A visible cutaneous scar develops from the excess formation of immature collagen in response to an inflammatory reaction. This study examined the role of epidermal growth factor (EGF) in the formation of cutaneous scars. Twenty Crl:CD-1 (ICR) mice were used and 2 full-thickness skin wounds were made on the dorsum of each mouse. One of the wounds was treated with recombinant human EGF by local application and the other was treated with saline for control until complete healing was achieved. The EGF-treated group's wounds healed faster than the control group's. The width of the scar was smaller by 30% and the area was smaller by 26% in the EGF-treated group. Inflammatory cell numbers were significantly lower in the EGF-treated group. The expression of transforming growth factor (TGF)-beta1 in the EGF-treated group was increased. It was observed that the amount of collagen in the EGF-treated group was larger than the control group. In the EGF-treated group, the visible external scars were less noticeable than that in the control group. These results suggest that EGF can reduce cutaneous scars by suppressing inflammatory reactions, decreasing expression of TGF-beta1, and mediating the formation of collagen.
Animals ; Cicatrix/pathology/*prevention & control ; Collagen/metabolism ; Epidermal Growth Factor/*pharmacology ; Humans ; Inflammation/metabolism ; Mice ; Recombinant Proteins/*pharmacology ; Skin/drug effects/metabolism/pathology ; Wound Healing/*drug effects

PURPOSE: Oral mucositis is a common toxicity of radiation or chemotherapy, which is used a treatment for head and neck cancer. We investigated effects of recombinant human epidermal growth factor (rhEGF) on radiation-induced oral mucositis in rat model. MATERIALS AND METHODS: Spraque-Dawley rats (7 per group) exposed to a single dose of 25 Gy (day 0) on their head, except for one group, were randomly divided into un-treated, vehicle-treated, and two rhEGF- treated groups. Rats were topically applied with rhEGF (15 or 30 microgram/oral cavity/day) or vehicle to their oral mucosa. Survival rate of rats, weight changes, and food intakes were examined from day 0 to 18 after radiation. Histology study was performed from oral mucosa of rats at day 7 and 18 after radiation. RESULTS: rhEGF-treated groups (15 or 30 microgram/day) showed all survival rate 33%, whereas un-treated and vehicle-treated groups showed all survival rate 0% at the end of experiment. rhEGF-treated groups statistically had less weight loss compared to vehicle-treated group from day 2 to 7 after radiation. Food intake of rats with rhEGF treatment turned to increase at day 14 after radiation. At 7 day after radiation, un-treated and vehicle-treated groups showed severe pseudomembraneous or ulcerative oral mucositis. On the other hand, rhEGF-treated groups had no more than cellular swelling and degeneration of epidermal cells in oral mucosa of rats. CONCLUSION: These results suggest that rhEGF has significantly positive effects on radiation-induced oral mucositis in rats. rhEGF display a therapeutic potential on a clinical level.
Animals ; Drug Therapy ; Eating ; Epidermal Growth Factor* ; Hand ; Head ; Head and Neck Neoplasms ; Humans* ; Models, Animal ; Mouth Mucosa ; Rats* ; Stomatitis* ; Survival Rate ; Ulcer ; Weight Loss

BACKGROUND: Sporotrichosis is a common deep cutaneous fungal disease caused by Sporothtix (S.) schenckii. The recent development of polymerase chain reaction (PCR) technology, in particular, arbitrarily primed PCR (AP-PCR) or random amplified polymorphic DNA (RAPD), has greatly enhanced the molecular detection and identification of various pathogenic agents, including fungi. OBJECTIVE: This study was aimed to differentiate Sporothrix schenckii, and related fungi such as S. schenckii var. luriei, S. flocculosa, S. nivea, Ophiostoma stenoceras, and clinical isolates on the basis of distinct DNA band patterns in the RAPD. METHODS: S. schendcii, S. schenckii var. luriei, S. flocculosa, S. nivea, Ophiostoma stenoceras from ATCC and KCCM, and clinical 10 isolates from Chonnam University Hospital were used for RAPD analysis. For RAPD, 3 random primers were used. Genomic DNA was extracted by Liu method. Amplification reactions were performed in volumes of 50 microL containing 10 mM Tris-HCI (pH 8.3), 50 mM KCl, 1.5 mM MgCl2, 0.1% Triton X-100, 200microM dNTP mixture, 40 pM primer, 1 U of Taq polymerase, DNA 20 ng. RESULTS: 3 decamers (5'-TGCCGAGCTG-3', 5'-AGTCAGCCAC-3', 5'-AATCGGGCTG-3') are generated in the RAPD, distinct DNA products from S. schenckii forming characteristic band patterns upon gel electrophoresis. Each random primer amplified characteristic same band patterns in DNA from clinical 8 isolates among 10 isolates, 2 isolates have different DNA band patterns. These results suggest of being a Sporothrix anamorph different from S. schenckii in Korea. CONCLUSION: With 2 random primers (5'-TGCCGAGCTG-3', 5'-AGTCAGCCAC-3') S. schenckii and related fungi investigated produced distinct DNA band patterns on gel electrophoresis. The RAPD was a very valuable laboratory method for identification of S. schenckii isolates.
DNA* ; Electrophoresis ; Fungi* ; Jeollanam-do ; Korea ; Magnesium Chloride ; Octoxynol ; Ophiostoma ; Polymerase Chain Reaction ; Sporothrix* ; Sporotrichosis ; Taq Polymerase

