Differences in foot skin microbiomes between patients with type 2 diabetes and healthy individuals.

Impaired immunity and changes in the microenvironment in patients with diabetes might influence the composition of the cutaneous microbiome. However, data on the cutaneous microbiome of these patients are scarce. This study compared the fungal and bacterial components of the skin microbiome between patients with type 2 diabetes mellitus (DM) and healthy individuals. We obtained skin swab samples from the plantar forefoot of 17 patients with DM and 18 healthy individuals to conduct a cross-sectional study. The samples were profiled with culture-independent sequencing of the V3 to V4 regions of the bacterial 16S rRNA gene and the fungal ITS2 region, followed by direct DNA extraction and molecular polymerase chain reaction (PCR). We observed a differential cutaneous microbiome, especially for fungi, in patients with type 2 diabetes compared to that in healthy controls. Trichophyton rubrum was more abundant in DM samples. The Shannon diversity index for fungi was lower in the DM patients. Principal coordinate analysis plots and permutational multivariate analysis of variance (PERMANOVA) tests based on Bray-Curtis distances between samples supported the association of the fungal microbiome with DM at the species level. The results suggest that clinicians should pay attention to both fungi and bacteria and provide appropriate prevention and therapeutic strategies for diabetic cutaneous complications including diabetic foot ulcers. These data also contribute to future research associated with diabetes and cutaneous microbiomes.

Assessment of Skin Physiology Change and Safety After Intradermal Injections With Botulinum Toxin: A Randomized, Double-Blind, Placebo-Controlled, Split-Face Pilot Study in Rosacea Patients With Facial Erythema.

BACKGROUND: Botulinum toxin (BTX) has been used cosmetically with good clinical efficacy and tolerable safety. OBJECTIVE: This randomized, double-blind, split-face clinical study aimed to investigate the efficacy and safety of intradermal BTX in patients with rosacea. MATERIALS AND METHODS: Twenty-four participants were enrolled and randomly given intradermal injections of BTX and normal saline in both cheeks. Clinician Erythema Assessment (CEA) score, Global Aesthetic Improvement Scale (GAIS) score, skin hydration, transepidermal water loss (TEWL), melanin content, erythema index, elasticity, and sebum secretions were evaluated at baseline and 2, 4, 8, and 12 weeks. RESULTS: On the BTX-treated side, the CEA score significantly decreased and the GAIS score significantly increased. The erythema index decreased at Weeks 4 and 8. Skin elasticity was improved at Weeks 2 and 4 and skin hydration, at Weeks 2, 4, and 8. However, TEWL and sebum secretion did not show significant differences. CONCLUSION: Intradermal BTX injections reduced erythema and rejuvenated the skin effectively and safely in patients with rosacea.

Analysis of the skin mycobiome in adult patients with atopic dermatitis.

With the recent availability of culture-independent sequencing methods, studies have been conducted to analyse skin micro-organisms present in patients with atopic dermatitis (AD). However, the database on the skin fungal communities, "mycobiome," has been relatively restrictive compared with the bacterial world. We aimed to comparatively analyse the overall skin mycobiome between patients with AD and healthy individuals in the Korean population. We analysed skin swab samples obtained from the antecubital fossae of 8 patients with AD and 8 healthy controls. Using sequencing method followed by direct DNA extraction and molecular PCR, taxonomic compositions of fungi at stepwise level ranks were analysed. The phylogenic marker used was internal transcribed spacer 2 regions of DNA. We observed the tendency of higher intra- and interpersonal taxonomic diversity at genus and species levels in AD samples. Non-Malassezia fungal diversity was also noticeable in the patient group compared with healthy controls. Malassezia globosa and Malassezia restricta were prevalent in all samples across both study groups, and some Malassezia species, including Malassezia sloofiae and Malassezia dermatis, characterized AD. Our data might provide a new insight into the mycobiome of adult AD, which contributes to building a systemic mycobiome database in AD.

TRIAD1 Is a Novel Transcriptional Target of p53 and Regulates Nutlin-3a-Induced Cell Death.

