Effect of apoptosis on G361 cells by Cimicifuga rhizoma extract / 대한구강보건학회지

OBJECTIVES: To investigate whether the cytotoxic effect of Cimicifuga rhizoma extract is associated with cell death in the human keratinocyte (HaCaT) and human melanoma cell lines (G361). METHODS: Apoptosis induced by Cimicifuga rhizoma extract was confirmed by water-soluble tetrazolium salts-1 (WST-1) assay, immunocytochemistry, and western blot. Additionally, the release of cytochrome c and apoptosis-inducing factor (AIF) was visualized by confocal laser scanning microscopy. RESULTS: The results showed that Cimicifuga rhizoma extract significantly reduced the viability of G361 cells with half-maximal inhibitory concentration (IC 50) of 200 µg/ml, and the apoptotic process was found to occur via the activation of caspase-3 and caspase-9 pathways. Besides, the release of cytochrome c and AIF was also detected. CONCLUSIONS: This study suggests that Cimicifuga rhizoma extract causes apoptosis of human melanoma cells through the intrinsic apoptotic pathway.

The Contributory Roles of Th17 Lymphocyte and Cytotoxic T Lymphocyte at the Hair Bulge Region as Well as the Hair Bulb Area in the Chronic Alopecia Areata Patients

BACKGROUND: Alopecia areata (AA) is a T cell-mediated autoimmune disease that targets hair follicles and interrupts hair regrowth. The microenvironment of the effector T cells and their related cytokines may affect immunopathogenesis around the hair bulb/bulge. OBJECTIVE: To determine the contributory roles of the effector T cell subsets and related cytokines to the pathogenesis of AA. METHODS: We investigated the correlation between histopathological grades and four clinical prognostic factors in 331 patients with AA, and analyzed the topography of T cell infiltrates and related cytokines around the hair bulb/bulge according to histopathological grades through immunohistochemical and double immunofluorescence studies on a subset of AA specimens. RESULTS: First, the groups with more severe histopathological grades were associated with earlier onset, longer duration, more hair loss, as well as poorer therapeutic outcomes. Second, the pattern of CD4 and CD8 expression around the hair bulb/bulge varied by histopathological grade, with staining density decreasing in the following order: type 1>type 2>type 3. In addition, interferon-γ and transforming growth factor-β1 expression appeared denser in the peribulbar area. Interestingly, the denser CCR6⁺ cells (Th17 cells) showed more infiltration than CCR5⁺ cells (Th1 cells) around the hair bulb/bulge as histopathological grade worsened. CONCLUSION: The insidious destruction of bulge stem cells and hair bulb matrix stem cells results in more severe hair loss in patients with chronic AA, which is mediated by Th17 lymphocyte and cytotoxic T lymphocyte infiltration. Furthermore, Th17 lymphocytes may play an even more important role than cytotoxic T cells in the development of AA.

An Evaluation of the Repetitive Tooth Bleaching with Nonthermal Atmospheric Pressure Plasma

This study was undertaken to achieve a high bleaching efficacy with plasma, through longer application and reparative bleaching processes, by different shade evaluation methods. Extracted human teeth were divided into 6 groups (n=10). All teeth were treated in pairs. Low concentration of 15% carbamide peroxide (CP) was applied, with and without plasma, for 10, 20, and 30-min tooth bleaching, respectively. The bleaching procedure was repeated once daily for four days. The teeth were maintained in a moist environment provided by artificial saliva. The Vitapan Classical shade guide and Commission Internationale de L'Eclairage (CIELAB) color system were collectively used to measure the bleaching efficacy. Color evaluation was statistically analyzed using Student t-test and one-way analysis of variance (ANOVA) complemented by Tukey's test. Combining the plasma with 15% CP showed significantly greater color changes compared to bleaching without plasma (p<0.05). A high bleaching efficacy with plasma is proportional to the repetitive application and the treatment time. A 30-min application with plasma provided the best bleaching. Repetitive bleaching showed lower probability of color relapse of the bleached tooth. The color change by shade guide correlated with the changes in CIELAB color system. A value of 1 color change units (CCU) conversion factor for overall color change (ΔE) values comparisons was 3.724 values. The two measuring methods provide a more accurate correspondence of color change. The repetitive and longer application for tooth bleaching, combined with plasma, has a strong bleaching effect and produces whiter teeth.

