Altered gene expression in squamous cell carcinoma arising from congenital unilateral linear porokeratosis.

Congenital unilateral linear porokeratosis (CULP) is a rare disorder of keratinization that shares clinical and molecular similarities with psoriasis. It also has an increased risk for malignant transformation to cutaneous squamous cell carcinoma (SCC). We investigated the expression of psoriasin, human beta-defensin-2, cathelicidin antimicrobial peptide/LL-37, e-cadherin, involucrin, p16(INK4a) , p53, cyclin D1 and microchromosome maintenance protein 7 in healthy skin and in lesions of psoriasis, CULP and SCC from the same patient. p16(INK4a) was overexpressed in CULP but not in the subsequent SCC. Psoriasin was overexpressed in psoriasis, CULP and SCC compared with healthy skin. Speculatively, p16(INK4a) and psoriasin could be involved in the pathogenesis of CULP. Moreover, psoriasin may play a role in the malignant transformation of CULP to SCC.

Prevalence of human polyomaviruses in common and rare types of non-Merkel cell carcinoma skin cancer.

BACKGROUND: Little is known about the association of human polyomaviruses (HPyVs) other than Merkel cell polyomavirus (MCPyV) with nonmelanoma skin cancer. OBJECTIVES: To evaluate the presence of HPyV6, HPyV7, trichodysplasia spinulosa-associated polyomavirus (TSV), also called HPyV8, and the recently discovered HPyV9 in basal cell carcinoma (BCC), actinic keratosis (AK), squamous cell carcinoma in situ (SCCis), squamous cell carcinoma (SCC), keratoacanthoma (KA), microcystic adnexal carcinoma (MAC) and atypical fibroxanthoma (AFX). METHODS: Archival paraffin-embedded samples (n = 193: 41 BCC, 31 AK, 8 SCCis, 52 SCC, 42 KA, 5 MAC and 14 AFX) were analysed for the presence of the respective HPyV by polymerase chain reaction (PCR). HPyV DNA loads (HPyV DNA copies per ß-globin gene copy) were determined in all HPyV-positive samples by quantitative real-time PCR. Immunohistochemical analysis of MCPyV large T-antigen (LTA) expression was performed using the monoclonal antibody CM2B4. RESULTS: MCPyV DNA was found in 29% of BCC, 19% of AK, 25% of SCCis, 27% of SCC, 29% of KA, 0% of MAC and 29% of AFX. MCPyV DNA loads never exceeded 0·3 MCPyV DNA copies per ß-globin gene copy (median 0·004). In the immunohistochemical analysis of MCPyV LTA expression, all evaluated samples (32 MCPyV DNA-positive samples) were LTA negative. HPyV6 DNA was found in 7% of BCC, 3% of AK, 12% of SCCis, 4% of SCC, 5% of KA, and 0% of MAC and AFX. HPyV6 DNA loads never exceeded 0·7 HPyV6 DNA copies per ß-globin gene copy (median 0·015). None of the 193 samples was positive for HPyV7, TSV or HPyV9 DNA. CONCLUSIONS: Our findings argue against a pathogenic role for MCPyV, HPyV6, HPyV7, TSV and HPyV9 in the analysed types of non-Merkel cell carcinoma skin cancer.

The expression of antimicrobial peptides is significantly altered in cutaneous squamous cell carcinoma and precursor lesions.

BACKGROUND: Antimicrobial peptides and proteins are not only effectors of the immune system but are also attributed important roles in tumour progression or tumour suppression in several malignancies such as oral squamous cell carcinoma (SCC). OBJECTIVES: These reports encouraged us to systematically investigate the expression of different classes of antimicrobial peptides and proteins in tissue samples of cutaneous SCC and its precursor lesions. METHODS: The protein expression of human beta-defensin (hBD)-1, -2, and -3, ribonuclease (RNase)-7 and the S100 protein psoriasin were analysed in 25 patients with actinic keratosis (AK), 30 with SCC in situ (SCCis), 23 with SCC, nine healthy skin controls and 10 healthy, chronically ultraviolet (UV)-exposed controls, by means of immunohistochemistry. RESULTS: Expression of hBD-1 was significantly reduced in SCC compared with UV-exposed healthy skin, AK and SCCis. RNase-7 expression was reduced gradually parallel to every step of malignant transformation, with the highest expression in healthy skin and the lowest expression in SCC. hBD-2 and psoriasin were significantly overexpressed in SCC and SCCis, compared with healthy controls. hBD-3 showed significantly more frequent expression in AK than in healthy controls, and in patients with SCCis and SCC. CONCLUSIONS: It is tempting to speculate that hBD-1 and RNase-7 might act as tumour suppressors while hBD-2 and psoriasin might act in the opposite way as promoters of tumour progression. Further investigations should clarify whether hBD-2 and hBD-3 could be potential targets for the development of pharmacological therapy.

