40 XXVI Congress of the IAP: Abstracts Dermatopathology was performed employing a heat-based antigen retrieval technique using the following monoclonal antibodies/to the following CT antigens: mAb MA454/MAGE-A1, M3H67/ 167 THE INTERPRETATION OF BENIGN BIOLOGIC BEHAVIOR MAGE-A3, 57B/MAGE-A4, CT7-33/CT7 (MAGE-C1), E978/NY-ESO-1, CT10#5/CT10 FOR KERATOACANTHOMA VERSUS CUTANEOUS SQUAMOUS CELL (MAGE-C2). In a subset of 8 Spitz nevi, the following mAbs were used for the analysis of CARCINOMA, USING A PANEL OF ANTIBODIES AGAINST: EGFR , P21 RAS, MDAs: HMB45/gp100, A103/Melan-A, T311/tyrosinase. P53 , CYCLIN D1 Results: No immunopositivity was seen with any of the tested anti-CT antigen mAbs Ahmad Zahi Al-Chawaf, Faculty of Medecine, Hama, Syria; Bassam Al-Hallani, Faculty (MA454, M3H67, 57B, CT7-33, E978, CT10#5) in any of the 32 Spitz nevi. However, all of Medecine, Homs, Syria three mAbs to MDAs (HMB45, A103, T311) showed strong and homogeneous staining in Background: The keratoacanthoma (KA)is a relatively common tumor which most often the subset of 8 Spitz nevi which were analyzed. occurs on the sun – exposed areas of light skinned individuals of middle age and older. Conclusion: The differential diagnosis of Spitz nevi versus malignant melanoma can pose This tumor may regress spontaneously, so recognition of the true nature of this tumor is of a diagnostic dilemma in surgical pathology. Markers such as mAb HMB45, A103 and T311 considerable practical and biological importance. Could it be named an aborted squamous indicate melanocytic lineage and their immunopositivity in the present lesions is consistent cells carcinoma? with the diagnosis Spitz nevus. However, they do not contribute to the distinction between Design: 24 cases of keratoacanthoma and 26 cases of SCC were immunostained for EGFR, benign and malignant. Previous analyses confi rmed the tumor-associated expression of P21 ras, P53, Cyclin D1 oncoprotein using a Dako Autostainer Plus. Lesions were scored CT antigen expression on a protein level. In melanomas, including so-called `Spitzoid for positive staining: diffuse basal and suprabasal staining and focal basal and suprabasal melanomas`, up to 50% were immunopositive for some CT antigen. The negative staining. immunoreaction of all our anti CT antigen reagents in the present series of Spitz nevi Results: The expression of P53(mutant gene)showed a great difference between SCC and confi rms the benign biology of this cutaneous lesion. Conversely, any expression of CT Keratoacanthoma (20/26 – 80% in SCC versus 4/24 – 16% in KA) (P<0.01) -2/4 of KA antigens on a protein level in a lesion regarded as Spitz nevus should raise serious doubts cases were reevaluated as SCC (KA –like SCC). The expression of P21 ras was more about its nature and diagnosis and rather speaks for the presence of a malignant tumor, evident in KA versus SCC (14/24 -58 % in KA versus 5/26- 19% in SCC (P < 0.05). EGFR particularly melanoma. was negative in all cases of KA and SCC but focally expressed in the neighboring reactive epithelium. Cyclin D1 showed more evident staining (focal expression) in SCC (10/26 170 PRIMARY MALE NEUROENDOCRINE NIPPLE ADENOCARCINOMA -38% in SCC versus 4/24 – 16 % in KA). SIMULATING MERKEL CELL CARCINOMA – A DIAGNOSTIC PITFALL Conclusion: Our results refer to the decisive role of P53 mutation in the development Klaus Busam, Memorial Sloan Kettering Cancer Center, New York, NY, United States of SCC and its invaluable role in setting down a convincing diagnosis of SCC. P21 ras Background: Male breast cancer is a rare entity accounting for <1% of all breast cancer was more defi nitive in KA. EGFR is expressed more in reactive and benign epidermal cases in the United States, but has a rate that has been rising over the last 25 years. Nipple growths and has no role in both KA and SCC. P53 mutation is critical and decisive in the skin/subcutaneous tumors in males are even rarer. Likewise, a true neuroendocrine development of cutaneous malignancy. Cutaneous malignancy may be ascribed more to carcinoma of the breast, defi ned as >50% of tumor cells staining for either chromogranin mutation in nuclear oncoproteins. or synaptophysin, is not a common entity, usually occurring in older females. Design: We present a case of a 70 year old male with a slowly growing nipple mass which 168 METASTATIC OVARIAN TUMOR ( KRUKENBERG`S TUMOR ) FROM had enlarged over the prior 1 ½ years. PRIMARY MALIGNANT MELANOMA OF THE SKIN: A CASE REPORT Results: The histology consisted of nests, trabeculae and sheets of basaloid cells with Zuhir Al-Shehabi, Department of Pathology ,Tishreen University, Lattakia, Syria; Lina rare abortive gland formation and a pushing edge. The case was originally misdiagnosed Al-Soufi , Department of Dermatology, Tishreen University, Lattakia, Syria as a Merkel Cell Carcinoma, based largely on histologic morphology. Strong staining for Background: Krukenberg tumor is classically defi ned as a metastatic signet ring cell synaptophysin (in greater than 50% of cells), CD56, keratins AE1:AE3 and Cam 5.2, as carcinoma arising in the stomach or gastrointestinal tract and metastasizing to the ovaries. well as ER and PR was noted. Myoepithelial cells within in-situ areas were identifi ed using Other primary sites have been described, and a few cases have no detectable primary. stains for calponin and 4A4, supporting a primary mammary duct origin. Additionally, a Design: We report a case of a 48 –year –old female with bilateral metastatic ovarian substantial portion of cells stained for GCDFP-15, confi rming some overlap with sweat tumors from primary malignant melanoma of the left thigh who underwent hysterectomy duct differentiation. & bilateral oophorectomy in the University Hospital of Tishreen University, Lattakia, yria Conclusion: To the best of our ability, although reported in the male breast, no case in November 2005. For confi rmation we used select immunostains, including cytokeratin of primary nipple neuroendocrine carcinoma in a male patient has been reported in the (CK), S-100 & Melan A. literature. The gender of the patient and association with the skin of the chest wall likely Results: The patient had an excisional biopsy in June 2003 for a pigmented tumor on contributed to the original misdiagnosis of Merkel Cell Carcinoma in this patient. the left thigh, followed by regional lymph node dissection. The pathological diagnosis was superfi cial spreading malignant melanoma (Clark level III). The regional lymph 171 DISTINCTION OF BENIGN SEBACEOUS PROLIFERATIONS FROM nodes were negative for metastasis. In November 2005 the patient presented with an SEBACEOUS CARCINOMAS BY IMMUNOHISTOCHEMISTRY acute abdomen. On laparotomy both right and left ovaries were enlarged, each measuring Erik Cabral, Stanford University Medical Center, Mountain View, CA, United States; 23x17x15 cm in maximum dimension. The histological examination revealed metastatic Aaron Aurbach, Armed Forces Institute of Pathology, Washington, DC, United States; malignant melanoma and the neoplastic cells were positive for S-100 and Melan –A and J. Keith Killian, National Cancer Institute, Bethesda, MD, United States; Terry Barrett, negative for CK . UT Southwestern Medical Center, Dallas, TX, United States; David Cassarino, Stanford Conclusion: To our knowledge, we report a rare case of Krukenberg`s tumor from University Medical Center, Palo Alto, CA, United States primary malignant melanoma of the skin. Review of the literature showed no cases of such Background: Sebaceous proliferations, including sebaceous adenomas, sebaceomas, and metastatic ovarian tumor. carcinomas, are histologically distinctive adnexal tumors with a spectrum of biological behavior ranging from benign to frankly malignant. The histologic distinction between 169 EXPRESSION OF CANCER TESTIS (CT) AND MELANOCYTE sebaceous adenomas and carcinomas may be challenging, especially in cases showing DIFFERENTIATION ANTIGENS (MDA) IN SPITZ NEVI atypical features and in small or partial biopsies. We studied multiple oncogenic and Nille Behrendt, Ackerman Academy of Dermatopathology, New York, NY, United States; therapeutic proteins by immunohistochemistry in order to identify differences in expression Achim A Jungbluth, Ludwig Institute for Cancer Research, New York, NY, United States; between benign and malignant sebaceous proliferations. Klaus J Busam, Department of Pathology Memorial Sloan-Kettering Cancer Center, New Design: A total of 27 cases, including 9 sebaceous adenomas, 4 sebaceomas, 8 sebaceous York, NY, United States; Sandra B Castelli, Ludwig Institute for Cancer Research, New carcinomas, and 6 cases of sebaceous hyperplasia, were examined by immunohistochemistry York, NY, United States; Geoffrey G. Gottlieb, Ackerman Academy of Dermatopathology, with antibodies directed against Ki-67, bcl-2, p53, p21WAF1, p27Kip1, c-erbB-2 (Her-2/ New York, NY, United States neu), CD117 (c-kit), cyclin D1, MDM2, CD99, MLH-1, and MSH-2. Background: Spitz nevi are rare melanocytic lesions primarily in children and young Results: We found that sebaceous adenomas and sebaceomas stained similarly to sebaceous adults, which can be diffi cult to differentiate from malignant melanoma. CT antigens such hyperplasia, while carcinomas had statistically increased levels of p53 (50% versus 11%, as the MAGE-gene family, CT7 and NY-ESO-1 resemble a group of recently identifi ed respectively, p < 0.05) and MIB-1 (average 30% versus 10%, p < 0.05). The carcinomas tumor-associated antigens, which are expressed in various malignant tumors and in normal also had signifi cantly reduced levels of bcl-2 (7% versus 56%, respectively, p < 0.05) and adult tissues solely in testicular germ cells. Extensive previous analyses have confi rmed p21 (16% versus 34%, p < 0.05) compared to the adenomas. that the protein expression of CT antigens is solely present in malignant tumors and not Conclusion: Thus, a combination of several of these markers may be diagnostically in any benign neoplasms or normal non-gametogenic tissue. Due to this strictly tumor- useful in challenging cases. In addition, we found little or no Her-2/neu and c-kit staining, associated expression pattern, CT Antigens have become a prime focus in the research indicating that immunotherapy with Herceptin or Gleevac would likely not be useful for for vaccine targets the immunotherapy of cancer. They may also be useful as a diagnostic sebaceous carcinomas. Moreover, these results, consistent with previous studies, show that marker in surgical pathology in order to differentiate malignant from benign lesions. Our sebaceous adenomas and carcinomas are distinct neoplasms, and provide no support for the group previously