Treponema strain PFB4G is a novel oral spirochete and one of the most frequently detected organisms in subgingival plaque samples from rapidly progressive periodontitis and adult periodontitis patients. In this study, a genomic library of Treponema strain PFB4G was constructed in lambdaZAP expression vector. One positive clone that carried a 2.6-kb fragment was identified by screening with chicken Ig Y (immunoglobulin yolk) antibody raised against whole bacterial sonicates. Nucleotide sequencing of the subclones revealed an open reading frame (ORF) lacking the 5'-end. This region was obtained by PCR amplification using a degenerative and a specific primer. A complete open reading frame of 1,770 bp was identified and the deduced polypeptide consisted of 590 amino acids with a molecular mass of 65 kDa. The polypeptide, designated as OmpTL, had a typical prokaryotic signal sequence (19 amino acids) with a potential cleavage site for signal peptidase I and showed a significant level of homology with the outer membrane proteins of other oral treponemes, especially with that of Treponema maltophilum. The isolation of the gene encoding an outer membrane protein may allow the study of their roles in future, possibly as adhesion, pore forming or induction of proinflammatory cytokine synthesis.
Amino Acids ; Chickens ; Chronic Periodontitis ; Clone Cells* ; Cloning, Organism* ; Genomic Library ; Humans ; Mass Screening ; Membrane Proteins* ; Membranes* ; Open Reading Frames ; Periodontitis* ; Polymerase Chain Reaction ; Protein Sorting Signals ; Sequence Analysis* ; Spirochaetales ; Treponema*

BACKGROUND: Ethyl crbmate(EC) has been identified at low microgram quantities in various fermented beverages, destiled products and tobacco smoke. EC has long been known as a carcinogen. Oxidation of the ethyl group of EC is followed by dehydration to yield the carcinogen vinyl carbamate (VC). This is further oxidized to vinyl carbamate epoxide(VCO). VC and VCO proved to be much more carcirogenic than EC. OBJECT: The objective of this study is to compare the potency of carcinogenic ability and histologic changes in skin tumors induced by EC, VC, or VCO. METHODS: In this exeriment, the tumor induction was performed by painting the mouse skin once a week for five weeks with EC, VC and VCO solution, and then 12-0-tetradecanoyl-phorbol-13-acetate(TP2) was treated in the same manner twice a week for 35 weeks. We biopsied the skin at B 0, 15, 25, 30 and 35 weeks and stained the specimens with hematoxylineosin. RESULTS: The time period for the first appearance of papilloma was 6 weeks in the VC-TPA and VCO-TPA group, but, 15 weeks in the EC TPA group. The average number of papilloma showed a statistically sign.ficant difference after 15 weeks between the EC-TPA, VC-TPA and VCO-TPA group. The occurrence of keratoacanthoma and squamous cell carcinoma was 16 and 21 weeks respectively. Histologic changes, such as epidermal layers, pseudohorncysts, degree of hyperkeratosis, hypergranulosis, dysplasia and dyskeratotic cells were more prominent in the papil loma than in the non-papilloma lesion. Dermal changes disclosed similiar findings, that is, increased dermal thicknes, proliferation of vessels and hair follicles, and fibrosis of the dermis. Squamous cell carcinomis and keratoacanthomas were produced only in the VCO-TPA group. CONCLUSION: From the above results it is concluded that VC and VCO have a more potent carcinogenic potential than EC. Various skin tumors, such as papilloma, keratoacanthoma or squamous cell carcinoma were prorduced by the above carcinogens.
Animals ; Beverages ; Carcinogens ; Carcinoma, Squamous Cell ; Dehydration ; Dermis ; Fibrosis ; Hair Follicle ; Keratoacanthoma ; Loma ; Mice ; Paint ; Paintings ; Papilloma ; Skin* ; Smoke ; Tobacco ; Urethane*