BACKGROUND: Sunscreens have been used widely to prevent the photosensitive skin diseases, skin cancer, and skin aging. However, no sunscreen blocks all kinds of effects caused by ultraviolet light(UVL), and the effect of sunscreens on the impairment of immune function by UVL irradiation is controversial. OBJECTIVE: We try to evaluate the efficiency of sunscreens for blocking the depletion of LC induced by UVB irradiation. METHOD: The ATPase positive LCs were observed in the skin of hairless mice(Hr+/Kud) irradiated by UVB with or without topical application of sunscreens. Two commercially available sunscreens with respective SPF 8 and SPF 30 were applied to the dorsal trunk skin. The mice were irradiated with different increasing doses of UVB at a single time. RESULTS: The ATPase positive LCs in the irradiated dorsal and ear skin were significantly de-creased in densities according to the dosage, and apparently revealed a loss of their dendrites, granulation, and clumping from a UVB dose of more than 60mJ/Cm2. With both sun-screen treatment on the dorsal trunk before irradiation, the densities of LCs on the dorsal skin were significantly higher compared to the un-treated groups at all ranges of UVB doses in spite of a dose dependent decrease in their density. However there was no significant difference on their preventive effect between both sunscreens(SPF 8 and SPF 30) except at high UVB dos-es of more than 240mJ/Cm². CONCLUSION: The LC depletion induced by UVB can be partially protected through the topical application of a sunscreen at a UVB dose dependent fashion. However SPF(sun protective factor) dose not appear to be a good indicator for evaluating sunscreens immunologically.
Adenosine Triphosphatases ; Animals ; Dendrites ; Ear ; Langerhans Cells* ; Methods ; Mice ; Mice, Hairless* ; Skin ; Skin Aging ; Skin Diseases ; Skin Neoplasms ; Sunscreening Agents*

BACKGROUND: Malignant skin tumor cells derived from epidemal keratinocyte penetrate the basement membrane to proliferate in dermal interstitial strozirarnd invade surrounding tissue and finally metastasize to distant organs. In this stage of invaiac a and metastsis, the existence of proteolytic enzymes, which are capable of degrading the tissue barrier composed primarily of collagen, elastin, glycoproteins and proteoglycans, is imagnant. One of theses enzymes, cathepsin B, a lysosomal thiol protease, has been reported to ie ound in association with plasma membrane in animal and human tumors and to be releasec b tumor cells. OBJECT: We assayed the cathepsin B activities of squamo is cell carcinoma and basal cell carcinoma in order to investigate the correlation between the degree of cathepsin B activities and invasiveness or metastatic potential of skin tumors. METHODS: Cathepsin B-like protease activity was measurec by the method of Hirao using a synthetic substrate, Z-Phe-Arg-MCA. The skin tissues (penie Koreskin for control, basal cell carcinoma and squamous cell carcinoma tumor masses) were homogenized and their subcellular organelles were fractionated by centrifugation. Each of the fractionated preparations were used as enzyme solution. RESULTS: Cathepsin B-lilke activities were found mainly in the membrane fractions in all the samples. The activities of squamous cell carcinoma (12.484+1.904) and basal cell carcinoma (10.598+1.926) were higber than those of the control skin (9.115+0.815). CONCLUSION: These results suggest that membrane-bound conrt epsin B-like protease participates in local dissolution of the extracellular matrices and were ar endothelial cells to be able to make metastasis to other remote organ during the invasivest ges of malignant skin tumors.
Animals ; Basement Membrane ; Carcinoma, Basal Cell ; Carcinoma, Squamous Cell ; Cathepsin B* ; Cathepsins* ; Cell Membrane ; Centrifugation ; Collagen ; Elastin ; Endothelial Cells ; Extracellular Matrix ; Glycoproteins ; Humans ; Immunohistochemistry ; Keratinocytes ; Membranes* ; Neoplasm Metastasis ; Oncogenes ; Organelles ; Peptide Hydrolases ; Proteoglycans ; Skin*