generated several monoclonal antibodies (mAbs) to various CT antigens. theory that all sebaceous adenomas are truly malignant. In the present study we analyze the usefulness of these anti-CT antigens in the differential diagnosis of Spitz nevi versus malignant melanoma. Since melanocyte differentiation 172 LOCALIZED AMYLOIDOSIS IN BASAL CELL CARCINOMA antigens (MDAs) are expressed in typical pattern in cells and lesions of melanocytic Hakan Cingil, Istanbul Military Hospital, Istanbul, Turkey lineage, we analyzed the presence of these antigens as a reference in a subset of Spitz nevi Background: Amyloid can be found in some diseaseS localized In lesionAL regions. One in our series. of these diseases is basal cell carcinoma (BCC). The existence of amyloid deposits in BCC Design: Formalin-fi xed paraffi n embedded tissue from 35 Spitz nevi were retrieved from the of the skin has been an established fact since 1930. archives of the Ackerman Academy of Dermatopathology. The lesions were re-evaluated by Design: In this study we aim to investigate the presence of amyloid in 60 BCC tissue an HE stain and the presence of a Spitz nevus was confi rmed in all cases; suffi cient amount specimens and the relationship between BCC histopathologic types, mitotic rate, solar of tissue for further analyses was available in 32 cases. Immunohistochemistry (IHC) elastosis, degree of apoptosis and the age of the patients. XXVI Congress of the IAP: Abstracts 41 Results: Patient ages ranged from 24 years to 92 years with a mean age of 56 years. The 175 A CLINICO-PATHOLOGICAL ANALYSIS OF PAPULAR FOLLICULAR male to female ratio was 1.14. Excisional biopsy specimens obtained from 60 patients were CUTANEOUS ERUPTIONS IN HUMAN IMMUNODEFICIENCY VIRUS fi xed in 10% buffered formalin and embedded in paraffi n. Five micrometer-thick sections INFECTED PATIENTS IN SOUTH AFRICA were stained with hematoxylin and eosin and Congo Red. The histologic distribution of Wayne Grayson, Division of Anatomical Pathology, University of the Witwatersrand & the amyloid deposits was fairly consistent. They were found in the fi brous stroma between NHLS, Johannesburg, South Africa; Judith Budavari, Division of Dermatology, University clumps of tumor cells and in the connective tissue abutting the advancing front of the of the Witwatersrand, Johannesburg, South Africa carcinoma. Amyloid was identifi ed as amorphous, eosinophilic extracellular material Background: Papular and follicular eruptions such as papulopruritic eruption (PPE), which stained rose-pink with Congo Red. eosinophilic folliculitis (EF) and infective folliculitis are relatively common disorders Conclusion: Amyloid was found in 55% of the BCC patients. The presence of amyloid in patients infected with the human immunodefi ciency virus (HIV), accounting for didn`t change with the age of the patients, the tumor histopathologic type or apoptosis. A approximately 6.9% of HIV-associated dermatoses in our academic centre. These relationship between solar elastosis and mitotic rate quantity and the presence of amyloid conditions may show considerable clinical overlap. was detected. Design: The aim of this study was to assess the relative proportion of pruritic papular cutaneous eruptions in South Africans with HIV-associated dermatoses, and to correlate 173 STROMAL/ENDOTHELIAL VEGF-C EXPRESSION RELATED TO the clinical and histopathological features of these lesions. Forty consecutive HIV-positive MELANOMA METASTASES IN SENTINEL LYMPH NODES patients who presented with papular follicular eruptions underwent skin biopsy. The Elena Gallego; Martina Alvarez; Luis Vicioso; Alfredo Blanes; Alfredo Matilla, University clinical and histopathological fi ndings were then reviewed. of Malaga, Malaga, Spain Results: Clinical features were similar in all patients and consisted of widespread papules Background: The prognosis of patients with melanoma depends on the tumour stage at and pustules involving the face, limbs and trunk. The most common histological fi nding diagnosis, and is based on microstaging and clinical/radiologic evaluation for metastases. was acute suppurative folliculitis, seen in 27 patients (67.5%). In most cases no cause Therefore, the early identifi cation of deposition of microscopic metastatic cells, by sentinel was found for the suppuration. PPE of HIV was diagnosed in six patients (15%), HIV- lymph node biopsy, is important as it determines the requirement for adyuvant therapy and associated EF in four (10%), Pityrosporum folliculitis in two (5%) and acne in one patient further management. Among several prognostic parameters, tumour thickness is currently (2.5%). Concordance between the initial clinical diagnosis and the fi nal histopathogical the most sensitive for predicting the metastatic risk of cutaneous melanoma. However, diagnosis was achieved in only 27.5% of cases. the prognosis for individual melanoma patients is still diffi cult to determine, since thin Conclusion: Skin biopsy remains an important aid to correct diagnosis and classifi cation melanomas can also develop into lethal metastases. More accurate prognostic indicators of papular and follicular eruptions in HIV-positive patients. Examination of multiple serial for melanoma metastasis are therefore urgently needed. Tumour angiogenesis is essential sections is essential for accurate diagnosis. for tumour growth; consequently, microvessel density has been used as a quantitative assessment of tumour vascularization in many human tumours, and also vascular endothelial 176 SYRINGOCYSTADENOMA PAPILLIFERUM - AN ATYPICAL EXAMPLE growth factor (VEGF-C) is an important player in some lymphatic metastasis. Therefore, WITH WORRYING FEATURES experimental studies have shown that some types of malignant tumours actively induce the James Harrison; A. J. Howat, Burnley General Hospital, Burnley, United Kingdom formation of lymphatic vessels, which promotes tumour spread to regional lymph nodes. Background:Syringocystadenoma papilliferum is a rare benign skin tumour usually Is the extent of tumour lymphangiogenesis a prognostic marker of lymph node metastasis presenting on the forehead. Syringocystadenocarcinoma papilliferum is the malignant in skin melanoma? counterpart which is characterised by similar morphological features but with defi nite Design: The purpose of our study was to assess the lymphatic vessel density (LVD) and malignant cytological features including Pagetoid spread into the surrounding epithelium. VEGF-C expression in 50 primary skin melanomas (10 positive sentinel lymph node and Design: We present a case report of an atypical syringocystadenoma papilliferum arising 40 negative sentinel lymph node) and investigate the relationship with lymph node status. on the back of a 54 year old man. LVD identifi ed by D2-40 immunostaining, was evaluated and quantifi ed within the tumor Results: Histological examination of the specimen excised by his GP showed a skin adnexal and in the peritumoral area, using computer-assisted morphometric analysis. VEGF-C tumour showing the overall morphology of a syringocystadenoma papilliferum, however, expression was evaluated by immunohistochemistry in tumor cells, endothelial cells and the epithelial cells showed nuclear pleomorphism and frequent mitotic fi gures. These stroma. features fell short of frank malignancy, and a diagnosis of an atypical syringocystadenoma Results: VEGF-C stromal and endothelial expression was positively associated with lymph papilliferum was made. node metastasis (p<0,005). No signifi cant difference in VEGF-C expression by neoplastic Conclusion: The lesion described does not appear to fulfi ll the criteria for a standard cells and LVD was shown between metastatic and non-metastatic melanomas. syringocystadenoma papilliferum or syringocystadenocarcinoma papilliferum. The lesion Conclusion: VEGF-C expression in the stromal/endothelial phase of the tumour appears to has been designated an atypical syringocystadenoma papilliferum and wider excision with be involved in lymph node metastasis of skin melanoma. close clinical follow up recommended.

174 MITF EXPRESSION DURING HUMAN CUTANEOUS EMBRYOGENESIS 177 B4 INTEGRIN SIGNALING IN SQUAMOUS CELL CARCINOMA Briana Gleason; Christopher Crum; George Murphy, Brigham and Women`s Hospital, Basil Horst; Susana Ortiz-Urda; Ngon Nguyen, Stanford University School of Medicine, Boston, MA, United States Stanford, CA, United States; Alan Russell, Cytokinetics, San Francisco, CA, United States; Background: The mechanism(s) whereby human melanocytes migrate from the neural Peter Marinkovich, Stanford University School of Medicine, Stanford, CA, United States crest to populate the epidermis and developing hair follicles during embryogenesis is Background: Squamous cell carcinoma (SCC) is the second most common malignancy incompletely understood. Murine studies implicate an intermediate mesenchymal stage overall, with an incidence of approx. 90/100.000 cases per year. Although SCC has a in this evolutionary process in which melanocyte precursors (`melanoblasts`) exist both in primarily invasive potential and metastasis is rare, these tumors nonetheless represent a intradermal as well as intraepithelial and intrafollicular compartments. A single study using signifi cant clinical problem. Recently, generation of malignant human epidermal tissue immunohistochemical staining for HMB45 suggested the existence of a similar intradermal resembling SCC via overexpression of constitutively active Ras/IkBa has been described. stage in human cutaneous embryogenesis. The melanocyte master transcriptional regulator, Formation of SCC-like neoplasms upon subcutaneous injection of Ras/IkBa transformed MITF, identifi es mature melanocytes as well as subpopulations of melanocyte precursor keratinocytes was found to be dependent on a6B4 integrin and laminin-5. The aim of stem cells that reside in the bulge region of the adult human hair follicle. this study was to identify the critical domains of laminin-5 and a6B4 integrin for tumor Design: To better defi ne the utility of MITF expression in evaluation of melanocyte formation, as well as to investigate possible mechanisms by which these molecules exert ontogeny, human embryonic skin (n=24) ranging from 6-8 weeks to 32 weeks gestational their effect on tumor progression. age was studied immunohistochemically for expression of MITF and Mart-1 on adjacent Design: Human keratinocytes isolated from patients with epidermolysis bullosa, which step sections. Localization of melanoblasts in the intradermal, intraepidermal, and lack endogenous expression of B4 integrin, were retrovirally transduced to stably re- intrafollicular compartments was evaluated at each gestational