Changes of Serum Cytokines ( TNF-alpha, IL-beta, IL-6 ) in Toxemic Mice Induced by Vibrio vulnificus Cytolysin / 대한피부과학회지

BACKGROUND: Cytokines such as TNF-a, and IL-1, 6, 8 are thought to mediate many host responses in patients with sepsis through the release of many kinds of inflammatory mediators. Vibrio vulnificus is a virulent, halophilic, marine, and Gram-negative bacterium causing wound infections or primary sepsis with fulminant course. It has been known to produce many exocellular factors, such as cytolysin and exozymes, which are possible causes of shock and tissue damage. OBJECTIVE: Our purpose was to investigate the changes of cytokines(TNF-a, Il-l p, and IL-6) after intravenous administration of V. vulnificus cytolysin which have known to be a major virulent factor for elucidating pathogenesis of septic shock. METHODS: After administration of V. vulnificus cytolysin(20 hemolytic units) and physiologic saline through a mouse tail vein, we obtained blood samples from the heart according to a time course(0, 30, 60, 90, 120, 240, and 360 minutes). We measured serum concentrations of circulating TNF- a, IL-1 p, and IL-6 using commercially available immunoassay kits. RESULTS: The serum concentration of TNF- a peaked at 60 min (1894 pg/ml, as compared with 829 pg/ml after saline administration), began to decrease at 90 min and fell to baseline at 360min. Concentration of IL-6 rose significantly at 90 min(>3,839 pg/ml) and began to decrease at 120 min, whereas the level of IL-1 0 increased slightly and the peaked around 90min(117 pg/ml) after the cytolysin injection. CONCLUSION: We conclude that the host response to cytolysin is associated with a brief pulse of circulating cytokines in the early stage of sepsis caused by V. vulnipcus infection.

The Extraction of Vibrio vulnificus Lipopolysaccharide by the Phenol-Chloroform-Petroleum Ether Method and Biological Activity of the LPS / 대한피부과학회지

BACKGROUND: Vibrio vulnificus is a pathogenic, marine, halophilic, gram-negative bacillus which causes fulminant infecticn in humans through the ingestion of raw seafood or skin wounds. V. vulnificus produces seveal kinds of virulent factors including cytolysin, endotoxin, exoenzymes, and siderphores. Among these, the lipopolysaccharide(LPS) of V. vulnificus has recently been purified, but the biological activity of this endotoxin is not well clarified as yet. OBJECTIVE: The purpose of this study was to extract LPS from Vibrio vulnificus and to test the biological activity of extracted LPS for the elucidation of the role in V. vulnificas septicemia. METHODS: V. vulnificus LPS was extracted by the Phenol Chloroform-Petroleurn ether(PCP) method. The biological activity of LPS was evaluated with a limulus amebocyte lysate assay and by assessment of lethality to ICR mice. RESULTS: Five hundreds mg of LPS were extracted from 10g of dried V. vulnificus. Lirnulus amebocyte lysate formed a hard gel in response to the extracted LPS. This LPS showed low level of protein contaminatior in SDS-PAGE electophoresis and spectrophotometry. A High dose of LPS(200 mg/ml body weight) was lethal to mice. CONCLUSION: The PCP extraction yield relatively large amounts of LPS from V. vnlnificus. with out significant protein contamination and e xtrated LPS has endotoxin activity. This extrated LPS can be used for further studies such as making antibody or characterizing pathogenic roles in the V. vulnificus infection.

A Case of Irritant Dermatitis due to Pulsatilla Koreana / 대한피부과학회지

Pulsatilla koreana is a member of the buttercup familly(Ranuculaceae) which contains the unsaturated lactone, protoanemonin(C3H4O2), formed after injury to the plant by the breakdown of the glycoside ranunculin. It has been prescribed as a herbal medicine or a folk medicine for antipyretic, analgesic, anti-inflammatory, astringent and hemostatic effects in Korea. A 20 year-old famale patient presented with confluent vesicobullae on the erythematous base with a burning sensation on her upper back. This was caused by contact with crushed Pulsatilla koreana, a folk medicine for the treatment of left facial nerve palsy, that had occurred 10 days before her visit. A provocation patch test with Pulsatilla koreana upon a heathy male volunteers upper back showed strong positive reactions with the stalk and leaf in fresh and frozen stat.e and after thaw ing resptively. However no reaction was seen when the folk medicine was used in a dried form or with the root in any state. This case indicates that misadvice and ignorant folk remedies could cause misfortunate results to innocent people.

A Case of Acquired Lymphangioma After Open heart Surgery / 대한피부과학회지

Lymphangiectasis(=acquirecl lymphangioma) ate dilated lymph caillaris and may be due to the obstruction of the lymphatic circulation from surgery, irradiation, chronic recurrent infection or trauma. The patient was a 5 years old girl who developed a straw-colcre, asymptomatic grouped vesicular mass resembling frog spawn measuring 9 cm in diameter the left axilla and arm, 2 months after the total excision of a previous lymphangiectatic mass. She had a history of previous open heart surgery at 2 years of age. Histopathologic findings showed large, irregularly shaped cysticalation lined by a single layer of endothelium in the superficial dermis. Immunohistochemical stainig showed vimentin positive and negative results in Factor VIII and HLA-DR antigen which was compatible with a lymphatic vessel.