[Show abstract][Hide abstract] ABSTRACT: Reflectance confocal microscopy (RCM) represents a noninvasive imaging technique that has previously been used for characterization of mycosis fungoides (MF) in a pilot study. We aimed to test the applicability of RCM for diagnosis and differential diagnosis of MF in a clinical study. A total of 39 test sites of 15 patients with a biopsy-proven diagnosis of either MF, parapsoriasis, Sézary syndrome, or lymphomatoid papulosis were analyzed for presence and absence of RCM features of MF. Cochran and Chi(2) analysis were applied to test the concordance between investigators and the distribution of RCM features, respectively. For selected parameters, the Cochran analysis showed good concordance between investigators. Inter-observer reproducibility was highest for junctional atypical lymphocytes, architectural disarray, and spongiosis. Similarly, Chi(2) analysis demonstrated that selected features were present at particularly high frequency in individual skin diseases, with values ranging from 73% to 100% of all examined cases.
Full-text · Article · Jan 2012 · Journal of Biomedical Optics
[Show abstract][Hide abstract] ABSTRACT: Histopathologically, basal cell carcinoma (BCC) is characterized by basaloid tumor nodules of varying size showing peripheral palisading of cells and nuclei, and separation from surrounding stroma by optically empty appearing clefts. These are usually regarded as an artifact, occurring during routine tissue processing. Recently, reflectance confocal microscopy (RCM) has been applied for noninvasive, in vivo evaluation of BCC. Besides other features, small areas of low refractility separating tumor islands from the surrounding tissue can be observed in vivo, suggesting that the presence of amorphous material like mucin might be the causal factor for these clefts.
A total of 13 BCCs were studied by RCM and histopathological techniques. Staining was performed with Alcian blue for the detection of peritumoral mucin. Correlation between RCM images and histopathological samples was studied, and the diameter of hyporefractile areas on RCM as well as the thickness of peritumoral mucin was measured.
Good correlation was seen between dark areas on RCM and thickness of peritumoral mucin with a mean diameter of 14 µm (RCM) and 11.44 µm (histopathology), respectively. Pearson correlation coefficient was 0.605 (p < 0.0001).
Our results show that the peritumoral cleft-like spaces seen in BCC on histopathology exist in vivo, and correspond to the peritumoral mucin deposition.
Full-text · Article · Oct 2010 · Journal of Cutaneous Pathology
[Show abstract][Hide abstract] ABSTRACT: In vivo reflectance confocal microscopy (RCM) has been used for evaluation of the morphologic features of nonmelanoma skin cancer. The application of RCM for diagnosis of basal cell carcinoma has been reported; however, the evaluation of actinic keratoses (AKs) has only been the subject of preliminary studies.
The goal of this study was to evaluate the applicability of RCM in the diagnosis of AK in correlation with routine histology.
Forty-four Caucasians with a minimum of one AK participated in this study. Evaluation consisted of clinical examination, RCM, and routine histology, including a total of 46 AKs in the final analysis. Ten normal skin sites served as controls. RCM features of AK included parakeratosis, architectural disarray, and keratinocyte pleomorphism. Following blinded evaluations, sensitivity/specificity, kappa analysis, and Spearman's correlation were performed on all parameters.
Sensitivity/specificity values of RCM features ranged from 80% to 98.6%. The presence of architectural disarray and cellular pleomorphism appeared to be the best predictor of AK.
In summary, RCM may be a promising technology for the noninvasive detection of AK and as adjunct tool to clinical diagnosis and monitoring. However, the preliminary nature of this study warrants further investigations.
No preview · Article · Jun 2008 · Dermatologic Surgery
[Show abstract][Hide abstract] ABSTRACT: The first choice treatment for vitiligo vulgaris is narrow-band UVB (NB-UVB), but no satisfactory treatment exists.
To investigate if Polypodium leucotomos, an antioxidative and immunomodulatory plant extract, improves NB-UVB-induced repigmentation.
Fifty patients with vitiligo vulgaris randomly received 250 mg oral P. leucotomos or placebo three times daily, combined with NB-UVB twice weekly for 25-26 weeks.
Repigmentation was higher in the P. leucotomos group vs. placebo in the head and neck area (44% vs. 27%, P = 0.06). Small repigmentation increases (P = n.s.) were observed for the trunk (6% increased repigmentation), extremities (4%), and hands and feet (5%) in the P. leucotomos group vs. placebo. Patients attending more than 80% of required NB-UVB sessions showed increased repigmentation in the head and neck area in the P. leucotomos group vs. placebo (50% vs. 19%, P < 0.002); no significant differences were seen in the other body areas. Patients with skin types 2 and 3 showed more repigmentation in the head and neck area in the P. leucotomos group vs. placebo (47% vs. 21%, P = 0.01), and no significant differences were seen in the other body areas. No conclusions could be drawn on skin types 4 and 5 due to low patient numbers.