age. express wt-B4 integrin or B4 integrin mutant constructs. After transformation with Results: At 6-8 weeks of gestational age, MITF and Mart-1-positive cells were primarily oncogenic Ras and IkBa, these cells were used in invasion assays, as well as for assessment round to ovoid and located within the dermis. By 12 weeks, many but not all of these cells of their tumorigenic capacity in vivo in animal models. Activity levels of critical signaling had migrated into the epidermis, primarily to the suprabasal layers. Between 13 and 18 intermediates were assayed in transformed keratinocytes lacking functional B4 integrin weeks, MITF and Mart-1-positive cells localized to the epidermal basal layer, developed expression and endogenous expression of laminin-5, the main binding partner of B4 dendritic processes, and gradually migrated into the basal layers of the outer root sheath integrin. (ORS) as well as the hair matrix of the developing follicles. Occasional intradermal Results: Lack of endogenous B4 integrin expression signifi cantly decreases the invasive collections of MITF and Mart-1 positive cells were documented as late as week 19, where and proliferative capacity of Ras/IkB transformed keratinocytes, re-expression of wtB4 they persisted in perifollicular and perivascular adventitia. In addition, scattered cells integrin restores their invasive as well as tumorigenic potential. Disruption of ligand strongly positive for MITF but not Mart-1 were present within the bulge region of the hair binding to laminin-5 with specifi c point mutations in the B4-integrin extracellular domain, follicle, consistent with melanocyte precursor cells. Unexpectedly, the keratinocytes of the as well as absence of endogenous laminin-5 expression, yields a comparable phenotype, bulge area also showed nuclear staining for MITF. with decreased invasion and impaired tumor proliferation in vivo. This phenotype may Conclusion: The in situ migratory fate of MITF/Mart-1-expressing cells in fetal skin refl ect the impaired ability of tumor cells to mount an adequate signaling response, involves a well-defi ned progression from intradermal to intraepidermal to intrafollicular involving activation of small Rho GTPases. Further rescuing studies are underway to localization. Occasional foci of intradermal melanocytes may persist after the intraepithelial assess the importance of these pathways in SCC tumor formation. stages are completed, a fi nding of potential signifi cance to melanocytic proliferations Conclusion: Ligation of a6B4 integrin to laminin-5 is a key event in the ability of that may arise de novo within the dermis. Because MITF is not a specifi c marker for transformed keratinocytes to proliferate and invade, impacting on two critical features melanocytes and may play a role in stem cell maintenance via induction of anti-apoptotic for malignancy. B4 integrin, distinct from its function as an adhesive device in normal mechanisms such as Bcl-2, its expression in bulge epithelial cells may represent a novel keratinocytes, may therefore be a potential therapeutic target in regard to tumor formation and functionally relevant marker for follicular stem cells of both epithelial and melanocytic and tumor survival signaling in human squamous cell carcinoma. lineage. 42 XXVI Congress of the IAP: Abstracts 178 DEEP VEGF UP-REGULATION IN ATYPICAL MELANOCYTIC 180 IMMUNOHISTOCHEMICAL ANALYSIS OF ANGIOTENSIN LESIONS: DIFFERENTIAL CORRELATION WITH APOPTOSIS IN SPITZ CONVERTING ENZYME, NEUTRAL ENDOPEPTIDASE AND SUBSTANCE P TUMORS AND ATYPICAL MELANOCYTIC NEVI IN ACNE PATIENTS Ehab Husein, Barts and The London Hospital, London, United Kingdom; Lucia Pozo, Nermine-Ehsan Ismail; Amira Gamal; Iman Selait, Faculty of Medicine, Shebin El Kom, Homerton University Hospital, London, United Kingdom; Alfredo Blanes, University Egypt of Malaga School of Medicine, Malaga, Spain; Salvador J. Diaz-Cano, King`s College Many clinical evidence suggests that the nervous system including psychological factors Hospital, London, United Kingdom can infl uence the course of acne vulgaris (AV). Angiotensin converting enzyme (ACE) and Background: The bases of the cell kinetics and microvessel profi les in cutaneous neutral endopeptidase (NEP) are involved in neuropeptide degradation and may modulate atypical melanocytic lesions (atypical melanocytic nevi, AMN, and Spitz tumors, ST) are neurogenic infl ammation. Moreover, previous studies have demonstrated that stress elicits poorly understood. No study has correlated cell kinetics, microvessel profi le, and VEGF the release of the neuropeptide substance P (SP) from peripheral nerves. However, there has expression by topographic compartments in AMN and ST to date. been little substantial evidence of specifi c participation of cutaneous neurogenic factors in Design: We selected low-grade AMN (93), high-grade AMN (31), ST (42), malignant the disease process of AV. Therefore, the aims of this study were: 1) to investigate the role melanomas (42, 15 radial growth phase MM-RGP and 27 vertical growth phase MM- of ACE, NEP and SP in the pathogenesis of AV, 2) to correlate between the expression of VGP), and conventional melanocytic nevi (15 junctional, 20 compound), the latter two ACE, NEP and SP and the clinical grading of the disease. We used immunohistochemistry groups used as controls. Immunostaining for Ki-67 and VEGF, and in situ end labeling to examine the expression of ACE, NEP and SP in skin biopsies from 20 patients (mean (ISEL) of DNA fragments (using Klenow fragment of DNA polymerase I) were scored age 21±2 y, 14 males, 6 females) and 6 normal control subjects of matching age and sex. by topographic compartments: junctional, superfi cial dermal (above 0.76 mm) and deep Immunohistochemical results were correlated with histological assessment of acne lesions dermal (below 0.76 mm), screening the whole compartment in each case. Appropriate and clinical grading of the disease. Results revealed intense immunoreaction of ACE in controls were run in each sample. CD-31-stained slides were used to estimate microvessel acne patients in endothelial cells lining blood vessels especially around infl ammatory cells. density. The results were statistically compared using analysis of variance and Student t- NEP and SP immunoreaction was pronounced in germinal cells of sebaceous glands, nerve test, and considered signifi cant if P<0.05. fi bres between sebaceous acini and in close vicinity to blood vessels and infl ammatory Results: Superfi cial-to-deep gradient was maintained for Ki-67 in all lesions, but was infi ltrate. Skin from control healthy subjects and uninvolved adjacent skin from acne signifi cantly higher in MM. From junctional to deep dermal compartments, ST showed a patients showed weak expression of ACE, NEP and SP. There was a high signifi cant progressive and statistically signifi cant increase of ISEL indices (4.38%, 4.73%, 8.35%), difference between ACE, NEP and SP expression in acne patients than from normal which correlate directly with microvessel density (4.38, 4.39, 7.41 vessels/HPF). Dysplasia healthy controls, p=0.001. There was also a high signifi cant difference between ACE, grading revealed an inverse correlation with apoptosis in AMN, but kept the superfi cial- NEP and SP immunoreaction in acne lesions than that of uninvolved adjacent skin from deep gradient. VEGF expression by compartments appears in the Table. acne patients, p=0.002. Whereas, there was no signifi cant difference between ACE, NEP Junctional/ Superfi cial/ Deep and SP immunoreaction and the clinical grading of the disease, p>0.05. Data from our Junctional Nevi/ 11.00/ --/ -- study demonstrate the involvement of neurogenic factors such as ACE, NEP and SP in the Compound Nevi/ 4.39/ 0.92/ 0.45 disease process of AV and thus explain a possible mechanism of the exacerbation of acne. Spitz/ 29.04/ 34.85/ 42.82 LG-AMN/ 18.57/ 8.76/ 0.53 181 CORRELATED EXPRESSION OF EGFR AND MATRIX HG-AMN/ 16.74/ 7.20/ 5.56 METALLOPROTEINASES IN SQUAMOUS CELL CARCINOMA OF THE SKIN MIS/ 5.52/ --/ -- Aeree Kim; Jung-suk An; Kayeun Chang; Nam-hee Won; Yang-seok Chae, Korea University MM-RGP/ 25.25/ 28.08/ 52.79 College of Medicine, Seoul, Korea, South MM-VGP/ 39.65/ 55.21/ 43.83 Background: Epidermal growth factor receptor(EGFR) is involved in the carcinogenesis Conclusion: Deep VEGF up-regulation characterizes the vascular reaction associated with of various human cancers, but its mechanisms in carcinogenesis of skin tumor requires high-grade AMN and inversely correlates with the apoptosis index. In contrast, VEGF elucidation. Matrix metalloproteinases(MMPs) are degradative enzymes which allow expression in ST directly correlates with the microvessel density and apoptosis index and breakdown of the extracellular matrix and tumor progression by angiogenesis, destroying inversely with the proliferation index, suggesting that this distinctive blood vessel pattern basement membrane and local tissue architecture. The purpose of this study is to assess is reactive to regressive cellular changes. the roles of EGFR and MMP2 and MMP9 in pathogenesis of basal cell carcinoma(BCC), squamous cell carcinoma(SCC) and actinic keratosis(AK). 