Nutlin-3a is a non-genotoxic, p53-activating, MDM2 inhibitor being investigated as an anticancer agent. Although Nutlin-3a selectively antagonizes the ubiquitin E3 ligase activity of MDM2, its efficacy is not entirely regulated by MDM2 levels in cancer cells. Here, we report that the cytotoxic effects of Nutlin-3a are regulated by TRIAD1 via a positive feedback loop with p53. We found that Nutlin-3a enhanced TRIAD1 transcription in a p53-dependent manner. Using in silico analysis and promoter luciferase assays, we demonstrated that p53-mediated transcription of TRIAD1 is mediated by a p53 consensus sequence in the TRIAD1 promoter region. Silencing TRIAD1 expression in wild-type p53 (p53WT ) cancer cells suppressed Nutlin-3a-mediated p53 activation and p53 target gene expression. These effects were enhanced in TRIAD1-overexpressing p53WT cancer cells, but not in p53-deficient cancer cells. Furthermore, TRIAD1 knockdown significantly reduced the growth inhibitory and cytotoxic effects of Nutlin-3a in p53WT cancer cells, as demonstrated by cell viability assays, cell cycle analysis, clonogenic growth, and soft-agar colony forming assays. Together, these data indicate that TRIAD1 regulates Nutlin-3a-mediated p53 activation and the cytotoxic activity of Nutlin-3a. J. Cell. Biochem. 118: 1733-1740, 2017. © 2016 Wiley Periodicals, Inc.

First isolation of Rickettsia monacensis from a patient in South Korea.

A Rickettsia sp. was isolated from the blood of a patient with an acute febrile illness using the shell vial technique; the isolate was named CN45Kr and was identified by molecular assay as Rickettsia monacensis, which was first recognized as a pathogen in Spain. Sequencing analysis showed that the gltA sequence of the isolate was identical to that of Rickettsia sp. IRS3. The ompA-5mp fragment sequence showed 100% identity to those of R. monacensis and Rickettsia sp. In56 and ompA-3pA In56 and 100% identity to that of Rickettsia sp. IRS3. The ompB sequence was found to have 99.9% similarity to that of R. monacensis IrR/Munich. This study confirms the pathogenicity of this agent and provides additional information about its geographic distribution.

Titrated extract of Centella asiatica increases hair inductive property through inhibition of STAT signaling pathway in three-dimensional spheroid cultured human dermal papilla cells.

Dermal papilla (DP) is a pivotal part of hair follicle, and the smaller size of the DP is related with the hair loss. In this study, we investigated the effect of titrated extract of Centella asiatica (TECA) on hair growth inductive property on 3D spheroid cultured human DP cells (HDP cells). Significantly increased effect of TECA on cell viability was only shown in 3D sphered HPD cells, not in 2D cultured HDP cells. Also, TECA treatment increased the sphere size of HDP cells. The luciferase activity of STAT reporter genes and the expression of STAT-targeted genes, SOCS1 and SOCS3, were significantly decreased. Also, TECA treatment increased the expression of the hair growth-related signature genes in 3D sphered HDP cells. Furthermore, TECA led to downregulation of the level of phosphorylated STAT proteins in 3D sphered HDP cells. Overall, TECA activates the potential of hair inductive capacity in HDP cells.

23-Hydroxytormentic acid protects human dermal fibroblasts by attenuating UVA-induced oxidative stress.

BACKGROUND: Ultraviolet A (UVA), one of the major components of sunlight, can penetrate the dermal layer of the skin and generate reactive oxygen species (ROS). It causes alterations in the dermal connective tissue and gene expression, inflammation, photoaging, and DNA damage. AIMS: Therefore, the harmful effects of UVA and strategies to reduce it have been consistently investigated. 23-Hydroxytormentic acid (23-HTA) has been demonstrated to improve drug-induced nephrotoxicity and exhibit several free radical scavenging effects with other molecules. Therefore, the aim of this study was to investigate the anti-inflammatory effects and extracellular matrix (ECM) reconstructive activity of 23-HTA in UVA-irradiated normal human dermal fibroblasts (NHDFs). MATERIALS AND METHODS: The antioxidant capacity of 23-HTA was determined by examining its scavenging activities against hydrogen peroxide, 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid), and diphenylpicrylhydrazyl in vitro. Its effect on cell viability was evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-tertazolium bromide, and 2,7-dichlorofluorescin diacetate was used to investigate intracellular ROS scavenging activity. The mRNA levels of antioxidant enzymes and pro-inflammatory cytokines were detected using quantitative real-time polymerase chain reaction. A senescence-associated ß-galactosidase (SA-ß-gal) staining kit was used to assess senescent cells. RESULTS: 23-HTA showed antioxidant capacity mediated by ROS scavenging and regulation of antioxidant-related gene expression. Further, the SA-ß-gal analysis and mRNA expression of matrix metalloproteinases and type I procollagen suggested that 23-HTA regulates the gene expression of ECM proteins and cellular senescence under UVA-irradiated conditions. CONCLUSION: In conclusion, 23-HTA protects against and attenuates UVA-induced oxidative stress in NHDFs likely via the nuclear factor erythroid-derived 2-like 2 signaling pathway.

