Anna Maria Cesinaro

In Vivo Confocal Microscopy for Diagnosis of Melanoma and Basal Cell Carcinoma Using a Two-Step Method: Analysis of 710 Consecutive Clinically Equivocal Cases

We describe two algorithms to diagnose basal cell carcinomas (BCCs) and melanomas (MMs) using in vivo reflectance confocal microscopy (RCM). A total of 710 consecutive cutaneous lesions excised to exclude malignancy (216 MMs, 266 nevi, 119 BCCs, 67 pigmented facial macules, and 42 other skin tumors) were imaged by RCM. RCM features were correlated with pathology diagnosis to develop diagnostic algorithms. The diagnostic accuracy of the BCC algorithm defined on multivariate analysis of the training set (50%) and tested on the remaining cases was 100% sensitivity, 88.5% specificity. Positive features were polarized elongated features, telangiectasia and convoluted vessels, basaloid nodules, and epidermal shadowing corresponding to horizontal clefting. Negative features were non-visible papillae, disarrangement of the epidermal layer, and cerebriform nests. Multivariate discriminant analysis on the training set (excluding the BCCs) identified seven independently significant features for MM diagnosis. The diagnostic accuracy of the MM algorithm on the test set was 87.6% sensitivity, 70.8% specificity. The four invasive MMs that were misdiagnosed by RCM were all of nevoid subtype. RCM is a highly accurate non-invasive technique for BCC diagnosis. Good diagnostic accuracy was achieved also for MM diagnosis, although rare variants of melanocytic tumors may limit the strict application of the algorithm.

In vivo assessment of melanocytic nests in nevi and melanomas by reflectance confocal microscopy.

In vivo reflectance confocal microscopy is a novel technique for the noninvasive study and diagnosis of the skin. The aim of this study was to describe and characterize the cytological and architectural aspects of cell clusters in melanocytic lesions observed by confocal microscopy, and to correlate them with routine histopathology. A total of 55 melanocytic lesions comprising 20 melanomas, 25 acquired nevi and 10 Spitz nevi were studied by means of reflectance confocal microscopy, dermoscopy and routine histopathology. Three different types of cell clusters at confocal microscopy observation (dense, sparse cell and cerebriform clusters) were identified and correlated with histopathology. Dense clusters appeared characteristic for benign lesions, although present in 13 out of 20 melanomas. Sparse cell clusters were more frequently observable in melanomas, but also sporadically present in one Spitz nevus. Moreover, cerebriform clusters were exclusively observed in five out of 20 melanomas. Confocal microscopy allowed the in vivo characterization of aspects of melanocytic nests and their exact correlation with histopathology.

Synchronous occurrence of epithelial and stromal tumors in the stomach: a report of 6 cases.

OBJECTIVE The synchronous development of epithelial and stromal tumors in the stomach has been reported rarely in the literature. A series of 6 such cases is described in this article. METHODS Clinical and pathologic data were recorded and the literature was reviewed. RESULTS Five cases featured the simultaneous occurrence of stromal tumors (1 benign, 3 borderline, 1 malignant) and adenocarcinomas, whereas the stromal tumor in the sixth case was found in association with a carcinoid. No collision tumors were observed. In 2 cases, tumors arose from the same site and were closely juxtaposed, but in 4 patients they developed from different areas of the stomach. A preoperative histologic diagnosis of both tumors was not achieved in any case. Two patients harbored occult infiltrative epithelial lesions (1 diffuse-type adenocarcinoma, 1 carcinoid), which were detected only at pathologic examination of the gastric mucosa adjacent to the stromal tumor. CONCLUSIONS The simultaneous occurrence of epithelial and stromal tumors in the stomach can be less rare than usually expected. Coincidence alone could account for such an association, particularly in areas with high incidence rates of gastric cancer. The hypothesis that a single carcinogenic agent might interact with two neighboring tissues in the stomach inducing the development of tumors of different histotype cannot be theoretically discarded.

Skin aging: In vivo microscopic assessment of epidermal and dermal changes by means of confocal microscopy

