A simple morphometric analysis method for dermal microstructure using color thresholding and moments.

BACKGROUND: Proper assessment of dermal collagen fibers by dermatologists and researchers is essential. Histologic evaluation methods have limitations. We present a simple method for measurement of collagen fibers in human skin using Masson's trichrome staining. MATERIALS AND METHODS: Normal skin specimens from a cadaver were processed with Masson's trichrome, which can effectively stain collagen fibers blue with aniline dye. Optical photomicrographs of these slides were analyzed using ImageJ software. Color image processing, a histogram-based function of ImageJ for image segmentation, was performed with color moments thresholding technique. We selected blue areas by adjusting the blue channel to include specific values. The selected areas were highlighted and evaluated. We divided the image into layers of 0.09-mm2 areas from the top to bottom of the dermis. Each area was cropped and evaluated. RESULTS: Quantitative assessment yielded the quantitative size occupied by collagen fibers in an area of 0.09 mm2 . Calculation of the percentage in each area can be used to determine the density of collagen fibers. CONCLUSION: Measurements obtained with our method can be applied to research on dermal collagen fibers. We present a convenient quantitative assessment method for the dermal constituents in Masson's trichrome-stained slides.

An assessment method for dermal structures using cross-polarized light imaging with a green light-emitting diode.

BACKGROUND: Several experimental methods for evaluating dermal structures exist; however, most of these are not used in dermatology clinics because of cost and functional limitations. OBJECTIVE: To propose a simple, non-invasive method for dermal structure evaluation using a green light-emitting diode (LED) with cross-polarized light (CPL) imaging and compare the quality of the images taken using either green or white LED. MATERIALS AND METHODS: Skin specimens were taken from fifteen cadavers. Images were captured using CPL photography with a green or white LED. The Commission International d'Eclairage L*a*b* (CIELAB) values were calculated for each image. The skin specimens were processed and stained with Masson's trichrome to visualize collagen fibers with major image scattering. The images were histologically analyzed, and correlational and regression analyses were performed to determine the relationship between the L* values and histologic measurements. RESULTS: The L* values for the green images were positively correlated with collagen fiber density, reticular collagen bundle diameter, and dermal thickness. They were effective for estimating dermal properties. The L* values for the white images were positively correlated with reticular collagen bundle diameter and dermal thickness. Correlational coefficients for white images were lower than those for green images. In regression analysis, green images showed a higher coefficient of determination (R2 ) for predicting reticular collagen bundle diameter than white images (0.1128 vs. 0.0827). CONCLUSION: Cross-polarized light imaging with a green LED is a simple, non-invasive method for evaluating dermal structures. The use of a green LED was also more effective for image analysis.

Retreatment using a dual mode of low-fluence Q-switched and long-pulse Nd:YAG laser in patients with melasma aggravation after previous therapy.

BACKGROUND: Aggravated melasma after treatment is vulnerable to stimulation, can easily deteriorate, and may be distressing without proper management. OBJECTIVE: To retrospectively assess the effectiveness and safety of combination therapy using low-fluence Q-switched neodymium-doped yttrium aluminum garnet (Nd:YAG) laser (QSNY) and long-pulse Nd:YAG laser (LPNY) (dual toning) in patients with rebound melasma. MATERIALS AND METHODS: A total of 30 patients with aggravated melasma after previous therapy who were treated with dual toning were enrolled. A total of 10 sessions were conducted at 1-week intervals, followed by maintenance treatment. The results were evaluated using the modified Melasma Area and Severity Index (mMASI) and the physician's global assessment (PGA) before and 2 months after completing the 10 treatment sessions. RESULTS: The baseline mMASI was 10.48 ± 3.64, which significantly decreased to 3.22 ± 1.45 2 months after completing the 10 treatment sessions (p < 0.001). Twenty-four patients (80%) had PGA grade 4 (76-100% improvement) and 6 patients (20%) had PGA grade 3 (51-75% improvement). CONCLUSION: Dual toning may be a safe and effective salvage treatment for patients with aggravated melasma after previous treatment. LPNY may stabilize melasma activity to prevent rebound hyperpigmentation via dermal remodeling.

