Clinical Characteristics of Patients with Erythema Nodosum and Risk of Relapse - a 17-Year Study.

Erythema nodosum (EN) is the most common type of septal panniculitis which causes inflammation of the subcutaneous fat, being the result of a hypersensitivity reaction to specific triggers. It usually presents with erythematous painful rounded lumps symmetrically on the anterior surface of the lower limbs. Rarely, it may occur in other areas such as thighs, neck and arms. This is a retrospective study describing a cohort of patients hospitalized in the University Hospital of Heraklion, Heraklion, Greece. The present research compares characteristics between patients with and without relapse and identifies independent factors associated with relapse. All patients with EN hospitalized during a 17-year period were included. Data regarding epidemiology, current or recent infections, symptoms, laboratory values and relapses were all recorded and evaluated. In total, 138 patients, of which 27 (19.6%) males, with a median age of 46.5 years, were evaluated. Clinical presentation involved multiple lesions in 115 (83.3%) patients, while 12 (8.7%) of them were febrile. Relapse was noted in 27 (19.6%) subjects. Multivariate logistic regression analysis showed that male gender was associated with a higher risk of relapse, while cases with multiple lesions were associated with a lower risk.

Ambivalent Effects of Tumor Necrosis Factor Alpha on Apoptosis of Malignant and Normal Human Keratinocytes.

INTRODUCTION: Tumor necrosis factor alpha (TNFα) is a pro-inflammatory cytokine that may paradoxically induce either apoptosis or cell survival. It mediates its activity through binding of TNF-receptor (TNFR) 1 or 2. TNFR1 is mainly responsible for transmitting apoptotic signals. The activation of apoptotic mechanisms can either be intrinsic (mitochondrial) or extrinsic (death receptors). Death ligands such as TNF-related apoptosis-inducing ligand (TRAIL) specifically induce extrinsic apoptosis, while cytostatic drugs such as 5-fluorouracil (5FU) induce intrinsic apoptosis. OBJECTIVES: To investigate the effects of TNFα on apoptosis in malignant and normal human keratinocytes. METHODS: Human cutaneous squamous cell carcinoma (SCC) cell line SCC-13 and immortalized human keratinocytes HaCaT as well as primary normal human keratinocytes (PNHK) were stimulated with TNFα and then treated either with TRAIL or 5FU. Cell viability and cell proliferation, DNA fragmentation, apoptosis, and cytotoxicity were determined by WST-1 proliferation assay, ELISA, flow cytometry, and colorimetric analysis of lactate dehydrogenase, respectively. In addition, Western blotting was performed for analysis of caspase-3. RESULTS: TNFα affected viability of SCC-13 and HaCaT cells in combination with 5FU or TRAIL. In contrast, TNFα did not influence cell viability of PNHK. It enhanced the apoptotic effects of both extrinsic and intrinsic stimuli in SCC-13 and HaCaT. In clear contrast, TNFα protected PNHK against TRAIL- and 5FU-induced apoptosis. The effects were dose-dependent and TNFα-specific; furthermore, the apoptosis pathway was caspase-dependent. CONCLUSIONS: In summary, opposing effects of TNFα in malignant versus normal human keratinocytes were observed with possibly relevant clinical implications, when patients are treated with TNFα inhibitors.

A Clinical Trial for the Identification of Metabolic Biomarkers in Hashimoto's Thyroiditis and in Psoriasis: Study Protocol.

Hashimoto's thyroiditis and psoriasis are inflammatory disorders that significantly impact patients' quality of life, stressing the need for novel biomarkers of early diagnosis. This randomized clinical trial (NCT04693936) aims to identify Hashimoto's thyroiditis' and psoriasis' metabolic biomarkers and to investigate the effect of environmental factors on the disease-related metabolic imprint and quality of life. Patients with Hashimoto's thyroiditis, patients with psoriasis, and healthy individuals aged 18-60 will be recruited, enrolled according to eligibility criteria (medical history, clinical thyroid markers and the PASI score) and randomized to two groups. The intervention group will receive a combination of nutraceuticals for 6 months as part of a Mediterranean diet, and the control group will follow their usual diet. Data will be collected at baseline and the end of the study, including metabolite levels, lifestyle and anthropometric measurements, adherence to the Mediterranean diet (through the Mediterranean Diet Score) and disease-specific quality of life (through the Thyroid Patient Report Outcome for Hashimoto's group, and the Dermatology Life Quality Index for the psoriasis group). This study will investigate metabolic biomarkers and related changes in Hashimoto's thyroiditis and psoriasis and evaluate the association of metabolic changes with dietary factors and quality of life.