A randomized, half-side comparative study of aminolaevulinate photodynamic therapy vs. CO(2) laser ablation in immunocompetent patients with multiple actinic keratoses.

BACKGROUND: Photodynamic therapy (PDT) and laser ablation (LA) are frequently used treatment options for multiple actinic keratoses (AK), yet they have not been compared head to head. OBJECTIVES: To compare PDT and carbon dioxide (CO(2) ) LA in the management of multiple AK using objective and subjective outcome measures. METHODS: A single-centre, randomized, two-treatment half-side comparative study of PDT vs. CO(2) LA was performed. Patients with at least four bilateral (e.g., scalp, forearms) AK were included. The primary outcome measure was the reduction of AK 3 months (v3) after therapy. Secondary outcome measures included the reduction of AK 4 weeks (v2) after therapy, decrease of epidermal p53 and Ki-67 protein expression, micromorphological changes as assessed by optical coherence tomography (OCT) in vivo, and investigators' and patients' satisfaction scoring. RESULTS: In total, 20 patients (18 men and 2 women) completed the study. Both treatments reduced AK quantity significantly. On v3, relative reduction of AK quantity was significantly higher following PDT (P = 0·0362). Ki-67 and p53 protein expression was reduced significantly from baseline (Ki-67, median 49·5%; p53, median 64·8%) to v2 by both procedures (PDT, median 18·5%, P < 0·0001; LA, median 16·2%, P < 0·0001). AK features as assessed by OCT imaging were also significantly reduced by both procedures. The investigators and patients rated the side-effects and inconveniences of PDT as more severe, but both overall preferred PDT due to the superior clinical outcome. CONCLUSIONS: CO(2) LA and PDT are both effective therapy options for multiple AK, yet PDT seems to be superior in terms of AK reduction and participants' and investigators' overall satisfaction.

Serum levels of antimicrobial peptides and proteins do not correlate with psoriasis severity and are increased after treatment with fumaric acid esters.

Overexpression of antimicrobial peptides and proteins (AMPs) such as human ß-defensin-2 (hBD2), LL37, and psoriasin has frequently been observed in lesional skin of psoriasis patients. We aimed to evaluate whether circulating AMP levels correlate with disease severity, and change under therapy with fumaric acid esters (FAE). We studied psoriasis patients who underwent systemic therapy using oral FAE (Fumaderm(®)). An enzyme-linked immunosorbent assay for the detection of serum protein expression of hBD2, LL37, and psoriasin was performed at baseline and after 12-week therapy. After 12-week FAE treatment of 28 patients, the median PASI significantly (P < 0.0001) decreased from 27.1 to 12.5. In psoriasis patients, mean ± SD serum hBD2, psoriasin, and LL37 levels at baseline were 295.6 ± 93.5 pg/ml, 79.4 ± 32.7 ng/ml, and 106.3 ± 90 ng/ml, respectively, which were significantly increased when compared to healthy controls (110 ± 53.7 pg/ml, P ≤ 0.0001; 3.1 ± 0.7 ng/ml, P ≤ 0.0001; 3.8 ± 0.9 ng/ml, P = 0.0004, respectively). After 12-week FAE treatment, a significant increase of serum hBD2 (339.7 ± 74.3 pg/ml; P = 0.0046), psoriasin (106 ± 58.9 ng/ml; P = 0.0014), and LL37 (136.6 ± 115.1 ng/ml; P = 0.0035) was observed. Correlation studies did not reveal significant relationships between serum AMP levels and PASI (r < 0.1; P > 0.05). In contrast to AMP expression in psoriatic skin serum, AMP levels seem not to correlate with disease severity. Increased serum AMP protein levels in psoriasis resolution are an unexpected observation that needs to be investigated more in detail in future studies.

Differential expression of connective tissue growth factor and extracellular matrix proteins in lichen sclerosus.

