The Distribution of Innervation and Immune Cell Infiltration Is Different in Genital and Extragenital Variants of Lichen Sclerosus.

Lichen sclerosus (LS) is a progressive skin disease that is characterized by chronic inflammation of either genital or extragenital skin, and it disproportionately affects women. We analyzed the distribution of nerve fibers, vanilloid receptors, cell proliferation, mast cells and macrophages in genital and extragenital LS samples, as well as in healthy skin, by using immunohistochemistry. The total amount of intraepidermal nerve fibers was lower in LS samples compared to healthy controls, while the total amount of subepidermal nerve fibers and calcitonin gene-related peptide (CGRP) positive fibers was higher in genital LS samples compared to both extragenital LS and healthy controls. Cell proliferation, macrophage and mast cell density were increased in LS samples compared to healthy controls. Genital LS had a higher macrophage density compared to the extragenital variant. Mast cell distribution significantly differed between genital and extragenital LS samples, even though their total mast cell densities were similar. These findings could explain the differences between pruritic symptoms of genital and extragenital LS and provide targets for the research of novel therapeutic strategies for LS management.

Single treatment of low-risk basal cell carcinomas with pulsed dye laser-mediated photodynamic therapy (PDL-PDT) compared with photodynamic therapy (PDT): A controlled, investigator-blinded, intra-individual prospective study.

BACKGROUND AND OBJECTIVE: To compare a single treatment of PDL-PDT with PDT for BCCs in terms of efficacy, aesthetic outcome, and pain in patients with multiple BCCs. STUDY DESIGN/MATERIALS AND METHODS: A prospective, controlled, intra-individual, investigator-blinded study was conducted on 15 patients with 62 BCCs. The BCCs on an individual patient were divided into two similarly-sized groups, and treated with PDT (630nm LED light source, fluence rate=30mW/cm2, total dose of 150J/cm2) and 585 nm-PDL-PDT (spot size=7mm, fluence=10J/cm2, pulse duration=10ms, 10% overlap, three passes, and cooling). Primary outcomes were complete BCC regression at months 3 and 12. Secondary outcomes were pain immediately after treatment, and aesthetic outcome evaluated by a blinded investigator. RESULTS: No significant difference was found in the therapeutic effect between the two treatments (P=0.285). Complete regression of BCCs at 3-months follow-up occurred in 79% of the PDT treated area and 74% of the PDL-PDT area. At month 12, complete regression using PDT was 75% (95% confidence interval (CI) 0.55-0.89) compared to 59% (95% CI 0.41-0.75) for the PDL-PDT treated areas. Both treatments had low mean pain scores: 1.7 for PDT and 2.6 for PDL-PDT (P=0.049) and the aesthetic appearance was similar (P=0.763). CONCLUSION: A single treatment with three passes of PDL-PDT is effective in clearing BCCs, but the recurrence rate is higher than in case of conventional PDT. PDL-PDT is associated with low treatment related pain, has similar cosmetic advantages as PDT but it requires less treatment time.

Aplasia Cutis Congenita in a Newborn Child Associated with Two Fetus Papyraceous.

Aplasia cutis congenita (ACC) is a rare inborn lesion, presenting with absence of skin. The etiology is unknown and is probably not attributable to a single cause but to a combination of genetic factors. Multiple causes have been suggested for ACC: syndromes and teratogens, intrauterine infection--varicella zoster virus, herpes simplex virus--fetal exposure to cocaine, heroin, alcohol, or antithyroid drugs. The most common site is the scalp. We report a case with multiple lesions on the trunk, resembling an instance with ACC group 5. This form of ACC occurs in association with the in utero death of a twin or more (in this case triple) fetus. Histological findings are available in very few reports. Therapy options depend on the characteristics of the lesion, but conservative treatment is usually chosen.

Matrix metalloproteinases and E-cadherin immunoreactivity in different basal cell carcinoma histological types.

