henk K KOSin!!" and MomlDnnsS• A A MulderS - H K Koenen'" M Ponees . sDcpanment of DennUlol Maria POoec· Labor.uory fo r Dcnu 45 46 THE NATURE OF THE COM PARTMENT FOR TH E INTRACELLULAR ULTRASTRUCT URA.L STU DIES OF CUTANEOUS LYMI'IIOMAS. E Beni BINDING OF PEPTIDE TO CLA SS II IN HUMA N EPIDERMAL S. Cavicchin i A. Bre7.zi and R Ca~ Insti tute of Dennatologic Sciences. IRCCS. LANGER HANS CE LLS CHANG ES DURING SHORT-TER M CULTURE. illIl Uni versity of Milan. It aly. a.. Mulder Coby Om-Lujtjng Frits Koning Bert J Vermeer A Mi eke MQl11l11aas Cutaneous lymphomas (CL) are a heterogeneous group of disorders In several Department of Dermatology. Il1ll1lu llohacmatology and Bl oodbank . and Laboratory cascs c1inico-path ological data must bc supponed by immunohistochemical and for Electron Microscopy. Uni versi ty J-I ospil . 47 48 STRUCTURAL IIAIR ABNO RMALITIES IN ECTODERMAL DYSI' LA SIA HYI'OMELANOS IS OF ITO. ULTRASTRUCT URAL AND LTRASTRUCTURAL OBSERVATIONS AN D DIAGNOSTIC ROL E OF IMMUNO HI STOCHEM ICA L INVESTIGATIO NS. SAGa," ti L Badiali-De SCANN ING ELE CTRON MICROSCOPY ANALYSIS. Bar ar chi S Giorgi (I) G Pas(Jui nelli (I) E Beni (2) G C Ma num (3). Department of 1 CVnhiaghi G.L. Tadi ni Institute of Dermatological Sciences, IRCCS, Uni versity of Dermatology. I)arma: ( I) Institute for Clinical Electron Microscopy. Bo logna, (2) I Milan. Ilal y. Depanment of Dermatology. Mi lan. (3) Private Denllalological Practicc. Il1Iola: Iialy The ectodermal dysplasias (E D) arc a group of congcnit al skin disorders that Hypotll eianosis of It o (HI ) is a vcry rare neuroClitaneous disorder characterized by • diffusely involve the epidcrmis and it s appendages. If th ere nrc, on one hand. Illany swirling hypopigmcntation along Blaschko's lines We rcpon a fivc-yea r-old female reports describing the classical hair shan abnormalit ies (bamboo hair. trichorhex is baby with typical sk in lesions noted at the age of one year Neurologic. musculoskeletal nodosa... ), few papers. on the other, have focused on hair involvemcnt in disorders - andlor ocular abnormalit ies were not detected. Sincc pathogenesis of 1-11 rema ins such as ED - wherc hair involvcment is not paradigmatic but very freq uent Our unclcar. we invesligated hYi>opigmented skin by means of uhrastnlctural as well as purpose was to study, by means of scanning electron microscopy, lit e alternt ions that immunohistochcmical (ICC) procedures. At the ultrastructural level l11eln nocytcs occur not o nly on th e hair shaH but also on its cu ticu lar arrangement s in ED syndromes showcd a pllllcit )' of I1l clanosomcs. mostly observed at stage IV. and short dcndri tic We reviewed specimens from a number of ED i c Olmsted Synd romc. Hydrotic ED processes Perinuclear rims of looscly textured intermediate filament s were observed. A (Clouston), Hypohydrolic X- linked ED, Oro-Facio-Digital Syndrome. Rapp Hodgkin mi ld dermal perivascular mononu clear ce ll infiltrate was present. ICC investigations ED, Tricho -Ocnt o-Osseous Syndromc. Tricho-Rhino-Phalangeal Synd ro me We di sp layed n low expression of thc c-kit protein on Illclanocytes HMB45 antibodies recorded hair shaft alterations bu t above alt , in ED, cuti cular derangcment s arc a failcd to stai n epidcrmal melanocytcs CD I a-positive cetl s appeared increased in number common denominator. We suggest that even if a single defect (shaft or cuticle) docs (lot within the epidem1is Finall y. numerous DR-positive cells were scatt ered in the dermis allow diagnosis, the paltern of both defccts may be of some aid Lo make the recognition Staining of the c-ki t protein has been cl aimed able to differenti ate hypopigmcntations of £D easier. provided with mela nin-synthesizing