Decreased expression of catechol-O-methyltransferase in the renal cortex of malignant spontaneously hypertensive rats.

Essential hypertension is a disease of unknown pathogenesis, although renal function has been implicated as an important factor in its cause. Stroke-prone spontaneously hypertensive (SHRSP) rats provide an animal model of essential hypertension. To understand the cause of hypertension, identifying proteins that are differentially expressed between hypertensive and normotensive rats may provide a key. Here, proteins in the renal cortex from SHRSP rats, malignant stroke-prone spontaneously hypertensive (M-SHRSP) rats, and Wistar Kyoto (WKY) rats as a normotensive control were subjected to two-dimensional difference gel electrophoresis (2D-DIGE). After in-gel digestion by trypsin, proteins were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). Several proteins showed differential expression patterns between hypertensive and normotensive rats. Among them, we focused on catechol-O-methyltransferase (COMT) because this enzyme inactivates catecholamines, possibly affecting blood pressure. To confirm the differential expression of COMT in each animal, we conducted Western blot analysis, which revealed that the expression of COMT is lower in M-SHRSP rats than in control and SHRSP rats, indicating that blood pressure and expression of COMT are related. In fact, the blood pressure of M-SHRSP rats was significantly higher than that of SHRSP rats at age of 10 weeks. Immunohistochemical and immunofluorescence studies showed that COMT in renal cortex is localized in tubular epithelial cells. The expression of COMT is lower in the renal cortex tubular epithelium of M-SHRSP rats than in normotensive rats. These results suggest that the decreased expression of COMT may be an important factor leading to the development of hypertension.

RAPD analysis of salt-tolerant yeasts from contaminated seasoned pickled plums and their growth inhibition using food additives.

Eight salt-tolerant yeasts were isolated from contaminated pickled plums which were seasoned with honey and "Umami" seasoning. They were classified into four main groups according to random amplified polymorphic DNA analysis, and three of ten kinds of food additives tested inhibited their growth. The type strains of each group were identified as Zygosaccharomyces bisporus, Pichia subpeliculosa, and two strains of Candida apicola based on the D1/D2 region sequence of the 26S rRNA gene. They were able to grow in medium containing 6% (w/v) NaCI. A number of yeasts were isolated from production lines by the swab method, but not from the salted plums used as raw materials. These results show that the production lines require washing with antimicrobial agents effective against salt-tolerant yeasts. Three commercial food additives, San-keeper 381, Sunsoft No.700P-2, and potassium sorbate inhibited the growth of Z. bisporus at 125 to 250 microg/ml. In particular, San-keeper 381 altered the morphology of this species at 125 microg/ml. C. apicola and P. subpelliculosa were inhibited by Sunsoft No.700P-2 and potassium sorbate at 250 microg/ml. These results indicate that the washing of production lines with disinfectant and the use of food additives that effectively prevent salt-tolerant yeast contamination are necessary.