Treatment of periorbital wrinkles using multipolar fractional radiofrequency in Korean patients.

A limited number of studies have evaluated the efficacy and safety of electrode pin fractional radiofrequency (FRF) for periorbital wrinkle treatment in Asian patients, but none have measured noninvasive methodological objective parameters such as periorbital wrinkle area. This study aimed to investigate the efficacy and safety of electrode pin multipolar FRF for the improvement of periorbital wrinkles in Korean patients by using a noninvasive methodological objective parameter. Seventy female subjects with periorbital wrinkles (age range, 40-60 years) participated in this study. Each patient underwent three sessions of FRF treatment to the periorbital region separated by 2-week intervals. The area of periorbital wrinkles was analyzed by using a Robo Skin Analyzer CS50 at 4 weeks after the final treatment session. Periorbital wrinkle area was significantly decreased at 1-month follow-up (75.77 ± 29.46 mm2) compared to baseline (94.74 ± 31.62 mm2). The improvement ratio of periorbital wrinkle area was 20.02 %. Side effects were limited to transient mild erythema, swelling, and crusts. Pain was tolerable without local anesthesia. Our findings suggest that the multipolar electrode pin FRF can be an effective and safe method for reducing periorbital wrinkles in Asian patients.

Isolation and identification of Malassezia species from Chinese and Korean patients with seborrheic dermatitis and in vitro studies on their bioactivity on sebaceous lipids and IL-8 production.

We investigated the distribution of Malassezia yeast in 120 Chinese (20 patients from each of six cities) and 20 Korean patients with scalp seborrheic dermatitis (SD) and dandruff (SD/D) using ITS1 and ITS2 polymerase chain reaction-restriction fragment length polymorphism. Bioactivity was studied by quantifying sebum lipid production by human primary sebocytes and inflammatory cytokine, interleukin-8 (IL-8) production was studied by exposing HaCaT keratinocytes with extracts of five standard Malassezia strains; M. globosa, M. restricta, M. sympodialis, M. dermatis and M. slooffiae. M. restricta and M. globosa were the most frequently encountered species from both Chinese and Korean patients. These two Malassezia species also promoted neutral lipid synthesis although the result was not statistically significant and induced significant increase in IL-8 production among the five Malassezia species studied. The study suggests a possible role of these organisms in the pathogenesis of SD/D.

Metaanalysis of BRAF mutations and clinicopathologic characteristics in primary melanoma.

BACKGROUND: BRAF mutations occur in some melanomas. We hypothesized that BRAF mutation rates may differ in melanomas found in Asian compared to white populations. OBJECTIVE: We performed a metaanalysis of BRAF mutations and their associations with the clinicopathologic characteristics of primary melanoma (PM), with a subgroup analysis to compare Asian and white patients with PM. METHODS: The PubMed, EMBASE, and Cochrane databases were searched up to November 2013. The incidence rates and odds ratios (ORs) of BRAF mutations were calculated using a fixed or random effects model. RESULTS: BRAF mutation was associated with younger age (OR = 1.734; P < .001), trunk location (OR = 2.272; P < .001), non-chronically sun damaged skin (OR = 2.833; P < .001), superficial spreading melanoma (OR = 2.081; P < .001), and advanced melanoma stage (OR = 1.551; P = .003). The incidence of BRAF mutations in Asian patients with PM was half that of white patients with PM, but it was linked to the same clinicopathologic characteristics. LIMITATIONS: Only a small number of studies have been conducted on Asian patients with PMs. CONCLUSIONS: The BRAF mutation in PM was associated with age, anatomic site based on ultraviolet radiation exposure, histologic subtype, and advanced stage of melanoma. The clinicopathologic associations with BRAF mutations were similar in Asian and white patients with PM.