Matrix metalloproteinases (MMPs) play an important role in the normal morphogenesis, maintenance, and repair of matrix and also have important functions in pathologic conditions characterized by excessive degradation of extracellular matrix, such as rheumatoid arthritis, osteoarthritis, periodontitis and in tumor invasion and metastasis. In this study, expression of MMP-1 and -2 mRNA in retrodiscal tissue of the temporomandibular joint (TMJ) was examined and compared with magnetic resonance imaging (MRI) and surgical findings. MMP mRNAs in the retrodiscal tissue samples were detected by reverse transcription - polymerase chain reaction. TMJ internal derangement (ID) was categorized as normal disc position, disc displacement with reduction, early stage of disc displacement without reduction (DDsR) and late stage of DDsR. TMJ osteoarthrosis (OA) was classified with normal, mild and advanced OA. The amount of synovial fluid collection was divided into not detected, small, large and extremely large amount on MR T2-weighted imaging. Perforation and adhesion were examined during open surgery of the TMJ. Six out of 37 samples were excluded because of little amount of extracted total mRNA. MMP-2 mRNA was detected whole joints, and so the MMP-2 mRNA seems to be expressed normally in retrodiscal tissue. However, MMP-1 mRNA was expressed in 8 of 31 joints. Frequencies of MMP-1 mRNA expression according to the TMJ IDs, amount of synovial fluid and surgical findings made no significant difference. MMP-1 mRNA was detected more frequently in OA groups (7/16 joints, 43.8%) than in normal bony structure group (1/15,joints, 6.7%). Expression of MMP-1 mRNA in retrodiscal tissue might be related with OA of the TMJ.
Arthritis, Rheumatoid ; Collagenases ; Extracellular Matrix ; Gelatinases ; Joints ; Magnetic Resonance Imaging ; Matrix Metalloproteinase 1* ; Matrix Metalloproteinases ; Morphogenesis ; Neoplasm Metastasis ; Osteoarthritis ; Periodontitis ; Polymerase Chain Reaction ; Reverse Transcription ; RNA, Messenger* ; Synovial Fluid ; Temporomandibular Joint*

No abstract available.
Arteriovenous Fistula* ; Humans ; Renal Dialysis*

No abstract available.
Arteriovenous Fistula* ; Humans ; Renal Dialysis*

The incidence of tumor and the time of expression, cellular localization and the molecular weight of tumor associated proteins of rat skin tumor induced by 7,12-dimethylbenz[a]anthracene (DMBA) with or without 12-O-tetradecanoyl-phorbol-13-acetate (TPA) were studied. The time of the development of skin tumors in 0.1% DMBA-TPA treated rats was significantly shorter than that in rats which were treated with DMBA alone. In the complete carcinogenesis case, papillomas developed more slowly and were less common and also squamous cell carcinomas appeared much later. From the analysis of the proteins of each experimental group by SDS-PAGE and two dimensional gel electrophoresis, at least three tumor associated proteins were identified (54kd, pl = 5.66; 27kd, pl = 5.85; 11kd, pl = 4.90). Also these proteins were found in rat dorsal skin from 14 days gestation to 21 days postpartum, and disappeared after 28 days. In conclusions, two stage skin carcinogenesis could be successfully demonstrated in Sprague-Dawley rats and abnormal proteins were produced in DMBA or DMBA-TPA induced skin tumor. The tumor associated proteins of skin tumor induced by DMBA or DMBA-TPA were appeared at the late initiation stage or early promotion stage, and they were localized in plasma membrane and were glycoproteins that are thought to be related to the epidermal differentiation process.
9,10-Dimethyl-1,2-benzanthracene ; Animal ; Antigens, Surface/*analysis ; Carcinoma, Squamous Cell/*analysis/chemically induced/pathology ; Cell Membrane/metabolism ; Comparative Study ; Glycoproteins/analysis ; Membrane Proteins/*analysis ; Papilloma/*analysis/chemically induced/pathology ; Rats ; Rats, Inbred Strains ; Skin Neoplasms/*analysis/chemically induced/pathology ; Support, Non-U.S. Gov't ; Tetradecanoylphorbol Acetate

The incidence of tumor and the time of expression, cellular localization and the molecular weight of tumor associated proteins of rat skin tumor induced by 7,12-dimethylbenz[a]anthracene (DMBA) with or without 12-O-tetradecanoyl-phorbol-13-acetate (TPA) were studied. The time of the development of skin tumors in 0.1% DMBA-TPA treated rats was significantly shorter than that in rats which were treated with DMBA alone. In the complete carcinogenesis case, papillomas developed more slowly and were less common and also squamous cell carcinomas appeared much later. From the analysis of the proteins of each experimental group by SDS-PAGE and two dimensional gel electrophoresis, at least three tumor associated proteins were identified (54kd, pl = 5.66; 27kd, pl = 5.85; 11kd, pl = 4.90). Also these proteins were found in rat dorsal skin from 14 days gestation to 21 days postpartum, and disappeared after 28 days. In conclusions, two stage skin carcinogenesis could be successfully demonstrated in Sprague-Dawley rats and abnormal proteins were produced in DMBA or DMBA-TPA induced skin tumor. The tumor associated proteins of skin tumor induced by DMBA or DMBA-TPA were appeared at the late initiation stage or early promotion stage, and they were localized in plasma membrane and were glycoproteins that are thought to be related to the epidermal differentiation process.
9,10-Dimethyl-1,2-benzanthracene ; Animal ; Antigens, Surface/*analysis ; Carcinoma, Squamous Cell/*analysis/chemically induced/pathology ; Cell Membrane/metabolism ; Comparative Study ; Glycoproteins/analysis ; Membrane Proteins/*analysis ; Papilloma/*analysis/chemically induced/pathology ; Rats ; Rats, Inbred Strains ; Skin Neoplasms/*analysis/chemically induced/pathology ; Support, Non-U.S. Gov't ; Tetradecanoylphorbol Acetate