BACKGROUND: Diagnosis of paucibacillary leprosy is difficult owing to lack of sensitive diagnostic tools. Recently, several investigators have studied the use of polymerase chain reaction(PCR) to detect Mycobacterium leprae. Using nested-PCR the sensitivity and specificity of DNA amplification is considerably improved. OBJECTIVE: The purpose of investigation is to assess the efficacy to nested-PCR which is applied to formalin-fixed, paraffin-embedded biopsies material of patients with leprosy. METHODS: Biopsy samples were taken from patients with lepromatous(11 patients) and tuberculoid (10 patients) leprosy, fixed in formalin, and embedded in paraffin. The DNA from samples was extracted and amplified through 25 cycles by using the outside pairs of primer(L1 and L2). The second amplification was allowed to proceed through 15 cycles using inside pairs of primer(L3 and L4). RESULTS: All twenty one samples showed 347-base-pair products. To confirm that the 347-bp product did correspond to the expected portion of the M. leprae groEL gene, the amplified product was digested with Pst I. Pst I digestion yielded 254-and 93-bp fragments, as predicted from the sequence of the M. leprae gene. The sensitivity was that a single organism was identified by nested-PCR. CONCLUSION: The nested-PCR is sensitive, specific, and simple diagnostic tool for leprosy.
Biopsy ; Diagnosis ; Digestion ; DNA* ; Formaldehyde ; Humans ; Leprosy ; Leprosy, Paucibacillary ; Mycobacterium leprae* ; Mycobacterium* ; Paraffin* ; Polymerase Chain Reaction* ; Research Personnel ; Sensitivity and Specificity

BACKGROUND: The various manifestations of intrinsic cutaneous aging may reflect the age-related changes of dermal ground aubstances, the important components of the dermis. OBJECTIVE: This study was undertaken to observe whether there is any age-relsted change, quantitatively and qualitatively, in dermal glycosaminoglycan(GAG) METHODS: The Senescence-A.ccelerated-Mice(SAM) strain "Prone" to develop senescence(P/8), aged 4 days(P-Y a group, n=5) and 4 months old (P-0 group, n=5) were used, with SAM strain Resistance to senescence (R/1), aged 4 montha old (R-0 group, n=5). The whole skin of SAM was incubated in 0.1 sodium phosphate buffer(NaPB), 2M guanidine-HC1, and 4M guanidine-HCl, sequencially, for the extractiop of GAG. The amount of GAG was measured by using alcian blue and by methylene blue metachrometic assay. Uronic acid(UA) was estimate l employing carbazole reaction. Extracted skin protein profiles were analyed by SDS-PAGE. RESULTS: 1. The total contents of GAG per wet weight of skin, as measured uairq, alcian blue, was highter in R-0 group than R-Y and P-Y group, which is statistically significant. 2. The methylene blue metaehromatic assay yielded highter absorbance values in 2M guanidine-HCl extract than NaPB ext:racts. 3. The total contents of UA decreased with aging in R strains, buttriking increase was noted in the P strain. 4. On SDS-PAGE, the protein profiles of NaPB extracts showed similarity to serum protein. 125 kDa protein band was noted only in guanidine-HCl extracts. 37.40kDa protein bands were appeared in 2M, 4M guanicline-HCl extract of R-0 group. But. there was no significant difference in both strains. CONCLUSION: These findings suggest that macromolecules, such as GAG, is one of the target molecules of the cutaneous eging process, and these change muy be related to the age-related changes of dermal water content.
Aging* ; Alcian Blue ; Animals ; Dermis ; Electrophoresis, Polyacrylamide Gel ; Epidermolysis Bullosa Acquisita ; Humans ; Infant ; Methylene Blue ; Mice* ; Pemphigoid, Bullous ; Skin* ; Sodium