The immunohistochemical staining of matrix metalloproteinases (MMPs) and E-cadherin in tumor epithelial and stromal cells was analyzed in a group of solid, superficial spreading and cystic tumors and in a group of morpheaform and recurrent basal cell carcinomas (BCC) in order to determine whether any of these factors possibly contribute to tumor therapy resistance. Tumor tissues of 64 patients were obtained by complete excisional or curettage biopsy of BCC and these were immunohistochemically stained for MMP-1, MMP-2, MMP-9, MMP-13 and E-cadherin. In the morpheaform and recurrent BCC, MMP-9 expression significantly increased in the stroma, while E-cadherin expression was negative in epithelial cells. Odds ratio for development of morpheaform and recurrent BCC was 6.2 for positive MMP-1 immunostaining in epithelial tumor cells, 5.8 for positive MMP-9 immunostaining in tumor stroma, 3.2 for positive MMP-13 immunostaining in tumor stroma, and 4.5 for negative E-cadherin in epithelial tumor cells. Our results suggest that MMP-1 immunostaining in tumor cells, MMP-9 expression in stromal cells, and absence of E-cadherin expression are associated with morpheaform and recurrent BCC.

[Prognostic value of ophthalmic artery color Doppler sonography for progression to glaucoma in vitiligo patients]. / Prognosticka vrijednost ultrazvucnog nalaza arterije oftalmike u bolesnika s vitiligom za nastanak glaukoma.

The aim of this study was to assess the prognostic value, i.e. the role of hemodynamic changes in the ophthalmic artery determined by color Doppler imaging in patients with vitiligo in relation to the occurrence of primary open-angle glaucoma (POAG), and to call attention to the importance of examination of the vascular component in routine examinations of patients by clinical dermatologists and ophthalmologists, due to the possible correlation of vitiligo and ophthalmologic changes indicating POAG. Clinical observations and research have shown that patients treated for vitiligo are often diagnosed with POAG or have confirmed diagnosis in their medical history. These findings motivated the research of the vascular component as a possible correlation between vitiligo and POAG. The vascular component of POAG and its role in the dynamics of glaucoma progression has not been fully explained and is still somewhat controversial, and the resulting ischemic disorders are primarily related to optical nerve damage. During the course of a continuous two-year research started in March 2008 and completed in March 2010, a total of 42 patients with vitiligo were examined in the specialist dermatovenereology unit of the Department of Skin Diseases and Sexually Transmitted Diseases. Following clinical examination and Wood's lamp examination, patients with suspected glaucoma were referred to the glaucoma unit of the Department of Ophthalmology to detect or check the progression of POAG. At the Department of Internal Medicine, Split University Hospital Center, color Doppler imaging of ophthalmic artery was performed in all patients on both eyes to examine its hemodynamics. All study participants underwent complete ophthalmologic examination: visual acuity, fundus of the eye, eye pressure measurement, anterior eye segment biomicroscopy with gonioscopy and visual field. In conclusion, our study showed color Doppler imaging and investigation of vascular parameters in ophthalmic artery to provide an insight into the vascular function in differential analysis and can be determined as a correlation or potential risk factor related to vitiligo and POAG. For this reason, we suggest that patients treated for vitiligo, irrespective of their age, sex, affected area, localization and duration of the disease, should regularly undergo complete ophthalmologic examination with special emphasis on POAG diagnosis. Considering the relatively limited sample and trial period, we can already substantiate and confirm that the correlation is not random. Future research in a larger number of vitiligo and POAG patients should provide definitive answer concerning correlation between the two diseases and lead to final conclusion, along with investigation of other hematologic factors related to blood hypercoagulability in order to reach better understanding of the vascular component function.

[Guidelines for the diagnosis and treatment of psoriasis]. / Smjernice za dijagnostiku i lijecenje vulgarne psorijaze.

Croatian dermatovenerological society of the Croatian Medical Association formed the working group which consists of leading experts for psoriasis in Croatia. After a critical analysis of relevant scientific papers, the working group has developed guidelines for the diagnosis and treatment of psoriasis.

Differences in epidermal thickness and expression of apoptosis regulatory proteins in the skin of patients with chronic renal failure and pruritus.