There is a clear trend towards an increase in repigmentation of vitiligo vulgaris affecting the head and neck area when NB-UVB phototherapy is combined with oral P. leucotomos. This effect may be more pronounced in light skin types.
No preview · Article · Aug 2007 · Journal of the European Academy of Dermatology and Venereology
[Show abstract][Hide abstract] ABSTRACT: Angiogenesis, resulting from an imbalance between angiogenic activator factors and inhibitors, is required for tumour growth and metastasis. The determination of the circulating concentration of all angiogenic factors (activators and inhibitors) is not feasible at present. We have evaluated diagnostic and prognostic values of the measurement of serum angiogenic activity in colorectal carcinoma (CRC) patients. Serum proliferative activity (PA) on human umbilical vein endothelial cells (HUVEC) in vitro, and serum vascular endothelial growth factor (VEGF) levels were determined by ELISA in 53 patients with primary CRC, 16 subjects with non-neoplastic gastrointestinal disease (SC) and 34 healthy individuals. Data were compared with clinical outcome of the patients. Although serum from CRC patients significantly increased the PA of HUVEC, compared to culture control (HUVEC in medium + 10% foetal bovine serum (FBS); P < 0.001); our results indicate that serum PA in CRC patients was similar to that of SC or healthy individuals. There was no correlation between serum PA and circulating VEGF concentrations. Surgery produced a decrease of PA at 8 hrs after tumour resection in CRC patients compared to pre-surgery values (186 +/- 47 versus 213 +/- 41, P < 0.001). However, an increase in serum VEGF values was observed after surgery (280 [176-450] versus 251 [160-357] pg/ml, P = 0.004). Patients with lower PA values after surgery showed a worse outcome that those with higher PA values. Therefore, this study does not support a diagnostic value for serum angiogenic activity measured by proliferative activity on HUVEC but suggests it could have a prognostic value in CRC patients.
Preview · Article · Mar 2007 · Journal of Cellular and Molecular Medicine
[Show abstract][Hide abstract] ABSTRACT: Lutein and zeaxanthin are xanthophyll carotenoids with potent antioxidant properties protecting the skin from acute photodamage. This study extended the investigation to chronic photodamage and photocarcinogenesis. Mice received either a lutein/zeaxanthin-supplemented diet or a standard nonsupplemented diet. Dorsal skin of female Skh-1 hairless mice was exposed to UVB radiation with a cumulative dose of 16,000 mJ/cm(2) for photoaging and 30,200 mJ/cm(2) for photocarcinogenesis. Clinical evaluations were performed weekly, and the animals were sacrificed 24 h after the last UVB exposure. For photoaging experiments, skin fold thickness, suprapapillary plate thickness, mast cell counts and dermal desmosine content were evaluated. For photocarcinogenesis, samples of tumors larger than 2 mm were analyzed for histological characterization, hyperproliferation index, tumor multiplicity, total tumor volume and tumor-free survival time. Results of the photoaging experiment revealed that skin fold thickness and number of infiltrating mast cells following UVB irradiation were significantly less in lutein/zeaxanthin-treated mice when compared to irradiated animals fed the standard diet. The results of the photocarcinogenesis experiment were increased tumor-free survival time, reduced tumor multiplicity and total tumor volume in lutein/zeaxanthin-treated mice in comparison with control irradiated animals fed the standard diet. These data demonstrate that dietary lutein/zeaxanthin supplementation protects the skin against UVB-induced photoaging and photocarcinogenesis.
No preview · Article · Feb 2007 · Skin pharmacology and physiology
[Show abstract][Hide abstract] ABSTRACT: This study was undertaken to investigate the ethnic susceptibility to irritant contact dermatitis induced by a common dishwashing liquid using noninvasive technologies.
A total of 30 participants (15 Caucasian, 15 African American) were patch tested to graded concentrations of a common household irritant and evaluated using clinical scoring, reflectance confocal microscopy, transepidermal water loss, and fluorescence excitation spectroscopy.
At 24 hours, the concentration thresholds for clinically perceptible irritancy were significantly higher for African American compared with Caucasian participants. Reflectance confocal microscopy showed stratum corneum disruption, parakeratosis, and spongiosis; these features were more severe in Caucasian participants (P < or = .002). Mean values for transepidermal water loss were significantly higher in the Caucasian group at comparable clinical scores (P < or = .005). Fluorescence excitation spectroscopy showed a broad excitation band at 300 nm (emission 340 nm) and values in both groups returned to baseline by day 7.
This pilot study was limited in scope and larger studies are needed to further evaluate ethnic differences in irritant contact dermatitis and to demonstrate the applicability of our findings for other irritants.
