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DNA-cytophotometry of benign compound and intradermal naevi, Spitz epithelioid naevi and malignant melanomas.

Single cell DNA cytophotometry was used to characterize seven compound, ten intradermal and six Spitz naevi as well as 23 primary cutaneous malignant melanomas. Compound and intradermal naevi were characterized by a smaller nuclear area than both Spitz naevi and malignant melanomas. Tumour ploidy could not be used as a single criterion of malignancy since both diploid and hyperdiploid melanomas were encountered. The very low mean optical density of Spitz naevi served to distinguish these lesions from malignant melanomas.

Adolescent↗

Cutaneous malignant melanoma: correlation between neovascularization and peritumor accumulation of mast cells overexpressing vascular endothelial growth factor.

To investigate the possible role of mast cells (MC) in the angiogenic process in cutaneous melanoma, we examined tissue samples from 35 adult patients with primary malignant melanoma and compared with 20 intradermal benign nevi. MC were identified by anti-tryptase, microvessels by anti-CD34, and vascular endothelial growth factor (VEGF) expression by standard immunohistochemical methods. Tryptase-positive MC expressing VEGF were identified by double immunostaining. The numbers of MC and microvessels around the tumor were determined by the point counting method. MC density was significantly greater in melanoma compared with benign nevi (197.6 +/- 19.4 v 95.7 +/- 5.0/mm2, P < .001). Vascular density was also significantly higher in melanoma than in benign lesions (3.6-fold, P < .001). Double immunostaining showed the presence of VEGF in the cytoplasm of tryptase-positive peritumoral MC. The percentage of this MC-subtype was significantly higher in melanoma than in nevus tissues (71.9 +/- 2.4% v 30.6 +/- 2.5%, P < .001). A strong significant correlation was shown between the number of VEGF+ MC and microvessel density (r = .811, P < .001). MC count and VEGF+ MC count, as well as microvessel density were significantly higher in aggressive (metastasizing) melanomas (P < .001). Our results suggest that peritumoral accumulation of MC and the subsequent release of potent angiogenic factor such as VEGF may thus represent a tumor-host interaction that may favor progression of this tumor.

Adolescent↗

Histochemical determinations of copper, zinc, and iron in pigmented nevi and melanoma.

Histochemical determinations of copper, zinc, and iron in intradermal pigmented nevi and melanomas revealed the presence of copper and iron in melanoma but not in nevi. Zinc was not detected in either melanomas or nevi. However, melanin was removed from the tissues prior to staining; therefore, it is possible that zinc was also removed by the procedure. Although the function of copper and iron in the melanoma cell is not known, they may be components of abnormal enzymes.

Copper↗

Nucleolar organizer regions in pigmented skin lesions. Value in the differential diagnosis of Spitz nevi.

Forty-one cases of typical melanocytic skin lesions (15 intradermal nevi, 14 Spitz nevi and 12 malignant melanomas) were used to investigate the value of staining of nucleolar organizer regions (NORs) in the differential diagnosis of such pigmented lesions. Histologic sections were stained by the silver colloid (Ag) method, with and without the prior use of a melanin blocking agent. There were statistically significant differences in the mean numbers of AgNORs per nucleus between the groups of lesions studied (1.658 for intradermal nevi, 3.0042 for Spitz nevi and 6.669 for malignant melanomas). Sections treated with potassium permanganate (melanin blocking agent) prior to staining showed an obvious increase in the AgNOR scores in all groups; this increase was highest for Spitz nevi. Although AgNOR staining allows a distinction to be made between intradermal nevi and malignant melanomas, the striking overlap between the counts for Spitz nevi and malignant melanomas precludes the use of this technique as the sole method for establishing the diagnosis of malignancy. Other clinical and morphologic data are especially required to make the diagnosis of Spitz nevi.

Analysis of Variance↗

Comparative distribution of S-100 protein and antigen HMB-45 in various types of melanomas and naevi.