Current and Future Trends in Molecular Biomarkers for Diagnostic, Prognostic, and Predictive Purposes in Non-Melanoma Skin Cancer.

Skin cancer represents the most common type of cancer among Caucasians and presents in two main forms: melanoma and non-melanoma skin cancer (NMSC). NMSC is an umbrella term, under which basal cell carcinoma (BCC), squamous cell carcinoma (SCC), and Merkel cell carcinoma (MCC) are found along with the pre-neoplastic lesions, Bowen disease (BD) and actinic keratosis (AK). Due to the mild nature of the majority of NMSC cases, research regarding their biology has attracted much less attention. Nonetheless, NMSC can bear unfavorable characteristics for the patient, such as invasiveness, local recurrence and distant metastases. In addition, late diagnosis is relatively common for a number of cases of NMSC due to the inability to recognize such cases. Recognizing the need for clinically and economically efficient modes of diagnosis, staging, and prognosis, the present review discusses the main etiological and pathological features of NMSC as well as the new and promising molecular biomarkers available including telomere length (TL), telomerase activity (TA), CpG island methylation (CIM), histone methylation and acetylation, microRNAs (miRNAs), and micronuclei frequency (MNf). The evaluation of all these aspects is important for the correct management of NMSC; therefore, the current review aims to assist future studies interested in exploring the diagnostic and prognostic potential of molecular biomarkers for these entities.

Successful treatment of cutaneous leishmaniasis with intralesional aminolevulinic acid photodynamic therapy.

Leishmaniasis is a protozoan infectious disease that often affects the skin and may acquire a chronic and difficult to treat course. Topical photodynamic therapy (PDT) is a novel treatment which involves the selective uptake of a photosensitizing agent. Exposure to an appropriate light source in the presence of oxygen leads to formation of reactive oxygen species and destruction of the target cells. We report on the successful treatment of a 69-year-old patient with a relapse of long-standing cutaneous leishmaniasis using intralesional aminolevulinic acid-PDT.

Identification of the M541L sequence variation of the transmembrane KIT domain in Merkel cell carcinoma.

BACKGROUND: Merkel cell carcinoma (MCC) is an aggressive KIT-positive cutaneous tumor. KIT mutations are considered to play a key role in the pathogenesis of various neoplasms, but have not been found so far in MCC. The aim of the present study was therefore to investigate the presence of KIT mutations in MCC. MATERIALS AND METHODS: The entire coding region of KIT in the MCC cell line MCC-1 was sequenced. KIT exon 10 was amplified from archival paraffin-embedded MCC specimens by PCR and sequenced. RESULTS: Exon 10 M541L KIT sequence variation, which confers increased sensitivity to KIT ligand stem cell factor, was detected in the MCC-1 cell line. Sequencing of KIT exon 10 in six archival MCC specimens revealed the wild-type sequence. CONCLUSION: The presence of the M541L KIT variation in MCC warrants further studies for its role in the pathogenesis of this tumor.

KIT receptor activation by autocrine and paracrine stem cell factor stimulates growth of merkel cell carcinoma in vitro.

The co-expression of KIT receptor and its ligand stem cell factor (SCF) has been reported in biopsy specimens of Merkel cell carcinoma (MCC). However, the functional role of SCF/KIT in the pathogenesis of this aggressive tumor has not been elucidated. The present study reports expression and effects of SCF and KIT in the Merkel cell carcinoma cell line MCC-1 in vitro. SCF and KIT were endogenously co-expressed in MCC-1 cells. Exogenous soluble SCF modulated KIT receptor mRNA and protein expression, stimulated growth of MCC-1 cells, upregulated endogenous activation of KIT, AKT, and of extracellular signal-regulated kinase (ERK) 1/2 signaling pathway. On the contrary, an inhibitory antibody that neutralized the KIT ligand binding site, reduced growth of MCC-1 cells, as did high doses of the KIT kinase inhibitors imatinib and nilotinib. Also, inhibitors of KIT downstream effectors, U0126 that blocks MEK1/2 as well as wortmannin and LY294002 that inhibit phosphatidylinositol 3-kinase-dependent AKT phosphorylation, inhibited the proliferation of MCC-1 cells. These data support the hypothesis that KIT is activatable by paracrine or autocrine tumor cell-derived SCF and stimulates growth of Merkel cell carcinoma in vitro. Blockade of KIT and the downstream signaling cascade at various levels results in inhibition of Merkel cell carcinoma growth in vitro, suggesting targets for therapy of this cancer.