Clinical evaluation, reflectance confocal microscopy, and transepidermal water loss showed significant differences in the cutaneous irritant response between both groups suggesting a superior barrier function of African American skin. Fluorescence excitation spectroscopy on the other hand demonstrated no differences in the hyperproliferative response after irritant exposure and indicated similar kinetics for the two groups.
No preview · Article · Mar 2006 · Journal of the American Academy of Dermatology
[Show abstract][Hide abstract] ABSTRACT: The validity, reproducibility, and specificity of patch-testing in the diagnosis of allergic contact dermatitis (ACD) have repeatedly been addressed. In vivo reflectance confocal microscopy (RCM) has been used for real-time evaluation of the histopathologic features of ACD. This pilot study was designed to determine the sensitivity and specificity of RCM in diagnosing ACD in reference to patch-testing. Sixteen participants were patch tested with allergens and control substances. Clinical scoring, digital photography, and RCM evaluation were performed at 72 hours, and RCM images were subjected to blinded evaluation. RCM evaluation parameters included stratum corneum (SC) disruption, parakeratosis, stratum spinosum (SS) and stratum granulosum (SG) spongiosis, and exocytosis. Overall, there was high specificity for all RCM features, ranging from 95.8% to 100%. Sensitivity ranged from 51.9% to 96.3%. Significant parameters with high sensitivity and specificity included spongiosis and exocytosis at the level of SS. Logistic regression analysis was performed on significant variables; P values were determined by chi2 analysis. RCM is a promising noninvasive technology for the evaluation of ACD. SC changes are not helpful in the diagnosis of ACD, although the presence of SG spongiosis and SS spongiosis shows high sensitivity in diagnosing ACD. Larger sensitivity and specificity studies are needed and the identification of ACD has to be based on a defined diagnostic algorithm. A limitation of this study is the small sample size; larger sensitivity and specificity studies are needed to confirm these findings. In addition, individual allergens have to be subjected to further evaluations in order to demonstrate the applicability of our findings for other contact allergens. In that regard, RCM may be considered as an adjunctive tool, rather than a substitute, to clinical evaluation.
No preview · Article · Jan 2006 · Journal of the American Academy of Dermatology
[Show abstract][Hide abstract] ABSTRACT: Laser treatment of port-wine stain (PWS) might be improved using a deeply penetrating wavelength.
PWSs in 17 patients were treated 3 times with a 595-nm pulsed dye laser (PDL) and a 1064-nm neodymium:yttrium-aluminum-garnet (Nd:YAG) laser. Fluences of 1.0, 0.8, and 0.6 times the minimum purpura dose (MPD) were used for Nd:YAG laser. Posttreatment biopsy specimens were taken. Blind assessment and quantitative analysis of PWS clearing were performed from digital photographs.
MPD for Nd:YAG laser varied widely, from 40 to 250 J/cm 2 . Purpura lasted longer after PDL. Treatment achieved similar 50% to 75% clearing with both PDL and Nd:YAG laser at 1 MPD. Nd:YAG caused greater perivascular and epidermal injury. Scarring occurred in the only patient treated with a Nd:YAG fluence greater than 1 MPD. Patients preferred Nd:YAG laser because of their faster recovery.
Nd:YAG laser used at MPD is as effective as PDL for treating PWS. Nd:YAG laser fluences higher than MPD may cause scarring.
Full-text · Article · Apr 2005 · Journal of the American Academy of Dermatology
[Show abstract][Hide abstract] ABSTRACT: Reflectance confocal microscopy (RCM) allows non-invasive visualization of human skin in vivo. It has been used to describe the histopathological features of acute contact dermatitis (CD). This work was designed to investigate the kinetics of both allergic and irritant CD (ACD and ICD) in vivo. Eighteen subjects with a prior diagnosis of ACD were patch tested with the specific allergen sodium lauryl sulfate as an irritant, and appropriate controls. RCM, transepidermal water loss (TEWL), and fluorescence excitation spectroscopy (FES) were performed at several time points within 2 wk after patch removal. After removal of the Finn chambers at 48 h, superficial epidermal changes, primarily involving the stratum corneum, and increased epidermal thickness were mainly present in ICD. ACD, on the other hand, showed microvesicle formation peaking at 96 h following patch removal. Both ACD and ICD showed exocytosis and similar degrees of spongiosis on RCM. TEWL and FES demonstrated a significant difference between ACD and ICD. RCM, TEWL, and FES are valuable non-invasive tools to quantitatively study the kinetics of the pathophysiology of acute CD reactions in vivo and monitor the changes at a cellular level.
Full-text · Article · Mar 2005 · Journal of Investigative Dermatology
[Show abstract][Hide abstract] ABSTRACT: The current standard diagnostic procedure for basal cell carcinoma (BCC) is histologic examination after invasive biopsy. Reflectance-mode confocal microscopy (RCM) offers noninvasive high-resolution imaging of human skin in vivo.
