Pemphigus vulgaris

Abstract: Pemphigus vulgaris is a chronic autoimmune bullous dermatosis that results from the production of autoantibodies against desmogleins 1 and 3. It is the most frequent and most severe form of pemphigus, occurring universally, usually between 40 and 60 years of age. It usually begins with blisters and erosions on the oral mucosa, followed by lesions on other mucous membranes and flaccid blisters on the skin, which can be disseminated. There is a clinical variant, pemphigus vegetans, which is characterized by the presence of vegetating lesions in the large folds of the skin. Clinical suspicion can be confirmed by cytological examination, histopathological examination, and direct and indirect immunofluorescence tests. The treatment is performed with systemic corticosteroids, and immunosuppressive drugs may be associated, among them azathioprine and mycophenolate mofetil. More severe cases may benefit from corticosteroids in the form of intravenous pulse therapy, and recent studies have shown a beneficial effect of rituximab, an anti-CD20 immunobiological drug. It is a chronic disease with mortality around 10%, and septicemia is the main cause of death. Patients need long-term and multidisciplinary follow-up.

Pemphigus foliaceus treated with rituximab monotherapy

In pemphigus foliaceus (PF), immunoglobulin G (IgG) autoantibodies directed against desmoglein-1 (Dsg-1), a cell adhesion molecule expressed mainly in the granular layer of the epidermis, are responsible for the intercellular widening between desmosomes resulting in intraepidermal blisters. Rituximab is a chimeric monoclonal antibody that by binding specifically to the transmembrane antigen CD20 found on the surface of normal and malignant B cells, leads to B-cell depletion. We report a 19-year-old Filipino woman with PF and controlled idiopathic thrombocytopenia purpura, initially treated with high-dose prednisone and azathioprine. Due to rapid PF progression with associated moderate reactive depression, rituximab was added to the treatment regimen with prompt improvement of lesions and clearance after five months. Five years later, lesions recurred with erythematous, dry, scaly plaques on both breasts, axillae, and on the scalp, associated with moderate to severe intermittent pruritus. After the first of a series of four weekly infusions, rituximab monotherapy resulted in immediate and sustained clearance up to 22 months. In parallel with skin clearance, serum CD19 and CD20 B cells decreased to almost zero after the first infusion, to zero after the second, while the decrease of Dsg-1 levels was more gradual, and down to normal after four months.We offer this case report to show that rituximab can be given as a first-line monotherapy option for indications similar to ours such as drug reactions (steroid-induced depression) or a history of recalcitrant PF to the usual medications; and to suggest using CD19 and CD20 in addition to the desmoglein levels to monitor disease activity and molecular change from which to learn how to continue to monitor for disease activity after clearance.

A Bama Minipig Model of Laryngopharyngeal Reflux and the Change of Laryngopharyngeal Mucosal Ultrastructure

BACKGROUND/AIMS: To establish an animal model of laryngopharyngeal reflux (LPR) and study the effect of LPR on the laryngopharyngeal mucosal ultrastructure. METHODS: Ten Bama minipigs were randomly divided into control group and stent group. Every pig underwent endoscope, and baseline pH was monitored for 4 hours at laryngopharynx and distal esophagus, then specimens from laryngopharyngeal mucosa were biopsied. For the control group, these procedures were repeated on the 14th day. In the stent group, a custom-designed esophageal stent suit was implanted into esophagus, laryngopharyngeal and distal esophageal pH monitoring lasted for 2 hours, then stent suit was removed 3 days later. At last, the same procedures were done as the control group on the 14th day. Specimens were observed under transmission electron microscope to measure the intercellular space and desmosome number. RESULTS: In the control group, there was no laryngopharyngeal reflux on the first day and 14th day. Before the stent was implanted, there was also no laryngopharyngeal reflux in the stent group. In both 2 hours and 14 days after stent implantation, the num -ber of reflux, reflux time, and percentage time of pH < 4.0 were significantly increased (P < 0.05) in the stent group. There was no difference in intercellular space and desmosomes in the control group between baseline and 14th day. In the stent group, intercellular space of laryngopharyngeal mucosa was significantly increased (0.37 mum vs 0.96 mum, P = 0.008), and the number of desmosomes was significantly decreased (20.25 vs 9.5, P = 0.003). CONCLUSIONS: A Bama minipig model of LPR was established by implanting a custom-designed stent suit. LPR might destroy the laryngophar yngeal mucosal barrier.