The content of S-100 protein and antigen HMB-45 in various types of melanomas and skin naevi was compared with respect to the frequency of two antigens, their distribution within lesions as well as diagnostic or prognostic relevance. The study material comprised 72 skin melanomas, 25 melanomas of the oral cavity and 63 non-malignant skin naevi. Formalin fixed, paraffin-embedded sections were used for immunohistochemical reactions with polyclonal rabbit anti-S-100 antibody (Dako) and monoclonal anti-HMB-45 antibody (Dianova) followed by PAP and APAAP Kits (Dianova), respectively. Detection of HMB-45 was preceded by a microwave treatment of sections. S-100 protein was found in all cases of melanomas irrespective of their location or histological type, but HMB-45 was missing in 8% of oral melanomas. Distribution of the antigens within tumors was heterogeneous and often mutually exclusive. S-100 protein was also present in all types of naevi, while HMB-45 was absent in intradermal naevi and strictly confined to epidermal component in the remaining ones. The intensity of staining for S-100 correlated inversely with the patients' survival. It was concluded that demonstration of HMB-45 might facilitate a differentiation of naevi from melanomas, whereas staining for S-100 protein might have some prognostic significance.

Adult↗

Nevoid malignant melanoma: morphologic patterns and immunohistochemical reactivity.

The term "nevoid malignant melanoma" (nevoid MM) is used here to describe rare nodular malignant melanomas that may escape detection in routine histological sections due to the lack of a prominent intraepidermal component, sharp lateral circumscription and evidence of partial maturation with descent in the dermis. Nevoid MM mimic ordinary compound or intradermal melanocytic nevi when the melanoma cells are small, or Spitz's nevi when the cells are large. The patterns of HMB-45 staining in 12 nevoid MM were compared with those in 107 melanocytic nevi. HMB-45 staining was strong in the dermal component of the nevoid MM, even in the absence of a junctional component. In common acquired and congenital nevi, the upper dermal component stained less than the junctional component of the lesion. The deepest components of these nevi were negative. Spitz nevi and cellular blue nevi had positive dermal cells, even without a junctional component. Additional staining for a proliferation marker, such as cyclin (PCNA) or Ki-67 (with the antibody MIB-1), can help further in distinguishing a nevoid MM from a Spitz's nevus. Melanoma has strong nuclear staining throughout the lesion. In contrast, Spitz's nevi have more staining at the top of the lesion than at the bottom. The patterns of HMB-45 and MIB-1 staining can be used along with standard histologic criteria for the diagnosis of nevoid MM. Clinicopathologic correlation is needed to distinguish some metastatic melanomas from primary nevoid MM.

Adult↗

HMB-45 staining of dysplastic melanocytic nevi in melanoma risk groups.

To evaluate the hypothesis that reactivity of the intradermal component of melanocytic nevi to the monoclonal antibody HMB-45 correlates with melanoma risk, dysplastic compound melanocytic nevi were examined for expression of the HMB-45 epitope in three subject groups differing in epidemiological risk for melanoma. The study groups consisted of 10 subjects with dysplastic nevi and a personal history of melanoma, 25 subjects with dysplastic nevi and a history of melanoma in one or more first degree relatives, and 15 population control subjects with sporadic dysplastic nevi. For each case, sections from one lesion, immunohistochemically processed for HMB-45 binding, were evaluated by two pathologists without knowledge of the clinical data. Of all dysplastic nevi, 98% showed diffusely positive cytoplasmic staining of the junctional nevomelanocytes and 90% had such positive staining of those cells within the superficial dermis. Nevus cells within the deeper dermis did not stain positively in any case. Furthermore, the data showed no differences in frequency, pattern, or intensity of HMB-45 reactivity between the subject groups. These observations indicate that evaluation of dysplastic nevi with the monoclonal antibody HMB-45, an apparent marker of proliferative or otherwise stimulated melanocytes, has no discriminating value for identifying subjects at increased historical risk for melanoma. The data, however, support the concept that so-called dysplasia within nevi, as defined by histologic criteria, actually represents the active or proliferative phase of melanocytic nevi.