Chronic renal failure is often associated with skin itching (pruritus) in dialysis patients. In order to investigate the possible causes of pruritus, the epidermis of the thigh of 12 dialysis patients and 4 controls from patients without renal disease were examined. The sections of the epidermis were measured and immunohistochemically analyzed using antibodies to Bcl-2, Bax, caspase-3 proteins and TUNEL method. While the mean thickness of normal epidermis was 53 µm, in dialysis patients it ranged between 23 and 34 µm during the 3-5 year period on dialysis. Compared to normal skin, the fine balance between the Bcl-2 and Bax proteins did not greatly change in the epidermis of dialysis patients during the three years of dialysis. Following five-year dialysis, the epidermis displayed increased Bax and decreased Bcl-2 expression in the basal and intermediate epidermal layers, as well as the presence of apoptotic cells (TUNEL and caspase-3 positive) both in the superficial and intermediate epidermal layers. Our study demonstrated the predominant expression of cell death Bax proteins over cell survival Bcl-2 proteins, and apoptotic cells in the deeper layers of the epidermis in patients on long-term dialysis. We speculate that the thinning of the epidermis might be associated with the appearance of dead cells in the deeper epidermal layers, while the changed internal milieu of epidermal cells could possibly affect the intra-epidermal nerve endings thus leading to the sensation of pruritus.

Descriptive epidemiology of cutaneous squamous cell carcinoma in Croatia.

The aim of the study was to investigate the squamous cell carcinoma (SCC) incidence in Croatia in the 2003-2005 period. The cases of SCC were retrospectively studied. Data were collected from University Department of Dermatology and Venereology, Zagreb University Hospital Center and National Cancer Registry. In the study period, there were 1,860 cases of SCC (934 men and 926 women). The crude incidence rate for the Croatian population of 100,000 was 14.6 for men and 13.4 for women. The age-standardized incidence rate (adjusted for the world standard population) was 8.9 for men and 5.2 for women. The head was almost exclusive localization of SCC in both sexes. The highest SCC incidence was recorded in Zadar County. These results will serve for the SCC trend monitoring in Croatia and Europe in the forthcoming years.

Modern approach to topical treatment of aging skin.

The main processes involved in skin aging are intrinsic and extrinsic. Apart from them, so called stochastic aging connotes cell damage caused by metabolic processes, free radicals and cosmic irradiation. The clinical expression of intrinsic aging include smooth, dry, and thinned skin with accentuated expression lines. It is inevitable and time dependent. Extrinsically aged skin shows signs of photodamage which include appearance of wrinkles, pigmented lesions, actinic keratoses and patchy hypopigmentations. Therapeutic modalities imply photoprotection with sunscreens that prevent sunburns and block ultraviolet irradiation. Other modalities include use of retinoids which regulate gene transcription with subsequent cellular differentiation and proliferation. The topical and peroral administration of network antioxidants, such as vitamin E and C, coenzyme Q10, alpha-lipoic acid and glutathione, enhance antiaging effect. The other antioxidants such as green tea, dehydroepiandrosterone, melatonin, selenium and resveratrol, have also antiaging and anti-inflammatory effects. Topical bleaching agents such as hydroquinone, kojic acid and azelaic acid can reduce signs of aging. Studies confirm the efficacy of these topical agents in combination with superficial and/or medium depth or deep peeling agents for photodamaged skin treatment. Indications for type of chemical peels according to various clinical diagnosis are done, as well as advantages and disadvantages of different types of chemical peels.

Robust demarcation of basal cell carcinoma by dependent component analysis-based segmentation of multi-spectral fluorescence images.

This study was designed to demonstrate robust performance of the novel dependent component analysis (DCA)-based approach to demarcation of the basal cell carcinoma (BCC) through unsupervised decomposition of the red-green-blue (RGB) fluorescent image of the BCC. Robustness to intensity fluctuation is due to the scale invariance property of DCA algorithms, which exploit spectral and spatial diversities between the BCC and the surrounding tissue. Used filtering-based DCA approach represents an extension of the independent component analysis (ICA) and is necessary in order to account for statistical dependence that is induced by spectral similarity between the BCC and surrounding tissue. This generates weak edges what represents a challenge for other segmentation methods as well. By comparative performance analysis with state-of-the-art image segmentation methods such as active contours (level set), K-means clustering, non-negative matrix factorization, ICA and ratio imaging we experimentally demonstrate good performance of DCA-based BCC demarcation in two demanding scenarios where intensity of the fluorescent image has been varied almost two orders of magnitude.