Long-Term Survival Analysis and Clinical Follow-Up in Acral Lentiginous Malignant Melanoma Undergoing Sentinel Lymph Node Biopsy in Korean Patients

BACKGROUND: In cutaneous malignant melanoma (MM) with clinically uninvolved regional lymph nodes, sentinel lymph node (SLN) status is the most powerful indicator of both overall survival (OS) and disease-free survival (DFS). However, no studies on the long-term survival and clinical follow-up of Korean patients with acral lentiginous MM (ALM) undergoing SLN biopsy (SLNB) have been published. OBJECTIVE: The purpose of this study was to investigate the clinical prognosis and long-term survival of Korean patients with ALM according to SLN status. METHODS: Thirty-four ALM patients undergoing SLNB were included in this study. We evaluated clinical and histopathological follow-up data such as the stage of disease, treatment, recurrence, and metastasis, and analyzed OS and DFS according to SLN status. RESULTS: The median follow-up time was 60.5 months (range 3~127 months). Positive SLNs were noted in 14 patients (41.2%). Patients with negative SLNs had better OS and DFS than those with positive SLNs (p<0.05). Increased Breslow thickness was associated with short OS and DFS (p<0.05), and female patients showed better DFS than male patients (p<0.05). CONCLUSION: To our knowledge, this is the first study on the long-term survival and clinical follow-up of patients undergoing SLNB for ALM in Korea. Our findings show that SLN status is an important prognostic factor for predicting OS and DFS.

Prognostic and Clinicopathologic Associations of BRAF Mutation in Primary Acral Lentiginous Melanoma in Korean Patients: A Preliminary Study

BACKGROUND: In the majority of melanomas, the RAS/RAF/MEK/ERK signaling pathway is constitutively activated, due to oncogenic mutations in the BRAF and NRAS genes. The BRAF mutation has been mainly described in Caucasian melanomas. However, there is a lack of study evaluating the status, and the clinical significance, of BRAF mutation in the Asian population. OBJECTIVE: This study was aimed to determine the frequency of BRAF mutation, and to evaluate the correlation of BRAF status with clinicopathologic features and outcomes, in Korean primary acral lentiginous melanoma (ALM) patients. METHODS: ALM samples (n=36) were analyzed for the BRAF V600E mutation, by dual-priming oligonucleotide (DPO) based real-time polymerase chain reaction. The clinicopathologic features and prognosis of the patients were analyzed with BRAF mutation status. RESULTS: The incidence of BRAF V600E mutation was 19.4% (7/36). The BRAF V600E mutations were not associated with clinicopathologic features, except for the age factor. All of the BRAF-mutant patients survived without recurrence or metastasis, and have a better clinical outcome than BRAF wild-type patients. CONCLUSION: In Korean primary ALM, a low frequency of BRAF mutation was shown; and BRAF mutation presented with a favorable prognosis. These results indicate that other distinctive genetic mechanisms may have more important roles in the development and progression of disease. Further multicenter study with large sample size is firmly needed, to confirm the results of our preliminary study.

Alteration of Hydroxyapatite Surface after Tooth Bleaching with Nonthermal Atmospheric Pressure Plasma

Nonthermal atmospheric pressure plasma has attracted great interest for biomedical applications. The plasma consists of charged particles, radicals, and a strong electric field as the fourth state of matter. This study evaluated the change in the surface roughness after tooth bleaching by plasma in combination with a low concentration (15%) of carbamide peroxide, specifically whether the application of plasma produced detriments, such as demineralization and structural change, with the goal of efficient and safe tooth bleaching. After being combined with plasma and 15% carbamide peroxide, the hydroxyapatite surface was significantly smoother with a low roughness average value. Tooth bleaching with 15% carbamide peroxide alone produced an irregular surface and increased the surface roughness with high roughness average value. Tooth bleaching with plasma resulted in no significant variations in hydroxyapatite in terms of change in surface roughness and surface topography. The application of tooth bleaching with plasma is not deleterious to dental hard tissue, implicating it as a safe tooth bleaching technique.