Arctiin blocks hydrogen peroxide-induced senescence and cell death though microRNA expression changes in human dermal papilla cells.

BACKGROUND: Accumulating evidence indicates that reactive oxygen species (ROS) are an important etiological factor for the induction of dermal papilla cell senescence and hair loss, which is also known alopecia. Arctiin is an active lignin isolated from Arctium lappa and has anti-inflammation, anti-microbial, and anti-carcinogenic effects. In the present study, we found that arctiin exerts anti-oxidative effects on human hair dermal papilla cells (HHDPCs). RESULTS: To better understand the mechanism, we analyzed the level of hydrogen peroxide (H2O2)-induced cytotoxicity, cell death, ROS production and senescence after arctiin pretreatment of HHDPCs. The results showed that arctiin pretreatment significantly inhibited the H2O2-induced reduction in cell viability. Moreover, H2O2-induced sub-G1 phase accumulation and G2 cell cycle arrest were also downregulated by arctiin pretreatment. Interestingly, the increase in intracellular ROS mediated by H2O2 was drastically decreased in HHDPCs cultured in the presence of arctiin. This effect was confirmed by senescence associated-beta galactosidase (SA-ß-gal) assay results; we found that arctiin pretreatment impaired H2O2-induced senescence in HHDPCs. Using microRNA (miRNA) microarray and bioinformatic analysis, we showed that this anti-oxidative effect of arctiin in HHDPCs was related with mitogen-activated protein kinase (MAPK) and Wnt signaling pathways. CONCLUSIONS: Taken together, our data suggest that arctiin has a protective effect on ROS-induced cell dysfunction in HHDPCs and may therefore be useful for alopecia prevention and treatment strategies.

Serum levels of LL-37 and inflammatory cytokines in plaque and guttate psoriasis.

Psoriasis is a chronic inflammatory skin disease. It is assumed that the plaque phenotype of psoriasis is associated with T helper (Th) 1 immune response activation, while the guttate phenotype is associated with the Th17 immune response. Previous investigations of differences in the serum levels of cytokines relative to the clinical psoriatic phenotype have yielded conflicting results. This study compared the levels of circulating inflammatory cytokines and LL-37 relative to the morphological phenotype in patients with psoriasis. Seventy-four age-matched patients with psoriasis (32 with guttate psoriasis and 42 with plaque psoriasis) and 12 healthy controls were included. A multiplex cytokine assay and enzyme-linked immunosorbent assay were used to measure levels of Th1- and Th17-derived cytokines and LL-37, respectively. Circulating levels of interferon- (IFN)-γ, interleukin- (IL)-1RA, IL-2, and IL-23, and LL-37 were significantly higher in patients with psoriasis than in healthy controls. However, the serum levels of inflammatory cytokines (IL-7, IL-22, and IL-23) and LL-37 did not differ significantly between the guttate and plaque phenotypes of psoriasis. There was a positive correlation between serum inflammatory cytokine levels and the Psoriasis Area and Severity Index score. The findings of this study suggest that the serum levels of inflammatory cytokines reflect the disease activity rather than determine the morphological phenotype.

The potential impact of systemic anti-inflammatory therapies in psoriasis on major adverse cardiovascular events: a Korean nationwide cohort study.

This nationwide population-based cohort study aimed to investigate the impact of systemic anti-inflammatory treatment on the major adverse cardiovascular events (MACE) risk in patients with psoriasis from January 2006 to December 2018, using a database provided by the Korean National Health Insurance Service. Patients were grouped based on the following treatment modalities: biologics, phototherapy, methotrexate, cyclosporine, and mixed conventional systemic agents. Patients who had not received any systemic treatment were assigned to the control cohort. The incidence of MACE per 1000 person-year was 3.5, 9.3, 12.1, 28.4, 39.5, and 14.5 in the biologic, phototherapy, methotrexate, cyclosporine, mixed conventional systemic agents, and control cohorts, respectively. During the 36-month follow-up, the cumulative incidence of MACE in the phototherapy and biologic cohorts remained lower than that of other treatment modalities. Cyclosporine (hazard ratio (HR) = 2.11, 95% confidence interval (CI) = 1.64-2.71) and mixed conventional systemic agents (HR = 2.57, 95% CI = 2.05-3.22) treatments were associated with increased MACE risk. Methotrexate treatment was not associated with MACE. Our finding demonstrates that treatment modalities may affect cardiovascular comorbidities in patients with psoriasis. Thus, an appropriate combination of anti-psoriatic therapies should be considered to manage patients with high cardiovascular risk.IRB approval status: Waiver decision was obtained by the institutional review board, Konkuk University Hospital, Seoul, Republic of Korea (KUH1120107).