Retrospective analysis of melasma treatment using a dual mode of low-fluence Q-switched and long-pulse Nd:YAG laser vs. low-fluence Q-switched Nd:YAG laser monotherapy.

BACKGROUND: Despite the effectiveness of low-fluence Q-switched Nd:YAG laser (QSNY) treatment in melasma, adverse events, including mottled hypopigmentation (MH) and rebound hyperpigmentation (RH) have been reported. OBJECTIVE: To compare the effectiveness and safety of combination therapy using low-fluence QSNY and long-pulse Nd:YAG laser (LPNY) (Dual toning), with low-fluence QSNY monotherapy (QS toning), in Asian melasma patients. MATERIALS AND METHODS: Patients were treated for 10 sessions at 1-week intervals with QSNY (6 mm spot); 2.5-3.0 J/cm(2) for QS toning (n = 177) or 2.1-2.5 J/cm(2) for dual toning (n = 183). The dual toning group was immediately treated with LPNY (7 mm spot, 15-17 J/cm(2)). The results were evaluated using the modified Melasma Area and Severity Index (mMASI) score and the physician's global assessment. RESULTS: MH or RH were significantly lower (1.1% vs. 14.1%) and the treatment efficacy was improved (median decrease of mMASI, 3.6 vs. 3.0) in the dual toning group compared with the QS toning group. Periorbital melasma showed distinctively high rates of adverse events in the QS toning group (23.9% vs. 5.7%), which were significantly reduced in the dual toning group (2.9%). CONCLUSION: Dual toning could represent a safe and effective treatment for Asian melasma patients, as it is associated with minimal adverse events and improved treatment efficacy compared with QS toning monotherapy.

Egr-1 is a key regulator of IL-17A-induced psoriasin upregulation in psoriasis.

The early growth response (Egr)-1 is a transcriptional factor which plays an important role in the regulation of cell growth, differentiation, cell survival and immune responses. Emerging evidences including our data demonstrate that the Egr-1 expression is up-regulated in the psoriatic skin lesions. The purpose of this study was to investigate the significance and regulatory mechanism of Egr-1 in the pathogenesis of psoriasis. Through microarray analysis, we found out that psoriasin (S100A7) expression was increased in the Egr-1 overexpressed cells. Our results showed that IL-17A increased Egr-1 expression in the skin of psoriatic patients and cultured human keratinocytes. We then investigated activation of mitogen-activated protein kinase as an upstream signal regulator of Egr-1 expression. IL-17A-induced Egr-1 expression was suppressed by ERK inhibitor. In addition, IL-17A induced psoriasin expression in cultured keratinocytes and the skin of IL-17A intradermally injected mouse. IL-17A-mediated psoriasin upregulation was reduced after treatment of small interfering RNAs against Egr-1. Furthermore, the results of chromatin immunoprecipitation assays demonstrated that Egr-1 directly binds the psoriasin promoter. Our findings present a novel signalling mechanism by which IL-17A can induce the Egr-1-dependent psoriasin expression via the ERK pathway in human keratinocytes. This study suggests that Egr-1 may be a novel and important modulator in IL-17A-mediated immune response in psoriasis.

3D reconstruction of skin pathological tissue: the understanding of microrelief pattern and dermal ridge.

AIM: Most studies about the structures of the human skin have been on the general histologic features of the skin. The purpose of this study was to understand the relationship between the microrelief (MR) pattern and histologic structure of the human skin, and to find the usefulness of the three-dimensional reconstruction (3DR) technique of pathological skin tissue. MATERIALS, SUBJECTS, AND METHODS: Human skin specimens were obtained from the medial forearm, abdomen, and volar aspects of the finger tips of cadavers. Two-dimensional surface-viewed images of the skin showing MR patterns were obtained using dry dermoscopy. Histologic structures of the skin were evaluated by 3D images obtained using the 3DR technique. RESULTS: MR was deeper and wider in the abdomen than in the medial side of the forearm. In the medial side of the forearm, 10.27 rete ridges were distributed in a plateau, on average. In the abdomen, about 14.37 rete ridges were distributed in a plateau. From 3DR technique, MR pattern thought to be a superordinate concept to rete ridges. CONCLUSION: The 3DR technique was useful and may allow a better understanding of the pathogenetic changes in the skin surface in aging skin.