IlIclnnocytes. from cut aneous di sorders devoid of IlIclanocytes Since bind ing of the c-kit liga nd sti mu la tes the production of melanin. the low expression of the c-kit protein on melanocytes may explain thc apparcntl y downregul atcd melanin syn th csis observed in our paticnt 664 ABSTltACTS THEJOUR.NAL OF INVESTIGATIVE DERMATOLOGY 49 50 PR URIGO PIGMENTOSA:CASE REPORT WITH IMMUNO AND ULTRASTRU CTURAL PRETIBI AL EPIDERMOLYSIS BULLOSA: A CASE REPORT WITH IMMUNO, OBSERVATIONS. *Christine H. Betts. Ombretta Calderoni Angela H Cos ta rENETIC AND ULTRA ST RUCT URAL OBSERVATIONS. Angela M. Costa . Dip. di Pat. Speri mental e* e Dip. di Ma d . Clinica Spec. e C Iri s line H. Betls Cluudj o Varotti. Di partimento di Patol. Speri mentale, Sez. di Clin. Darmatal . Uni v. of Bologna, Italy. Sperimentale* e Diparlimento di Medicina Cl inicu Specialist. e Prurigo pigmentosa ; s a rare inflammatory dermatosis, Spcdmentale, Sez ione di Cli ni cll Dermatologica, University o f Bologna, first described by Nagas hima at a 1 i n 197 1 . characterized by I t.aly. recurrent pruritic erythematous papul es t hat evolve into Pretibiul epidermolysis bullosu ( PEB) is a rare variant reti culate hyperpigmentation. We desc ribe a case of a 52 year of dominant dystrophic epidermolysis bullosu (DDEB) with old woma n with a s udde n onset d i ff u se p ruritic reddish papular di s Linct clinicopathological featu res. We describe a case r a s h on the back, chest and l i mbs. Papul es coales ced into of a 37 year old Italian man with PEB . Pretibial s k in b iopsy marked reticulate hyper pigmented plaques. Skin biopsies f r om s pecimens were examined by light a n d electron microscopy. r e ddi s h papules and pigmented lesions were examine d by light lI ist-ology revealed u s ubepidermal blister wi th moderate and e l ectron mi c r oscopy (EM). Hi stol ogy showe d hy perker atosis , edema and a p erivascular lymph ocytic i n flammatory acanthosis, exocytosis and foca l s pongios i s; in the upper infiltration of l h e upper d ermis . Immunofluorescence or dermi s moderate pigme n tary incontinence. mi l d i nflammatory immunocy Loch emical s Laining for bullous pemphigoid Ag, perivascu lar lympho- hi stiocyte infiltration a nd papillary laminin, type IV collagen, GD3 a n d type VII collagen (LH 7:2 edema were seen. Immunostaining for C04+/C08+ cell s a nd ICAM-l mo noclonal a n t ibody) were performed, Trans mission electron was performed; direct immunofluorescence was negative. By EM microscopy of perilesion al and lesional skin confirmed marked i ntercell ul ar edema was observed wi th injury of the h istological f i ndings. Ultrastructu ral alteration of basal cell l ayer i nc luding degeneration of the cytomemb r ane a nc horing fibril(AF) distribution a nd appearunce was a nd agg r egation of tonof; l amen ts. Ke r ati nocyte des mosomal observed . Phenol Ichloro form ex truc ted lymph ocyte genomic connections we re generall y preserved . Me l anin conten t was DNA has b een examined ( or mutation s of the COL7i\1 gene. s l igh tly increased; granules appeared large a nd separate with Con c lusion s: PED was confirmed by i mmuno, ultrastructural litt l e agg r egation. No f r agmen tation o r thi cken ing of the and gen etic s tudies whic h demons trated altered structural bas al memb r a n e was observed. Dermal perivascul ar inflammatory integ rity or the APs at. the derma-epidermal junction. cell; nfi 1 trat ion was composed of 1 ymphocytes and macr ophag6s. with no po l ymorphonu c l eocytes. Some macrophages were heavi ly l oaded with phagocytized me l anin . 