The relation between the amount of sunscreen applied and the sun protection factor in Asian skin.

BACKGROUND: The measurement of the sun protection factor (SPF) is the usual method in the examination of the effectiveness of sunscreen. The declared SPF is based on the use of a sunscreen layer of 2 mg/cm(2). However, only around a quarter (0.5 mg/cm(2)) of this amount is generally used in real life. Theoretical calculations have suggested that the effectiveness of SPF is related to sunscreen quantity in an exponential way but this was not confirmed in Asian skin. OBJECTIVES: This study was performed to investigate the change in SPF values when less than the recommended amount of sunscreen was applied. METHODS: A board divided into 10 areas measuring 7 x 4 cm was placed on the back of 15 healthy volunteers. Low- and high-SPF standard reference sunscreens, and two types of sunscreen (SPF 30 and 35) were each applied on 4 areas, 0.5, 1.0, 1.5, and 2.0 mg/cm(2), respectively, and were left to dry for 20 minutes. The irradiation was conducted at a distance of 50 cm using a template (1 x 1 cm) placed directly on the skin with 10 windows allowing ultraviolet (UV) radiation to pass through with a dose increment of 20%. Erythema was evaluated 20 to 24 hours after exposure to UV radiation. RESULTS: Sunscreen showed its expected SPF value when 2.0 mg/cm(2) was applied. The SPF values of the different amounts were significantly different from each other and decreased when less was applied (P < .05). The relation between the amount of sunscreen applied and the SPF provided was most likely to follow exponential growth. LIMITATIONS: Spectral differences between our solar simulator and the UV sources of commercial laboratories are likely to be important. In addition, differences in sunscreen application techniques may have influenced the ultimate SPF values. CONCLUSIONS: This study concludes that to get the expected SPF value, it is important to apply the UV protective sunscreen precisely in the amount of 2.0 mg/cm(2) on Asian skin as recommended by the Food and Drug Administration. In addition, it was difficult to predict the SPF values when the usual amount of 0.5 mg/cm(2) was applied.

Clinical characteristics of psoriatic patients with latent tuberculosis infection.

BACKGROUND: Psoriasis is an immune-mediated inflammatory disease in which imbalance of the immunological response may be associated with disease severity and comorbidities. Latent tuberculosis infection (LTBI) is a growing concern for treatment of psoriasis, as the use of biologics has recently increased. OBJECTIVES: To investigate the clinical and immunological influence of LTBI on the features of psoriasis. MATERIALS AND METHODS: We conducted a retrospective analysis of 300 patients with psoriasis using clinical information, including severity, comorbidities, and presence of LTBI. Serum cytokine levels were measured for immunological analysis. RESULTS: The prevalence of psoriatic arthritis (p = 0.001) and nail psoriasis (p = 0.014) in patients with LTBI was significantly higher than in those without LTBI, although other data including the Psoriasis Area Severity Index showed no association. The serum levels of interleukin (IL)-6, IL-8, and IL-23A in the LTBI-positive group were higher than those in the LTBI-negative group (p = 0.014, p = 0.025, and p = 0.004, respectively). CONCLUSION: LTBI may be a risk factor for the development of psoriatic arthritis during chronic inflammatory conditions induced by tuberculosis infection.

Flavonoid Silibinin Increases Hair-Inductive Property Via Akt and Wnt/ß-Catenin Signaling Activation in 3-Dimensional-Spheroid Cultured Human Dermal Papilla Cells.