With the development of antenatal diagnostic tools such as ultrasonography, some congenital anomalies or diseases can be detected in early fetal life. Routine serial antenatal check-ups have made it possible to predict the prognosis of these problems, and a few life-threatening single anatomic malformations have been treated by open fetal surgery. The experience of fetal surgery revealed that the human fetus appears to heal without any scarring. In contrast to adult animals, the response to tissue injury in the fetus is conspiciously devoid of acute inflammation. Indeed, the absence of neutrophils is perhaps the most consistent observation in fetal wounds and seems to be followed by absent or scanty fibroblast infiltration, which results in healing with sparse and well organized collagen deposition. Actually, the amount and quality of the collagen deposition were decided by the fibroblasts which infiltrated the wound. It is well known that fetal wounds have sparse collagen deposition, however, the mechanisms are still unclear. This study was designed to evaluate the role of fibroblast activity in the differences of the scar formation between the fetus and neonate. Fibroblast activity such as the cell growth rate, the amount of collagen synthesis and the synthesized collagen types of fetus(IUP 18-22 weeks) was compared with that of neonate. The amount of collagen synthesis was measured by H-proline uptake and the amount of collagen type III was measured by Western blot using antihuman procollagen type III. The cell growth rate as determined by cell proliferation from the initial cell count of 5x10(5) to cell confluence was 3.6 x 10(6) in the fetal fibroblasts compared to 2.5x10(6) in neonatal fibroblasts. Fetal fibroblast synthesize 16.9 x 10(4) cpm of collagen and neonatal fibroblasts synthesize 2.7 x 10(4) cpm of collagen. The synthesized amount of type III collagen was 2.1x10(4) ug/ml, and 1.5x10(4) ug/ml by fetal and neonatal fibroblasts, respectively. In conclusion, fetal fibroblasts grow faster and synthesize a smaller amount of collagen, but produce more type III collagen than neonatal fibroblasts.
Adult* ; Animals ; Blotting, Western ; Cell Count ; Cell Proliferation ; Cicatrix ; Collagen Type III ; Collagen* ; Fetus* ; Fibroblasts* ; Humans ; Infant, Newborn ; Inflammation ; Neutrophils ; Prognosis ; Ultrasonography ; Wounds and Injuries

A keratinolytic proteinase secreted by Microsporum canis in a broth containing human hair was purified 134-fold from the culture filtrate by ion-exchange chromatography using DEAE-Sephacel, CM-Sephadex C-50, and by Sephadex G-75 gel filtration. The purified enzyme was electrophoretically homogeneous with a molecular weight of 33,000. The enzyme had an optimum pH of 8.0, and the activity was stable in the alkaline pH range. Enzyme activity increased with temperature up to 35 degrees C and was stable up to 45 degrees C. The keratinolytic activity was not affected by the addition of nonionic detergents, was activated by Mg2+, but inhibited by Zn2+. The purified enzyme was used to obtain guinea pig antiserum. The antiserum tested by double diffusion against the purified enzyme showed a single line of precipitation and completely neutralized the proteinase activity. This study reaffirms that the proteinase from M. canis may be a biochemical mechanism for the invasion of keratinized tissue, and could possibly play a role in the hypersensitivity reactions arising from superficial infections of this fungus.
Microsporum/enzymology* ; Peptide Hydrolases/isolation & purification*

A 76-year-old female presented with a 2-month history of skin lesions on both her buttocks. The skin lesions showed diffuse, erythematous scattered papules and macules with brown to black crusts. The patient believed that she had skin cancer and that the lesions had to be scrapped. She thus soaked her buttock in sikhye, a traditional Korean beverage, to treat the lesions. Based on patient history, she was diagnosed with dermatitis artefacta at the initial hospital visit. KOH examination was positive for hyphae and fungal cultures of Saccharomyces cerevisiae. She was treated with oral and topical antifungal agents for 6 months. The skin lesions improved at that time; however, she continued to scratch her buttock, and the skin lesions aggravated. Repeat fungal culture of the lesions revealed Candida glabrata, which was confirmed by genetic polymerase chain reaction sequencing. Consequently, she was treated with antifungal agents for 6 months. S. cerevisiae is widely used in baking, brewing, wine making, and biotechnology. S. cerevisiae and C. glabrata are considered relatively non-pathogenic fungal organisms. Here, we report a rare case of dermatitis artefacta combined with superficial infection of non-pathogenic fungal organism.