Flagellate dermatitis after consumption of Shiitake mushrooms.

Flagellate dermatitis occurs in patients who have eaten Shiitake mushrooms. We are reporting on a 55-year-old man, who developed whiplash-striped, severely itching efflorescences on the trunk 3 days after eating Lentinula edodes. Flagellate dermatitis is also known as a cutaneous side effect of bleomycin therapy.

RASSF10 promoter hypermethylation is frequent in malignant melanoma of the skin but uncommon in nevus cell nevi.

The Ras association domain family (RASSF) consists of several tumor suppressor genes, which are frequently silenced in human cancers. We analyzed the epigenetic inactivation of RASSF2 and RASSF10 in malignant melanoma (MM) of the skin, including 5 MM cell lines, 28 primary MM, 33 metastases of MM, 47 nevus cell nevi (NCN), and 22 control tissues. The RASSF2 promoter was epigenetically downregulated in two MM cell lines only, but not in any of the investigated tumor samples. In contrast, hypermethylation of the RASSF10 promoter was found in all investigated cell lines, 19/28 (68%) of the primary MM and 30/33 (91%) of the MM metastases, 2/18 (11%) of the dysplastic NCN, and 0/29 (0%) of the non-dysplastic NCN (difference between MM and all nevi, P<0.001). RASSF10 promoter hypermethylation correlated with a reduced RASSF10 mRNA expression in 3/4 MM cell lines, and treatment with a DNA methylation inhibitor reactivated RASSF10 transcription. Furthermore, immunohistological RASSF10 expression corresponds negatively to its promoter methylation state. In summary, RASSF10 proved to be a characteristically epigenetically silenced tumor suppressor in melanomagenesis, and analysis of RASSF10 methylation status represents a new candidate tool to assist in discrimination between MM and NCN.

B7-h4 expression in human melanoma: its association with patients' survival and antitumor immune response.

PURPOSE: Cancers have developed a number of strategies to escape immune responses including the differential expression of costimulatory molecules of the B7 family. B7-H3 and B7-H4 have recently been described in different tumor entities but the relevance for melanoma has not yet been studied so far. EXPERIMENTAL DESIGN: Using immunohistochemistry, B7-H3 and B7-H4 expression was studied on 29 melanoma lesions. Survival curves and log-rank tests were used to test the association of protein expression with survival. Cell lines were evaluated for B7-H3 and B7-H4 expression by PCR and flow cytometry. Functional T-cell-tumor coculture assays were carried out with in vitro generated tumor transfectants. RESULTS: B7-H3 and B7-H4 expression was detected in primary tumor lesions (29 of 29 and 28 of 29) and in metastases (28 of 29 and 26 of 29). The numbers of CD68(+) macrophages were significantly lower in patients with low B7-H4 expression, whereas CD8(+) T-cell infiltrates were independent of expression levels. Furthermore, a survival benefit for patients with B7-H4 low expressing melanoma was found, whereas B7-H3 was not associated with any clinical parameter. All 23 melanoma cell lines analyzed expressed B7-H3 and B7-H4 mRNA and protein, but B7-H4 was restricted to intracellular compartments. On silencing of B7-H3 by specific shRNA tumor-associated antigen-specific T cell responses were unaltered. Overexpression of B7-H4 on melanoma cells did not alter the cytotoxicity of different CD8(+) effector cells, but drastically inhibited cytokine production. CONCLUSIONS: Our study provides for the first time evidence of B7-H4 expression on melanoma cells as a mechanism controlling tumor immunity which is associated with patients' survival.

Frequent occurrence of RASSF1A promoter hypermethylation and Merkel cell polyomavirus in Merkel cell carcinoma.

Merkel cell carcinoma (MCC) is one of the most aggressive cancers of the skin. It has recently been reported that integration of a Merkel cell polyomavirus (MCPyV) in receptor tyrosine phosphates type G (PTPRG) gene occurs in MCC, and that viral infections are associated with epigenetic silencing of tumor suppressor genes (TSG) in cancer. To examine whether a correlation between TSG inactivation and viral infection can be found in MCC, we investigated the promoter hypermethylation of RASSF1A, TP73, PTPRG, FHIT, and CDKN2A and the presence of MCPyV and SV40 in 98 MCC by PCR. Hypermethylation of RASSF1A was frequently found in 42 of 83 (51%) of MCC. Methylation of CDKN2A was present in 9 of 41 (22%) of MCC. Hypermethylation of TP73 (0%), PTPRG (4%), and FHIT (0%) was infrequent in MCC. Interestingly, MCPyV was found in 90 of 98 (92%) MCC, however, no SV40 signal was detected. No correlation between TSG hypermethylation and viral infection was found. Our results show frequent hypermethylation of RASSF1A and the presence of MCPyV in primary MCC, and that these events may contribute to the pathogenesis of MCC.

