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PubMed1999

Skin flap storage with intracellular and extracellular solutions containing trehalose.

To evaluate and compare the effects of IT-Kyoto (IT-K) solution (intracellular type) and ET-Kyoto (ET-K) solution (extracellular type) on long-term skin flap preservation (from 24-120 hours), Japanese white rabbits were used as an animal model and Euro-Collins (EC) solution was used as a comparative solution. A total of 180 animals were divided randomly into 15 groups (N = 12) for 24, 48, 72, 96, and 120 hours of flap storage in three solutions. The survival rate, weight change, and histological appearance of flaps were compared among the three solutions. The survival rates in the IT-K and ET-K solutions were the same: 100%, 100%, 100%, 92%, and 50% in flaps preserved for 24, 48, 72, 96, and 120 hours, which were higher (p < 0.01 in the 72-, 96-, and 120-hour groups) than those in the EC solution group: 100%, 75%, 33%, 8.3%, and 0%. The flap weight gain in the IT-K and ET-K solution groups was less than that in EC solution group (p < 0.01). Histologically, flaps in the IT-K and ET-K solution groups showed a better appearance than those in the EC solution group. It was concluded that in a rabbit model IT-K and ET-K solutions are superior to EC solution for long-term skin flap preservation, and the safe preservation period for IT-K and ET-K solutions is approximately 96 hours. It was also concluded that the effects of intracellular (IT-K)-type solution and extracellular (ET-K)-type solution on rabbit skin flap preservation are similar.

Animals↗
PubMed2001

Screening and characterization of trehalose-oleate hydrolyzing lipase.

Various soil samples were collected to screen the presence of microorganisms which have ability to degrade TOE. One strain (AKU-883) with good TOE degrading activity was isolated and identified as Burkholderia cepacia and the extracellular enzyme was purified to homogeneity. The purification was achieved by ultrafiltration, Super Q anion-exchange chromatography and Superdex 200HR gel-filtration in the presence of Triton X. The enzyme was purified to 85-fold, and specific activity of 4.910 kU mg protein(-1). The peak preparation on gel filtration showed a single band of 34 kDa on SDS-PAGE and native PAGE which indicate the monomeric nature of the enzyme. The pI of the enzyme was 6.3. The enzyme showed the maximum activity at pH 9 and 65 degrees C, and was stable in the range of pH 5--10 and up to 60 degrees C. Almost all the activity (92%) was kept after incubation for more than 1 week at 50 degrees C (pH 7.3). High activities remained even in water-miscible solvents such as ethanol, dimethyl formamide, diisopropyl ether, and dioxane. The N-terminal 16 amino acid residues were determined as A-N-G-Y-A-A-T-R-Y-P-I-I-L-V-G-G, which showed a consensus sequence for lipases from Burkholderia species. Thus the enzyme was concluded to be a kind of lipase.

Amino Acid Sequence↗
PubMed1973

Granulomatous hypersensitivity to trehalose-6,6'-dimycolate (cord factor) in mice infected with BCG.

Cord factor in the form of emulsion is unable to sensitize mice to react with a more extensive granulomatous response to a subsequent challenge with the same substance. Mice infected with BCG bacilli are sensitized to cord factor. Such animals react to administration of cord factor with a very extensive granulomatous response, much stronger than normal ones. This hypersensitivity seems to be specific and is distinct from delayed hypersensitivity to PPD. PPD, administered to mice sensitized with BCG bacilli, has no effect on the granulomatous response. Our findings and their importance are discussed.

Animals↗
PubMed1974

Suppression of growth of Ehrlich ascites tumor cells in mice by trehalose-6,6'-dimycolate (cord factor) and BCG.

Growth of Ehrlich ascites tumor cells in mice pretreated with cord factor was compared to growth of the tumor cells after pretreatment with Calmette-Guérin bacilli. Growth of Ehrlich ascites cells was strongly inhibited in the peritoneal cavities of mice pretreated with 80 mug of cord factor. The median survival time of the animals was prolonged (70 versus 17 days), and 40% of the mice survived more than 90 days. Tumor suppression was still detectable 36 days after administration of cord factor. The effect of cord factor was local. Comparable results were obtained with living or killed Calmette-Guérin bacilli. The results are discussed.

Animals↗