Vitiligo and glaucoma - an association or a coincidence? A pilot study.

Our aim was to draw attention of clinicians, dermatovenereologists and ophthalmologists to the possible association of vitiligo and ocular findings characteristic of primary open angle glaucoma (POAG). According to our clinical experience, POAG is relatively often diagnosed or previously diagnosed in patients treated for vitiligo. We found no literature report directly connecting vitiligo and POAG. The articles mentioning glaucoma, i.e. increased intraocular pressure (IOP), are mainly based on secondary type of glaucoma as a direct consequence of vitiligo treated by corticosteroids. On the other hand, there is a substantial number of articles elaborating the association of vitiligo with ocular findings in known clinical syndromes. During a 2-year period, from May 2007 to May 2009, 42 patients with vitiligo were examined at University Department of Dermatology and Venereology and referred for previously appointed ophthalmologic examination at University Department of Ophthalmology, Split University Hospital Center. All patients with vitiligo underwent complete ophthalmologic examination including visual field analysis, in order to treat and follow up or diagnose newly discovered POAG. Of 42 patients with vitiligo suspect of glaucoma, POAG was confirmed in 24 (57%) patients. Age median of all vitiligo patients was 56 (range 19-82) years. In patients with vitiligo and glaucoma, the duration of vitiligo was twofold that recorded in patients with vitiligo alone, the difference being statistically significant (z=3.3; P<0.001). The risk of developing glaucoma in vitiligo patients was 4.4-fold in >56 age group versus <56 age group and 3.5-fold in patients with >13 year versus patients with <13 year duration of vitiligo. Multivariate logistic regression for glaucoma development according to vitiligo duration (< or =13 years vs. > or =13 years) and patient age (< or =56 years vs. > or =56 years) pointed to the association of glaucoma development and age and yielded a 92% probability for the association of glaucoma development and vitiligo duration. Therefore, we believe that patients treated for vitiligo should regularly undergo complete ophthalmologic examination with special attention paid to POAG irrespective of age, sex, severity, localization and duration of the disease. Although performed in a relatively small sample and over short period of time, the results of this pilot study demonstrated that this association was not accidental. Additional studies in a greater sample of vitiligo patients and POAG are expected to provide definitive answers and conclusion on the association of these two diseases.

A rare case of pulmonary tuberculosis with simultaneous pulmonary and skin sarcoidosis: a case report.

BACKGROUND: Tuberculosis and sarcoidosis are chronic diseases that rarely occur concomitantly. Sarcoidosis is a multisystem granulomatous disorder characterized pathologically by the presence of non-caseating granulomas in involved tissues. Tuberculosis is infectious disease caused by Mycobacterium tuberculosis characterized by granulomas with caseous necrosis. CASE PRESENTATION: We present a case of 43-year-old female refugee from Kosovo with microbiological confirmation of pulmonary tuberculosis and pulmonary and skin sarcoidosis at the same time. Three weeks after corticosteroid therapy for pulmonary sarcoidosis was introduced, positive finding of mycobacterium culture of bronchial aspirate was observed. Based on these results, corticosteroid therapy was excluded and antituberculous therapy was introduced for six months. In the meantime, new nodes on face and nose appeared and skin sarcoidosis was diagnosed. The patient was given corticosteroids and colchicine according to the skin and pulmonary sarcoidosis therapy recommendation. CONCLUSION: The authors of this study suggest that in cases when there is a dilemma in diagnosis between tuberculosis and sarcoidosis we should advance with corticosteroid therapy until we have microbiological confirmation of mycobacterium culture. This case is remarkable because this is a third described case of sarcoidosis and tuberculosis together (the first reported in Asia, the second in South Africa), and to authors knowledge, this is a first case report in Europe.