179 DEEP DERMAL UP-REGULATION OF HIF-1 IS DIRECTLY CORRELATED Design: The study groups consisted of 75 BCCs, 28 SCCs and 14 cases of AK in skin. WITH APOPTOSIS IN SPITZ TUMORS Immunohistochemical studies using antibodies against EGFR, MMP2 and MMP9 were Ehab Husein, Barts and The London Hospital, London, United Kingdom; Lucia Pozo, performed. Information regarding patient age, gender and site were obtained from Homerton University Hospital, London, United Kingdom; Alfredo Blanes, University pathology reports. Chi square and Fisher`s exact tests were used for comparison of data. of Malaga School of Medicine, Malaga, Spain; Salvador J. Diaz-Cano, King`s College Results: Sixteen patients (21.3%) with BCC and 20 patients (47.6%) with SCC or AK Hospital, London, United Kingdom expressed EGFR (p=0.003). MMP2 in tumor cells was found in 1 (1.3%) case of BCC and Background: The microvessel profi le bases in cutaneous atypical melanocytic lesions 7 (16.7%) of SCC or AK (p=0.003). MMP2 in endothelial cells of stroma was found in 46 (atypical melanocytic nevi, AMN, and Spitz tumors, ST) are poorly understood. No study (61.3%) cases of BCC and 36 (85.7%) of SCC or AK (p=0.004). Twenty-seven patients has correlated microvessel profi le and HIF-1 expression with cell kinetics by topographic (36.0%) of BCC and 39 patients (92.9%) of SCC or AK expressed MMP9 in tumor cells compartments in AMN and ST to date. (p<0.0001). Tumors with EGFR expressed MMP9 more frequently (69.4%) than tumors Design: We selected low-grade AMN (93), high-grade AMN (31), ST (42), malignant without EGFR (50.6%). However this had borderline statistical signifi cance (P=0.058). melanomas (42, 15 radial growth phase MM-RGP and 27 vertical growth phase MM- Expression of MMP2 was not correlated with expression of EGFR. VGP), and conventional melanocytic nevi (15 junctional, 20 compound), the latter two Conclusion: EGFR, MMP2 and MMP9 are more frequently expressed in squamous groups used as controls. Immunostaining for Ki-67 and HIF-1, and in situ end labeling cell carcinoma than in basal cell carcinoma. The expression underlies the presence of (ISEL) of DNA fragments (using Klenow fragment of DNA polymerase I) were scored invasiveness of SCC and progression of AK to invasive disease. EGFR may be involved in by topographic compartments: junctional, superfi cial dermal (above 0.76 mm) and deep the expression of MMPs in tumor or stroma. dermal (below 0.76 mm), screening the whole compartment in each case. Appropriate controls were run in each sample. CD-31-stained slides were used to estimate microvessel 182 ZOONOTIC DEEP CUTANEOUS FILARIASIS: 3 CASES FROM QUEBEC, density. The results were statistically compared using analysis of variance and Student t- CANADA test, and considered signifi cant if P<0.05. Victor Kokta, Dermatopathology, Sainte-Justine Hospital, Montreal, QC, Canada; Chantal Results: Superfi cial-to-deep gradient was maintained for Ki-67 in all lesions, but was Buteau, Infectious Diseases, Sainte-Justine Hospital, Montreal, QC, Canada; Pascal signifi cantly higher in MM. From junctional to deep dermal compartments, ST showed a Savard, Dermatology, Sainte-Justine Hospital, Montreal, QC, Canada progressive and statistically signifi cant increase of ISEL indices (4.38%, 4.73%, 8.35%), Background: Zoonotic deep cutaneous fi lariasis in Québec, Canada is a rare fi nding. and no correlation with microvessel density. Dysplasia grading revealed an inverse Several species cause fi lariasis in mammals including racoons, rabbits, and possibly correlation with Ki67/ISEL index in AMN, but kept the superfi cial-deep gradient. HIF-1 bobcats and mink. Humans are believed to be infected through the bite of a mosquito and expression by compartments appears in the Table. are accidental, dead-end hosts. In North America, only about 30 Brugia cases have been Junctional Superfi cial Deep reported, mostly from rural and suburban areas in the north eastern United States. No cases Junctional Nevi/ 2.48/ --/ -- have been reported from Québec. Compound Nevi/ 2.59/ 2.41/ 0 Design: Three rare cases of Québec-based zoonotic fi larial nematode deep skin infections Spitz/ 16.32/ 15.09/ 14.12 were reviewed. Patient age, travel information, lesional characteristics, systemic fi ndings, LG-AMN/ 2.59/ 0.86/ 0 serology, histopathology, treatment, and follow-up were gathered from the submitting HG-AMN/ 3.42/ 0.55/ 0 specimen and the treating physicians. Species identifi cation was performed by the MIS/ 0.76/ --/ -- Parasitic Disease Branch, Division of Infectious and Tropical Diseases Pathology, AFIP, MM-RGP/ 5.81/ 11.63/ -- Washington, DC. MM-VGP/ 10.58/ 18.78/ 18.08 Results: All patients developed cutaneous deep seated nodules 3-12 weeks following a Conclusion: Deep dermal upregulation of HIF-1 characterizes Spitz tumor and vertical presumed mosquito bite. The patients travelled within Québec within the North Laurentian, growth phase MM, with preservation of the superfi cial-deep gradient and direct correlation Abitibi or Gaspé regions. Histopathological evaluation revealed a granulomatous with apoptosis in Spitz tumors only. This over-expression represents an additional eosinophilic deep reticular and-or subcutaneous dermatitis with evidence of a fi larial mechanism of neovascularization, which does not correlate directly with the vascular nematode. Based on morphological characteristics, two cases were classifi ed as zoonotic density level. North American Brugia and one case as zoonotic Dirofi laria immitis. No patients had circulating microfi lariae. One patient had blood eosinophilia. In all cases, the nodules XXVI Congress of the IAP: Abstracts 43 disappeared within one month of biopsy. One patient was treated medically. neoplasm or even complete dedifferentiation. In the most diffi cult cases, for differential Conclusion: Firstly, cutaneous fi larial nematodes are not uniquely restricted to tropical diagnosis histochemical and immunohistochemical reactions can be used. regions. Canadian cases are typically transmitted through mosquito bites from infected mammals. Secondly, in the context of a deep cutaneous granulomatous eosinophilic 185 C16 LAMININ PEPTIDE INCREASES MELANOMA ANGIOTROPISM dermatitis, one must search extensively for a fi larial nematode. The worm is sometimes AND EXTRAVASCULAR MIGRATION IN THE SHELL-LESS CHICK CAM diffi cult to identify and typically presents variable states of degeneration. Also various ASSAY secondary skin lesions may be associated with the primary focus of infection and multiple Claire Lugassy, University of Miami Miller School of Medicine, Miami, FL, United States; biopsies might be necessary for diagnosis. Lastly, in most cases, the human is the dead-end Hynda Kleinman, National Institutes of Health, Bethesda, MD, United States; Stephen host, and spontaneous resolution follows an excisional biopsy. Vernon, University of Miami Miller School of Medicine, Miami, FL, United States; Danny Welch, University of Alabama at Birmingham, Birmingham, AL, United States; Raymond 183 EXPRESSION OF WILMS TUMOR 1 GENE (WT-1) IN CONGENITAL Barnhill, University of Miami Miller School of Medicine, Miami, FL, United States HEMANGIOMAS Background: The fundamental problem of human cancers, such as cutaneous melanoma, Victor Kokta, Dermatopathology, Sainte-Justine Hospital, Montreal, QC, Canada; Julie is their ability to metastasize. As distinct from intra-vascular dissemination, extravascular Powell, Dermatology, Sainte-Justine Hospital, Montreal, QC, Canada migratory metastasis (EVMM) has been described as a potential additional mechanism Background: Congenital or post-natal hemangiomas present a therapeutic challenge of melanoma spread in which tumor cells migrate along the outside of vessels. We have for the clinician. Rapid post-natal growth followed by slow involution in the fi rst few previously demonstrated a tumor-endothelial cell interaction, the angio-tumoral complex, years of life characterize the common infantile hemangioma (IH). Rapid growth and rapid in which melanoma cells occupy a pericytic location on the outside of the endothelium involution within the fi rst year caracterize the rapidly involuting congenital hemangioma without of intravasation, suggesting EVMM. In addition, we have shown that angiotropism (RICH), whereas growth followed by no regression are characteristic of the non-involuting in human melanoma biopsy tissue (the histopathological counterpart of the angio-tumoral congenital hemangioma (NICH). Cutaneous biopsy specimens from these hemangiomas complex) could be a prognostic factor predicting risk for metastasis. The melanoma cells demonstrate overlapping histological characteristics, especially in the early proliferative are linked to the endothelium by an amorphous matrix confi rmed to contain laminin. Given phases. Glut-1 staining is positive is all phases of IH. No known marker readily the role of specifi c laminins in angiogenesis, protease induction, migration, and metastasis, distinguishes NICH from RICH. we have investigated whether laminin plays a role in this extravascular mechanism of Design: Since WT-1 expressions have been found in proliferative vascular neoplasms tumor spread. Biologically active sites on laminin-1 have been identifi ed using proteolytic but not in vascular malformations, we hypothesized that the non-involuting congenital fragments and synthetic peptides. The C16 (KAFDITYVRLKF) peptide from the ƒ×1 hemangioma (NICH) would present the same profi le as vascular malformations. laminin chain has previously been shown to signifi cantly enhance pulmonary metastases Moreover, radiological studies show overlap between NICH and vascular malformations. of B16-F10 mouse melanoma cells. Using human melanoma cells stably expressing Green An immunohistochemistry study for expressions of the Wilms tumor gene (WT-1) was Fluorescence Protein (GFP), we have tested the effect of the C16 laminin peptide on performed on 8 infantile hemangiomas (IH), 3 rapidly involuting congenital hemangiomas melanoma angiotropism and extravascular migration in a shell-less chick chorioallantoic (RICH), 5 non-involuting congenital hemangiomas (NICH) , 5 pyogenic granulomas, 5 membrane (CAM) model. angiokeratomas, and 4 vascular malformations. The specimens were evaluated for intensity Design: Chicken eggs were opened after day 4 of incubation into a glass dish. At day 8, and distribution of cytoplasmic staining in the studied endothelial cells. 105 C8161 GFP melanoma cells were applied to the surface of the CAM in the presence of Results: The immunohistochemical study revealed a positive cytoplasmic staining in the 1 ƒÝg of the C16 peptide. Non treated melanoma cells and cells treated with a scrambled endothelial cells for WT-1 in all infantile hemangiomas (8/8), all RICH (3/3), all NICH peptide (SC16) were used as negative controls. The GFP expressing cells were observed (5/5), and all pyogenic granulomas (5/5). The intensity of the staining was strong and did after 3 days with a stereo fl uorescence microscope. At day 11, the CAM was dissected for not differ in the various phases of IH, RICH or NICH. Vascular malformations (0/4) as well histopathology. as angiokeratomas (0/5) were negative for this marker. Results: After 3 days, melanoma cells were observed spreading along or in the immediate Conclusion: WT-1 expression distinguishes vascular proliferations from vascular proximity of vessels. C16 laminin peptide signifi cantly enhanced angiotropism and the malformations and telangiectatic lesions (angiokeratomas). However, WT-1 expression distance of extravascular migration of C8161 cells compared to controls, whereas the does not distinguish IH and RICH from NICH. Moreover, the intensity does not change scrambled peptide did not. Histopathology confi rmed the angiotropism of melanoma cells in the different phases of the latter lesions. Defects in WT-1 signaling do not seem to be without intravasation. implicated in the inability of endothelial cells in NICH to undergo involution. Conclusion: Taken together, these results suggest that 1) the use of GFP tumor cells in the shell-less CAM assay is a relevant model for studying tumor dissemination and its 184 METASTATIC CUTANEOUS DISEASE