Lipoblastoma: A Rare Lesion in the Elderly / 대한피부과학회지

Lipoblastoma is a rare, benign tumor arising from embryonic fat tissues that continue to proliferate in the postnatal period. So, lipoblastoma occurs almost exclusively in infants and children younger than 3 years of age. Published reports showed that 70~90% of cases occur before the age of 3. Histopathologically, lipoblastoma, mimicking liposarcoma, was composed of well-defined lobulated fat tissue with multilobulated lipoblasts, undifferentiated mesenchymal cells, such as stellate or spindle cells and minimal myxoid stromas. The diagnosis of lipoblastoma is important because its management differs from that of lipoma and liposarcoma. To our knowledge, there are only a few reports of lipoblastoma in the Korean dermatologic literature. Herein, we present an interesting and atypical case of lipoblastoma in 68-year-old woman.

Pityriasis Rotunda Developed during Pregnancy / 대한피부과학회지

Pityriasis rotunda is an uncommon dermatosis characterized by asymptomatic, multiple, widely distributed, round or oval-shaped, hyperpigmented or hypopigmented, fine, scaly patches. They typically involve the abdomen, the trunk and extremities. Histopathologic findings are consistent with ichthyosis vulgaris, such as hyperkeratosis, hypogranulosis or agranulosis, hyperpigmentation of the basal layer in epidermis, and perivascular lymphohistiocytic infiltration in the dermis. Although the etiology of the disease remains unknown, it has been associated with a variety of underlying systemic diseases including infectious diseases, hormonal disorders, malignancies, and chronic disorders. One clinical case has been reported about the occurrence of the disease during pregnancy of a 24-year-old african woman, but there is only one reported case of exacerbation of the disease during pregnancy in Korea. Herein, we report a case of pityriasis rotunda occurring during pregnancy.

Erythema Annulare Centrifugum Associated with Candida guilliermondii / 대한피부과학회지

Erythema annulare centrifugum (EAC), first described by Darier in 1916, is characterized by annular, circinate, gyrate or serpiginous lesions. Although the pathogenesis of EAC is not fully understood, it is considered to be a hypersensitivity reaction to a wide variety of etiological factors, including autoimmune diseases, internal malignancies, drugs, infections, and pregnancy. To the best our knowledge, no case of EAC associated with Candida guilliermondii infection has ever been published in the Korean medical literature, and there have been a few reported cases associated with Candida albicans in other countries. Herein, we report on a rare and interesting case of EAC in a 62-year-old female patient with Candida guilliermondii infection.

Frequency of BRAF Mutation and Clinical Relevance for Primary Melanomas

BACKGROUND: This study was conducted to clarify the frequency of the BRAF mutation in primary melanomas and its correlation with clinicopathologic parameters. METHODS: We analyzed the frequency of BRAF mutation in patients with primary cutaneous melanoma (n=58) or non-cutaneous one (n=27) by performing dual priming oligonucleotide-based multiplex real-time polymerase chain reaction to isolate and to purify the DNA from the formalin-fixed and paraffin-embedded tumors. RESULTS: The BRAF mutation was found in 17.2% (10/58) of patients with primary cutaneous melanoma and 11.1% (3/27) of those with non-cutaneous melanoma. The frequency of BRAF mutation was not correlated with any clinicopathologic parameters with the exception of the patient age. The frequency of the BRAF mutation was significantly higher in patients younger than 60 years as compared with those older than 60 years (p=0.005). CONCLUSIONS: Compared with previous reports, our results showed that the frequency of the BRAF mutation was relatively lower in patients with primary cutaneous melanoma. Besides, our results also showed that the frequency of the BRAF mutation had an inverse correlation with the age. Further studies are warranted to exclude methodological bias, to elucidate the difference in the frequency of the BRAF mutation from the previous reports from a Caucasian population and to provide an improved understanding of the molecular pathogenesis of malignant melanoma.