Antibodies, Monoclonal↗

Cyclin D1 expression in melanocytic lesions of the skin.

BACKGROUND: Progression through the cell cycle is controlled by cyclins, cyclin-dependent kinases, and cyclin-dependent kinase inhibitory proteins. The role of cyclin D1 in the development and progression of melanomas is controversial. The goal of this study is to evaluate the role of cyclin D1 in benign and malignant melanocytic lesions of the skin. METHODS: A total of 126 pigmented lesions of the skin including compound nevi (21), intradermal nevi (18), melanoma in situ (28), primary invasive melanomas (30), and metastatic melanoma (29) were evaluated for cyclin D1 expression. The following tiered system was used for scoring: 0% nuclear staining (score 0), 1% to 19% nuclear staining (score 1), 20% to 49% nuclear staining (score 2), and 50% or greater nuclear staining (score 3). RESULTS: Average scores were significantly higher for primary melanomas compared with nevi and for in situ melanomas compared with primary invasive melanomas. The average score for metastatic melanomas was not significantly different compared with primary invasive melanomas. Scores for primary invasive melanomas did not correlate with depth of invasion or presence of metastases. Compound nevi exhibited a slightly higher level of cyclin D1 expression compared with intradermal nevi. CONCLUSION: Although primary melanomas show a higher level of cyclin D1 expression compared with nevi, cyclin D1 appears to have little role in development of a metastatic phenotype. It is not clear why lesions localized near the dermal-epidermal junction express higher levels of cyclin D1. Further studies are indicated to ascertain the biologic role and practical utility of cyclin D1 in melanocytic lesions of the skin.

Biomarkers, Tumor↗

Can HLA-DR expression help in the routine diagnosis of malignant melanomas?

Two monoclonal antibodies, TAL-IB5 and LN3, directed at epitopes of HLA-DR were applied to a variety of benign naevi and malignant melanomas. Of the 31 naevi examined, 28 were completely unreactive. Weak staining was confined to a few cells in two intradermal naevi and there was more widespread reactivity in a halo naevus. Seven of the 25 primary cutaneous malignant melanomas were reactive with one or both of the antibodies and this reactivity was more marked in the deeper lesions. These positive cases were not felt to present diagnostic difficulties and it was concluded that TAL-IB5 and LN3 were not of use in distinguishing between benign and malignant melanocytic proliferations.

Diagnosis, Differential↗

Patterns of staining for neurone specific enolase in benign and malignant melanocytic lesions of the skin.

Neurone specific enolase (NSE) is a useful marker for melanoma independent of the criterion of melanogenesis. A range of benign and malignant melanocytic skin lesions has been studied to assess the diagnostic potential of staining for NSE. The detailed features of the presence and distribution of NSE in 47 skin lesions are presented. These include 3 junctional, 7 compound, 15 intradermal and 8 SPitz naevi; 6 superficial type, 2 lentigo maligna and 6 nodular melanomata. A progressive increase of staining for NSE from benign to malignant is seen. Spitz naevi could easily be differentiated from melanoma. An advantage of this stain is the simultaneous demonstration of the innervation of these lesions. We conclude that NSE is a useful aid to the diagnosis and assessment of melanocytic skin tumours and may prove of value in clarifying the nature of these enigmatic lesions.

Humans↗

[Immunohistochemical study of nevocellular nevi].

Immunohistochemical analyses were performed on 64 nevocellular nevi (12 compound nevi and 52 intradermal nevi). S-100 protein and its alpha- and beta-subunits were almost always demonstrated in type A, B and C cells, and the staining intensity tended to increase in the type C cells. Neuron-specific enolase was detected in each type of cell; however, the population of positive cells was smaller among type C cells. Beta 2-microglobulin was occasionally demonstrated, but only in type A cells. Vimentin was frequently revealed in every type of cell. Neither myelin basic protein nor glial fibrillary acidic protein was observed in any type of cell. In contrast, normal epidermal melanocytes were positive for vimentin, but negative for S-100 protein and its subunits and neuron-specific enolase. Schwann cells were positive for S-100 protein and its beta-subunit, but negative for the alpha-subunit. Thus, the nevus cells shared a common nature with epidermal melanocytes and Schwann cells which originate from the neural crest; however, the former cells were somewhat different from the latter two kinds and from benign and malignant tumors derived from these cells in the expression of these antigenic substances. Such differences in the expression of antigenic substances may be due to dysontogenic manifestations in nevus cells.