FROM INTERNAL CARCINOMA relationship with the vascular network, 2) the migration of melanoma cells along the Eva Labropoulou, Department of Pathology, Hatzikostas General Hospital of Messolonghi, abluminal surfaces of vessels supports extravascular migratory metastasis, and 3) the C16 Messolonghi, Greece; Miltiadis K. Gerolymos, Department of Internal Medicine, Hatzikostas laminin ƒ×1 chain peptide has angiotropic, extravascular migration-promoting activity General Hospital of Messolonghi, Messolonghi, Greece; Georgia Charalambopoulou, on human melanoma cells, and might be a molecular target for preventing melanoma Department of Pathology, General Hospital of Agrinio, Agrinio, Greece; Catherine metastasis. Labropoulou, Patras University, Medical School, Patras, Greece; Polymnia Morfaki, Department of Pathology, General Hospital of Agrinio, Agrinio, Greece 186 LACK OF EXPRESSION OF C-KIT EXPRESSION IN DIFFERENT STAGES Background: Cutaneous metastases from internal carcinoma are not common, with a OF MYCOSIS FUNGOIDES reported incidence between 0.7% and 9.0%. Cutaneous tumor metastasis may be the fi rst Mandana Mahmoodi; Sadia Salim; Haider Asad, Drexel University College of Medicine, manifestation of cancer, but more often there are a prior known cancer and, frequently, Philadelphia, PA, United States; Franklin Sedarat; Drazen Jukic, University of pittsburgh, other known sites of metastasis as well. In this report, we aimed to present our experience Pittsburgh, PA, United States; J Steve Hou, Drexel University College of Medicine, on cutaneous metastases from internal carcinoma and discuss the related problem of Philadelphia, PA, United States differential diagnosis. Background: MF is typically an indolent disorder ,but the disease may progress toward Design: A search of the histopathological archival fi les of a 6-year period (2000-2005) more advanced forms. One of the histological indicators of poor prognosis is the presence of was performed for histopathological evidence of metastatic spread of internal carcinoma large CD 30 cells suggestive of large cell transformation of the tumor. Traditional systemic to the skin. chemotherapy has not resulted in durable remissions in MF. As a consequence, emerging Results: Seven patients could be identifi ed including four females (67, 72, 75 and 78 year therapeutics efforts have focused upon targeted biological agents and manipulation of old) and three males (45, 53 and 73 year old). In every case, routine hematoxylin-eosin the host immune response. Introduction of a new kinase inhibitor termed STI571 -with stains were available. Immunostains were carried out when necessary. In one case, the an inhibitory effect on C-KIT -created a new outlook in cancer therapeutics. C-KIT cutaneous metastasis was the fi rst indication of internal cancer. A poorly differentiated expression is an attractive target for an innovative approach enrolling STI571. There has adenocarcinoma was found, positive for cytokeratins AE1/AE3 and 7. A further clinical been multiple reports in literature pointing to C-KIT expression in CD 30 lymphomas . and laboratory examination revealed the lung as the primary tumor. In the rest cases, Goos et al reported expression of CD 117 in up to 50% of cases of mycosis fungoides the primary tumor was known before the biopsies. Primary tumors were invasive ductal with blast transformation. This study although very interesting was designed as a screening breast carcinoma (2 cases), colon adenocarcinoma (1), rectal adenocarcinoma (1), tool as such lacked the precision and detail in cases of mycosis fungoides. Due to potential adenocarcinoma of the ampulla of Vater (1), and adenocarcinoma of the stomach (1). The therapeutic implications, we decided to study expression of CD 117 in a larger group with diagnosis was simple in all cases with the exception of the case of gastric adenocarcinoma. different stages of cutaneous and lymphatic mycosis fungoides . In this case, the metastatic tumor was consisted exclusively of neoplastic cells, isolated Design: A total of sixty paraffi n embedded skin specimens from sixty patients with or in small clusters, with a signet ring appearance in contrast with the primary tumor that different stages of MF and Fifty nine positive lymph node from different stages were was a moderately differentiated adenocarcinoma of intestinal type. Immunohistochemical obtained . A distinction was made between tumors with and without blastic transformation, studies for cytokeratins AE1/AE3 and 20 confi rmed the stomach as the origin of metastatic Immunostaining was performed with monoclonal antibodies against CD 4 , CD 117 and spread. CD 30.Toluidine Blue stain was used to highlight mast cells. Consequently, high power Conclusion: The recognition of cutaneous metastases from internal carcinoma has a very fi eld counting of C-KIT-positive cells and Toluidine Blue -positive cells was performed important impact on the future treatment and prognosis of these patients. The diagnosis and these two values were subtracted to determine the C-KIT score: a new variable that of cutaneous metastasis from adenocarcinoma is not always simple; diagnostic problems was representative of non-mast cell CD117 positive cells. The relationship between C-KIT arise when skin lesions are the fi rst manifestation of the disease or when the clinical history score and clinicohistological stage of disease was assessed by statistical analysis . is unknown. Furthermore, the histological picture may simulate other primary cutaneous Results: Staining lymph nodes revealed scattered CD117 positive cells in subcapsular, adnexal neoplasms, including apocrine gland carcinoma, eccrine gland adenocarcinoma paracortical and medullary areas . The number of cells dwindled at higher stages of of classic type, mucinous carcinoma of eccrine glands, and primary cutaneous signet ring the lymph nodes where normal structures were replaced by tumor cells. Large cell cell carcinoma. Another diagnostic problem arises when there is a known prior malignancy transformation did not increase expression of CD117. Cutaneous specimens presented and the metastatic tumor shows a lesser degree of differentiation than those of the primary only scattered expression of C-KIT within the lymphatic infi ltrate. There was no signifi cant 44 XXVI Congress of the IAP: Abstracts difference in CK117 expression between different stages in skin or in cases with large the positive cells expressed as percentage of tumor cells. Appropriate controls were run cell transformation. in each sample. Cases were stratifi ed according to patient’s age in <50 years (group A, Conclusion: In this study, we were able to document absence of a relationship between 10 cases), 50-70 years (group B, 30 cases) and >70 years (group C, 10 cases). The results progression of disease in skin or lymph nodes and C-KIT expression. Neither there was were statistically compared using analysis of variance and Student t-test, and considered an association between large cell transformation and expression of CD117.Most of the signifi cant if P<0.05. positive cells appear to be mast cells. These fi ndings are in contrast to previous reports of Results: The average age in each group was 34 (group A), 59 (group B) and 78 years presence of CK 117 in CTCLs specially with CD 30 cells and in concert with recent paper (group C). All neoplasms were revealed positive for mlh1 and msh2, regardless of the age by Medeiros et al –which contributes previous reports to use of oversensitive anibody. We group. Proliferation and the percentage of FISH-detectable telomere revealed and inverse are in process of performing PCR to confi rm our fi ndings. correlation, being proliferation signifi cantly higher in group A than in group C; telomere revealed the opposite pattern. Apoptosis and the global kinetic index Ki67×Telomere/ 187 STROMAL INVASION AND NODAL METASTASIS IN EXTRAMAMMARY TUNNEL (%) were signifi cantly higher in group C as compared with A and B (P=0.0003 PAGET’S DISEASES and P=0.0006). Group B was heterogeneous, cases being able to stratifi ed by group A and Yasuka Miyakuni; Toshiharu Matsumoto; Yuki Yamada; Astushi Arakawa; Hiroshi Sonoue; C patterns. Koichi Suda, Juntendo University School of Medicine, Tokyo, Japan Group A/ Group B/ Group C Background: In extramammary Paget’s disease (EMPD), stromal invasion and nodal <50/ 50-70/ >70 metastasis are occasionally seen. However, a detailed study focusing these phenomena has Ki67/ 19.74/ 16.75/ 15.40 not been published. Thus, the purpose of the present study is to clarify stromal invasion Telomere FISH/ 41.66/ 53.97/ 53.25 and nodal metastasis in EMPD. Ki67×Telomere (%)/ 3.42/ 7.35/ 7.56 Design: The examined cases consisted of 35 cases, in which wide resection was performed. TUNNEL / 0.50/ 2.82/ 10.17 The examined materials, consisted of scrotum, penis and inguinal lesion from 27 males, Ki67×Telomere/TUNNEL (%) / 31.64/ 26.49/ 43.94 and vulva from 8 females. Sections for histological study were obtained from resected Conclusion: In the absence of mismatch repair abnormalities, malignancies in older patients materials, and histological sections were stained with conventional stains. are defi ned by signifi cantly increased apoptosis, related with a telomere-independent Results: Stromal invasion was found in 16 cases (46%); superfi cial stromal invasion in accumulation of genetic alteration that is facilitated by lower cellular turnover. 