Expression of nucleoside diphosphate(NDP) kinase, which is homologous to the nm23 gene product in a variety of species, has been found to be inversely associated with metastatic potential. However, the relationship remains controversial according to the tumor cell types and experimental system, with conflicting results from different research groups. In order to determine whether NDP kinase expression serves as a marker for metastatic potential in human skin cancer, we assessed the levels of NDP kinase expression in 9 keratoacanthomas (KAs), 26 squamous cell carcinomas (SCCs), and 25 basal cell carcinomas (BCCs) using immunohistochemistry. The expression of NDP kinase was intense in KA and SCC compared with BCC. However, the difference of NDP kinase expression between KA and SCC was not statistically significant. And there was no statistically significant difference in NDP kinase expression between SCC with metastasis and SCC without metastasis. Our results contradict the hypothesis concerning the possible role of nm23 gene as a metastatic suppressor gene in human skin cancer. The mechanism of overexpression in various tumor cell types and its biological significance in cutaneous carcinogenesis remain to be determined.
Antibodies, Monoclonal ; Carcinoma, Basal Cell/enzymology/secondary ; Carcinoma, Squamous Cell/enzymology/secondary ; Comparative Study ; Erythrocytes/enzymology ; Human ; Immunohistochemistry ; Keratoacanthoma/enzymology ; Nucleoside-Diphosphate Kinase/*analysis/blood ; Skin Diseases/enzymology ; Skin Neoplasms/*enzymology/secondary ; Transcription Factors/analysis

OBJECTIVESTo detect telomerase activity in patients with mycosis fungoides (MF) and to study the role of telomerase in the tumorigenesis of MF.

METHODSThe technique of PCR-ELISA was employed to detect telomerase activity in 35 patients with various stages of MF.

RESULTS92.3% tumor stage of MF, 78.6% plaque stage of MF and 75.0% patch stage of MF had positive telomerase activity. The control samples had no telomerase activity. Telomerase activity in tumor stage of MF was significantly higher than that in plaque stage, while the latter was higher than that in patch stage. Telomerase activity was correlated with the stage of MF.

CONCLUSIONHigh level of telomerase activity frequently occurred in patients with MF, suggesting that telomerase might play an important role in the tumorigenesis of MF and is a useful marker for the diagnosis of MF possibly.

Humans ; Mycosis Fungoides ; enzymology ; pathology ; Neoplasm Staging ; Skin Neoplasms ; enzymology ; pathology ; Telomerase ; metabolism

Cell Line, Tumor ; Humans ; Matrix Metalloproteinase 2 ; analysis ; Matrix Metalloproteinase 9 ; analysis ; Melanoma ; enzymology ; Skin Neoplasms ; enzymology

Telomerase activity is usually detected in most tumor tissues but not in normal tissues. Recently, there is increasing evidence that telomerase activity is associated with cell proliferation without malignancy, whereas there is little information about telomerase activity and its relationship with cell proliferation in chronic hyperproliferative skin diseases. Thus, we studied telomerase activity in skins from 10 patients with psoriasis and compared telomerase activity with the expression of Ki-67, a proliferation marker, using immunohistochemical staining. The effect of retinoic acid on the telomerase activity in HaCaT cells was also evaluated. Telomerase activity was detected in 7 (70%) of 10 lesional skins of psoriasis and none of the nonlesional skin. Telomerase activity in lesional skin was significantly associated with Ki-67 labelling index. Retinoic acid treatment on HaCaT cells inhibited telomerase activity, which correlated with inhibition of cell proliferation by the agent. The results of our study represent another example that shows telomerase activity correlates with cellular proliferation. Further studies on the regulation of the telomerase are needed to understand the cellular factors involved in controlling telomerase activity.
Cell Division/drug effects ; Cell Line ; Enzyme Inhibitors/*pharmacology ; Human ; Ki-67 Antigen/*analysis ; Psoriasis/*enzymology ; Skin/*enzymology ; Telomerase/antagonists & inhibitors/*metabolism ; Tretinoin/*pharmacology

OBJECTIVETo evaluate the diagnostic value of the changes in serum CK and its isozymes in the muscular infection and necrosis in electrical injured patients.

