Peter Kaskel

Antiapoptotic bcl-2 and bcl-xL in advanced malignant melanoma

Abstract Apoptosis is an important cofactor in the pathogenesis of a plethora of malignancies. However, little is known about modulation of the expression of bcl gene family in melanocytic tumors. To determine the role of bcl-2, bcl-x and bax in melanocytic tumors we investigated the differential expression of these genes via RT-PCR in tissue samples from human ¶benign nevi, primary melanomas and melanoma metastases in comparison with normal skin. Bcl-2 was strongly expressed in 14/16 metastases (87.5%), whereas only 7/13 primary melanomas (53%), 7/15 nevi (46%) and 7/16 normal tissue samples (43%) showed expression of bcl-2 ( P < 0.05). There was a strong indication of a correlation between tumor thickness and bcl-2 expression in nodular malignant melanomas. Expression of bcl-x was found in 16/16 melanoma metastases (100%), 11/13 primary melanomas (84%), 12/15 nevi (80%) and 10/16 normal tissue samples (62%) ( P < 0.05). Bcl-xL expression increased from primary melanoma to melanoma metastases, whereas bcl-xS showed a decreasing expression level during melanoma progression. No differences in bax expression were seen between melanoma metastases, primary melanoma, nevi and normal tissue. Immunohistochemical investigations of another 53 tissue samples showed similar results. Our results strongly indicate that bcl-2 and bcl-xL gene expression increases with progression of malignant melanoma. Bcl-2 and bcl-xL expression could reflect an increased malignant potential caused by an inhibition of apoptosis and growth advantage for metastatic melanoma cells.

Extra c-myc oncogene copies in high risk cutaneous malignant melanoma and melanoma metastases

Amplification and overexpression of the c-myc gene have been associated with neoplastic transformation in a plethora of malignant tumours. We applied interphase fluorescence in situ hybridization (FISH) with a locus-specific probe for the c-myc gene (8q24) in combination with a corresponding chromosome 8 α-satellite probe to evaluate genetic alterations in 8 primary melanomas and 33 advanced melanomas and compared it to 12 melanocytic nevi, 7 safety margins and 2 cases of normal skin. Additionally, in metaphase spreads of 7 melanoma cell lines a whole chromosome 8 paint probe was used. We investigated the functionality of the c-myc gene by detecting c-myc RNA expression with RT-PCR and c-myc protein by immunohistochemistry. 4/8 primary melanomas and 11/33 melanoma metastases showed additional c-myc signals relative to the centromere of chromosome 8 copy number. None of the nevi, safety margins or normal skin samples demonstrated this gain. In 2/7 melanoma cell lines (C32 and WM 266–4) isochromosome 8q formation with a relative gain of c-myc copies and a loss of 8p was observed. The highest c-myc gene expression compared to GAPDH was found in melanoma metastases (17.5%). Nevi (6.6%) and primary melanomas (5.0%) expressed the c-myc gene on a lower level. 72.7% of the patients with c-myc extra copies had visceral melanoma metastases (UICC IV), patients without c-myc gain in 35.0% only. The collective with additional c-myc copies also expressed the gene on a significantly higher level. These results indicate that a c-myc gain in relation to the centromere 8 copy number might be associated with advanced cutaneous melanoma.

Mycophenolate mofetil: A new therapeutic option in the treatment of blistering autoimmune diseases

BACKGROUND Mycophenolate mofetil (MMF), an ester of mycophenolic acid (MPA), was approved by the Food and Drug Administration in 1995 and is currently primarily indicated for the prophylaxis of rejection in renal transplant patients. The drug seems also to be of value in the treatment of psoriasis and rheumatic arthritis. Recently there have been 6 reported cases of successful treatment of blistering autoimmune diseases with MMF in combination with high dose prednisone therapy. OBJECTIVE On the basis of these reports we administered this new treatment regimen to several patients with blistering autoimmune diseases. Besides using a combination of MMF and high-dose prednisone we wanted to evaluate whether MMF monotherapy is also effective in the treatment of blistering autoimmune diseases. METHODS We administered MMF to 5 patients who had severe pemphigus vulgaris or bullous pemphigoid. Two patients received MMF in combination with high-dose prednisone therapy and 3 patients received MMF monotherapy. To our knowledge, this is the first report of successful treatment of pemphigus vulgaris and bullous pemphigoid with MMF monotherapy. RESULTS All patients were completely free of symptoms within 8 to 11 weeks of therapy. Patients who had received MMF monotherapy responded as well to treatment as those who received a combination of MMF and high-dose prednisone. CONCLUSION Our experiences strongly suggest that MMF monotherapy may be effective for patients even with severe pemphigus vulgaris and bullous pemphigoid. In addition, MMF monotherapy, at least over the short term, offers the advantage of fewer side effects in comparison to immunosuppressive combination therapy and was well tolerated by our patients.