12 cases and deep stromal invasion in 4 cases. In 15 cases, inguinal lymph nodes were dissected, and nodal metastasis was noted in 5 cases. Nodal metastasis with or without 190 ECCRINE SYRINGOFIBROADENOMA OF THE SKIN. CASE REPORT stromal invasion group consisted of 0% (0/9) in non-invasion group, 80% (4/5) in mild AND LITERATURE REVIEW invasion group and 100% (1/1) in severe invasion group. Danilo Odashiro, LAC-Laboratorio, Ocular Pathology Laboratory McGill, Ophthalmology Conclusion: In EMPD, stromal invasion is a frequent event, and patients with stromal UNIFESP, Campo Grande, Brazil; Jeferson Cavalcante, Hospital do Cancer, Campo invasion relates to nodal metastasis. Grande, Brazil; Luciana Miiji; Macanori Odashiro; Neuza Odashiro; Cristina Katayama; Patricia Pereira, LAC-Laboratorio de Anatomia Patologica e Citopatologia, Campo 188 QUANTITATIVE ANALYSIS OF CCL21, CCL19, AND CCR7 MRNA Grande, Brazil; Alexandre Odashiro, Lac-Laboratorio, Ocular Pathology McGill, EXPRESSION IN PRIMARY AND METASTATIC CUTANEOUS MALIGNANT University for Pantanal Region and State Development, Campo Grande, Brazil MELANOMA Background: Eccrine syringofi broadenoma (ES) is a rare benign neoplasm arising from Carlos Monteagudo; David Ramos, University of Valencia, Valencia, Spain; Vicent Alonso, the intraepidermal portion of eccrine ducts. There are approximately 60 cases of ES Hospital Clinico Universitario, Valencia, Spain; José Antonio López-Guerrero, Fundacion reported in the English literature, and in recent years the lesion has been described as Instituto Valenciano de Oncologia, Valencia, Spain; Esperanza Jorda, Hospital Clinico occurring in association with other skin conditions. Universitario, Valencia, Spain; Antonio Llombart-Bosch; Antonio Pellin, University of Design: To report one case of ES on the face. Valencia, Valencia, Spain Result: A 71 year-old male complained of a lesion on his left cheek that had been presented Background: A role for the CCR7 chemokine receptor in melanoma lymph node metastasis for 5 years with occasional bleeding without major changes. Physical examination revealed has been proposed. The fact that CCR7 ligands, CCL21 and CCL19, are expressed in a 1,5 cm well-delimitated reddish lesion on his left cheek. A few seborrheic keratosis lymph node and lymphatic vessels supports the hypothesis that a gradient concentration of were also present. The clinical differential diagnosis included Dermatofi broma and these chemokines might facilitate lymph node metastasis. Hemangioma. The lesion was totally excised with free margins. The microscopic exam Design: Our goal was to quantify mRNA expression of CCR7 and its ligands, CCL21 showed a tumor with thin anastomosing epithelial cords and strands forming a net and and CCL19, in primary and metastatic human cutaneous malignant melanomas in order connected to the undersurface of the epidermis. The cells were smaller and more basophilic to determine if there is a gradient expression of both chemokines between primary and than the epidermal keratinocytes. The epithelial strands showed ductal differentiation, often metastatic lesions which could be responsible for the metastatic dissemination of human associated with a well-formed cuticle. Between the strands there was a rich fi brovascular melanoma. CCR7, CCL21 and CCL19 mRNA expression was evaluated by Real-Time stroma with adipocytes. No cellular atypia was present. The diagnosis of ES was made. Quantitative PCR in 62 frozen tissue samples from primary and metastatic melanoma: Conclusion: We presented a case of a rare benign eccrine tumor with approximately 60 <=1 mm, (n=15), >1mm (n=15), “in transit” metastases(n=14), lymph node metastases cases reported in the English literature to date. (n=10), and distant metastases (n=8). In order to evaluate the cell types responsible for chemokine ligand and receptor expression, an immunohistochemical study (avidin-biotin 191 ISOFORM-SPECIFIC REGULATION OF THE ACTIN-ORGANIZING immunoperoxidase technique) was also performed in adjacent frozen sections. PROTEIN PALLADIN DURING TGF-β1-INDUCED MYOFIBROBLAST Results: CCL21 and CCL19 mRNA levels were signifi cantly higher in thin primary DIFFERENTIATION melanomas than in thick tumors (p=0,019; p=0,016), in transit metastases (p=0,000, Mikko Rönty, University of Helsinki, Helsinki, Finland; Suvi-Katri Leivonen, University p=0,000), lymph node metastases (p=0,007, p=0,016) , and distant metastases (p=0,000, of Turku, Turku, Finland; Boris Hinz, Swiss Federal Institute of Technology, Lausanne, p=0,000). CCR7 mRNA levels were also higher in thin tumors than in “in transit” and Switzerland; Andrew Rachlin; Carol Otey, University of North Carolina, Chapel Hill, NC, lymph node metastases (p=0,022). CCL21 and CCL19 immunoreactivity was found in United States; Veli-Matti Kähäri; Olli Carpen, University of Turku, Turku, Finland endothelial cells of lymphatic vessels and high endothelial venules of lymph nodes as well Background: Contractile myofi broblasts are responsible for remodeling of extracellular as in some tumor cells. CCR7 immunostaining was present in dendritic and tumor cells in matrix during wound healing. However, their continued activity results in various primary and metastatic lesions. fi brocontractive diseases. Conversion of fi broblasts into myofi broblasts is induced by Conclusion: The fact that the highest CCL21 and CCL19 chemokine levels are present in transforming growth factor-β1 (TGF-β1) and is hallmarked by the neo-expression of thin primary melanomas suggests that these chemokine ligands might retain CCR7+ tumor α-smooth muscle actin (SMA), a commonly used myofi broblast marker. Moreover, cells in the primary site in these patients. In contrast, the lower levels of both chemokines myofi broblast differentiation and acquisition of the contractile phenotype involves in thick melanomas support its potential implication in lymph node metastasis of CCR7+ functionally important alterations in the expression of actin-organizing proteins. Palladin tumor cells. *Performed with FIS-PI030512 grant, from Fondo de Investigación Sanitaria, is a recently identifi ed cytoskeletal protein that controls the integrity of actin-containing Spain. stress fi bers. It is expressed as several isoforms, including major 3Ig (90 kDa) and 4Ig (140 kDa) forms that differ in their N-terminal sequence. 189 UPREGULATION OF TELOMERE-INDEPENDENT APOPTOSIS Design: Our purpose was to investigate, using cultured fi broblasts, a rat skin wound model CHARACTERIZES CUTANEOUS MALIGNANCIES IN OLDER PATIENTS and human tissue specimens, (1) whether myofi broblast differentiation is accompanied Jane Moorhead; Salvador J. Diaz-Cano, King`s College Hospital, London, United by changes in the expression of palladin, and (2) which signaling pathways regulate Kingdom expression of palladin. Background: There are controversial results on the infl uence of age on malignancy Results: Cultured untreated fi broblast express only palladin 3Ig isoform. However, TGF- prognosis, being the patient age used as staging criteria (e.g., thyroid neoplasms). To β1 induces neoexpression of palladin 4Ig isoform within 24 h. The expression of palladin test biologic features that can explain differences in tumors by age, the skin model was precedes upregulation of SMA, which is upregulated only after 48 h. Palladin was located selected for two main reasons: tumors tend to be detected relatively early due to their easy to the dense regions of robust actin stress fi bers and to focal adhesions. Elucidation of access and a whole variety of common neoplasms (carcinomas, melanomas, sarcomas and the TGF-β1 signaling pathways showed that Smad signaling, in particular via Smad3, lymphomas) can be analyzed. mediates the TGF-β1-induced upregulation of palladin and SMA in fi broblasts. Further Design: We selected 50 cases of nodular basal cell carcinomas, well-differentiated studies implied additional role for ERK1/2 and p38 pathways in regulating palladin gene squamous cell carcinoma, superfi cial spreading malignant melanoma, dermatofi brosarcoma expression. Analysis of the rat skin wound model showed that palladin 4Ig isoform is protuberans, and patch stage mycosis fungoides that have appropriate archival material. not expressed in fi broblasts of early (3d) wound granulation tissue that do not exhibit Representative samples were evaluated by standard immunohistochemistry for Ki67, microfi lament bundles. However, day 6 post-wounding granulation tissue fi broblasts telomerase, mlh1, msh2, In situ end labeling of DNA fragments (TUNNEL for apoptosis de novo express palladin 4Ig isoform in conjunction with the development of stress detection), and FISH-PNA of telomere. The tests were assessed in the whole lesion and fi bers (i.e. acquisition of the proto-myofi broblast phenotype). Thus, palladin expression XXVI Congress of the IAP: Abstracts 45 precedes SMA also in vivo. Finally, immunohistochemical analysis of conditions rich in 194 HISTOPATHOLOGIC ANALYSIS OF EPIDERMAL SKIN TUMOURS AND myofi broblasts (fresh human scar tissue, nodular fasciitis and myofi broblastic tumor of TUMOUR LIKE LESIONS IN ZARIA the breast) demonstrated a clear correlation between staining of SMA and 4Ig palladin in Modupeola Samaila; Abdulmummin Rafi ndadi, Ahmadu Bello University, Teaching myofi broblasts. On the other hand, normal connective tissue fi broblasts did not express 4Ig Hospital, Zaria, Kaduna., Nigeria palladin isoform or SMA. Background: Skin tumours are common worldwide. This is a documentation of the Conclusion: These results identify palladin 4Ig isoform as a novel marker of myofi broblast pattern of distribution in a referral laboratory in a Nigerian Teaching Hospital. AIM: To conversion in vitro and in vivo. They also provide for the fi rst time information about the study the histopathological pattern of epidermal tumours and tumour-like lesions seen in signaling cascades involved in the regulation of palladin expression. the Department of Pathology, Ahmadu Bello University Teaching Hospital ,Zaria over a ten year period. 192 CELL KINETICS IN SUPERFICIAL SQUAMOUS CELL CARCINOMA OF Methods: All epidermal tumours and tumour like lesions seen in the Department of THE SKIN: ROLE OF MISMATCH REPAIR PROTEINS AND TELOMERASE Pathology Ahmadu Bello University Teaching Hospital, Zaria over a ten-year study Inmaculada Ruiz; Juan J. Sanchez-Carrillo; Alfredo Blanes, University of Malaga School period [1991-2000] were reviewed. The age, sex and anatomical site of the lesion were of Medicine, Malaga, Spain; Salvador J. Diaz-Cano, King`s College Hospital, London, obtained from patients’records. These tumours were classifi ed according to World Health United Kingdom Organization’s International Histological Classifi cation for Skin Tumours.The data was Background: The cell kinetics of superfi cial squamous cell carcinoma (SCC) has revealed analysed and tabulated into frequency tables. controversial results due to the heterogeneity of lesions considered, making it more Results: A total of 350 such lesions comprising 9.9% of all cutaneous neoplasms seen diffi cult to assess key elements in squamous cell tumor progression. This study analyzes within the study period were collected. Overall, they have a male to female ratio of 1.7:1. kinetics, cell survival and mismatch repair in a series of high-grade intraepithelial and Malignant tumours constituted 72.5%; benign tumours 18.3% and tumour-like lesions microinvasive SCC. 9.2%. The commonest malignant lesion was squamous cell carcinoma, which constituted Design: We selected bowenoid actinic keratosis (HG-AK, 22 cases), SCC in-situ (37 68.3% of all the lesions with a male to female ratio of 1.7:1. Epidermal cyst comprised cases) and microinvasive SCC (<3mm depth, 36 cases) that had appropriate archival 16.3% with a male to female ratio of 1.5:1. material. Representative samples were evaluated by standard immunohistochemistry Conclusion: Epidermal tumours and tumour-like lesions