BACKGROUND Skin aging is thought to be a complex biological process that is traditionally classified as intrinsic and extrinsic aging. Several clinical score and instrumental devices have been applied to obtain a precise assessment of skin aging. Among them, confocal microscopy has emerged as a new technique capable of assessing cytoarchitectural changes with a nearly histopathologic resolution. OBJECTIVE We sought to determine the microscopic skin changes occurring on the face in different age groups by means of confocal microscopy. METHODS The skin of the cheek in 63 volunteers belonging to distinct age groups was analyzed by confocal microscopy. In 4 cases, routine histopathology was performed on site-matched surplus areas from routine excisions for obtaining a better comparison with confocal findings. RESULTS Young skin was characterized by regular polygonal keratinocytes and thin reticulated collagen fibers. With aging, more irregularly shaped keratinocytes and areas with unevenly distributed pigmentation and increased compactness of collagen fibers were observed. In the elderly, thinning of the epidermis, marked keratinocyte alterations, and huddles of collagen and curled fibers, corresponding to elastosis, were present. A side-by-side correlation between confocal descriptors and histopathologic aspects has been provided in a few cases. LIMITATIONS Reticular dermal changes cannot be assessed because of the limited depth laser penetration. CONCLUSIONS Confocal microscopy was successfully applied to identify in vivo skin changes occurring in aged skin at both the epidermal and dermal levels at histopathologic resolution. This offers the possibility to test cosmetic product efficacy and to identify early signs of sun damage.

Microsatellite instability in multiple colorectal tumors.

Tumor multiplicity is a hallmark of hereditary cancers: in the colon‐rectum multiple tumors represent 5–10% of all colorectal cancer cases. A portion of these cases belongs to hereditary non‐polyposis colorectal cancer (HNPCC), a genetic cancer syndrome due to mismatch repair (MMR) gene mutations, phenotypically expressed as microsatellite instability (MSI); the majority of multiple tumors, however, is apparently without any family history. We analyzed 78 (38 synchronous and 40 metachronous) neoplasms from 37 patients with multiple tumors of the large bowel, both HNPCC and sporadic, with the aim of identifying a common genetic basis in multiple tumors. DNA was extracted from normal and cancerous formalin‐fixed tissue and was analyzed for MSI using 6 markers. Tumors showing MSI in at least 2 of 6 microsatellite loci were defined as MSI(+). The overall number of MSI(+) tumors was 22 (28.2% of the total). A significant difference in the rate of MSI(+) between HNPCC and sporadic tumors was observed (85% vs. 17%). In the same patients, the MSI phenotype of synchronous tumors (both HNPCC and sporadic) tended to be more concordant than that of the metachronous ones. The higher frequency of MSI in HNPCC than in sporadic tumors, even when multiple, suggests that the involvement of MMR genes in the pathogenesis of the sporadic cases may be uncommon, thus confirming that screening for MSI in multiple colorectal tumors could be a useful tool in the identification of HNPCC in the general population. Int. J. Cancer 81:1–5, 1999.

Is confocal microscopy a valuable tool in diagnosing nodular lesions? A study of 140 cases

Nodular lesions pose diagnostic challenges because nodular melanoma may simulate all kinds of melanocytic and nonmelanocytic lesions. Reflectance confocal microscopy (RCM) is a novel technique that allows visualization of the skin at nearly histological resolution although limited laser depth penetration hampers visualization of the deep dermis.

New insights into nevogenesis: In vivo characterization and follow-up of melanocytic nevi by reflectance confocal microscopy

BACKGROUND Development of melanocytic nevi is a complex process. OBJECTIVE The aim of the study was to characterize the in vivo confocal microscopy patterns and histopathologic correlates of melanocytic nevi. In addition, for the first time, confocal follow-up of characteristic nevi was performed documenting histologic changes in nevi. METHODS For the correlation study, 33 melanocytic nevi showing characteristic dermatoscopic patterns were studied by confocal microscopy. For the follow-up study 20 nevi were monitored for 12 to 18 months. RESULTS Reticular nevi showed two different confocal patterns, ringed and meshwork, mostly corresponding to lentiginous and nested junctional patterns, respectively. Globular nevi presented large junctional clusters, whereas cobblestone nevi were constituted by dermal dense melanocytic clusters. Homogeneous nevi did not show distinctive confocal and histopathologic findings. Nevi with a rim of globules presented a meshwork pattern with junctional clusters at the periphery. At the confocal follow-up study all lesions showed limited dynamic changes resulting in stable dermatoscopic and confocal patterns, but 3 globular nevi with junctional nests at baseline evolved into reticular-meshwork pattern nevi with peripheral rim of globules-junctional nests. LIMITATIONS Longer confocal follow-up of more melanocytic nevi is required to confirm this theory and to validate our preliminary findings. CONCLUSIONS A model explaining the nevus classification and patterns of evolution of nevi observed in the study was proposed.