Analysis of epidemiology, clinical features and management of erysipelas.

BACKGROUND: Erysipelas is a superficial form of cellulitis affecting the upper dermis and superficial lymphatics. The widespread use of antibiotics may affect clinical findings and response to therapy of infectious disorders. The purpose of the study was to investigate the epidemiological, clinical, and laboratory features of erysipelas and to compare the results of treatment with penicillin vs. other antibiotic regimens. METHODS: All charts of erysipelas patients treated at the University Hospital of Heraklion, Crete, Greece from 1994 to 2002 were retrospectively studied. RESULTS: Median age of the 99 patients was 54.5 years; 59% were females. The most frequent site involved was the lower extremity (76%), followed by the face (17%) and upper extremity (6%). In 61 patients (62%), a possible entry portal was identified. The most common manifestation of erysipelas was local symptoms and signs (pain, erythema, and swelling) in all patients, together with elevated erythrocyte sedimentation rate (ESR) (60%). Fever was present in 25% of patients. The most commonly used antibiotic was intravenous penicillin G (64%). In the penicillin group, mean duration of fever after treatment initiation was shorter than in the nonpenicillin group (1.7 vs. 4.5 days, P = 0.002). Both treatment failures and recurrences were the same between the two groups. DISCUSSION: The diagnosis of erysipelas can be based on careful examination for local signs and symptoms. The role of ESR in primary diagnosis needs further investigation. Penicillin seems to preserve its fundamental role in the treatment of disease.

Co-expression of KIT receptor and its ligand stem cell factor in Merkel cell carcinoma.

BACKGROUND/AIMS: KIT receptor has been implicated in the pathogenesis of cancer, either by mutation or autocrine activation. Merkel cell carcinoma (MCC) is a rare KIT-positive cutaneous tumor. We investigated the co-expression of KIT and its ligand stem cell factor (SCF) in MCC. METHODS: Sixteen specimens from 13 MCC patients of various tumor stages were examined by immunohistochemistry for SCF, KIT, Ki67/MIB-1 and cleaved caspase 3 expression, and for apoptosis by TUNEL. RESULTS: KIT was expressed in 13 of 16 tumors, and SCF in 15 of 16 specimens. Co-expression of KIT and SCF was detected in 12 of 16 tumors. KIT and SCF immunoreactivity scores were independent of tumor stage. Ki67/MIB-1 proliferation rates were high, whereas apoptosis rates were low, and did not depend on KIT or SCF expression. CONCLUSION: Co-expression of KIT and SCF in a high percentage of MCC tumors hints to an autocrine mechanism. KIT and SCF expression in primary tumors and in metastases suggests an early event in Merkel cell transformation.

Interferon-alpha inhibits proliferation and induces apoptosis of merkel cell carcinoma in vitro.

Merkel cell carcinoma is a tumor with aggressive biological behavior and limited response to chemotherapy. The present study investigated the effect of interferon (IFN)-alpha on growth and apoptosis of Merkel carcinoma cells in vitro. Proliferation of MCC-1 cell line was reduced dose-dependently by IFN-alpha and diminished when higher IFN-alpha concentrations were used. Additionally, IFN-alpha potently decreased DNA-synthesis and Ki67/MIB-1 proliferation index of MCC-1 cultures. Furthermore, IFN-alpha induced dose-dependently apoptosis of MCC-1 cells as shown by caspase-3 activation, and detection of apoptotic DNA strand breaks and fragmented nuclei. These findings suggest that IFN-alpha may have antitumor activity against Merkel cell carcinoma.

A rare case of leishmaniasis recidiva cutis evolving for 31 years caused by Leishmania tropica.