are not uncommon in Zaria and for MCM2 (minichromosome maintenance-2), telomerase, mlh1, msh2 and TP53. The they show a male preponderance. Squamous cell carcinoma was the commonest epidermal immunoexpression was assessed in the whole lesion and the positive cells as percentage tumour and it also predominantly affects males. of tumor cells. Appropriate controls were run in each sample. The results were statistically Correspondence: Dr M.O.A SAMAILA, FMCPath. Department of Pathology Ahmadu compared using analysis of variance and Student t-test, and considered signifi cant if Bello UniversityTeaching Hospital,Zaria Email: [email protected] Mobile no P<0.05 08035891007 Results: Both MCM2 and TP53 expression increased from HG-AK to microinvasive SCC, revealing no statistically signifi cant differences (Table). Telomerase showed signifi cantly 195 PIGMENTED SQUAMOUS CELL CARCINOMA OF THE SKIN: REPORT upregulated expression in microinvasive SCC when compared with intraepidermal OF TWO CASES AND LITERATURE REVIEW lesions (P=0.0037) and mlh1 expression was signifi cantly down-regulated in HG-AK Alaa Samkari; Samih Salama, Department of Anatomical Pathology, St.Joseph`s Hospital, (P=0.0022). McMaster University, Hamilton, ON, Canada AK-HG/ SCC IN-SITU/ SCC MICRO Background: Pigmented squamous cell carcinoma (PSCC) is a rare variant of squamous MCM2, %Cellularity/ 9.00/ 19.17/ 28.58 cell carcinoma (SCC), which contains numerous pigmented melanocytes. This type TELOMERASE, %Cell/ 55.75/ 48.08/ 72.39 of SCC may be diffi cult to distinguish clinically and microscopically from pigmented MLH1/MSH2, %Cellularity/ 7.73/ 19.04/ 20.15 basal cell carcinomas (BCC), and cutaneous melanoma, especially those associated with % NULL/ 58.00/ 12.70/ 10.86 pseudoepitheliomatous hyperplasia. TP53, %Cellularity/ 30.50/ 32.78/ 36.63 Design: We studied two cases of PSCC of skin by histological, Immunohistochemical Conclusion: 1) The invasion capacity in superfi cial SCC is marked by up-regulation of and ultrastructural analysis, and reviewed the literature. The possible mechanisms of telomerase in lesions preferentially expressing abnormal TP53. 2) The differential growth pigmentation in such unusual lesions were elucidated. pattern of intraepidermal SCC is kinetically related with down-regulation of mismatch Results: The fi rst case is of an 83-year-old male, presented with a pigmented lesion on repair proteins. his forehead, clinically diagnosed as melanoma. The second case is of a 76-year-old female, presented with a pigmented lesion on her neck, clinically diagnosed as dysplastic 193 RELIABILITY OF NAIL CLIPPING HISTOLOGY FOR DIAGNOSING nevus. Histological examination of the excisional biopsies showed keratinizing SCC ONYCHOMYCOSIS with numerous pigmented cells containing melanin interspersed among the neoplastic Dussadee Sakonlaya, Division of Pathology, Faculty of Medicine, Thammasat University, squamous cells. These cells expressed S100 protein and HMB-45 indicating melanocytic Pathumthani, Thailand; Mali Achariyakul, Division of Internal Medicine, Faculty of origin. Ultrastucturally cytoplasmic melanosomes in the neoplastic epithelial cells were Medicine, Thammasat University, Pathumthani, Thailand confi rmed. There is no evidence of disease after 6 month of follow-up of the fi rst case. Background: Onychomycosis, fungal infection of nails, is a common nail disease that affects Conclusion: The literature and our fi ndings suggest that the pigment is derived from the patients’ emotion, occupation and consumes considerable health care expense. Routine non-neoplastic melanocytes mixed between the tumor cells. Recent review of the literature laboratory techniques for diagnosing onychomycosis are direct microscopy of potassium revealed only 28 cases of PSCC in skin and mucous membranes. Long-term follow-up hydroxide (KOH) preparation and fungal culture. Both techniques have limitations, such showed no evidence of local recurrence or metastasis after complete excision, indicating as time-consuming and low sensitivity. Recent studies have suggested that histological that PSCC of the skin appears to behave similarly to SCC of the usual type. A larger study examination of nail clipping specimens may be a very sensitive method. Aim of this study comparing cases of each type that are matched for thickness of tumor, site, extent of is to compare sensitivity, specifi city, and predictive value of nail clipping histology with precursor lesions, and other factors are however needed to confi rm this fi nding. those of KOH preparation and fungal culture for diagnosing onychomycosis. Design: Nail specimens from patients with suspected onychomycosis were evaluated 196 THE EVALUATION OF DIFFERENT DIAGNOSTIC METHODS FOR using KOH preparation, fungal culture and histological examination of the nail clippings, ONYCHOMYCOSIS which were clipped from the most proximal point of abnormal nail plate that separated Johann Schneider, Anatomical Pathology, Stellenbosch University and National Health from the nail bed using sterile standard nail clipper. Subungual keratinous debris, if Laboratory Services Tygerberg, Parow, South Africa; Joanie Maré, Medical Microbiology, any, was collected by scalpel blade scraping. Histological samples were fi xed in 10% Stellenbosch University, Parow, South Africa; Francois Jordaan, Dermatology, formalin and processed for routine histological examination. H&E, and special stains for Stellenbosch, University and Tygerberg Hospital, Parow, South Africa; Elizabeth fungus: periodic acid-Schiff (PAS) and Gomeri methenamine silver (GMS) were applied. Wasserman, Medical Microbiology, Stellenbosch University and National Health Slides were examined under light microscope. Presence of pathogenic fungi by at least Laboratory Services Tygerberg, Parow, South Africa; D. Hayward, Molecular Biology and one diagnostic technique was considered as true positive (the gold standard). Sensitivity, Genetics, Stellenbosch University, Parow, South Africa specifi city, positive, and negative predictive value of different techniques were compared Background: Onychomycosis is a common clinical problem that presents signifi cant using Z test with statistical signifi cance level of .05. challenges in terms of accurate diagnosis and appropriate treatment. Objectives: To Results: 75 nail samples were included in this study. 31 out of 75 specimens (41%) had assess the sensitivity and specifi city of mycological culture and histopathology in the at least 1 of the 3 diagnostic methods positive for pathogenic fungi. Among these, 30 diagnosis of onychomycosis. specimens (40%) were positive by histology. Histologically, PAS and GMS demonstrated Design: This retrospective study included one hundred patients who presented at Tygerberg fungal elements lying between nail lamina. Sensitivity of each technique was: histology 97 Hospital with clinically suspicious onychomycosis. The study comprised relevant clinical %; KOH 84 %; and culture 74 %. Histology was more sensitive than culture (p = .0059) data, microscopic review of paraffi n-embedded and PAS stained sections from nail and KOH (p = .0427). Specifi city was: histology 100 %; KOH 100 %; and culture 55 %. clippings (PATHPAS), and correlation with mycological culture reports. All data was Histology and KOH were more specifi c than culture (p < .0001). Positive predictive value captured according to predesigned protocols. was: histology 100 %; KOH 100 %; and culture 54 %. Negative predictive value was: Results: The study group included 74 females and 26 males with an age range of 2 to histology 98 %; KOH 90 %; and culture 75 %. 84 years. The PATHPAS method confi rmed fungal infection of the nails in 64 patients, Conclusion: Of the 3 diagnostic techniques, histology is not only the most sensitive comprising 20 with Candida albicans, 35 with dermatophytes, and 9 with both. method, but also highly specifi c with highly negative and positive predictive value in Mycological cultures were done in 39 cases. Of the 33 negative cultures, 21 showed diagnosing onychomycosis. Procedure of nail clipping is painless and can be performed fungi in nail clippings, leaving 12 cases without confi rmation of onychomycosis. Of the 6 with simple equipment. Nail clipping histology can be used as a screening test, especially positive fungal cultures, 3 correlated with negative histology, one with Candida albicans, in cases of KOH-negative, clinically suspected onychomycosis while waiting for culture and one with dermatophytes. One case showed dermatophytes on histology, but Candida results. albicans was cultured. Conclusion: Mycological culture of nail clippings shows a low yield. The PATHPAS 46 XXVI Congress of the IAP: Abstracts method is most effective to demonstrate fungi, but poor correlation with culture results signifi cantly lower than either HISPOS group (p<0.0001). Multivariate analysis of clinical suggests that identifi cation of fungal species may not be accurate. Alternative methods such data and pathologic characteristics of the primary tumour identifi ed Breslow thickness as as polymerase chain reaction may improve the diagnostic accuracy of onychomycosis. the only other independent prognostic factor. Conclusion: Our fi ndings support previous observations that histologic evidence of 197 AUTOCRINE TUMOR SUPPRESSION EFFECTS OF CXCL10 GENE ON metastatic melanoma in SLN is an independent positive predictor of disease recurrence. HUMAN MALIGNANT MELANOMA CELLS: RECEPTOR DEPENDENCY In contrast, detection of tyrosinase mRNA without concurrent histologic positivity does Steven Tahan; Shuping Zhao, Beth Israel Deaconess Medical Center and Harvard Medical not increase the likelihood of developing short-term disease recurrence when compared to School, Boston, MA, United States patients with negative SLN evaluation by both methods. Further studies are necessary to Background: CXCL10 (formerly IP-10) was originally identifi ed as an IFN-gamma- validate these fi ndings and to determine whether the observed outcome patterns alter with inducible gene in monocytes, fi broblasts and endothelial cells. It has been shown to exert longer duration of follow-up. tumor suppression activity in experimental models principally through immunomodulatory and anti-angiogenesis mechanisms. Little work has been done on the effect of CXCL10 199 A CD-ROM TO AID CLINICOPATHOLOGICAL ANALYSIS OF gene on human melanoma cells. In this study, we investigate the effects of CXCL10 gene CUTANEOUS MELANOCYTIC LESIONS product on human malignant melanoma cells by transfecting CXCL10 gene into cell Beatrice Vergier; Jean Marc Dubois, Medical University of Bordeaux (France), Pessac lines and observing its effect on cell proliferation