BACKGROUND: The histopathology of lichen sclerosus (LS) suggests abnormalities in extracellular matrix (ECM) composition. OBJECTIVES: We aimed to investigate the expression pattern of ECM proteins and related growths factors and Smad signal transducers in LS as compared with healthy skin. METHODS: To assess the expression of decorin, biglycan, versican, perlecan, fibronectin, dermatopontin, extracellular matrix protein 1 (ECM-1), matrix metalloproteinase 1, tissue inhibitor of metalloproteinase 1, connective tissue growth factor (CTGF), transforming growth factor ß1, and Smad-3 protein, real-time RT-PCR and immunohistochemistry were performed on skin specimens obtained from the genital region of healthy subjects (n = 10) as well as LS patients (n = 26). RESULTS: Median mRNA as well as mean protein expression of biglycan, versican, fibronectin, and ECM-1 was significantly higher in LS when compared with healthy controls. Both mRNA and protein CTGF expression observed in LS was significantly higher than in controls. CTGF mRNA expression significantly correlated with mRNA expression of biglycan, versican and fibronectin. CONCLUSIONS: Expression of ECM proteins (e.g. proteoglycans, ECM-1) and CTGF is altered in LS. TGF-ß/Smad-3 independent up-regulation of CTGF may induce accumulation of ECM proteins and maintain fibrosis in chronic LS.

[Acquired reactive perforating collagenosis following herpes zoster as isotopic response?]. / Erworbene reaktive perforierende Kollagenose nach Herpes zoster als isotopische Antwort?

Reactive perforating collagenosis is a disease whose pathogenesis is still not fully understood. Histological findings are degenerated collagen bundles which are arranged in vertical direction penetrating the epidermis into a dome-shaped crater. Usually diabetes mellitus and renal failure can be found among patients with reactive perforating collagenosis. To date, there have been five cases described where the eruption of reactive perforating collagenosis followed herpes zoster infection. This could be a form of Wolf's isotopic response, a term that is used for dermatoses which arise after the healing of a preexisting dermatosis. We report the sixth case of a herpes zoster-associated reactive perforating collagenosis and discuss the current literature.

Etanercept plus narrowband ultraviolet B phototherapy of psoriasis is more effective than etanercept monotherapy at 6 weeks.

BACKGROUND: A substantial portion of patients with psoriasis does not achieve a satisfactory response under antitumour necrosis factor (TNF)-α biological therapies. OBJECTIVES: We aimed to evaluate whether etanercept plus narrowband ultraviolet B (NB-UVB) phototherapy is superior to etanercept monotherapy in the management of psoriasis. METHODS: In this prospective study, patients with psoriasis were treated with etanercept 25 mg twice weekly. Two marker lesions were selected for determination of the modified Psoriasis Area and Severity Index (M-PASI). NB-UVB was administered thrice weekly whereby one marker lesion was covered as nonirradiated control. Skin biopsies for histology and immunohistochemistry were performed in both marker lesions after a 6-week treatment course. RESULTS: After 6 weeks of therapy, the relative M-PASI reduction (mean ± SD) in etanercept-treated sites (53·7 ± 36·9%) was significantly lower than the reduction in etanercept plus NB-UVB-treated lesions (64 ± 27·8%; P = 0·011). At the end of treatment, histology scores of etanercept-treated psoriatic plaques were significantly higher than scores of etanercept plus NB-UVB-treated sites (4·6 ± 2·7 vs. 3·7 ± 2·4; P =0·045). Epidermal immunoreactivity for CD1a, CD4 and CD8 was significantly lower in etanercept plus NB-UVB-treated lesions when compared with etanercept monotherapy. CONCLUSIONS: Etanercept combined with NB-UVB is more effective than etanercept monotherapy at 6 weeks as demonstrated at a clinical, histological and immunohistological level. However, as there is an increased risk for malignancy by treatment with TNF-α blockers alone or in combination with phototherapy, we recommend to restrict this highly effective combination to short periods of time, for instance to obtain a quicker response, and to avoid long-term treatment.

Expression of antimicrobial peptides and proteins in etanercept-treated psoriasis patients.