Dynamic assembly of intercalated disc during postnatal development in the rat myocardium / 生理学报

The intercalated disc (ICD) complex of cardiomyocyte consists of fascia adherens, desmosomes and gap junctions which are mainly constructed by their transmembrane proteins: N-cadherin (N-cad), desmoglein-2 (DSG2) and connexin 43 (Cx43), respectively. The aim of this study was to observe the dynamic changes in colocalization of N-cad, DSG2 and Cx43 with each other in the rat left ventricular myocardium at 1, 7, 14, 28 and 90 day(s) after birth (P1, P7, P14, P28 and P90) using immunofluorescent staining. The results showed that, N-cad, DSG2 and Cx43 located all around the plasma membrane at the P1. These proteins accumulated to the long ends of cardiomyocytes, indicating preliminary formation of the ICD at the P7. The localization of three proteins at the ICD increased progressively, but their lateral localization showed an inverse trend from the P14 to P90. However, Cx43 still kept a certain amount of lateral localization in cardiomyocytes even at the P90 as compared with N-cad and DSG2. Quantitative colocalization of proteins was analyzed by the stereological method. Total percentage of colocalization of N-cad with DSG2 was 33.5% at the P1, and increased to 38.6% at the P7, 9.4% in ICD and 29.2% in lateral side. The total percentage of colocalization of N-cad with DSG2 increased to 65.7% at the P90, ICD colocalization increasing to 60.5% and lateral colocalization decreasing to 5.2%. Total percentage of colocalization of N-cad with Cx43 increased from 10.3% at the P1 to 37.1% at the P90, and only ICD colocalization increased, but lateral colocalization kept about 5%. The colocalization pattern of DSG2 with Cx43 was similar to that of N-cad with Cx43. Total percentage of colocalization of N-cad with DSG2 was higher than those of N-cad or DSG2 with Cx43. The above results suggest that the formation of mechanical junctions at the ICD of cardiomyocyte is prior to that of electrochemistry junctions during postnatal development. In other words, cardiomyocyte growth needs a stable mechanical environment at first.

Imprint Cytology of Soft Tissue Myoepithelioma: A Case Study

Soft tissue myoepithelioma is a rare neoplasm composed of myoepithelial cells. Here, we describe the cytologic features of soft tissue myoepithelioma arising on the right forearm in an 18-year-old man. The excised tumor (3.0x1.8x1.5 cm) was well-demarcated, yellow-gray, soft, and myxoid. The cytologic smears showed round to spindle, epithelioid, and plasmacytoid cells in the myxoid background. The nuclei were uniform, round to ovoid, with finely distributed chromatin and eosinophilic or pale cytoplasm. The tumor cells demonstrated immunoreactivity for cytokeratin (AE1/AE3), epithelial membrane antigen, S100 protein, and glial fibrillary acidic protein. Electron microscopy showed intermediate filaments, desmosomes, and basal lamina.

Immunohistochemical Study on the Expression of Desmocollin 1 during Skin Development / 대한피부과학회지

BACKGROUND: Desmocollins (Dsc) are calcium-dependent transmembrane glycoproteins of desmosomes that are important in the junction complex of epidermis and maintain structural integrity of the skin from external stressors. Among three Dscs (Dsc 1, 2, 3), Dsc 1 and 3 are distributed on skin. OBJECTIVE: The purpose of this study was to observe the Dsc 1 distribution pattern on the skin and oral mucosa during fetal development. METHODS: Skin was obtained from the sole and scalp of 33 fetuses, ranging from 10 to 37 weeks of gestational age. Immunohistochemical staining was performed on the paraffin-embedded tissue using a Dsc 1 monoclonal antibody. RESULTS: Dsc 1 was expressed in the suprabasal layer but not in the basal layer of the epidermis of the sole at the 10th week of gestation. Thereafter, Dsc 1 expression further increased in the suprabasal layer with initiation of stratification and increased gradually in the granular layers of the sole and scalp epidermis. Dsc 1 was strongly expressed in the superficial layer of the infundibulum and inner root sheath of the hair follicle but was not expressed in the sebaceous cells or other hair components. The eccrine duct epithelium was focally and weakly positive for Dsc 1 expression. Furthermore, Dsc 1 was not expressed in oral mucosa, although the oro-cutaneous portion was strongly expressed in the superficial layer. CONCLUSION: Dsc 1 was strongly expressed in the suprabasal cells of the epidermis during fetal skin development, and expression increased gradually in the granular layer and inner root sheath of the hair follicle. However, Dsc 1 was not expressed in basal cells or in oral mucosa. Dsc 1 may play a role in the maintenance of epithelial integrity as part of desmosomes.