METHODSSeventeen patients were divided into A and B groups according to the causes, i.e. electrical injury as A and electrical arc flame burn as B groups. Obvious muscle necrosis was identified in A but not in B groups. The serum CK-MM concentration was determined after injury, before and after the operations. Simultaneously, the blood and urine routine, the hepatic and renal function and wound bacterial counting were determined and compared with those in 20 healthy people.

RESULTS1. The serum CK-MM in A group increased evidently after injury and 1 day after wound debridement to the 6 times that in normal control. The enzyme decreased to normal at 3 post-operative days in 15 cases and remained at relative high level in 2 cases due to the wound infection and lowered down to normal level after wound re-debridement. 2. The serum CK-MM in B group increased slightly before and after skin grafting.

CONCLUSIONCK-MM could be employed as the index for the infection and necrosis of the muscle in electrical injured patients due to its high specificity and sensitivity.

Adolescent ; Adult ; Burns, Electric ; blood ; enzymology ; Creatine Kinase ; blood ; Electric Injuries ; blood ; enzymology ; Humans ; Isoenzymes ; blood ; Male ; Middle Aged ; Skin Transplantation

AIMTo study the stereoselectivity of skin carboxylesterase metabolism and its molecular biological foundation for improving drug percutaneous absorption.

METHODSKetoprofen ethyl ester was used as a model drug, and skin homogenate was applied for studying the stereoselectivity of carboxylesterase metabolism. Human liver L02 cell was used as control of carboxylesterase expression, and RT-PCR was used for studying the expression of carboxylesterase.

RESULTSThe main metabolite of ketoprofen ethyl ester in human skin homogenate was R-ketoprofen. Human carboxylesterase-2 was highly expressed in skin and its cells. However, the expression of human carboxylesterase-1 was very weak or not detectable.

CONCLUSIONHuman carboxylesterase-2 is the main hydrolytic enzyme of prodrugs in percutaneous absorption, and shows metabolic stereoselectivity to prodrugs with chiral esters.

Adult ; Carboxylesterase ; genetics ; metabolism ; Cell Line ; Cells, Cultured ; Humans ; Ketoprofen ; metabolism ; Liver ; cytology ; enzymology ; Prodrugs ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Skin ; enzymology ; Stereoisomerism

In a previous search for the differentially expressed genes in keratinocyte differentiation, we identified neutrophil gelatinase-associated lipocalin (NGAL) as a calcium- induced gene. In this study, we further verified the expression of NGAL in cultured keratinocytes as well as in several skin diseases. Reverse transcription-polymerase chain reaction (RT-PCR), Western blot, and ELISA clearly showed that NGAL expression was markedly increased in calcium-induced keratinocyte differentiation in vitro. However, in our previous report, NGAL expression was not detected in normal skin tissue except for hair follicle by in situ hybridization and immunohistochemistry, indicating the difference of cell status between in vitro and in vitro conditions. Interestingly, NGAL expression was highly increased in psoriasis-like inflammatory disorders (lichen planus and pityriasis rubura pilaris) and skin cancers (keratoacanthoma and squamous cell carcinoma), implying that NGAL may be related with the epidermal hyperplasia. Collectively, these results reveal the potential importance of NGAL in the maintenance of skin homeostasis.
Acute-Phase Proteins/*biosynthesis ; Calcium/*metabolism ; Cell Differentiation ; Culture Media ; Culture Media, Conditioned ; Enzyme-Linked Immunosorbent Assay ; *Gene Expression Regulation ; Homeostasis ; Humans ; Keratinocytes/enzymology ; Lipocalins/*biosynthesis ; Models, Biological ; Proto-Oncogene Proteins/*biosynthesis ; Psoriasis/enzymology ; Reverse Transcriptase Polymerase Chain Reaction ; Skin/*metabolism ; Skin Neoplasms/enzymology