Humans↗

Histopathologic evaluation of melanocytic nervi in oculocutaneous albinism.

This is a report of the histopathologic findings of melanocytic nevi in patients with oculocutaneous albinism. Seven albino patients, four tyrosinase positive and three tyrosinase negative, with a total of 34 melanocytic lesions were available for examination. The specimens were examined histopathologically for the presence of intranuclear pseudoinclusions, giant melanocytic cells, eosinophilic globules, features of congenital nevi, and atypical changes. A statistically significant increased number of giant melanocytic cells were found in intradermal nevi (p less than 0.0025), compound nevi (p less than 0.01), and junctional nevi of the patients with albinism compared with controls. Eosinophilic globules were found in 17% of the 34 nevi, and congenital features in 8.8%. All lesions showed adequate maturation of cells with no atypical changes. Marked solar elastosis was found in the dermis around and beneath the nevi, even though most lesions were in covered areas.

Adult↗

Malignant melanoma originating from intradermal nevi. A clinicopathologic entity.

Two cases of malignant melanoma arising in preexisting intradermal nevi without a junctional element are described. These cases and the two known to have been reported previously demonstrate similar clinical and morphologic findings. These include an innocuous gross appearance, a nodular growth pattern, and deep levels of invasion. It appears that the origin of the tumor in the dermis conceals malignant transformation. The consequent time lapse and the dermal position of the lesion favor deep penetration of the tumor prior to removal.

Adult↗

Proliferative activity of cutaneous melanocytic neoplasms defined by a proliferating cell nuclear antigen labelling index.

To evaluate the proliferative activity of benign, borderline and malignant cutaneous melanocytic neoplasms, 30 cases of malignant melanoma (MM) and 41 cases of naevi were studied by immunostaining using a monoclonal antibody against proliferating cell nuclear antigen (PCNA). PCNA is a nuclear antigen expressed in the late G1 and S phase and serves as a marker of proliferating cells. Invasive MM and MM in situ showed much higher PCNA positivity rates than melanocytic naevi (invasive MM, 18.0%; MM in situ, 11.3%; ordinary melanocytic naevi, 2.6%). The PCNA positivity rate did not increase significantly with the thickness of MM. Among ordinary melanocytic naevi, junctional naevi had a higher PCNA positivity rate than compound or intradermal naevi. Mean PCNA positivity rates for Spitz's naevi and sporadic dysplastic naevi were within the range for ordinary melanocytic naevi, indicating the benign nature of both types of naevus. Contrary to some previous studies, MM in situ showed high proliferative activity, indicating that cells of MM in situ are actively proliferating. This study clearly demonstrates that MM and various types of naevi can be separated according to differences in proliferative activity defined by the PCNA labeling index.

Antibodies, Monoclonal↗

Giant congenital pigmented naevus with unusual presentation and early malignant transformation in a Nigerian infant: case report and review of the literature.

A six month-old female infant with a giant congenital pigmented naevus is reported because of the rarity and unusual presentation and early malignant transformation. It was the first case to be documented in our centre. There was an extensive giant, darkly pigmented naevus of the "bathing trunk" distribution noted at birth, associated with multiple darkly pigmented smaller naevi scattered over the trunk and limbs. Bv the third month of life large pigmented lesion began to demonstrate small papillomatous projections studding the entire surface and at age six months nodular and warty excrescences appeared including a large pedunculated spherical mass attached to the anal verge. The labia majora became anatomically disfigured by these nodular, warty lesions of various sizes associated with the large pigmented hairy naevus. Histological examination of the excised large perianal and vulvar nodules showed the typical picture of an intradermal naevus consistent with giant congenital pigmented (melanocytic) naevus. The deeper portions of the lesion showed features of malignant transformation. The rapidity of evolution of the lesions including histologic features of early malignant transformation and the gross disfiguring of the genitalia and perineum are uncommon presentation of this melanocytic naevus. In view of the early malignant transformation immediate surgical treatment was indicated. Unfortunately the patient was lost to follow-up. The case report is complimented with a review of the literature.