Inmunopatogenia del pénfigo vulgar y el pénfigo foliáceo / Immunopathogenesis of pemphigus vulgaris and pemphigus foliaceus

El pénfigo vulgar y el pénfigo foliáceo son enfermedades ampollosas autoinmunes mediadas por autoanticuerpos dirigidos contra proteínas de los desmosomas, las desmogleínas 1 y 3. Están asociadas con moléculas del complejo mayor de histocompatibilidad (HLA) que por su estructura tienen la capacidad de presentar péptidos antigénicos de las desmogleínas. En los individuos afectados se han descrito la presencia de linfocitos T y B autorreactivos y alteraciones en la regulación del sistema inmune con desequilibrio de las respuestas Th1/Th2. No se conocen con precisión los mecanismos de daño pero la investigación actual indica que los anticuerpos tienen un papel patogénico, inician diferentes cascadas de señalización que provocan la acantólisis y apoptosis de los queratinocitos. El conocimiento de la inmunopatogenia de las enfermedades ampollosas autoinmunes ha permitido el desarrollo y la puesta en práctica de nuevas alternativas terapéuticas.

Pemphigus vulgaris and pemphigus foliaceus are autoimmune blistering diseases mediated by antibodies against desmosomal proteins. They are strongly associated with major histocompatibility complex alleles with the ability to present antigenic peptides of desmogleins. In the affected individuals the presence of auto-reactive T and B lymphocytes, and alterations in the immune system regulation with imbalance of the Th1/Th2 responses have been described. Damage mechanisms are not yet precisely known but current investigation indicates that antibodies play an important pathogenic role: they start different signaling cascades that lead to acantholysis and apoptosis of keratinocytes. Better knowledge of the pathogenesis of autoimmune blistering diseases has been the basis for the development and implementation of new therapeutic approaches.

Expression of Desmoglein-1 in Fetal Skin Development / 대한피부과학회지

BACKGROUND: Desmosomes are cell-cell adhesion complexes that provide mechanical integrity to keratinocytes by linking them to keratin intermediate filaments. Desmosomes are composed of two major transmembrane proteins, desmoglein and desmocollin. In humans, four desmoglein isoforms have been identified: Dsg1, Dsg2, Dsg3, and Dsg4. Desmogleins are Ca2+-dependent adhesion molecules and play important parts in the formation and maintenance of desmosomes. Desmoglein-1 is the main skin-associated desmosomal cadherin. It is expressed throughout the epidermis, but most prominently in the differentiated layers. OBJECTIVE: The purpose of this study was to observe the distribution pattern of desmoglein-1 in the skin and oral mucosa during fetal development. METHODS: Skin was obtained from the sole and scalp of 35 fetuses, ranging from 10 to 37 weeks of gestational age. Immunohistochemical staining was performed on paraffin embedded tissue using anti-human monoclonal antibody against desmoglein-1. RESULTS: Expression of desmoglein-1 in the epidermis appeared in the upper layer of the sole, but the basal layer was negative at the 10th gestational age. Thereafter, stratification began with stronger expression in the middle layer than in the basal layer of the sole and scalp epidermis. Expression in the middle spinous layer is stronger in the fetal period than in other layers of the epidermis. Expression in the superficial layer seemed to increase in later stages. Expression of desmoglein-1 in hair was strong in the infundibulum, inner root sheath, sebaceous glandular epithelium, and eccrine duct epithelium. Expression of desmoglein-1 in oral lip mucosa was very weak or negative in the upper half of the mucosal epithelium, though the lower half was strongly positive, while the skin side of the mucosa was similar with the sole skin. CONCLUSION: Desmoglein-1 may play a complementary role in the maintenance of epithelial integrity along with other desmogleins, because desmoglein-1 distribution is slightly different from that of desmoglein-3 in epidermis, hair and mucosa in fetal skin development.