In Korea, common whelk (Buccinum undatum) is a popular edible shellfish. The aim of this study was to observe the sensitization rate to common whelk and to characterize its allergens. We carried out skin prick test (SPT) in 1,700 patients with various allergic diseases. Specific IgE were detected by ELISA in the patient sera and ELISA inhibition tests were conducted. IgE-binding components were identified by means of SDS-PAGE and IgE-immunoblotting. The effects of diges-tive enzymes were evaluated in both raw and thermally treated extracts. SPT to common whelk was positive (> or =2+) in 83 (4.9%) patients studied. Twenty-four (38.7%) out of 62 SPT positive patients had high serum specific IgE to common whelk. ELISA inhibition test showed significant inhibitions by abalone as well as by common whelk. IgE-immunoblotting demonstrated three IgE-binding components (40, 71, 82 kDa), which were digested by simulated intestinal fluid and moderately digested by simulated gastric fluid, and the digestibility of allergens remained unchanged after thermal treatment. In conclusion, IgE-sensitization rate to com-mon whelk was 4.9% in allergy patients. IgE-immunoblotting demonstrated three IgE-binding components, which were degraded by digestive enzymes. Further studies are needed to evaluate the clinical significance of the sensitized patients to common whelk.
Allergens/immunology ; Animals ; Comparative Study ; Cookery ; Digestion/*physiology ; Food Handling/methods ; Food Hypersensitivity/diagnosis/*epidemiology/*immunology/metabolism ; Heat ; Humans ; Immunoglobulin E/*immunology/metabolism ; Intestines/enzymology ; Korea/epidemiology ; *Mollusca ; Research Support, Non-U.S. Gov't ; Shellfish/*adverse effects ; Skin Tests ; Stomach/enzymology

Proteases in the skin are essential to epidermal permeability barrier homeostasis. In addition to their direct proteolytic effects, certain proteases signal to cells by activating protease-activated receptors (PARs), the G-protein-coupled receptors. The expression of functional PAR-2 on human skin and its role in inflammation, pruritus, and skin barrier homeostasis have been demonstrated. Atopic dermatitis (AD) is a multifactorial inflammatory skin disease characterized by genetic barrier defects and allergic inflammation, which is sustained by gene-environmental interactions. Recent studies have revealed aberrant expression and activation of serine proteases and PAR-2 in the lesional skin of AD patients. The imbalance between proteases and protease inhibitors associated with genetic defects in the protease/protease inhibitor encoding genes, increase in skin surface pH, and exposure to proteolytically active allergens contribute to this aberrant protease/PAR-2 signaling in AD. The increased protease activity in AD leads to abnormal desquamation, degradation of lipid-processing enzymes and antimicrobial peptides, and activation of primary cytokines, thereby leading to permeability barrier dysfunction, inflammation, and defects in the antimicrobial barrier. Moreover, up-regulated proteases stimulate PAR-2 in lesional skin of AD and lead to the production of cytokines and chemokines involved in inflammation and immune responses, itching sensation, and sustained epidermal barrier perturbation with easier allergen penetration. In addition, PAR-2 is an important sensor for exogenous danger molecules, such as exogenous proteases from various allergens, and plays an important role in AD pathogenesis. Together, these findings suggest that protease activity or PAR-2 may be a future target for therapeutic intervention for the treatment of AD.
Anti-Infective Agents/pharmacology ; Dermatitis, Atopic/*enzymology ; Endopeptidases/metabolism ; Homeostasis ; Humans ; Hydrogen-Ion Concentration ; Inflammation ; Models, Biological ; Models, Genetic ; Peptide Hydrolases/*metabolism ; Receptor, PAR-2/*metabolism ; Serine Proteases/metabolism ; Signal Transduction ; Skin/enzymology/pathology ; Treatment Outcome

OBJECTIVE: To assess the role of p38 in fibroblast orientation and to explore the cell signal transduction mechanism of cyclic strain induced cell orientation. METHODS: Fibroblasts were seeded onto collagen coated flexible membranes. Membranes were then deformed at 10 cycles per minute under 135 mmHg subatmospheric pressure. Orientation angles of cells treated with or without SB203580 were measured with inverted microscope. P38 phosporylation was analyzed with Western blot. RESULTS: Eighty percent cyclic strain induced cells rotated from 60 degree to 90 degree perpendicular to stretch direction after 4 h strain exposure. P38 phosphorylation reached the peak at 5 min. Fibroblast orientation was inhibited after SB203580 treatment. CONCLUSION Fibroblast orientation in response to cyclic strain is mediated by p38 phosporylation.
Cell Movement ; Cells, Cultured ; Enzyme Inhibitors ; pharmacology ; Fibroblasts ; cytology ; enzymology ; Humans ; Imidazoles ; pharmacology ; Male ; Mechanotransduction, Cellular ; physiology ; Phosphorylation ; Pyridines ; pharmacology ; Skin ; cytology ; enzymology ; Stress, Mechanical ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; metabolism