Mismatch repair proteins expression and microsatellite instability in skin lesions with sebaceous differentiation : A study in different clinical subgroups with and without extracutaneous cancer

Muir-Torre syndrome (MTS) is defined as the association of a sebaceous tumor or keratoacanthoma and an extracutaneous neoplasm, mainly from the gastrointestinal or genitourinary tracts. MTS is related to hereditary non-polyposis colorectal cancer (HNPCC), a syndrome with germline mutations in the mismatch repair (MMR) gene(s), leading to microsatellite instability (MSI). In this study, using immunohistochemistry and a microsatellite instability assay, we analyzed the incidence of MMR gene abnormalities in 79 sebaceous lesions from 70 patients, 26 of whom also had an extracutaneous visceral neoplasm. We were unable to investigate the family histories of our patients regarding other tumors in order to assess which of our cases met the Amsterdam criteria. Defective MMR protein expression (MMR-) was found in 18/70 (25.7%) patients, with an identical distribution between those having an isolated skin tumor (11/44, 25.0%) and those with an extracutaneous cancer (7/26, 25.4%). In the sporadic group, MMR negative lesions were significantly more frequent in extrafacial areas (P = 0.03). High concordance was found between MMR expression in sebaceous lesions and the extracutaneous neoplasm in the same patient (20/23, 86.9%), as well as between MMR expression and microsatellite status (18/20, 90%). In conclusion, this study confirms the value of immunohistochemistry to identify MMR defective tumors. However, since only a minority of sebaceous neoplasms in patients who also have an extracutaneous cancer display MMR defects, these techniques are of limited value for the identification of “clinically defined” MTS.

In vivo confocal microscopy for detection and grading of dysplastic nevi: A pilot study

BACKGROUND Dysplastic nevi are thought to be precursors of melanoma during a stepwise process. However, this concept is still controversial and precise correlation between clinical and histopathologic features is lacking. In vivo confocal microscopy represents a noninvasive imaging technique producing horizontal sections at nearly histopathologic resolution. OBJECTIVE We sought to determine whether specific histologic features in dysplastic nevi have reliable correlates on confocal microscopy and to develop an in vivo microscopic grading system. METHODS Sixty melanocytic lesions with equivocal dermatoscopic aspects, corresponding to 19 nondysplastic nevi, 27 dysplastic nevi, and 14 melanomas, were analyzed by confocal microscopy and histopathology, using the Duke grading criteria. RESULTS All architectural and cytologic features of the Duke grading score had significant reflectance confocal microscopy correlates. Confocally, dysplastic nevi were characterized by a ringed pattern, in association with a meshwork pattern in a large proportion of cases, along with atypical junctional cells in the center of the lesion, and irregular junctional nests with short interconnections. A simplified algorithm was developed to distinguish dysplastic nevi from melanoma and nondysplastic nevi. The contemporary presence of cytologic atypia and of atypical junctional nests (irregular, with short interconnections, and/or with nonhomogeneous cellularity) was suggestive of histologic dysplasia, whereas a widespread pagetoid infiltration, widespread cytologic atypia at the junction, and nonedged papillae suggested melanoma diagnosis. LIMITATIONS A small number of cases were evaluated because of the necessity to analyze numerous histopathologic and confocal features. CONCLUSION The possibility to detect dysplastic nevi in vivo may lead to an appropriate management decision.

Atypical Spitzoid melanocytic tumors: A morphological, mutational, and FISH analysis

BACKGROUND Identification of the clinical behavior of atypical Spitzoid tumors with conflicting histopathologic features remains controversial. OBJECTIVE We sought to assess whether molecular findings may be helpful in the diagnostic and prognostic assessment of atypical Spitzoid tumors. METHODS A total of 38 controversial, atypical Spitzoid lesions (≥ 1 mm in thickness) were analyzed for clinicopathological features, chromosomal alterations by fluorescence in situ hybridization (FISH) analysis (RREB1/MYB/CCND1/CEP6), BRAF(V600E) mutation by allele-specific real-time polymerase chain reaction confirmed by sequencing, and H-RAS gene mutation by direct sequencing. RESULTS Atypical Spitzoid lesions developed in 21 female and 17 male patients (mean age 22 years). Nine patients underwent sentinel lymph node biopsy and a sentinel lymph node micrometastasis was detected in 4 of these 9 cases. Four additional patients, who did not receive a sentinel lymph node biopsy, experienced bulky lymph node metastases and one experienced visceral metastases and death. Lesions from patients with lymph node involvement showed more deep mitoses (P < .01), less inflammation (P = .05), and more plasma cells (P = .04). FISH analysis demonstrated the presence of chromosomal alterations in 6 of 25 cases. Correlation with follow-up data showed that the only case with fatal outcome showed multiple chromosomal alterations by FISH analysis. BRAF(V600E) mutation was detected in 12 of 16 cases (75%) and H-RAS mutation on exon 3 was found in 3 of 11 cases (27%). LIMITATIONS Our results require validation in a larger series with longer follow-up information. CONCLUSIONS FISH assay may be of help in the prognostic evaluation of atypical Spitzoid tumors. Diagnostic significance of BRAF(V600E) and H-RAS mutations in this setting remains unclear.