The objective of this study was to explore the sensitivity and specificity of RCM for diagnosis of BCC.
This was a retrospective study of RCM images from 4 institutions of 152 skin lesions representing a variety of benign and malignant diagnoses. These 152 lesions were examined clinically, with biopsies recorded for all the 83 BCCs detected. Based on a previous study, a set of 5 histologically correlated confocal imaging criteria for diagnosing BCC was established, eg, the presence of elongated monomorphic nuclei. Blinded retrospective analysis of the images from the 152 lesions was carried out by a single novice reviewer to determine the sensitivity and specificity of these 5 RCM criteria for diagnosing BCC. The accuracy of combining the probability of BCC based on examination of clinical photographs with the predicted probability of BCC based on confocal criteria was also evaluated.
The presence of two or more criteria is 100% sensitive for the diagnosis of BCC, and with 4 or more RCM criteria present the specificity was 95.7% and sensitivity was 82.9%. These results were found to have little variability across study sites and across BCC subtypes. The combination of RCM with photography-based predictions of clinical probability of BCC significantly improved the accuracy for noninvasive diagnosis of BCC.
RCM offers a sensitive and specific tool for the noninvasive diagnosis of BCC in vivo.
No preview · Article · Jan 2005 · Journal of the American Academy of Dermatology
[Show abstract][Hide abstract] ABSTRACT: UV radiation induces damage to human skin. Protection of skin by an oral photoprotective agent would have substantial benefits. Objective We investigated the photoprotective effect of oral administration of an extract of the natural antioxidant Polypodium leucotomos (PL).
A total of 9 healthy participants of skin types II to III were exposed to varying doses of artificial UV radiation without and after oral administration of PL (7.5 mg/kg). At 24 hours after exposure the erythema reaction was assessed and paired biopsy specimens were obtained from PL-treated and untreated skin.
A significant decrease in erythema was found in PL-treated skin (P < .01). Histologically, PL-treated biopsy specimens showed less sunburn cells (P < .05), cyclobutane pyrimidine dimers (P < .001), proliferating epidermal cells (P < .001), and dermal mast cell infiltration (P < .05). A trend toward Langerhans cell preservation was seen.
Oral administration of PL is an effective systemic chemophotoprotective agent leading to significant protection of skin against UV radiation.
Full-text · Article · Jan 2005 · Journal of the American Academy of Dermatology
[Show abstract][Hide abstract] ABSTRACT: Acute irritant contact dermatitis (ICD) and allergic contact dermatitis (ACD) may be difficult to distinguish by clinical or histologic assessment. Reflectance confocal microscopy (RCM) enables real-time, high-resolution skin imaging in vivo.
We sought to image, characterize, and distinguish acute ACD and ICD in vivo.
Volunteers with ACD were patch tested with an allergen and the irritant, sodium lauryl sulfate. RCM imaging and transepidermal water loss measurements were performed at 24 and 72 hours. Biopsy specimens were correlated with RCM images.
Spongiosis, epidermal inflammatory cell infiltrate, and vesicle formation were observed in ACD and ICD. Compared with ACD, ICD showed greater disruption of the stratum corneum, and more parakeratosis. There was a significantly greater increase in transepidermal water loss for ICD compared with ACD.
RCM is a promising tool for dynamic, noninvasive assessment and may help to differentiate acute ACD and sodium lauryl sulfate-induced ICD.
Full-text · Article · Mar 2004 · Journal of the American Academy of Dermatology
[Show abstract][Hide abstract] ABSTRACT: The use of psoralen-UVA (PUVA) in patients of skin phototype I to II is limited by side effects of acute phototoxicity and possible long-term carcinogenesis.
We sought to assess oral Polypodium leucotomos (PL) extract in decreasing PUVA-induced phototoxicity of human skin on a clinical and histologic level.
A total of 10 healthy patients with skin phototypes II to III were exposed to PUVA alone (using 0.6 mg/kg oral 8-methoxypsoralen) and to PUVA with 7.5 mg/kg of oral PL.
Clinically, phototoxicity was always lower in PL-treated skin after 48 to 72 hours (P<.005), and pigmentation was also reduced 4 months later. Histologically, PL-treated skin showed a significant numeric reduction of sunburn cells (P=.05), preservation of Langerhans cells (P< or =.01), decrease of tryptase-positive mast cell infiltration (P<.05), and decrease of vasodilation (P< or =.01). No differences were found in Ki-67+ proliferating cells.
PL is an effective chemophotoprotector against PUVA-induced skin phototoxicity and leads to substantial benefits of skin protection against damaging effects of PUVA as evidenced by histology.
Full-text · Article · Jan 2004 · Journal of the American Academy of Dermatology