The incidence of basal cell carcinoma in Croatia: an epidemiological study.

The aim of the study was to investigate the basal cell carcinoma (BCC) incidence in Croatia in the 2003-2005 period. Data were collected from University Department of Dermatology and Venereology, Zagreb University Hospital Center and National Cancer Registry. The age-specific incidence rate and age-standardized incidence rate were calculated per 100,000 inhabitants according to the latest population census in Croatia from 2001. In the study period, there were 7,244 BCC cases (3,519 men and 3,725 women) in Croatia. The crude incidence rate for the Croatian population of 100,000 was 54.9 for men and 53.9 for women. The age-standardized incidence rate (adjusted for the world standard population) was 33.6 for men and 24.5 for women. The head and neck were almost exclusive localizations of BCC. The highest BCC incidence was recorded in Zadar County. The incidence of BCC was high in both littoral and inland counties of Croatia. Study results will serve as reference figures on studying the trend of BCC incidence in Croatia and Europe in the forthcoming years.

Skin aging.

There are two main processes that induce skin aging: intrinsic and extrinsic. A stochastic process that implies random cell damage as a result of mutations during metabolic processes due to the production of free radicals is also implicated. Extrinsic aging is caused by environmental factors such as sun exposure, air pollution, smoking, alcohol abuse, and poor nutrition. Intrinsic aging reflects the genetic background and depends on time. Various expressions of intrinsic aging include smooth, thinning skin with exaggerated expression lines. Extrinsically aged skin is characterized by photo damage as wrinkles, pigmented lesions, patchy hypopigmentations, and actinic keratoses. Timely protection including physical and chemical sunscreens, as well as avoiding exposure to intense UV irradiation, is most important. A network of antioxidants such as vitamins E and C, coenzyme Q10, alpha-lipoic acid, glutathione, and others can reduce signs of aging. Further anti-aging products are three generations of retinoids, among which the first generation is broadly accepted. A diet with lot of fruits and vegetables containing antioxidants is recommended as well as exercise two or three times a week.

An overview of Bcl-2 expression in histopathological variants of basal cell carcinoma, squamous cell carcinoma, actinic keratosis and seborrheic keratosis.

The Bcl-2 protein has been shown to suppress cell death and protects cell against apoptosis induced by different death-inducing signals. In this study the authors have analyzed imunohistochemically the expression of Bcl-2 protein in the histopathological variants of the most common malignant tumors of the skin--basal cell carcinoma (BCC) and squamous cell tumor (SCC), as well as in the precancerous lesion actinic keratosis (AK) and in benign tumor seborrheic keratosis (SK). Bcl-2 expression in solid, adenoid and cystic variants of BCC exhibited immunoreactivity of tumor stroma with more intense staining among peripheral palisading cells. Morphoeic variant demonstrated reduced amount of Bcl-2 expression. Among SCC in all samples, tumor tissue lack to express Bcl-2 positivity. In cases of hypertrophic and atrophic variants of AK, Bcl-2 expression was confined to basal cell layer, as well as in one case of hypertrophic variant in suprabasal cells. In three histological variants of SK expresseion of Bcl-2 protein was in areas of basaloid proliferation, while in areas of squamous differentiation was negative. In clonal variant immunostaining was positive among cells in characteristic "nests" Distribution of Bcl-2 protein expression in solid, adenoid and cystic variant of BCC showed that peripheral proliferating cells are protected against apoptosis what permits tumor growth. In morpheaform variant reduced amount of Bcl-2 expression indicated that this variant of BCC has increased cell proliferation, and in practice shows tendency for recurrence and difficulties to eradicate. Bcl-2 expression supports the observation that tumor cells are derived from basal keratinocytes. In SCC, lack of Bcl-2 expression indicates that origin of tumor cells is from more differentiated suprabasal keratinocytes. In AK results suggest that immunoreactivity is regulated with respect of the keratinocyte's differentiation status, but not closely correlate with proliferative rate.