and invasiveness. We then evaluate the (Bordeaux), France; Alistair Cochran, David Geffen School of Medicine at UCLA, Los dependency of these actions on expression of its receptor, CXCR3. Angeles, CA, United States; Christiane Bailly, Centre Leon Berard, Lyon, France Design: The 294 base-pair full-length coding sequence for human CXCL10 gene was Background: Melanocytic lesions may be diffi cult to analyze and a simple and readily cloned into a protein expression vector (PcDNA3) and transfected into human malignant accessible reference system would be of value. melanoma cells (WM2664, A375) using FuGENE 6 transfection reagent. Empty PcDNA Design: We have conceived a CD-Rom not to replace available books on melanoma but 3 vector was transfected as vector control. Stable transfection cell lines were generated by to provide a tool that could be used as an diagnostic aid to facilitate day to day evaluation G418 screening. CXCL10 mRNA was highly expressed, resulting in a 10-fold increase in of microscopic slides from patients with melanocytic lesions or as an accessible tool for protein product in both of the CXCL10 gene transfected melanoma cell lines compared to education (including self evaluation on the basis of 40 exemplary cases). We have chosen control cells. Cell proliferation and matrigel invasion assays were carried out in triplicate to prepare a CD-Rom because this is currently the most adaptable medium permitting on these stably transfected human malignant melanoma cell lines at 72 hrs of culture. The the use of algorithms with numerous readily accessible critical links to the full range of expression of the CXCL10 receptor gene and its protein, CXCR3, was then evaluated using melanocytic tumors. RT-PCR and western blot analysis in the two transfected lines. An Affymetrix GeneChip Results: First part of this CD-Rom contains three algorithms: - the fi rst based on “clinical array and RT-PCR assays were performed to explore the downstream gene regulatory information” (age of patient, site and size of lesion, its duration and clinical appearance), effects of CXCL10 gene tranfection. - the second on “lesional silhouettes” evaluating slides without magnifi cation against Results: Cell proliferation rate was reduced by nearly 40% in CXCL10 transfected good light, - and the third on “microscopic examination” emphasizing lesional location WM2664 melanoma cells compared with control cells, as measured by an OD reading in the skin: epidermis alone, epidermis and dermis, dermis alone or dermis and subcutis. at 490 nm of 0.61 +/- 0.04 compared with 0.90 +/- 0.02 for control cells, p = 0.025. Cell These algorithms assist pathologists to sharpen their diagnostic focus. The second part of invasiveness was similarly reduced in CXCL10 transfected WM2664 melanoma cells this CD includes more traditional chapters: - the relative signifi cance of different criteria compared with control cells (18.5 +/- 10.1 vs. 64.8 +/- 4.3, p = 0.005). No difference was in determining “benign versus malignant” - technical aspects of the management and observed in CXCL10 transfected A375 melanoma cells compared with controls for cell evaluation of melanocytic tumors (including immunohistochemistry and sentinel lymph proliferation rate (0.55 +/- 0.04 vs. 0.55 +/- 0.04) and for cell invasiveness (33 +/- 3.9 vs node analysis) - preparation of optimized reports that include macroscopic and microscopic 34 +/- 3.5). CXCR3 receptor mRNA and protein expression were observed in the CXCL10 information essential for clinical management and accurate assessment of likely clinical transfected WM2664 melanoma cells, but not in CXCL10 transfected A375 melanoma outcome (including the TNM classifi cation). At least the third part of the CD-Rom is an cells. Affymetrix GeneChip array (12,000 genes) and RT-PCR assays showed at least four encyclopedic section detailing the clinical and histopathological features of 130 different fold upregulation of 13 genes and down-regulation of 14 genes in CXCL10 transfected melanocytic entities. This CD-Rom comprises more than 2000 color photomicrographs, WM2664 melanoma cells compared with vector control. 350 clinical pictures and 200 line diagrams. Seven french pathologists with different levels Conclusion: This study demonstrates that CXCL10 gene can exert autocrine tumor of experience in dermatopathology evaluated this CD and its algorithms. All found the CD suppression effects on human melanoma cells in addition to its previously described easy to use. The best improvement in diagnostic accuracy was reported by less experienced paracrine (immunomodulatory and antiangiogenesis) anti-tumor activity. These autocrine pathologists. effects are dependent upon the expression of its receptor, CXCR3. CXCL10 gene expression Conclusion: The tests confi rmed that this CD is a valuable education tool that can also is associated with up-regulation or down-regulating of specifi c genes. Further study is facilitate routine evaluation especially in case of less experienced pathologists. The French required to elucidate the mechanism and signal pathways involved in these actions. version of this CD-Rom is available from march 2006 and is distributed by the french division of International Academy of Pathology (IAP). We hope the English version will 198 SENTINEL LYMPH NODE EXCISION FOR MELANOMA: MOLECULAR be published in 2007/2008 (discussion in progress with publishers). DETECTION OF TYROSINASE MRNA ALONE IS NOT A PREDICTOR OF SHORT TERM DISEASE RECURRENCE, IN CONTRAST TO HISTOLOGIC Education EVIDENCE OF METASTASIS Cuneyt Tatlidil; Winston Parkhill; Carman Giacomantonio; Wenda Greer; Steven Morris; 200 A CASE BASED, STUDENT RUN COMPUTER ASSISTED MINICOURSE Noreen Walsh, Capital District Health Authority and Dalhousie University, Halifax, NS, IN DIAGNOSTIC AND MOLECULAR PATHOLOGY Canada Alain Borczuk; Eugene E. Marcantonio; Jay H. Lefkowitch, Columbia University, New Background: Examination of sentinel lymph nodes (SLN) by histology and molecular York, NY, United States methods has become standard practice in staging patients with intermediate thickness Background: Clinical applications of pathology (diagnostic and molecular pathology) melanoma and clinically negative regional lymph nodes (LN). The goal is to improve are underrepresented in medical school curricula. Multidisciplinary sessions are ideal prognostic accuracy and identify those patients who might benefi t from elective regional demonstrations of the role of pathology in medical decision-making, but present diffi culties lymph node dissection (ERLND). Our current practice is to proceed to ERLND only in in recruitment and scheduling of clinician contributors. The creation of multidisciplinary patients with histologic evidence of SLN metastasis. The proper approach to patients with video “expert” clips could obviate this problem. Using such clips in conjunction with only positive molecular fi ndings is unclear. To further complicate the issue, incidental textbook and scientifi c literature resources could allow for preparation of student-run benign nodal inclusions such as capsular nevi can yield RT-PCR positivity in the absence multidisciplinary teaching sessions. of true metastatic disease. Design: Two pilot sessions on the role of diagnostic and molecular pathology were Design: Our study consisted of a retrospective review of all patients at our institution with developed: (1) Lung adenocarcinoma (screening CT, pre-operative risk assessment for cutaneous melanoma who underwent SLN excision between October 1998 and December malignancy, pathology diagnosis, immunohistochemistry, EGFR FISH, EGFR mutation 2004. 315 SLN biopsies from 170 patients were evaluated by methodology used in the analysis) and (2) chronic myelogenous leukemia (clinical presentation, pathologic Multicentre Selective Lymphadenectomy Trial at the John Wayne Cancer Institute. Briefl y, diagnosis, cytogenetics, bcr-abl FISH, RT-PCR and realtime PCR). Experts in the three quarters of each LN was examined by routine histology and immunohistochemistry disciplines of oncology, radiology, pathology, molecular pathology, pulmonary medicine while the remainder was frozen for RNA extraction and RT-PCR analysis for tyrosinase and hematology were videotaped to address relevant topics for these pilots. Students were mRNA. SLN status (histologic and molecular) was then correlated with clinical outcome. assigned to subgroups representing each discipline and asked to use the videos and resource Results: Complete pathologic and clinical data was available for 139 patients. In 70 material to independently create 10-15 minute presentations to teach the entire group about patients (50.4%), SLN were negative by both histology and RT-PCR (HISNEG/ PCRNEG). how their discipline related to the disease entity. A multi-item questionnaire was created Histologic evidence of melanoma was identifi ed in 34 cases (24.5%), of which 26 asking for student responses using a scale from 1-5 (disagree strongly, disagree somewhat, demonstrated concurrent RT-PCR positivity (HISPOS/ PCR2POS). Detection of tyrosinase neutral, agree somewhat, agree strongly) and for free-text comments. mRNA in the absence of histologic metastasis (HISNEG / PCRPOS) was seen in 35 patients Results: A session to assess student preparedness for the scientifi c material was held prior (25.2%). Capsular nevi were identifi ed in 5 of these cases and in 20 cases (14.3%) overall. to Pilot 1. A post-session examination consisting of 5 questions showed a mean score of RT-PCR positivity was the sole fi nding (HISNEG/ PCRPOS/ CNNEG) in the remaining 30 4.6 (+/- 0.6), with 108 of 151 students achieving 5 of 5 questions correct. Survey questions cases (21.6%). With a median follow-up of 25 months, local and/or systemic recurrence regarding interest in the topic and their fi rst-year preparation for this subject scored 4.7 and had developed in 31 patients (22.3%). Recurrence rates were similar among patient 4.04 of 5.0, respectively. After the fi rst student-run multidisciplinary session (12 students) groups with positive histology, irrespective of RT-PCR status (HISPOS/ PCRPOS: 62%; on lung carcinoma, survey questions regarding success in teaching goals and success of HISPOS/PCRNEG: 75%). Only 10% of HISNEG/ PCRNEG patients developed recurrence, session format scored 4.0 and 4.0, respectively. Questions regarding clinical decision- signifi cantly less than either HISPOS group (p<0.0001). The lowest recurrence rate was in making and probability that journal articles would encourage future continuing education the HISNEG / PCRPOS/ CNNEG group (7.7%), comparable to HISNEG/ PCRNEG patients but received lower scores of 3.6 and 3.7, respectively.