Hair loss, also known as alopecia, is a common dermatological condition of psychosocial significance; development of therapeutic candidates for the treatment of this condition is, hence, important. Silibinin, a secondary metabolite from Silybum marianum, is an effective antioxidant that also prevents various cutaneous problems. In this study, we have investigated the effect of silibinin on hair induction using three-dimensional (3D) cultured, human dermal papilla (DP) spheroids. Silibinin was found to significantly increase viability through AKT serine/threonine kinase (AKT) activation in 3D DP spheroids. This was correlated with an increase in the diameter of the 3D DP spheroids. The activation of the wingless and INT-1 (Wnt)/ß-catenin signaling pathway, which is associated with hair growth induction in the DP, was evaluated using the T cell-specific transcription factor and lymphoid enhancer-binding factor (TCF/LEF) transcription factor reporter assay; results indicated significantly increased luciferase activity. In addition, we were able to demonstrate increased expression of the target genes, WNT5a and LEF1, using quantitative real-time PCR assay. Lastly, significantly elevated expression of signature genes associated with hair induction was demonstrated in the 3D DP spheroids treated with silibinin. These results suggest that silibinin promotes proliferation and hair induction through the AKT and Wnt/ß-catenin signaling pathways in 3D DP spheroids. Silibinin can be a potential candidate to promote hair proliferation.

Synergistic Effect of H1-Antihistamines on Topical Corticosteroids for Pruritus in Atopic Dermatitis: A Systematic Review and Meta-Analysis.

BACKGROUND: Although oral antihistamines (H1-histamine receptor antagonists) are the main treatment option for pruritus in general skin dermatosis, their effect in treating pruritus of atopic dermatitis (AD) has not yet been established. OBJECTIVE: We conducted a systematic review and meta-analysis to evaluate the effectiveness of combined therapy of H1-antihistamines and topical steroids. METHODS: We systematically searched MEDLINE, Embase, and CENTRAL databases for articles published from 1967 to 2015. We identified 1,206 studies and assessed their titles, abstract, and full-text. Random effects meta-analysis was used to calculate mean differences (MD) with 95% confidence intervals (CI). RESULTS: Two studies satisfying the inclusion criteria of antihistamine therapy with mandatory topical steroid use were selected. Comparing antihistamine monotherapy with combination therapy, patients treated with the addition of antihistamine to topical corticosteroids showed a statistically significant clinical improvement (standard MD, -0.24; 95% CI, -0.42 to -0.05; p=0.01). CONCLUSION: H1-antihistamines may have a synergistic effect when combined with topical steroids by influencing various associative factors of chronic pruritus in AD.

The IL17F His161Arg polymorphism, a potential risk locus for psoriasis, increases serum levels of interleukin-17F in an Asian population.

Interleukin 17 (IL-17) plays pivotal role in the pathogenesis of psoriasis. In a previous study, we identified a locus in the IL17F gene that is associated with psoriasis, the IL17F rs763780 (His161Arg) T/C variant. The current study aimed to elucidate the association between this polymorphism and psoriasis, and to determine its effect on serum levels of cytokine. A total of 116 psoriasis patients and 97 healthy volunteers were recruited. Genotyping analysis was performed using quantitative polymerase chain reaction, and serum levels of cytokine were measured using a multiplex immunoassay. The IL17F His161Arg polymorphism was significantly associated with psoriasis based on the genotype and allele analyses. Psoriasis patients harbouring the mutant allele had significantly increased serum levels of IL-17F. Our results suggest that this polymorphism is a potential risk locus for psoriasis and that it results in a direct increase in IL-17F production.

Isolation of 19 strains of Malassezia dermatis from healthy human skin in Korea.

This research was conducted on the cultured samples of 160 healthy men and women aged 0-80 years without any skin disease. Nineteen clinical isolates of Malassezia dermatis showed positive in a catalase test and all grew in 0.5% Tween-60 and 0.1% Tween-80 added to 2% glucose/1% peptone culture medium. Round and ellipsoidal yeast cells and budding of the yeast cells were observed by microscopy, resembling Malassezia sympodialis, Malassezia furfur and Malassezia nana. The 26S rDNA polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) pattern was the same as for M. dermatis (JCM 11348), the standard strain. 26S rDNA and ITS1 sequencing were performed for exact identification, showing 99% accordance with M. dermatis (AB070361), M. dermatis (AB070356), confirming the species to be new and first to be reported in Korea. Taking a molecular biological classification approach by analyzing the 26S rDNA PCR-RFLP patterns, we have successfully isolated 19 cases of M. dermatis- the first in Korea.