A comparison of the scar prevention effect between carbon dioxide fractional laser and pulsed dye laser in surgical scars.

BACKGROUND: The use of ablative lasers based on the fractional approach is a novel strategy for the treatment of postoperative and acne scars in addition to wrinkles. OBJECTIVE: To evaluate and compare the efficacy of carbon dioxide ablative fractional laser (AFL) and the pulsed dye laser (PDL) for the improvement of surgical scars. MATERIALS AND METHODS: Fourteen Korean patients with surgical scars were enrolled for this study. Half of each scar was treated with a 10,600-nm AFL and the contralateral half with the 595-nm PDL. For early intervention of the postoperative scar, the laser treatments were begun after 2 weeks from the Mohs micrographic surgery. RESULTS: Both PDL and AFL produced statistically significant improvements. However, comparatively, there was no statistical difference between them. In each variable, AFL was more effective than PDL in the improvement of pliability and thickness. In contrast, PDL was superior to AFL in the improvement of vascularity and pigmentation. CONCLUSION: Pulsed dye laser and AFL treatments for surgical scar provide significant improvement. Pulsed dye laser was more effective in color of scar compared with AFL, which showed marked improvement in the contour of scar. Overall improvement was not statistically different in the Vancouver Scar Scale.

The basic chromospheres of skin that might not be differentiated visually: observations on aneurysmal fibrous histiocytomas and malignant melanomas.

AIM: Aneurysmal fibrous histiocytoma (AFH) is a variant of fibrous histiocytoma, which has a cleft-like cavernous blood-filled space in the tumor. It appears as a single reddish black tumor with variable levels of pain and size from its bleeding. And, it must be differentiated from other similar looking malignant conditions such as malignant melanoma. The visual mimicry of AFH to melanoma was raised by some careful dermatologists, but never be confirmed objectively by colorimetric analysis. MATERIALS, SUBJECTS, AND METHODS: In this study, we simply analyzed conventionally photographed digital images of thirty-seven cases of fibrous histiocytomas, including three AFH cases into colorimetrically useful color space, CIELAB, of which coordinates are L*, a*, and b* representing lightness, red to green, and yellow to blue axis, respectively. In addition, we also analyzed the clinical digital images of seven cases of malignant melanomas. Using statistical package, each coordinates of CIELAB were compared using Wilcoxon rank sum test between AFH and melanomas. The CIELAB coordinates of AFH and non-aneurysmal fibrous histiocytomas were compared statistically as well. RESULTS: Comparing with banal fibrous histiocytomas, the colors of AFH showed significantly smaller a* and b* coordinates (P = 0.008, 0.008, respectively), which implies more green and blue hue of AFH lesion. Rather, they were more like melanomas (P = 0.2839, 0.2040, respectively). As for L*, there were no significant differences for all comparisons. CONCLUSIONS: As a result, more objective analysis of the digital images using colorimetric color space confirmed the visual mimicry of AFH to melanoma.

Tinea incognito in Korea and its risk factors: nine-year multicenter survey.