Role of the Chinese herbal medicine xianhuayin on the reversal of premalignant mucosal lesions in the golden hamster buccal pouch / 国际口腔科学杂志·英文版

<p><b>AIM</b>To investigate the role of the Chinese herbal medicine Xianhuayin on the reversal of 7,12-dimethylbenz[a]anthracene (DMBA)-induced premalignant mucosal lesions in the oral buccal pouch of golden hamsters.</p><p><b>METHODOLOGY</b>The animals were randomly divided into a non-diseased control group (n=5) and an experimental group including 50 animals in which the buccal mucosa had been painted with DMBA (0.5% in acetone) to generate an oral mucosa premalignant lesion. Animals in the experimental group were further divided into Xianhuayin-treated group (n=30), untreated premalignant lesion group (n=10) and normal saline (NS)-treated group (n=10). The cheek (buccal) pouch mucosa of the golden hamsters in each group was observed with light and electron microscopy eight weeks after intragastric administration with NS or Xianhuayin.</p><p><b>RESULTS</b>In the non-diseased control group, the buccal mucosa was keratinized and stratified squamous epithelium under a light microscope. In the untreated premalignant lesion group, variable degrees of epithelial dysplasia was observed. The irregular epithelial mucosa gradually became distinct in the Xianhuayin-treated group. Scanning electronic microscopic (SEM) analysis showed that surface of the cells exhibited honeycomb structures in the hamster of untreated-group. The cells were morphologically irregular, overlapped and loosened in the untreated premalignant lesion group. Most of the cell surface exhibited honeycomb structure in the Xianhuayin-treated group. Transmission electronic microscopic (TEM) analysis showed that buccal mucosal epithelial cells were morphologically regular in the non-diseased control group. Desmosomes and tonofibrils were reduced and the nucleus was morphologically irregular in the untreated premalignant lesion group. In the Xianhuayin-treated group, the widening intercellular gap was gradually reduced, desmosomes and the cells becoming morphologically regular. No significant difference was observed between the hamsters in NS-treated group and those in the untreated premalignant lesion group. Significant therapeutic efficacy was observed in the group receiving Xianhuayin.</p><p><b>CONCLUSION</b>Xianhuayin is effective in the reversal of DMBA-induced premalignant lesions in the buccal pouch of golden hamsters.</p>

Immunohistochemical Study on the Expression of Desmoglein-3 in Fetal Skin Development / 대한피부과학회지

BACKGROUND: Desmogleins are calcium-dependent transmembrane glycoproteins of the desmosome that form an import component of the junction complexes of epithelial cells. Desmogleins are involved in maintaining the structural integrity of tissues. So far, four different desmogleins (Dsg1, Dsg2, Dsg3 and Dsg4) have been identified. OBJECTIVE: The purpose of this study was to observe the distribution pattern of desmoglein-3 in the fetal skin during development. METHODS: Skin was obtained from the sole, scalp and lip of 34 fetuses that ranged in age from 10 to 39 weeks of gestational age. Immunohistochemical staining was performed on the paraffin embedded tissue using anti-human monoclonal antibody against the desmoglein-3. RESULTS: The expression of desmoglein-3 in the epidermis appeared in the basal layer of the sole at the 10th week of gestation age. Thereafter, a stronger expression was noted in the middle layer of the sole and scalp epidermis. The basal layer had a stronger expression than did the other layers of the epidermis, followed by the middle and superficial layers. A stronger expression of desmoglein-3 in hair was noted in the outer root sheath, the bulge cells and the eccrine duct cells. The expression of desmoglein-3 in the lip mucosa was strong in both the basal and middle layers, while the skin side of the mucosa showed a stronger expression in basal layer. CONCLUSION: These results suggested that desmoglein-3 plays an important role in the development and differentiation of the epidermis and skin adnexa in the fetal stage, and especially in basal and suprabasal layers.