Recent papers highlight the role of dysregulated expression of antimicrobial peptides and proteins (AMPs) in the pathogenesis of psoriasis. Etanercept, a blocker of the pro-inflammatory cytokine tumour necrosis factor-α (TNF-α), is effective in the treatment of psoriasis. We aimed to evaluate the expression profiles of AMPs in psoriatic skin before and after a 6-week course of etanercept therapy. We included 12 psoriasis patients who underwent medium-dose etanercept treatment for 6weeks. At baseline and at the end of therapy immunohistochemistry from lesional skin was performed for psoriasin, LL-37, and human ß-defensin 2 (hBD-2). After 6-week treatment, the modified psoriasis area and severity index significantly decreased from 37.5±5.9 to 14±13.4. Lesional immunoreactivity scores of psoriasin, LL-37, and hBD-2 also significantly decreased after a 6-week course of etanercept. We have demonstrated that etanercept-induced improvement of psoriasic lesions is associated with a significant decline of AMP protein expression.

Impact of etanercept treatment on ultraviolet B-induced inflammation, cell cycle regulation and DNA damage.

BACKGROUND: Current studies indicate that treatment with tumour necrosis factor (TNF)-α blockers plus ultraviolet (UV) B phototherapy results in higher relative Psoriasis Area and Severity Index reduction as compared with TNF-α monotherapy. OBJECTIVES: This study aimed to investigate the acute impact of etanercept on UVB-induced inflammation, cell cycle regulation and DNA damage. METHODS: Eleven subjects diagnosed with psoriasis who fulfilled the indication criteria for etanercept treatment were studied. A healthy skin site on the upper back was treated with UVB at 2 minimal erythema doses (MED). After 1, 24 and 72 h punch biopsies were taken from this site. Following the 72 h biopsy etanercept 50 mg was administered subcutaneously. After 48 h, 2 MED was given on healthy skin adjacent to previously treated skin sites. Again, after 1, 24 and 72 h punch biopsies were taken from this site. UVB- as well as UVB plus etanercept-treated skin was assessed by means of colorimetry and immunohistochemical studies for caspase 3, cyclin D(1), interleukin-12, Ki-67, p16, p53, survivin, thymine dimers and TNF-α. RESULTS: Erythema formation did not differ significantly between UVB- and UVB plus etanercept-treated sites. Comparisons between UVB- and UVB plus etanercept-treated sites at a given time (1, 24, 72 h) did not result in significant differences in immunoreactivity of the markers investigated, except for cyclin D(1), p53 and survivin. Immunoreactivity of cyclin D(1) and p53 was significantly decreased in UVB plus etanercept-treated sites at 24 h. Survivin expression was significantly higher in UVB plus etanercept-treated skin as compared with UVB monotherapy. CONCLUSIONS: Our data indicate that combined treatment with broadband UVB and TNF-α blockers might increase the risk of photocarcinogenesis by influencing apoptotic as well as antiapoptotic pathways.

Immunophenotyping of inflammatory cells in subacute prurigo.

BACKGROUND: Subacute prurigo (SP) is a relatively common disease of papular cutaneous lesions that itch intensely. However, there is a lack of systematic clinical and histological studies on SP. OBJECTIVES: We aimed to immunophenotype inflammatory cells in SP using immunohistochemistry and flow cytometry. METHODS: Lesional and non-lesional skin of 21 patients with SP was investigated. Immunohistochemical staining for CD1a, CD3, CD4, CD8, CD15, CD34, CD68, and anti-human tryptase (AHT) was performed. In order to evaluate the absolute counts and percentages of CD4+ and CD8+ lymphocytes in the peripheral blood of SP patients, flow cytometric methods were used. RESULTS: Compared to non-lesional skin, there was a significant increase of the median percentage of CD3+, CD4+, and CD8+ cells in the lesional dermis (12.6% vs. 19.7%, P=0.044; 0.8% vs. 3.7%, P=0.016; and 1% vs. 15.6%, P=0.0039, respectively). The mean ± SD CD4+/CD8+ ratio was 0.58 ± 0.6. Median percentage of CD15+ cells was significantly increased in lesional skin as compared to non-lesional skin (11.7 vs. 1%, P=0.027). Median percentage immunoreactivity of CD68+ cells was significantly increased in lesional dermis as compared to non-lesional skin (32.5% vs. 9.4%, P=0.0005). CD1a, CD34, and AHT positive cells did not significantly differ between lesional and non-lesional skin. T lymphocyte subsets in the peripheral blood of SP patients did not show significant pathologies. CONCLUSIONS: We observed that the inflammatory cell infiltrate in SP mainly consists of T lymphocytes, particularly CD8+ cells, CD15+ neutrophils, and CD68+ macrophages.