Tinea incognito (TI) is a dermatophytic infection which has lost its typical clinical appearance because of improper use of steroids or calcineurin inhibitors. The incidence of TI is increasing nowadays. We conducted retrospective review on 283 patients with TI from 25 dermatology training hospitals in Korea from 2002-2010 to investigate the demographical, clinical, and mycological characteristics of TI, and to determine the associated risk factors. More than half (59.3%) patients were previously treated by non-dermatologists or self-treated. The mean duration of TI was 15.0 ± 25.3 months. The most common clinical manifestations were eczema-like lesion, psoriasis-like, and lupus erythematosus-like lesion. The trunk and face were frequently involved, and 91 patients (32.2%) also had coexisting fungal infections. Among 67 isolated strains, Trichophyton rubrum was the most frequently detected (73.1%). This is the largest study of TI reported to date and the first investigational report concerning TI in Korea. We suggest that doctors should consider TI when a patient has intractable eczema-like lesions accompanied by tinea pedis/unguium. Furthermore, there should be a policy change, which would make over-the-counter high-potency topical steroids less accessible in some countries, including Korea.

A nationwide study of acne treatment patterns in Korea: analysis of patient preconceived notions and dermatologist suggestion for treatment.

Patients often have preconceived notions about acne treatments before visiting dermatologists. The aim of this study was to explore the association between patients' beliefs regarding acne and physicians' suggestion for treatment modality in dermatology clinics. A cross-sectional, nationwide multicentre study was conducted. A total of 1,370 patients completed questionnaires about beliefs about acne treatment before seeking medical care, and 101 dermatologists assessed their acne severity and proposed treatment methods. We found that patients had preconceptions in understanding disease characteristics, assessing subjective acne severity and preferring specific treatment modalities. Dermatologists' determination of topical agents as first-line treatment was affected by disease severity and patients' preferences. They were also more likely to prescribe isotretinoin even in moderate acne compared to oral antibiotics and topical agents. Selections of physical treatments and light-based therapies were affected by patients' preferences, subjective self-evaluation and dermatologists' assessments. Thus, we suggest that acne treatment strategies should incorporate both patients' subjective perceptions and objective clinical practices into a management paradigm.

A case of spitzoid melanoma with lymph node metastasis in a child.

The distinction of a spitz nevus from a melanoma can be difficult and in some cases, impossible. A misdiagnosed spitz nevus can metastasize and lead to fatal outcomes, especially in children. A 5-yr-old girl presented with a 1-yr history of a solitary pinkish nodule on her left hand. On physical examination, she had a palpable left axillary lymph node. We performed biopsy and checked 3 sentinel lymph nodes (SLN) on her axillary area. The biopsy specimen showed multiple variably sized and shaped nests with large spindle or polygonal cells and SLN biopsy showed 3 of 3 lymph nodes that were metastasized. Under the diagnosis of spitzoid melanoma, she was treated with excision biopsy and complete left axillary lymph nodes were dissected. She received interferon-α2b subcutaneously at a dose of 8 MIU per day, 3 times weekly for 12 months, and shows no recurrence.

Recalcitrant cutaneous ulcer of comorbid patient treated with platelet rich plasma: a case report.

The platelet-rich plasma (PRP) has been advocated as a way to introduce increased concentrations of growth factors and other bioactive molecules to injured tissues in an attempt to optimize the local healing environment. A 94-yr-old woman with various comorbidities presented with a two-week history of severe cutaneous ulcer on the left dorsum of foot. It was caused by recurrent mechanical trauma and did not respond to several wound debridement and simple dressings. However, after she was completed on seven times of autologous PRP treatments, we observed complete healing of the skin lesion within 3 months. Herein, we report a case of recalcitrant cutaneous ulcer with various comorbidities and discuss about the promising possibility of autologous PRP as an effective alternative therapeutic modality.

Phacomatosis pigmentokeratotica without extracutaneous abnormalities: a case study involving a preterm baby.

Phacomatosis pigmentokeratotica (PPK) is a rare syndrome defined by the association of an organoid nevus occasionally with sebaceous differentiation, a speckled lentiginous nevus, and other extracutaneous anomalies. A preterm male infant of only 830 g at 27 week gestational age had an organoid nevus showing sebaceous differentiation. Also, he had multiple speckled-lentiginous nevus. Correlating the observed clinical presentation with the histopathological findings, the diagnosis of PPK was established. There have been less than 10 cases of PPK without extracutaneous manifestation. We present an uncommon case of a preterm patient with PPK who had no extracutaneous abnormalities.