S-100 protein in peripheral blood : A marker for melanoma metastases. A prospective 2-center study of 570 patients with melanoma

BACKGROUND S-100 protein, commonly used in the immunohistochemical diagnosis of malignant melanoma and melanoma metastases, has recently been introduced as a tumor marker in peripheral blood. OBJECTIVE This prospective, observational, 2-center study evaluates S-100 in peripheral blood of patients with melanoma as a marker for metastasis. METHODS With application of an immunoluminometric assay, S-100 levels in 1396 samples of 570 patients with melanoma and 53 control subjects were measured in a blinded manner. RESULTS The cut-off level for patients with melanoma without medical history of metastases versus patients with newly occurring lymph node, visceral, and/or brain metastases was 0.114 microg/L, with a sensitivity of 94% (95% confidence interval, 86.4%-98.5%) and a specificity of 91% (95% confidence interval, 87.7%-93.6%). False-negative results included patients with unknown primary melanoma and those with amelanotic melanoma metastases. CONCLUSION The data suggest that S-100 in the peripheral blood of patients with melanoma could serve as a marker indicating new melanoma metastases and could help to monitor the course of the disease.

Psoralen plus ultraviolet-A-bath photochemotherapy as an adjunct treatment modality in cutaneous chronic graft versus host disease.

Background: Cutaneous chronic graft versus host disease (GVHD) is a severe complication following allogeneic stem cell (PBSCT) and bone marrow transplantation (BMT). Immunosuppressive therapy consists of prednisone, cyclosporine‐A, azathioprine or mycophenolate mofetil (MMF). Treatment of patients refractory to immunosuppression represents a major problem.

Calcipotriol vs. tazarotene as combination therapy with narrowband ultraviolet B (311 nm): efficacy in patients with severe psoriasis.

Background Phototherapy has been shown to be one of the most effective treatment modalities for patients with psoriasis. Nevertheless, photocombination therapies capable both of reducing cumulative ultraviolet (UV) doses and of accelerating clearance of skin lesions are important and of high interest. There have been no published studies comparing the effect of narrowband UVB irradiation in combination with topical application of tazarotene vs. calcipotriol.

Expression of c-myc and bcl-2 in Primary and Advanced Cutaneous Melanoma

Apoptosis is an important co-factor in the pathogenesis of a plethora of malignancies. Enhanced c-myc activation can result either in proliferation or apoptosis. Coexpression with antiapoptotic bcl-2, which abrogates the apoptotic function of c-myc might lead to an enormous growth advantage of cells. In order to elucidate the role of c-myc and bcl-2 as well as the coexpression of both genes in human melanoma, their expression was studied in four samples of normal skin (SK), 15 surgical margins (SM), 20 benign melanocytic nevi (MN), 20 primary melanomas (MM), and 30 melanoma metastases (MMET) by RT-PCR. These results were compared with immunohistochemistry (IH) in 7 SK, 7 SM, 26 MN, 50 MM, and 34 MMET. Similar results were found with both methods. However, MMET expressed c-myc (PCR 28/30, IH 23/34) as well as bcl-2 (PCR 27/30, IH 24/34) more frequently. Primary melanomas showed a similar expression pattern as SM and nevi. Moreover, in contrast to SK, SM, MN, and MM coexpression of bcl-2 and c-myc was found more frequently in MMET (PCR 25/30, p<0.01, IH 19/34, p<0.01). These results indicate that coexpression of c-myc and bcl-2 appears to be associated with advanced melanoma and contributes to the malignant phenotype.