Non-invasive imaging of mid-dermal elastolysis.

Mid-dermal elastolysis (MDE) is a rare disease that is characterized histopathologically by selective loss of elastic tissue in the mid dermis. We aimed to assess MDE using noninvasive skin imaging techniques in vivo. We examined both the lesional and adjacent healthy skin of a woman with the reticular variant of MDE, using confocal scanning laser microscopy, optical coherence tomography (OCT) and high-frequency ultrasound (HFUS). The median diameter of blood vessels at the top of dermal papillae was significantly increased in erythematous lesional skin compared with healthy skin. The mid-dermal signal intensity detected by OCT was higher in healthy skin than in lesional skin. With HFUS, mid-dermal density values were significantly higher in healthy skin than in lesional skin. Our preliminary findings indicate that noninvasive skin imaging methods such as OCT and HFUS might be suitable techniques for the evaluation and monitoring of elastolytic skin disorders such as MDE.

[Positive strip patch test reactions with coexistent negative patch test reactions. Relevance for the assessment practice of occupational skin disease]. / Positive Testreaktionen im Abrissepikutantest bei negativem Standardepikutantest. Bedeutung für die berufsdermatologische Begutachtung.

The strip patch test is recommended whenever a patch test is presumed to be false negative. The aim of this technique is to increase skin sensitivity to test substances by removing the upper layers of the stratum corneum prior to patch testing. We report on a 57-year-old former heavy-current electrical worker, who suffered from hyperkeratotic fissured hand eczema. Sensitization to nickel sulphate was demonstrated in both 2001 and 2009 with strip patch testing even though conventional patch testing was negative. To our knowledge, this is the first description of an allergy to nickel with a pension entitlement according to no. 5101 of the German ordinance on industrial disease based solely on positive strip patch tests.

Expression of extracellular matrix proteins in reticular variant of mid-dermal elastolysis.

BACKGROUND: Mid-dermal elastolysis (MDE) is a rare disorder of the elastic tissue that is characterized histopathologically by selective loss of elastic fibres in the mid dermis. OBJECTIVE: We aimed to investigate the protein and mRNA expression of extracellular matrix-related proteins and growth factors in the skin (lesional and non-lesional) of a female patient with the reticular variant of MDE. METHODS: Real-time RT-PCR and immunohistochemistry was performed for matrix metalloproteinase-1 (MMP-1), tissue inhibitor of metalloproteinase-1, decorin, biglycan, versican, fibronectin, elastin, extracellular matrix protein 1, cathepsin G, transforming growth factor ß1 and connective tissue growth factor. RESULTS: Although protein expression of decorin, biglycan and versican was reduced in the mid dermis of lesional skin, mRNA expression did not differ between lesional and non-lesional skin. As expected, elastin expression was significantly diminished in the mid dermis of lesional skin, whereas mRNA expression levels of elastin were equal to non-lesional skin. Immunoreactivity of MMP-1 was increased in lesional upper and mid dermis. Accordingly, MMP-1 mRNA was also significantly higher expressed in MDE when compared with non-lesional skin. CONCLUSIONS: The results of this study confirm data of the previous investigations indicating that increased MMP-1 activity followed by elastin degradation seems to constitute the pathogenetic background of MDE.

Differential regulation of MAPK (JNK 3) gene expression in human head and neck squamous cell carcinomas.

BACKGROUND: The JNK 3 gene encodes a protein which belongs to the mitogen-activated protein (MAP) kinases, a group of molecules involved in signaling pathways. MATERIAL AND METHODS: Messenger RNA extracted from head and neck squamous cell carcinoma (HNSCC) cells and normal upper aerodigestive tract mucosa keratinocytes was reversely transcribed. The resulting cDNA populations were subjected to an arbitrarily primed mRNA fingerprint. After electrophoresis, the band pattern was detected by autoradiography. RESULTS: A 107 bp mRNA fragment was detected in HNSCC cells showing considerable repression in comparison with the benign phenotype. After cloning of this fragment, a database search revealed an exact homology with sequences belonging to the c-jun N-terminal MAP-10 kinase (JNK 3). Northern hybridization confirmed the distinctly reduced expression of this gene in HNSCC biopsies in contrast to adjacent normal mucosa. CONCLUSION: The results show evidence that the expression of the JNK 3 gene is strongly repressed, suggesting that JNK 3 is implicated in carcinogenic processes in head and neck cancer.