Cigarette smoke-induced early growth response-1 regulates the expression of the cysteine-rich 61 in human skin dermal fibroblasts.

Tobacco smoke is known to be an element contributing to accelerate premature skin ageing. Cysteine-rich 61 (Cyr61) is a member of the connective tissue factor CCN (Cyr61, CTGF and Nov) family, and early growth response-1 (Egr-1) is a generally expressed member of the zinc-finger family of transcription factors. To investigate the regulatory potential of Egr-1 on expression of Cyr61 by smoking, this study examined the hypothesis that cigarette smoke-induced Egr-1 induces expression of Cyr61 in human skin dermal fibroblasts (HSDF). HSDF were exposed to different concentrations of cigarette smoke extract (CSE) for 24 h; a cytotoxicity assay was then performed for the detection of cell proliferation. Results of Western blot and reverse transcription-polymerase chain reaction (RT-PCR) showed that CSE induces a transient synthesis of Egr-1 in HSDF. Cyr61 mRNA and protein levels showed a marked increase in a time-dependent manner after CSE exposure. Following transfection with an Egr-1 overexpression vector, HSDF showed increased activity of the Cyr61 promoter in a dose-dependent manner. Using Egr-1 interfering RNA, we confirmed that CSE-induced Cyr61 expression was dependent on Egr-1 expression. Findings of this study indicate that Egr-1-dependent induction of Cyr61 may contribute to premature skin ageing by smoking.

Analysis of familial factors using the basic and specific (BASP) classification in Korean patients with androgenetic alopecia.

BACKGROUND: The mechanisms and inheritance of androgenetic alopecia (AGA) have yet to be elucidated. Several clinical studies suggest that a predisposition to AGA is affected by a variety of paternal and/or maternal hereditary factors. No previous study has addressed the association of AGA with family history based on the pattern of hair loss. OBJECTIVES: The purpose of this study was to investigate paternal and/or maternal genetic influences in each type of hair loss pattern using the basic and specific (BASP) classification and to explore whether the morphology of AGA tends to be inherited in family members. METHODS: Between October 2007 and September 2008, 1220 Korean participants, 998 male and 222 female, were classified according to the BASP classification at 13 university dermatologic centers. Information was collected using a standard questionnaire and BASP classification sheet. RESULTS: Parental influences on anterior hairline shape in men were predominantly from the paternal side, whereas these effects were less notable in women. In the absence of a family history, statistical analysis showed a higher frequency for early-onset AGA than late-onset AGA (Pearson χ(2)P < .05). Basic types of hair loss had a higher degree of inheritance from the paternal side of the family, regardless of specific type. LIMITATIONS: The evaluation of hair loss pattern and family history was done by the patients. CONCLUSIONS: Familial factors affecting the morphology of AGA differ between male and female individuals, and for each type of BASP classification.

Better understanding of digital photography for skin color measurement: With a special emphasis on light characteristics.

BACKGROUND/AIM: Digital photography can be used to measure skin color colorimetrically when combined with proper techniques. To better understand the settings of digital photography for the evaluation and measurement of skin colors, we used a tungsten lamp with filters and the custom white balance (WB) function of a digital camera. MATERIALS AND METHODS: All colored squares on a color chart were photographed with each original and filtered light, analyzed into CIELAB coordinates to produce the calibration method for each given light setting, and compared statistically with reference coordinates obtained using a reflectance spectrophotometer. They were summarized as to the typical color groups, such as skin colors. We compared these results according to the fixed vs. custom WB of a digital camera. RESULTS: The accuracy of color measurement was improved when using light with a proper color temperature conversion filter. The skin colors from color charts could be measured more accurately using a fixed WB. In vivo measurement of skin color was easy and possible with our method and settings. CONCLUSION: The color temperature conversion filter that produced daylight-like light from the tungsten lamp was the best choice when combined with fixed WB for the measurement of colors and acceptable photographs.