Matrix Metalloproteinases and Autoimmune Bulbous Diseases: Expression of MMP-2, -3, and -9 in Bullous Pemphigoid, Pemphigus Vulgaris, and Pemphigus Foliaceus / 대한피부과학회지

BACKGROUND: Matrix metalloproteinases (MMPs) participate in extracellular matrix degradation and may play an important role in basal membrane damage in many dermatologic diseases. Recent studies implicated the importance of MMP-9 in the pathogenesis of bulla formation of bullous pemphigoid (BP). Various autoimmune bullous diseases are strongly associated with desmosome or hemidesmosome pathologies, and show an increased level of lesional MMP and exposed autoantigens from these structures. OBJECTIVE: This study evaluated the level of MMP-2, -3, and -9 in three types of autoimmune bullous disease [BP, pemphigus vulgaris (PV), pemphigus foliaceus (PF)] with the aim of investigating the role of MMPs in the pathogenesis of autoimmune bullous diseases. METHODS: Sample specimens were obtained from skin lesions of patients with BP (n=12), PV (n=10), and PF (n=12), and from normal controls (n=8). The immunohistochemical expression of MMP-2, -3, and -9 was analyzed and serum levels of MMP-2, -3, and -9 were measured by enzyme-linked immunosorbant assay (ELISA). The results were analyzed with reference to graded levels of clinical severity. RESULTS: Expression of dermal MMP-2, -3, and -9 were increased in BP, PV, and PF (p=0.036, 0.022, and 0.015, respectively). However, decreased expression of the three MMPs in the epidermis of skin lesions may have resulted from epidermal destruction. ELISA-determined serum levels of MMP-2, -3, and -9 increased in BP, PV and PF. Interestingly, MMP-2 was significantly increased in the sera of BP patients (p=0.015), consistent with the previous studies concerning the role of gelatinase (MMP-2 and -9) in the pathogenesis of BP. In BP patients, clinical severity was proportional to increased levels of MMP-2 in both skin lesions and and sera. CONCLUSION: The increased expression of MMP-2, -3, and -9 in skin lesions and sera may reflect the involvement of these enzymes in the mechanism of bulla formation in autoimmune bullous diseases including BP. In addition, expression of MMP and clinical severity may be closely connected.

Transmission electron microscopy of the preclinical phase of experimental phytophotodermatitis

OBJECTIVE: To examine the epidermis in induced phytophotodermatitis using transmission electron microscopy in order to detect histologic changes even before lesions are visible by light microscopy. INTRODUCTION: In the first six hours after the experimental induction of phytophotodermatitis, no changes are detectable by light microscopy. Only after 24 hours can keratinocyte necrosis and epidermal vacuolization be detected histologically, and blisters form by 48 hours. METHODS: The dorsum of four adult rats (Rattus norvegicus) was manually epilated. After painting the right half of the rat with the peel juice of Tahiti lemon, they were exposed to sunlight for eight minutes under general anesthesia. The left side was used as the control and exposed to sunlight only. Biopsies were performed immediately after photoinduction and one and two hours later, and the tissue was analyzed by transmission electron microscopy. RESULTS: No histological changes were seen on the control side. Immediately after induction, vacuolization in keratinocytes was observed. After one hour, desmosomal changes were also observed in addition to vacuolization. Keratin filaments were not attached to the desmosomal plaque. Free desmosomes and membrane ruptures were also seen. At two hours after induction, similar changes were found, and granular degeneration of keratin was also observed. DISCUSSION: The interaction of sunlight and psoralens generates a photoproduct that damages keratinocyte proteins, leading to keratinocyte necrosis and blister formation. CONCLUSIONS: Transmission electron microscopy can detect vacuolization, lesions of the membrane, and desmosomes in the first two hours after experimental induction of phytophotodermatitis.