A 64-year-old woman presented with erythematous, infiltrative plaques with a central atrophic area on both zygomatic regions. Several yellow-reddish papules were seen at the periphery of the plaques and showed an "apple-jelly" color on diascopy (Fig. 1). No visceral involvement was detected. The past medical history revealed that, at 3 years of age, she had developed an "Oriental sore" on both cheeks that healed with permanent scars. Thirty years later, she noticed an erythematous patch around the scars. She reported a hospital admission 22 years earlier for cutaneous leishmaniasis (CL); this was treated with pentavalent antimonial therapy for 10 days with partial improvement, when she refused further treatment. The lesions worsened in the summer and gradually became disfiguring, which prompted her to seek medical consultation. Laboratory findings were normal. Leishmania antibody titers were negative. Tissue samples were obtained by biopsy from the border of the lesion for culture, polymerase chain reaction (PCR), and histopathologic examination. Histology revealed a dermal infiltrate with tuberculoid granulomas surrounded by lymphocytes, histiocytes, and some plasma cells, but no caseation necrosis. A few Leishmania organisms were found on careful searching (Fig. 2). Leishmania tropica was identified by culture and PCR. A diagnosis of leishmaniasis recidiva cutis (LRC) was made on the basis of the anamnestic data together with the clinical, histopathologic, biologic, and molecular findings. Complete regression was achieved with meglumine antimoniate (Glucantime) given intramuscularly (15 mg Sb(V)/kg/day for 15 days) and cryosurgery with liquid nitrogen. No recurrence was noted during a 12-month follow-up period.

From the protein to the graph: how to quantify immunohistochemistry staining of the skin using digital imaging.

Quantitative immunohistochemistry is needed in order to reliably and accurately assess the expression of cellular proteins in tissue. Skin is a difficult tissue for automated image analysis due to its heterogeneous composition and its architecture. In the present study we used a psoriatic skin model to compare the expression of p53 and bcl-2 before and after treatment with anti-tumor necrosis factor-alpha using digital image analysis. Digital photomicrographs were acquired and analyzed with Scion image software in order to obtain the fraction of p53 and bcl-2 immunoreactive cells' area out of the total area investigated. Statistical analysis with ANOVA revealed a significant increase of p53 expression and a decrease of bcl-2 expression in all 3 epidermal layers during the course of therapy (p<0.001). The results were in line with the conventional histopathological evaluation using an arbitrary scale to grade the extent and intensity of the staining. So, the estimation of volume fraction of immunohistochemically labelled cells in skin tissue can be performed easily and rapidly using commonly available image analysis software and provides reproducible and unbiased numerical estimations of the amount of cell labelling.

Morphological analysis of integrin-mediated adhesion of immature human mast cells to extracellular matrix proteins.

Specific heterodimers of alpha and beta integrins are implicated in mediating adhesion and functional activation of mast cells to extracellular matrix (ECM) proteins, determining thus homing, secretion and tissue distribution of these cells. In the present study, we have examined integrin expression and associated morphological features of mast cells adhering to ECM, also depending on cell activation and under the influence of protein kinase C (PKC) inhibitors. Unstimulated and PMA-activated human leukaemic mast cells (HMC-1 line) were allowed to adhere to fibronectin or vitronectin-coated surfaces. Cells were specifically stained for actin, beta(1, )alpha(1)-alpha(6), alpha(v) and alpha(v)beta(5 )integrins and were evaluated by fluorescence microscopy and confocal laser scan microscopy. Spontaneously adhering cells rapidly assumed an oblong shape, with pronounced formation of filopodia, whereas PMA-stimulated cells were round in shape. Clustering of integrins on filopodia and on comma-like shapes at the cell circumference in rounded cells was noted only for alpha(4), alpha(5) and beta(1 )chains in fibronectin-adhering cells, and for alpha(v) and alpha(v)beta(5) chains in vitronectin-adhering cells. On double staining, clustered integrins co-localized with each other and with actin at the cell membrane and along intracellular tension lines of actin filaments. PKC inhibitors affected the shape of cells, but adhesion was maintained. These data provide a morphological correlate to previously reported functional studies, demonstrating clustering of selected integrins during ECM adhesion at the cell membrane. This was associated with alignment of integrins along actin filaments within the cytoplasm, PKC signalling and changes in shape and activation of mast cells.

The incidence of cutaneous melanoma on Crete, Greece.