Radiotherapy in multilocalized lymphedema-associated angiosarcoma.

We report an 80-year-old woman, suffering from a recurrence of a multilocalized lymphedema-associated angiosarcoma of the right arm. The tumor consisted of solid tumor cell formations and "classical" spongiform tumor complexes. In the tumor periphery, pathological endothelial cell proliferates on pre-existing dilated lymphatic capillaries were detectable, which, together with immunohistology (CD 31+/Desmoplakin-1-2.17+/CD 34-), supported the diagnosis of lymphangiosarcoma. Complete remission was achieved under radioimmunotherapy (54 Gy/Interferon beta). A further recurrence 3 months later outside the primary therapy fields was successfully treated with radiotherapy alone. During a follow-up observation period of 3 years, there was neither local recurrence nor metastasis. This case demonstrates for the first time the long-lasting efficacy of photon radiation in a case of histologically-defined lymphangiosarcoma. Further studies should elucidate the suitability of radio monotherapy as first-line therapy in lymphedema-associated angiosarcoma with lymphatic endothelium-like immunohistology.

Paratumoral epidermal hyperplasia: a novel prognostic factor in thick primary melanoma of the skin?

There is a lack of histopathological factors to sub-stratify prognosis in pT3/4 melanoma primaries. In the presented pilot study, the prognostic significance of different clinical and histopathological parameters was studied in thick primary melanoma taking paratumoral epidermal hyperplasia (PTEH) into consideration. Of 1632 melanoma patients in the melanoma register of the Martin Luther University Halle-Wittenberg in the years 1980 to 1987, 16 cases with tumor thickness (TT) of the primary > or = 3 mm, documented metastasis-free follow-up of 10+ years after primary therapy and available histologic sections were compared with an adequate recurrence control group (n = 62) by PTEH and standard prognostic parameters. PTEH was demonstrable in 15 of 16 patients of the metastasis-free group (PTEH penetration depth 1.42 +/- 0.82 mm/mean +/- SD) and 27 of 62 of controls (0.29 +/- 0.46 mm), P < or = 0.001. Of the standard prognostic parameters, TT, sex, location, and lack of nevus association also correlated with metastasis. In multivariate analysis, PTEH > or = 1 mm was the single independent parameter with the highest (negative) association to recurrence (odds ratio 52.3). Occurrence of PTEH might predict a more moderate course of disease in thick melanoma. Thus, it might become an easily determinable and effective tool to sub-stratify prognosis in thick primary melanoma of the skin. Further studies are necessary to prove these findings.

Skin function parameters in intensive-care patients.

BACKGROUND/PURPOSE: The study was performed to investigate the transepidermal water loss (TEWL) and pH-value in patients in intensive care. METHODS: Forty intensive-care patients (22 men, 18 women) were included in the study. TEWL and pH-values were measured at admission, and after 24, 96 and 168 h. The areas of measurement were the forehead, the volar forearm, paraumbilical and the ventral thigh. The measurements were made under standardized environmental conditions according to the recommendations of the EMCO Group. RESULTS: Elevated values were found on the forehead compared with the other skin areas examined. There was no significant change in mean TEWL-values in any patient in the course of the study. There was also no significant influence of TEWL at the time of admission on the prognosis. The course analysis of the mean pH-values, however, showed that patients who developed a systemic inflammatory response syndrome (SIRS) or sepsis during the further course had a higher pH-value over the entire study period. CONCLUSION: TEWL and the pH of the skin surface could be measured at bedside in the intensive-care unit and delivered reproducible results. These parameters appear, however, to be relevant only for subgroups of patients under intensive care.

Intratumoral DNA stem-line heterogeneity in superficial spreading melanoma.

BACKGROUND: In primary melanomas, data on the degree of intratumoral heterogeneity to date have been lacking. OBJECTIVE: Our purpose was to investigate intratumoral DNA stem-line heterogeneity in superficial spreading melanoma (SSM). METHODS: Multiple measuring fields of 54 SSMs (tumor thickness median 1.60 mm) were studied by DNA image cytometry to obtain data on the number of DNA stem lines per tumor, their ploidy characteristics, and intratumoral distribution. Results were compared with standard histopathological criteria. RESULTS: Twenty-three of 54 SSMs were found to have two or three distinct proliferating tumor cell stem lines (1.46 +/- 0.57 per tumor). Stem lines appeared spatially separated in 22 of 23 SMMs. At least 3 measuring fields per tumor were necessary to identify all stem lines with a likelihood of 95%. DNA heterogeneity correlated with tumor thickness, but occurred in 5 of 19 cases of pT1 melanoma. CONCLUSIONS: Primary SSMs can be regarded as potentially clonally unstable with a tendency for spatial separation of tumor cell stem lines.