Effect of ionizing radiation on rat parotid gland

A common side effect of radiotherapy used in the treatment of oral cancer is the occurrence of structural and physiological alterations of the salivary glands due to exposure to ionizing radiation, as demonstrated by conditions such as decreased salivary flow. The present study evaluated ultrastructural alterations in the parotid glands of rats receiving a fractionated dose (1,500-cGy) of radiation emitted by a Cesium-137 source and rats that were not subjected to ionizing radiation. After sacrifice, the parotid glands were removed and examined by transmission electron microscopy. Damage such as cytoplasmic vacuolization, dilatation of the endoplasmic reticulum and destruction of mitochondria, as well as damage to the cellular membrane of acinar cells, were observed. These findings lead to the conclusion that ionizing radiation promotes alterations in the glandular parenchyma, and that these alterations are directly related to the dose level of absorbed radiation. Certain phenomena that appear in the cytoplasm and nuclear material indicate that ionizing radiation causes acinar cell death (apoptosis).

Um efeito colateral comum da radioterapia usada no tratamento de câncer na cavidade oral é a ocorrência de alterações estruturais e fisiológicas sobre as glândulas salivares por exposição à radiação ionizante, como demonstrada em situações com decréscimo do fluxo salivar. O presente estudo teve por objetivo avaliar as alterações ultra-estruturais de glândulas parótidas de ratos que receberam uma dose fracionada (1500 - cGy) de radiação emitida por uma fonte de Césio 137 e ratos que não receberam a radiação ionizante. Após o sacrifício, as glândulas parótidas foram removidas e examinadas por microscopia eletrônica de transmissão. Lesões das organelas citoplasmáticas, como dilatação do retículo endoplasmático, destruição das mitocôndrias e formação das vacuolizações citoplasmáticas, além de lesão da membrana celular das células acinares foram observadas. Portanto, a radiação ionizante promove alterações no parênquima glandular, o que está diretamente relacionado com a dose de radiação absorvida. Determinados fenômenos que surgem no citoplasma e material nuclear são indicadores de que a radiação ionizante leva a célula acinar a morte programada (apoptose).

Mutación de la proteína de Desmosoma Plakofilina-2 en una familia guanacasteca / Mutation of the protein Desmosoma plakofilina 2 in a family guanacasteca

Se reporta una mutación en la plakofilina-2, proteína del Desmosoma miocárdico, encontrada en dos pacientes de una misma familia procedente de Guanacaste, Costa Rica, portadores de Displasia/cardiomiopatía arritmogénica del ventrículo derecho sintomática (ARVC) y tratados con cariodesfibrilador implantable.

Electron microscopic study of desmosomes and tonofilaments in Lichen planus

Lichen planus [L.P] is an idiopathic inflammatory disease of the skin and mucous membrane. Desmosomes are responsible for the adhesion of keratinocytes. Tonofilaments are one of the major cytoskeleton structure in mammalian epidermis. Six patients complained from Lichen planus are included in this study for the study of ultrastructure of both desmosomes and tonofilaments. Revealed an increase in the size and numbers of both of them which can be explained as a one of the defensive mechanism of the cells against frequent rubbing which accompanied L.P. which is considered as a resistance mechanism of keratinocytes

Localization of protein kinase C [PKC] alpha at the cell junction desmosomes during epidermal wound healing in mice

Desmosomes are intercellular junctions that provide strong adhesion between epidermal keratinocytes. This function is rapidly modulated from calcium independent to calcium dependent upon wounding of an epithelial cell sheet. This modulation was found to be signaled by an isoenzyme named protein kinase C alpha [PKC alpha] which alters the adhesive state of desmosomes at the wound edges. Specimens from intact and wounded mice skin were taken and prepared for immunohistochemical studies and the immune reactions of both desmosomes and PKC alpha were examined, recorded and discussed. It is hypothesized that wounding triggers a stimulus that activates PKC alpha which through a phosphorylation of certain desmosomal proteins modulates desmosomal adhesion. This breaks intercellular contacts enabling keratinocytes to migrate and reepithelialize the wound

The Expressions of Desmoglein 1 and 3 according to Chronologic Skin Aging / 대한피부과학회지