BACKGROUND: The various manifestations of intrinsic cutaneous aging may reflect the age-related changes of dermal ground aubstances, the important components of the dermis. OBJECTIVE: This study was undertaken to observe whether there is any age-relsted change, quantitatively and qualitatively, in dermal glycosaminoglycan(GAG) METHODS: The Senescence-A.ccelerated-Mice(SAM) strain "Prone" to develop senescence(P/8), aged 4 days(P-Y a group, n=5) and 4 months old (P-0 group, n=5) were used, with SAM strain Resistance to senescence (R/1), aged 4 montha old (R-0 group, n=5). The whole skin of SAM was incubated in 0.1 sodium phosphate buffer(NaPB), 2M guanidine-HC1, and 4M guanidine-HCl, sequencially, for the extractiop of GAG. The amount of GAG was measured by using alcian blue and by methylene blue metachrometic assay. Uronic acid(UA) was estimate l employing carbazole reaction. Extracted skin protein profiles were analyed by SDS-PAGE. RESULTS: 1. The total contents of GAG per wet weight of skin, as measured uairq, alcian blue, was highter in R-0 group than R-Y and P-Y group, which is statistically significant. 2. The methylene blue metaehromatic assay yielded highter absorbance values in 2M guanidine-HCl extract than NaPB ext:racts. 3. The total contents of UA decreased with aging in R strains, buttriking increase was noted in the P strain. 4. On SDS-PAGE, the protein profiles of NaPB extracts showed similarity to serum protein. 125 kDa protein band was noted only in guanidine-HCl extracts. 37.40kDa protein bands were appeared in 2M, 4M guanicline-HCl extract of R-0 group. But. there was no significant difference in both strains. CONCLUSION: These findings suggest that macromolecules, such as GAG, is one of the target molecules of the cutaneous eging process, and these change muy be related to the age-related changes of dermal water content.
Aging* ; Alcian Blue ; Animals ; Dermis ; Electrophoresis, Polyacrylamide Gel ; Epidermolysis Bullosa Acquisita ; Humans ; Infant ; Methylene Blue ; Mice* ; Pemphigoid, Bullous ; Skin* ; Sodium

BACKGROUND: The human sebaceous glands have long been known to change their activity with aging. Downing and his co-workers state that the ratio of wax ester/cholesterol+cholesterol ester in the skin surface lipids might be a good index for sebaceous gland acti ity. OBJECTIVE: Our purpose was to evaluate the effects of aging on the sebaceous gland activity and relative skin surface lipid composition by using thin layer chromatography. METHOD: Skin surface lipids of anterior chest from 65 healthy Korean indivisuals were collected by using the cup method. Skin surface lipid were separated and meaurd by thin layer chromatographic analysis. RESULTS: The sebaceous gland activity, vrhich was expressed by tlie ratio wax ester/[cholesterol+cholesterol ester] showed a ilistinct change from infancy to senescenc. The curve of the ratio makes a peak in the third decade and decreases with advancing age. CONCLUSION: This result disclosed that sebaceous gland activity iaifected by advancing age in Koreans and can be used as one of the biologic markers of aging.
Aging* ; Biomarkers ; Chromatography, Thin Layer ; Humans ; Sebaceous Glands* ; Skin* ; Thorax

Epidermal growth factor(KGF) and transforming growth factor arpha(TGF-a) are polypeptides of 53 amd 50 amino acid residuies. Both bind to epidermal growth factor receptor (EGR-R) leading to phosphorylation of the receptor, enhancement of its tyrosine-specific kinase activity and ultimately to stimulation of cell growth. To study he role of EGF, TGF-a, and EGF-R in differentiation and hyperproliferation of cell, we se lected psoriasis vulgaris, because the affected keratinocyte may house both an abnormally increased proliferative capacity and an abnormally differentiated state. The biopsy specimens were taken from involved and uninvolved skin of 20 patients with psoriasis and immunoperoxidase studies with formalin-fixed paraffin-embedded tissues were performed with EGF, TGF-a and EGF-R useing the Vectastain ABC irnmunoperoxidase stain system. The antibodies were used at a concentration of 6 ug/ml In involved psoriatic skin, distributions of TGF-a and EGF-R were increased in all layers of epidermis as compared to normal, uninvolved psoriatic skin, in which chev were showed to the basal and parabasal layers. However, distribution of EGF was weekly positive in the basal layers of epidermis in both involved and uninvolved skin with no difference between toem. These results suggest that increased distribution of TGF-a and EGF-R may be involved in hypoproliferative state of epidermal keratinocytes in psoriatic lesion.
Antibodies ; Biopsy ; Epidermal Growth Factor ; Epidermis* ; Humans ; Keratinocytes ; Peptides ; Phosphorylation ; Phosphotransferases ; Psoriasis ; Receptor, Epidermal Growth Factor ; Skin ; Transforming Growth Factors