51 52 T il E III' I XII ANTIGEN AND TYPE VII COLLAGEN A RE NOT STRUCTURALLY AND BIOCHEMICALLY NORMAL PERMEABILITY EX PRESSED IlY II UMAN MELANOCYTES I N V I VO. M..i.l:.lJ..tl BARRIER OF HUMAN EPIDERMIS RECONSTITUTED IN CHEMICALLY HELLER PjlIII TS I ANAKAS Frl'd('rir rAUX Cjltht:rint> PROST Centre DEFINED MEDIUM . M. Rosdy M. Fartasch M. Ponec, Laboratoire Skin ethic. d 'ctud es tt de dia!;llostic d cs maladies hlilicuscs. lIopit al Sain i · Louis • Paris - Fr:.mcc Nice. France, Dept.of Dermatology. University of Erlangen.Germany. Dept. of The I3P 230 and 131> I XU ;!nti,;cns (ilg) are two con stilllcnt s of hcmidcs mosol1lcs (HI)) Dermalology. University Leiden. The Netherl ands. whi r.: h arc rer.:og nizcd by th e sc ra of patil:llts suffcring from bullous pemphigoid A fully differentiated ep id ermis is produced routinely in vitro by culturing (BP).The first is loc ated within the plaque while the second Olll: is a transmembrane normal human adult keratinocytes (NHK) on inert polycarbonate filter substrates at protein and probably one or th e constitu:lllIs of :LI1 choring filament s. T ype V II collage n th e air·liquid interface in modified and supplemented chemically defined medium is the target of :ltLto:.Lrllibodics in epidermolysis bullosa ar.: quisi ta (EBA). which is MCOB 153. Vertical sections stained for histology and indirect immunofluorescence loc atl:d within anchoring fibri ls (AF). Recently Giudice ct al have shown that BP 180 ag studies show a proper stratification and expression of major differentiation markers was ex pressed by 1llelanocytcs (Me) in cuhurc. while 13P 230 wa s not ex pressed. As aft er 2 weeks of air·exposure. Up to 12 weeks of culture. the stratum corneum is HI) and Ar: an: not idcllIificd in human Me. wc have ex amined if M e in vivo arc the continously produced, and the thickness of th e living layers stabilizes at a more targe t of alltn:lIlIihodics in 9 paticllIs (N° I ·9) with a I3P or cicatricial pemphi goid (CP), reduced level. Electron microscopical studies (using ruthenium tetroxide·post whose se ra rer.:og nizcd the lJP/CP I XO ag by Western blot on epidermal ex tracts. and I fixation) show that th e structural organization and distribulion of the stratum corneum patienl (N° lO) with a clinir.:: lily typical EllA. lipids in keratinocytes cu ltured for 18 days at the ai r·liquid interface is similar to that A peri\csion:d hiopsy W:I~ processed for imllllllloe lectron microscopy lIsin g the seen in the native epidermis. Supplementation of defin ed med ium with linoleic and preelllbl:th.lin g Icclllliqllc with peroxida se labelling prev iously described by liS. Und er palmitic acids perturbed the structural organization of stratum corn eum lipids and declron mir.: l'o!<.l:o pe, al lcast Dlle Mc ha s been obsl:rved in c:lr.:h ultrathin section. The induced formation of trig lycerid e containing droplets. Biochemical analysis of deposits were prese nt in fronl of keratinocYH:s (Ke) but stopped in front of Me in all 10. epidermal li pid s show thal the lipid profile in th e reconstituted epidermis is close to They wc re IOC:Hcd in the lamina lucida (1..1..) in 4 patient s (N° 1-4). bot h in the LL and in the in vivo situation. with appearance of free fally acids and most of th e ceramides the LI) in 5 (N°5-9) and in the AF zonc in the l:t st one (pa tient N°H}). in keeping wi th a responsible for th e barrier function. Generation of living human epidermiS in the diagnostic with a diagnosis of BP. CP'und EBA res pec ti vely. minimal presence of growth factors and other supplements in the culture medium In r.:onclu sion. it appea rs dearly that the 9 patiellts whose sera rl:cogni zcd exclusively enables creation of an in vitro model with hi gh reproducibility and pharmacological the I3P/CP I HO 53 54 ATOPIC DERMATITIS