BACKGROUND: Desmogleins are transmembrane glycoproteins of the desmosome which provide mechanical strength to epithelial tissue. Desmogleins have so far, been implicated in several diseases such as pemphigus, striate palmoplantar keratoderma, 4S and squamous cell carcinomas. Skin cancer usually occurs in old age. And there are reports that the expression of desmogleins are increased in squamous cell carcinoma. However the role of desmogleins in skin aging has not yet been reported. OBJECTIVE: The purpose of this study was to investigate the expression of desmoglein 1 and 3 according to chronologic skin aging. METHODS: A total of 6 normal tissue samples from sun-protected skin of different age groups (from 34-year-old to an 84-year-old) and 1 squamous cell carcinoma tissue from a 72-year-old patient were taken. Western blotting and immunohistochemical staining were performed with anti desmoglein 1 and 3 antibodies. The expression of desmoglein 1 and 3 by Western blotting were calculated semiquantitatively by a densitometer. RESULTS: The expression of desmoglein 1 was 0.382 in the 34-year-old, 0.450 in the 45-year-old, 0.369 in the 56-year-old, 0.761 in the 65-year-old, 1.035 in the 77-year-old and 1.329 ODu/mm2 in the 84-year-old. The expression of desmoglein 3 was 0.830 in the 34-year-old, 0.984 in the 45-year-old, 1.029 in the 56-year-old, 1.534 in the 65-year-old, 1.714 in the 77-year-old and 1.878 ODu/mm2 in the 84-year-old. In immunohistochemical staining, the expression of Dsg1 increased from the basal layer to the granular layer and Dsg3 was expressed in the basal and suprabasal layers. CONCLUSION: The expression of desmoglein 1 and 3 were increased according to chronologic skin aging.

Sarcomatoid Carcinoma of the Pancreas: A Case Report

We report here on an unusual case of a 73-year-old Korean female with sarcomatoid carcinoma. This tumor was composed of pancreatic ductal adenocarcinoma and a malignant fibrous histiocytoma-like stroma. The CT imaging revealed a multiseptated heterogenous hypodense mass that was 15 cm in size. The mass was located in the body and tail of the pancreas, spleen and gastrosplenic area. The pathologic examination showed that the carcinomatous component was negative for vimentin, and the sarcomatous component was positive for vimentin and CD 68. The ultrastructural examination showed that both the carcinomatous and sarcomatous components had desmosomes at the cell-cell contact sites. The patient refused postoperative adjuvant chemotherapy and she died of cachexia with generalized tumor extension about 3 months later. This report presents special data that can clarify the clinicopathological features and pathogenesis of this rare neoplasm.

Application of Human Dermal Fibroblast and Keratinocyte on Allogenic Dermis(AlloDerm(R))

PURPOSE: Large skin defect by various causes, should be covered by autologous skin graft. But, the donor site of autologous skin graft is limited and leaves permanent donor scar and contracture. There have been our trial to engineer artificial skin using allogenic dermis (AlloDerm) with basement membrane. METHODS: Dermal and epidermal layer were separated by immersing in dipase solution for 30 minutes, and the separated layers were treated with 0.05% trypsin for 10 minutes. And then each layer was cultivated to fibroblasts and keratinocytes on a culture medium. Fibroblasts were first penetrated into basement membrane of allogenic dermis facing down, then allogenic dermis was flipped over to face up and keratinocytes were transplanted to allogenic dermis. RESULTS: Observing artificial skin fabricated in vitro, we found following: 1) The artificial skin opened in air for 5 days formed epidermal layer. In dermal layer, fibroblast was distributed evenly among all. 2) The artificial skin opened in air for 30 days formed thicker and thicker, and it formed basement membrane, spinous and granular layers. PAS stain to confirm existence of basement membrane showed positive reaction. 3) Cytokeratin 10 stain to confirm the formation of epidermal layer showed positive reaction. 4) The formation of thick keratin, lamellar body and desmosome similar to human skin were observed in result of an electron micrograph. CONCLUSION: As a result of research, the structure seen in normal skin such as rete ridge, is found in reproduced artificial skin. This type of artificial skin can be used as a useful model for investigating skin disease and for clinical application also.