Expression of epidermal N-methyl-D-aspartate receptors (NMDAR1) depends on formation of the granular layer--analysis in diseases with parakeratotic cornification.

Ionotrope glutamate receptors of the N-methyl-D-aspartate (NMDA) receptor type are expressed on keratinocytes and influence the intracellular calcium concentration. The importance of NMDA receptors in pathophysiological processes in the skin is, however, still unclear. Epidermal distribution patterns of NMDA receptors were investigated in dermatoses with parakeratotic cornification (psoriasis vulgaris and verrucae vulgares) and compared to the expression of filaggrin. The expression of NMDA receptors (R1 component) in paraffin-embedded normal epidermis (n = 22), psoriasis vulgaris (n = 21) and verrucae vulgares (n = 23) was examined and evaluated by means of digital image analysis. For quantitative characterization of the distribution patterns, a quotient was formed of the expression in the stratum granulosum and stratum basale ("NMDA ratio"). The distribution of NMDAR1 was compared to the immunohistochemical expression of filaggrin. Additionally the expression of filaggrin was investigated in HaCaT cells after treatment with the NMDA receptor antagonist MK-801. NMDA receptors were demonstrated in the epidermis of all preparations. In healthy skin, the highest receptor density was found in the stratum granulosum. This distribution pattern was basically also present in the dermatoses examined. Thus, the occurrence of parakeratosis in psoriasis vulgaris, but not in verrucae vulgares, was characterized by a significant reduction in the NMDA ratio (reduced expression of NMDAR1 in the upper epidermis). The immunohistochemical distribution of filaggrin was similar to that of NMDAR1. In HaCaT cells MK-801 suppressed the expression of filaggrin. NMDA receptors are expressed in human epidermis under physiological conditions especially in the stratum granulosum. Their reduced expression within parakeratotic epidermis in psoriasis vulgaris may be evidence of impaired intracellular calcium influx in this disease.

N-methyl-D-aspartate receptors influence the intracellular calcium concentration of keratinocytes.

In the present study, the distribution of ionotropic glutamate receptors of the N-methyl-D-aspartate (NMDA)-receptor type was immunohistochemically demonstrated in healthy human skin (n = 22) and healthy buccal mucosa (n = 20). Moreover, the intracellular calcium concentration of HaCaT-cells and native human keratinocytes were studied under the influence of the selective agonist NMDA and the selective NMDA-antagonist MK-801. Immunohistochemical imaging of NMDA receptors in healthy epidermis showed a positive reaction in the stratum basale, spinosum and granulosum, whereby the greatest expression was observed in the granular layer. In the mucosal preparations, the distribution of NMDA receptors was observed to be equal in all cell layers. In the cell culture (HaCaT-cells), NMDA concentrations between 25 microM and 1 mM resulted in a significant increase in the number of cells showing elevated intracellular calcium concentration. This effect could be significantly reduced by prior application of MK-801 (100 micro M). In supplementary tests on HaCaT-keratinocytes, blockade of the keratinocytic NMDA receptors with MK-801 suppressed the differentiation of the cells (expression of cytokeratin 10). The proliferation of cells was not influenced by NMDA. The investigations showed that glutamate receptors of the NMDA type have an influence on keratinocytic calcium concentration. This appears especially important for the differentiation of keratinocytes.

Hyperplasia of dermal microvascular pericytes in scleroderma.

BACKGROUND: Pericytes (PCs) are smooth muscle-like mural cells of capillaries and venules, which can synthesize matrix components and fibroblast-activating cytokines, and are thus potential mediators of pathological changes in scleroderma. In this study, alterations in microvessels were quantitatively imaged, taking PC into account for the first time. METHODS: Skin biopsies from systemic (12) and localized (14) scleroderma forms as well as age-, sex-, and body location-matched controls were examined with respect to capillary and venular densities as well as endothelial cell (EC) and PC counts using a newly developed (in respect of PC and EC) indirect collagen IV immunostaining-based method. RESULTS: Hyperplasia of the PC that doubled the microvascular PC density was the most conspicuous characteristic. In the capillaries of the upper dermal plexus of the periphery of the sclerotic zones, median ratios of PC : EC were 0.23 (controls 0.10) or 0.18 (controls 0.11) in systemic or localized scleroderma, respectively. Furthermore, an increase in capillary density in the upper dermal plexus could be demonstrated in the marginal zones of both types of disease. CONCLUSIONS: The observed PC increase in the peripheral zones of active disease supports the hypothesis of a vascular pathogenesis of scleroderma and directs the focus to microvascular PC.