Frequency of BRAF Mutation and Clinical Relevance for Primary Melanomas

BACKGROUND: This study was conducted to clarify the frequency of the BRAF mutation in primary melanomas and its correlation with clinicopathologic parameters. METHODS: We analyzed the frequency of BRAF mutation in patients with primary cutaneous melanoma (n=58) or non-cutaneous one (n=27) by performing dual priming oligonucleotide-based multiplex real-time polymerase chain reaction to isolate and to purify the DNA from the formalin-fixed and paraffin-embedded tumors. RESULTS: The BRAF mutation was found in 17.2% (10/58) of patients with primary cutaneous melanoma and 11.1% (3/27) of those with non-cutaneous melanoma. The frequency of BRAF mutation was not correlated with any clinicopathologic parameters with the exception of the patient age. The frequency of the BRAF mutation was significantly higher in patients younger than 60 years as compared with those older than 60 years (p=0.005). CONCLUSIONS: Compared with previous reports, our results showed that the frequency of the BRAF mutation was relatively lower in patients with primary cutaneous melanoma. Besides, our results also showed that the frequency of the BRAF mutation had an inverse correlation with the age. Further studies are warranted to exclude methodological bias, to elucidate the difference in the frequency of the BRAF mutation from the previous reports from a Caucasian population and to provide an improved understanding of the molecular pathogenesis of malignant melanoma.

Vitiligo Developed in a Patient with Discoid Lupus Erythematosus / 대한피부과학회지

Vitiligo has been associated with various disorders, including pernicious anemia, diabetes, hyperthyroidism, alopecia areata, and Addison's disease. Discoid lupus erythematosus (DLE), the most common form of chronic cutaneous lupus erythematosus, is an autoimmune disease characterized by skin involvement. A 64-year-old man presented with multiple DLE lesions on the face and the neck, as well as photosensitive eczema on the nape of the neck. After treatment, the DLE lesions improved, but poliosis and several depigmented patches appeared on the vertex, the nape of the neck, and on the right hand. Few reports are available about the concurrence of DLE and vitiligo in other countries, and only one case, which showed coexisting systemic lupus erythematosus and vitiligo, has been reported in the Korean literature. The pathogenesis of concurrence of these two diseases has not been fully elucidated. Herein, we present a case of vitiligo that developed in a patient with DLE.

Chilblain Lupus Erythematosus: A Case Report and Review of Literature / 대한피부과학회지

Chilblain lupus erythematosus (LE) is a rare, chronic form of cutaneous LE (CLE), which presents mostly in women as erythematous to violaceous plaques on the acral areas and face, precipitated by cold and damp climates. It may be accompanied by discoid LE (DLE) lesions or other forms of CLE. Up to 20% of patients develop systemic LE (SLE). Although two missense mutations in TREX1, encoding the 3'-5' repair exonuclease 1, were described in familial chilblain LE, the pathogenesis of sporadic chilblain LE remains unknown. To our knowledge, there are a few reports of chilblain LE in the Korean dermatologic literature. Herein, we present a rare and interesting case of sporadic chilblain LE in 71-year-old man and review the Korean literatures.