BACKGROUND: For Greece, no data regarding the incidence of cutaneous melanoma (CM) have been reported. In this report, we present epidemiologic data for CM on Crete, an island in southern Greece, during the years 1999-2002. We attempt a comparison with corresponding data reported for the Italian population. METHODS: One hundred and two CM patients of Cretan origin with primary CM first diagnosed between the years 1999-2002 were interviewed and underwent complete skin examination by the same two experienced dermatologists. Crude and/or age-standardized incidence rates were calculated for Crete as a whole, as well as for each one of the four prefectures of the island. RESULTS: The age-standardized incidence rate according to the Greek population was 4.6 per 100,000 person-years for men and 4.7 per 100,000 person-years for women. The crude incidence rates did not differ significantly between the four prefectures. Significant differences between Cretan and Italian CM patients were found in terms of gender, age at diagnosis, anatomic site and histogenetic type of CM, hair color, skin reaction to sun exposure, history of sunburn before the age of 15 years, presence of solar lentigines, and total common nevus count. CONCLUSIONS: The incidence of CM on Crete is higher than that estimated for the whole of Greece and comparable with the incidence reported for other southern European countries.

Skin symptoms and work-related skin symptoms among grape farmers in Crete, Greece.

BACKGROUND: Grape farmers are exposed to a variety of agents capable of inducing occupational skin disease. We conducted a study to measure the prevalence of skin symptoms and work-related skin symptoms among grape farmers in the Malevisi region of Crete and to provide data on associated risk factors. METHODS: One hundred twenty grape farmers and 100 controls participated in the study. The protocol consisted of a questionnaire, skin prick tests for 16 common allergens, and measurement of specific IgE antibodies against 8 allergens. RESULTS: Self-reported itchy rash (OR, 2.31; 95%CI, 1.10-4.84, P<0.05) within the last 12 months, and work-related itchy rash (OR, 4.08; 95%CI, 1.01-20.33, P<0.05) were significantly higher in grape farmers than in controls, after adjusting for age and sex. Sensitization to pollens (OR, 4.20; 95% CI, 1.41-12.82, P<0.01) and allergic rhinitis (OR, 3.06; 95% CI, 1.21-8.28, P<0.05) were found to be significantly associated with self-reported itchy rash in the grape farmers group. CONCLUSIONS: Grape farmers reported skin symptoms more frequently than non-exposed controls, and IgE-mediated sensitization to pollens was found to be significantly associated with the reported symptoms. Further studies are needed to evaluate the impact of specific occupational agents on skin diseases among grape farmers.

Epidemiological differences for cutaneous melanoma in a relatively dark-skinned Caucasian population with chronic sun exposure.

The aim of this study was to reveal differences in the epidemiology and to identify significant risk factors for cutaneous melanoma (CM) in a relatively dark-skinned, chronically sun-exposed Caucasian population. This group is considered to have a low risk for this tumour. One hundred and ten newly diagnosed patients with primary CM and 110 age- and gender-matched controls, all of Cretan origin, were interviewed and underwent a complete skin examination. Solar keratoses odds ratio (OR) 6.2 and lentigines (OR 2.2), common and atypical naevi (OR 5.4 and 3.0, respectively), blonde or red hair colour (OR 3.1), skin phototypes I/II (OR 1.8), as well as total sun exposure (weeks per year) (OR 1.03), were all significantly associated with CM risk in a multivariate logistic regression analysis. In the relatively dark-skinned Cretan population, sun exposure indices represent the most important risk markers for CM which contrasts with data from fair-skinned Caucasian populations where melanocytic naevi are the main risk factors.

Human skin mast cells express H2 and H4, but not H3 receptors.

Mast cells generate and release histamine during anaphylactic reactions, and there is pharmacological evidence that histamine regulates this process via specific receptors. Therefore, we examined human leukemic (HMC-1) and normal skin mast cells for the expression of all four currently known histamine receptors. Both cell types expressed H2 and H4 receptors at mRNA and protein levels, whereas H3 receptor specific mRNA and receptor protein was undetectable. Similarly, immunohistochemistry of cutaneous tissue showed an absence of H3 receptor in these cells. Despite transcription of mRNA, H1 receptor protein was only moderately expressed in HMC-1 cells and was virtually absent in skin mast cells. Furthermore, only H1, H2, and H4 receptors were detectable by Western blot analysis of HMC-1 cells. Radiolabeled histamine binding was strongly inhibited only by H2 (ranitidine)- and H3/H4 (FUB 108)-specific antagonists. Histamine-induced increase of cAMP was inhibited by the H2 receptor antagonist famotidine, whereas induction of IP3 was not observed, making signaling via the H1 receptor unlikely. These data show that human mast cells constitutively express primarily H2 and H4 receptors and that H2 receptors are functionally linked to cellular processes. They provide new insights into the mechanisms that govern auto- and paracrine histamine-induced mast cell functions.