Up-regulation of TNF-alpha secretion by cigarette smoke is mediated by Egr-1 in HaCaT human keratinocytes.

Many epidemiologic studies have pointed to a significant association between cigarette smoking and inflammatory skin disease such as psoriasis. Cigarette smoke induces expression of regulators of inflammation such as interleukin (IL)-1, IL-6, IL-8 and tumor necrosis factor (TNF)-alpha. It was recently demonstrated that early growth response-1 (Egr-1) transcription factor is significantly up-regulated in the skin lesions of patients with psoriasis. The mechanism by which cigarette smoke extract (CSE) regulates inflammatory cytokine expression in keratinocyte was still unknown. The aim of this study was to investigate the signalling of CSE-induced Egr-1 expression and the role for Egr-1 in CSE-induced TNF-alpha expression. Cytotoxicity of CSE in HaCaT cells was measured by thiazolyl blue tetrazolium bromide (MTT) assay. CSE-induced Egr-1 expression was investigated by western blot, luciferase reporter assay and confocal microscopy. TNF-alpha expression was measured by reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay. Involvement of Egr-1 in CSE-induced TNF-alpha secretion was determined by using Egr-1 specific siRNA. CSE increases the Egr-1 expression, promoter activity and its nuclear translocation in human HaCaT keratinocytes. CSE activates mitogen-activated protein kinase (MAPK) pathways including extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK). Up-regulation of Egr-1 expression by CSE stimulation was found to be inhibited by an ERK and JNK but not p38 inhibitor. CSE increases TNF-alpha expression and secretion. This increase is mediated by CSE-induced Egr-1 expression. Our results showed that CSE induces Egr-1 expression via MAPK pathway in human keratinocytes and TNF-alpha expression by Egr-1. This pathway may contribute to the development of inflammatory disease such as psoriasis.

A quantitative assessment of the human skin surface using polarized light digital photography and its dermatologic significance.

BACKGROUND: Quantitative and objective investigations of parallel-polarized light (PPL) photography for dermatologic purposes are scarce. OBJECTIVE: To determine the significance of quantitative analysis of PPL photography to develop objective measurement methods of skin surface characteristics. METHOD: For PPL photography, a digital camera, a light-emitting diode illuminator, and polarizing filters were arranged and kept constant. We analyzed the PPL photography images of the glabella, the medial aspect of the forearm, and the posterior aspect of the lower leg, and measured the CIELAB values. A dermatologist evaluated the clinical severity of the actual photographed sites with respect to glossiness and dryness. These clinical severities were compared statistically with the CIELAB values. RESULTS: According to the correlation analysis, the L* value was negatively correlated with dryness (r=-0.29869, P=0.0047) and glossiness (r=-0.35367, P=0.0185). CONCLUSION: The method used in this study is applicable to the quantitative evaluations of other dermatologic diseases or conditions, especially in relation to skin surface changes.

Treatment of Becker's nevi with a long-pulse alexandrite laser.

BACKGROUND: Becker's nevus (BN) can be a distressing cosmetic handicap for patients and a treatment challenge for physicians. Various treatment modalities have been used, but repigmentation commonly occurs after treatment. OBJECTIVE: To evaluate the efficacy of long-pulse alexandrite laser in the treatment of BN. MATERIALS AND METHODS: Eleven Korean patients with Fitzpatrick skin type III to V were included in this study. A long-pulsed alexandrite laser with a wavelength of 755 nm and a pulse duration of 3 ms was used. Patients were treated with a fluence of 20 to 25 J/cm(2) and a spot size of 15 to 18 mm. Cryogen spray cooling was not used. RESULTS: Two patients had excellent responses, five had good responses, and four had fair responses. Hair density simultaneously decreased with treatment in all patients. Although mild hypopigmentation was observed in some patients, and partial hypertrophic scarring was observed in one patient, the outcomes were cosmetically acceptable. No repigmentation was noted during the follow-up period. CONCLUSION: A long-pulsed alexandrite laser without cryogen spray cooling is an effective and safe alternative in the treatment of BN.