Progressive Zosteriform Macular Pigmented Lesion / 대한피부과학회지

A progressive zosteriform macular pigmented lesion (PZMPL) is a chronic pigmentary dermatosis similar to progressive cribriform and zosteriform hyperpigmentation (PCZH), but characteristically accompanied by pruritus as a prodromal symptom and histologic findings, such as pigmentary incontinence. PZMPL was described by Simoes in 1980 and manifests a uniformly tanned macular pigmented lesion in a zosteriform distribution. PZMPL is usually preceded by multiple pruritic macular pigmentation in a part of the dermatome for a period of time. Despite the difficulty in differentiation among other pigmentary disorders with unilateral distribution, such as linear and whorled nevoid hypermelanosis, partial unilateral lentiginosis, and Becker's melanosis, PZMPL can be diagnosed by clinical symptoms and signs, the pattern of lesions, and histologic features. A 6-year-old Korean girl presented with unilaterally located brown macules and patches on the left side of the chest, arm, and back. From childhood, the pigmented macules appeared and coalesced into patches on the left back. The arm and chest lesions extended along the Blaschko's line in a zosteriform distribution. She had no history of previous skin diseases, injuries, or inflammation. The histologic findings revealed increased melanin pigment in the basal layer and focal pigmentary incontinence in the upper dermis. We report a rare case of PZMPL thought to be the same case reported by Simoes.

Clear Cell Syringoid Eccrine Carcinoma / 대한피부과학회지

Syringoid eccrine carcinoma (SEC) is a rare adnexal tumor with some controversy surrounding its correct definition. It may also be difficult to differentiate from its benign counterpart (syringoma), other adnexal carcinomas, and cutaneous metastasis from adenocarcinomas. Histologically, the tumor is characterized by syringoma-like tadpole morphology composed of basaloid cells with ductular differentiation. However, the tumor cells are deeply invasive and often extend to subcutaneous tissue, which distinguishes this malignancy from syringoma. Moreover, the tumor cells may rarely show striking vacuolization due to intracytoplasmic glycogen accumulation, called clear cell syringoid eccrine carcinoma. To our knowledge, no case of clear cell SEC in Korea has ever been reported. Herein, we present a case of a SEC (of the clear cell variant) complemented with an immunohistochemical study, the latter revealing cytoplasmic accumulation of glycogen and the presence of intercellular and intracellular lumina in clear tumor cells, as well as diverse hallmarks of SEC.

S Phase Cell Cycle Arrest and Apoptosis is Induced by Eugenol in G361 Human Melanoma Cells

Eugenol is an essential oil found in cloves and cinnamon that is used widely in perfumes. However, the significant anesthetic and sedative effects of this compound have led to its use also in dental procedures. Recently, it was reported that eugenol induces apoptosis in several cancer cell types but the mechanism underlying this effect has remained unknown. In our current study, we examined whether the cytotoxic effects of eugenol upon human melanoma G361 cells are associated with cell cycle arrest and apoptosis using a range of methods including an XTT assay, Hoechst staining, immunocytochemistry, western blotting and flow cytometry. Eugenol treatment was found to decrease the viability of the G361 cells in both a time- and dose-dependent manner. The induction of apoptosis in eugenol-treated G361 cells was confirmed by the appearance of nuclear condensation, the release of both cytochrome c and AIF into the cytosol, the cleavage of PARP and DFF45, and the downregulation of procaspase-3 and -9. With regard to cell cycle arrest, a time-dependent decrease in cyclin A, cyclin D3, cyclin E, cdk2, cdk4, and cdc2 expression was observed in the cells after eugenol treatment. Flow cytometry using a FACScan further demonstrated that eugenol induces a cell cycle arrest at S phase. Our results thus suggest that the inhibition of G361 cell proliferation by eugenol is the result of an apoptotic response and an S phase arrest that is linked to the decreased expression of key cell cycle-related molecules.

Leukocytoclastic Vasulitis Induced by Gefitinib / 대한피부과학회지

Gefitinib (Iressa(R), AstraZeneca) is an oral form of an anticancer drug called epidermal growth factor receptor-tyrosine kinase inhibitor. It is widely used for various solid cancers, including lung cancer. Cutaneous adverse reactions induced by gefitinib have recently been reported with an incidence ranging from 49% to 100%, and they include acneiform skin rash, hyperpigmentation, xerotic skin, pruritus, skin fissures, nail change and disorders of the mucous membranes, eyes and hair. To the best of our knowledge, no cases of leukocytoclastic vasculitis associated with gefitinib have ever been published in the Korean medical literature, and there have been only four such reported cases in other countries. Herein, we report on a case of leukocytoclastic vasulitis induced by gefitinib in a patient with lung cancer.