IN CHILDREN: EXP RESS ION OF CD30 IN SKIN LESIONS ASSOCIATED WITH f-lERPESVIRUS INFECTION IN FROGS CUTANEOUS BIOPSIES ANO EVALUATION OF SERUM LEVELS OF (IlANA IJA/MA 1'!NA) Antonio Lnva7.za* Daniela Gclmetti* Rolando Bennatio. SOLUBLE CD30 . Giovanni C:w 55 56 MORPHOFUNCTIONAL ALTERATIONS OF KERATINOCYrE ADHESION IN PSORIASIS. BASIC FUCHSIN AND ELECTRON MICROSCOPY FOR THE DIAGNOSIS MO£Q lldda L Mom/clio MR ' Ya cono M p"r£olizzj S ' ea/ullwQ sr "C!I(mrrri R S{/1I/oro A OF EPIDERMOLYSIS BULLOSA SIMPLEX (DOWLlNG·MEARA). Department of Uiol1lorphology and · Institute of Oermatology· Univers ity of Messina. 01 MUZIO Marcello'. PROIETTO Gianluca, FELICIANI Claudio, AMERIO Paolo, TOTO Paola, COSCIONE Giulia, MOHAMMAD POUR Samano Recent s ludies h 57 58 A NEW PROCESS FOR SCANN ING ELECTRON MICROSCOPY WITHOUT PHENOTYPICAL CHA RA CTERIZATION AND STRUCTURAL ANY TREATMENT EXCEPT FREEZING. APPLIED TO HUMAN SK IN STUDY. SPECIFIC ITY IN SK IN CAIlCINOl\'IAS. G, Tucci · A Pu 'na1011 1 "'G Luca rini ~ Sll rl ~yc- njlz c ill c"'. M... ~+. ~ ~ +"' . Y:.. .G.n.Ll"' :. A Omdani · 5. Dubini He. Tietz "s Carraro · 0 Biagini Institute of fuctr':>n 1'Z~~? ·0 n ;v~~rt~~tc~oCr~~a~~ P.loj~yl~r s~ C ~ ~ If;:rt i hJr:l~~~lr.o31°J(:5 Dcrmatology, "' Institutc of 1-llIlllan Morphology or Ancona, H lnstittllc or HistOlogy Talence. France. ++ Ins litut de recherche Pierre Fahrc. C H U de and Embriology of Bologna, Italy Rangueil, 31400 Tou l ollse. France. , The ultrast ru ctural study of skin discases is II wide and rerti lc field of !"escnreh In Electr~n microscopy illl.plic:" prc li min.ury ph x~.i ca l . and{or ~hcrn ~ ~al the light of ever increasing bi omolecu lar information, howcver, it beco mes ncces:>ary to pre p~rallvc pro~cdu:c s which Induce maJo r l1 ~o~IiI,I Ca110 n S III hlo log,l.ca l cc interpret submi croscopic data also on the b • 59 ,., CASE OF HEREDITARY RETARDED WOUND HEALING RECOVERED BY DEXAMETHASONE. A Ch id!n! 01 N zoppl (21 5 Grnlfembergbi III u.r1at f (21 Q De panfilis III M colomb! /21. (1) Dept Dermatology, Spedali Civili , Brescia, Italy; (2) Div Biology and Genetics, Biomed Sciences and Biotechnol, Brescia Univ, Ita ly. OUt" previous studies reported that skin fibroblasts, derived fro m patients affected by different t ypes of Ehlers Danlos syndrome (EDS), lack a n organized extracellular matrix (ECH) of fibroncctin (FN) and chat , after a dexamethasone (dex) treatment, a rapid restoration of tN-ECM ie observed. A 22-year-old man rocently came to our attention for three chronic wounds, rosistant to the s tllnda.rd thorapies, o n his rL-gh t leg. Since ths patient ' s fathor was affected by retarded .carring and aince r etarded wound healing io one of tho typical oigna o f EOS, we hypothesized that he might have been affected by a defect of Ptl-ECM organization oimilar to that observed in EDS. Patient's akin fibroblasts , obtaine d from a perileoional biops y, completely l lacked, when cultured in vitro, the FN-ECM, which could be restored by lO-7M dex treatment . An ultrastructural control on a furthe r okin biopsy WaB also planned . since FN deposition is a fundame ntal Btep in wound-healing, the patient was submitted to two suboequent s ystemic creatmentB with dex (8 mgjday for 15 days Decodron in t he first and 4 =1/day for 15 days in the s econd treatment) combined with topic therapy (dex cream), and a repair of the leBionD was observed . TheBe data suggest that de)( treatment can be effective in favouring tho repair of wound-healing defects in patients s howing 8 lack of FN-ECH organization. It is likely that the molecular basis o f the a1:l.41ulatl.on of the repair proc~ss might be r o lated to the i nductio n .-: of f'1I transcription exerted by dex.