Ultraviolet exposure and risk of melanoma and basal cell carcinoma in Ulm and Dresden, Germany

There is a perpetuating increase in melanoma and basal cell carcinoma (BCC) incidence in Europe. Few studies are evaluating various risk factors for both tumours.

Why ultraviolet protection with clothing makes sense.

Sir, Bowenoid papulosis is a disease presenting with fleshy, usually pigmented papules involving the genitalia of both sexes. In vulvar lesions, the term vulvar intraepithelial neoplasia (VIN3) is also used to describe the same condition. Histological examination reveals a picture consistent with carcinoma in situ, although the course of the disease appears to be benign. Association with oncogenic human papillomavirus (HPV 16, 18, 33) has been demonstrated. In the past, excisional surgery, electrocoagulation, cryotherapy and 5-fluorouracil have been used in the treatment of the condition with varying degrees of success. A 38-year-old woman was referred to our clinic with a 4-year history of pruritic skin lesions on her labia majora. She had previously been treated with podophyllin without any benefit. The molecular biological examination of the vaginal mucosa had been negative in terms of HPV typing (groups 6/11, 16/18, 31/33). However, HPV 31/33 was detected in the cervical smear (Hybrid Capture II test). Previous treatment with erbium laser, KTP 532 mm laser and podophyllin had been unsuccessful. On examination, erythema and numerous small papules were detectable on the labia minora and majora as well as in the clitoral and perianal area, each papule 1±3 mm in diameter (Fig. 1). The dermatohistopathological examination of several biopsies taken from these sites showed a uniform pattern with bowenoid epithelial changes, thus supporting the clinical diagnosis of bowenoid papulosis (Fig. 2). Treatment with imiquimod was initiated with a regimen different from that recommended for the treatment of genital warts. Imiquimod cream 5% was applied to affected areas on alternate days for 10 days until the skin became visibly irritated. The cream was now applied once daily for another 10 days, but washed off after 2 h each time. This treatment led towards complete clinical resolution within 8 weeks (Fig. 3). Histology performed 1 month, 6 months and 18 months after treatment demonstrated the absence of any precancerous epidermal changes (Fig. 4). The lympho histiocytic dermal infiltrate observed 1 month after treatment decreased in density in the control biopsies taken 6 and 18 months after treatment. Colposcopy performed 10 months after treatment showed minor acetic-white areas around the portio. No low-risk or high-risk HPV material was detected on the cervix (Hybrid Capture II test). More than 18 months after treatment the patient continues to be clinically clear. Imiquimod is a topical immune response modifier so far registered for the treatment of condylomata acuminata. Imiquimod directly induces antiviral and immunomodulating cytokines such as interferon-a, tumour necrosis factor-a and different interleukins from monocytes, macrophages and dendritic cells. Its effect on the innate immune response and the activation of Th1-immunity are responsible for its acute

Can the immersion time of PUVA bath therapy be shortened

Up to now, there are only a few data available concerning the influence of bathing time on skin phototoxicity. We compared the erythemal responses of normal skin to bath PUVA with 8-methoxypsoralen (8-MOP) after 5, 10 and 20 min immersion time. Currently, 20 min is the routinely performed immersion time in many European countries, including Germany, while in other countries bathing times are shorter. The minimal phototoxic dose (MPD) following immersion times of 5 min and 10 min in a warm water bath (37 degrees C) containing 1 mg/l 8-MOP was compared to the MPD following 20 min immersion time in a half-sided manner in a total of 24 patients. Our results revealed that an immersion time of 5 min did not yield a detectable erythema after 72 h. In contrast, both 10 and 20 min PUVA baths induced visible erythemas with a significantly higher median MPD following 10 min immersion (2.25 J/cm2) compared to 20 min baths (1.5 J/cm2). As an erythemal response of 8-MOP PUVA bath seems reduced after shorter immersion times, comparative studies on the clinical efficacy using shorter time regimens have to be conducted before conclusive recommendations for clinical PUVA-bathing time can be given.