Cell Transformation, Neoplastic↗

[Clinical and histopathological observations of pigmented spots on the feet].

Malignant melanomas, originating in the foot are said to have a higher frequency in Japan compared to Western countries, accounting for almost one third of all cutaneous malignant melanomas. In this context, pigmented spots on the feet which can be thought of as precursor lesions of malignant melanomas attract a great deal of attention from clinical dermatologists. In this study, 159 pigmented spots, consisting of 110 on the soles as well as 49 on the dorsal lesion have been studied clinically and histopathologically to clarify their relationship with the development of malignant melanomas. The results were as follows: 1) Junction nevi were most commonly observed at both sites. 2) Lesions tended to be darker in color and bigger in size as they proceeded from the simple lentigo to the junctional and compound nevi. 3) Most of the lesions were flat, but the elevated lesions were seen more frequently in the compound nevi and intradermal nevi which contained more dermal components. Also, lesions on the soles appeared to be more elevated. 4) Among the 21 cases which showed some cutaneous change, one case showed histopathological evidence of a premalignant lesion. 5) In the pigmented spots on the feet, the initial signs of premalignant transformation were skin elevation, more pronounced spot growth, measuring up to 5 mm in diameter, and dark black coloring (pigmentation). Therefore preventive excision of the lesion is recommended in cases where any of the three above features are in evidence.

Adolescent↗

Doppler ultrasound as an adjunct to the differential diagnosis of pigmented skin lesions.

One hundred and forty-one pigmented skin tumours were examined with a 10 MHz ultrasound Doppler flowmeter to investigate the value of detecting blood flow in the diagnosis of raised pigmented skin lesions. Most of the benign lesions except for those with an angiomatous basis were devoid of blood flow signals while all basal cell carcinomas and 96 per cent of thick melanomas (greater than or equal to 0.9 mm) were associated with detectable Doppler frequency shift signals. While the characteristics of analysed Doppler waveforms are of research interest, the simple detection of blood flow by a hand held instrument gives sufficient information for clinical purposes. In practice this simple test has been sufficiently reliable to prove a useful adjunct in the diagnosis of raised pigmented skin lesions. It has been found particularly helpful in the common clinical problem of differentiation of nodular melanoma from basal cell papilloma and benign intradermal naevus. If such a lesion is thought to be benign, and has detectable flow signals, one should reconsider the diagnosis. Absence of flow signals is strong confirmation of a benign clinical diagnosis. The test should be regarded as an adjunct to clinical diagnosis since, in common with all noninvasive diagnostic techniques, false negative and false positive cases will be encountered. The technique is not appropriate to macular lesions since these are usually flow negative, irrespective of the pathology.

Adolescent↗

[The feasibility of the razor-blade shave technique in the diagnosis and treatment of benign and malignant skin lesions on the face].

The razor-blade shave technique uses a slightly-curved classic razor blade to shave and remove the exophytic part of a skin lesion and part of the intradermal structure down to the stratum papillare. It can then be sent for pathology investigations. This treatment is curative for many benign skin conditions. The cosmetic result ofthe razor-blade technique is superior to that of surgical excision. The razor-blade technique is useful as a biopsy instrument in the diagnosis of skin rumours of unknown nature, especially in keratoacanthomas. The technique is very easy and cheap. However, if the skin lesion is a suspected malignant melanoma, traditional surgical excision should be performed. The shave technique is not suitable for malignant skin tumours.

Biopsy↗