Fractionated illumination improves the outcome in the treatment of precancerous lesions with photodynamic therapy.

Photodynamic therapy (PDT) is a noninvasive therapy for non-melanoma skin cancer. The aim of this study was comparison of efficacy between fractioned versus single dose illumination in photodynamic therapy (PDT) of actinic keratosis (AK) and Bowen's disease (BD). Fifty-one patients (36 AK and 15 BD) were treated with PDT They were randomly arranged in two treatment groups. Group one included 26 patients (20 AK and 6 BD) that, after five hours of incubation with 20% 5-ALA, were treated with a single illumination of 100 Jcm(-2) at fluence rate of 30 mWcm(-2). Group two included 25 patients (16 AK and 9 BD) that, after 16 hours of incubation with 20% 5-ALA, were treated with two light fractions (50 plus 50 Jcm(-2)) at same fluence rate with dark interval of two hours between fractions. Twenty-four weeks later, a treated area was incubated for four hours again with 5-ALA in order to detect occult areas of abnormal skin with possible remaining tumor tissue. In case of fluorescence, histological examination was performed. In the group one, fluorescence at the end of the session was absent in 19 (73%) or very weak in 7 (27%). Residual tumor was found in 15 (75%) AK and in 4 (66.6%) BD. In the group two, fluorescence at the end of second session was more intense; in one patient (4%) was absent, very weak in 5 (20%) and weak in 19 (76%) of patients. In this group histology revealed remaining tumor tissue in only 2 (12.5%) AK and 2 (22.2%) BD. Among the patients in the first group, the remaining tumor tissue was significantly bigger (p=0.005). The treatment response with clearing of tumor tissue was significantly higher in fractionated illumination than in a single dose illumination group. Fractionated illumination scheme with 16 hours of incubation separated by two hours dark interval significantly improves the therapeutic outcome in tumor eradication.

De novo NEMO gene deletion (delta4-10)--a cause of incontinentia pigmenti in a female infant: a case report.

Incontinentia pigmenti (IP) is a rare, inherited, multisystem genodermatosis. It is transmitted as an X-linked dominant trait. The disorder is a consequence of mutations in the NEMO gene (Xq28) that completely abolish expression of the NF-kappaB essential modulator. Here we present a female infant of healthy nonconsanguinous, young parents with a clinically evident first phase of IP. PCR analysis of patient's peripheral blood lymphocytes DNA was done for detection of NEMO delta4-10 deletion. Skin changes present at birth appertain to first inflammatory stage. However, a pathohistological feature of the skin biopsy showed second phase of disease. Genetic testing of diseased child revealed delta4-10 in NEMO gene. However, the assumption that the female child has familial IP was rejected as PCR performed on the mother's leukocytes did not record the presence of the same mutation. Moreover, the existence of a healthy male infant of the same mother as well as the lack of any phenotypic signs of the disease in other family members additionally support that IP was not inherited, but it was a consequence of de novo NEMO gene mutation. In conclusion, here we describe a Croatian female with clinical IP phenotype having de novo genomic rearrangements in the NEMO gene.

Hla-Cw7 allele as predictor of favorable therapeutic response to interferon-alpha in patients with chronic hepatitis C.

AIM: To evaluate the association between human leukocyte antigens (HLA) class I and therapeutic response to interferon-alpha in Croatian patients with chronic hepatitis C. METHODS: HLA-A, -B, and -C genotyping was performed in 55 patients with sustained virological response and in 57 patients without sustained virological response to interferon-alpha therapy. Patients were treated in the period from 1998-2001 with interferon-alpha at a dose of 3 million units three times a week. Patients who became negative for hepatitis C virus RNA after 12 weeks of therapy completed 48 weeks of therapy. RESULTS: There was no association between therapeutic outcome and frequency of HLA-A, as well as of HLA-B alleles. HLA-Cw7 was significantly more frequent in patients with than those without sustained virological response (27.0% vs 6.7%; P=0.011). CONCLUSION: In Croatian patients with chronic hepatitis C, HLA-Cw7 is the predictor of sustained virological response to interferon-alpha therapy.