A Case of Fabry's Disease with Congenital Agammaglobulinemia

Fabry's disease is an X-linked lysosomal storage disorder caused by abnormalities in the alpha-galactosidase A (GLA) gene, which leads to a GLA deficiency and to the intracellular deposition of globotriaosylceramide (Gb3) within vascular endothelium and other tissues. It manifests as progressive multiple organ dysfunctions caused by the deposition of Gb3. On the other hand, congenital agammaglobulinemia is usually caused by mutations in Bruton's tyrosine kinase (Btk) gene with X-linked dominence, suppresses B cell maturation, and causes recurrent pyogenic infections. In former reports, the distance between the loci in the Xq22 region of the human X chromosome was found to be about 69 kilobases. A 23-yr-old man diagnosed with congenital agammaglobulinemia at age 5, showed typical clinical and laboratory and histopathological findings of Fabry's disease. The genetic basis of this combination of the two syndromes was studied in this patient. Here, we report a case of Fabry's disease with congenital agammaglobulinemia.

Malassezia folliculitis Resembling Erythema Annulare Centrifugum / 대한피부과학회지

Malassezia yeasts are lipophilic fungi that are regarded as normal flora of the skin. They are associated with a number of diseases affecting human skin, such as pityriasis versicolor, Malassezia folliculitis, and seborrheic dermatitis, and less commonly with other dermatologic disorders, such as confluent and reticulated papillomatosis and transient acantholytic dermatosis. Clinically, Malassezia folliculitis shows erythematous papules, sometimes pustules. Here, mutiple erythematous papules and pustules over the trunk and upper arms developed in a 37-year-old man. The skin lesions became enlarged and confluent, resulting in the formation of an annular configuration. They spread gradually to form a large plaque with central hyperpigmentation, with the edge of the lesions showing multiple papules and pustules. Microspores were found in potassium hydroxide preparations of these lesions, and large-sized circular, cream colored, raised, smooth, and dentate colonies were found in culture media. Histologically, sectioning showed mild acanthosis of the epidermis and perivascularinflammation in the superficial dermis. PAS-stained sections showed spheric to oval yeast organisms in the follicularepithelium. In PCR analysis, Malassezia sympodialis was identified. The patient was treated with oral itraconazle, resulting in complete clearing of the lesions. Here, we report an interesting case of Malassezia follicultis that showed clinical configuration of erythema annulare centrifugum.

CD20 Positive T Cell Lymphoma Involvement of Skin

CD20 positive T cell lymphoma is a rare condition that is associated with the coexpressions of CD20 and T cell markers, such as, CD3, CD5, or UCHL-1. Positivity for CD20 in this tumor represents an aberrant immunophenotype, but the presence of monoclonal T cell receptor (TCR) gene rearrangements and negativity for immunoglobulin heavy chain gene rearrangement indicate that this tumor is a T cell lymphoma. The majority of cases of CD20 positive T cell lymphoma have been reported as immature peripheral T cell lymphoma not otherwise specified. However, we believe that this disease is likely to be re-listed as a new disease entity after its pathogenesis has been elucidated and more cases have been evaluated. Here, we present a case of peripheral T cell lymphoma coexpressing CD20 and T cell markers with a demonstrable TCR gene rearrangement, in a patient who had been misdiagnosed as having B cell type lymphoma 4 years previously. We hypothesize that in this case initially circulating normal CD20+ T cell subsets underwent neoplastic transformation and CD20 positive T cell lymphoma subsequently developed in the lymph node, and then recurred in the skin due to systemic disease or metastasized from the nodal disease.