Identification of reaction products of acylated anthocyanins from red radish with peroxyl radicals

Red radish anthocyanin extract, which consists of 12 known acylated anthocyanins, was reacted with 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH) to generate peroxyl radicals under acidic pH conditions at 37 degrees C. The reaction products were isolated using preparative HPLC, and their chemical structures were determined to be p-hydroxybenzoic acid (1), 6-O-(E)-p-coumaroyl-2-O-beta-d- glucopyranosyl-alpha-d-glucopyranoside (3), p-coumaric acid (4), 6-O-(E)-feruloyl-2-O-beta-d-glucopyranosyl-alpha-d-glucopyranoside (5), and ferulic acid (6). Some products were not identified. HPLC analyses of the mixture of acylated pelargonidin isolated from red radish and AAPH revealed that the acylated pelargonidins possess the radical scavenging ability on some common sites even if the characteristics of the intramolecular acyl units are different. Degradation rates of acylated pelargonidins and the formation rates of the resulting reaction products were found to be quite different.     

Down regulation of a novel protein,PSP, in rat hepatoma cdRLh 84-beared tumor

Perchloric acid-soluble protein (PSP) is a members of a new hypothetical family (YER057c/YJGF family) of small proteins with presently unknown function. The high degree of evolutionary conservation of these proteins reflect an involvement in basic cellular regulation. The expression of PSP was examined in rat hepatoma cell dRLh 84-beared rats. The tumor weight increased to 4.24 g at 3 weeks after the transplantation of hepatoma cells and hepatoma which has less differentiated characteristics were observed in rat liver. The expression of PSP in rat hepatoma was down regulated as compared with intact tissue. Thus the expression of PSP seems to be associated with the differentiation process in these transformed cells. On the other hand, some positive cells against the PSP-antibody were observed in the central region of tumor tissue by immunohistochemistry. These cells were shown to be the lymphocytes and the macrophages. The involvement of PSP to cellular growth and differentiation is discussed.     

Experimental Final Hosts of Metagonimus Hakubaensis (Trematoda: Heterophyidae) and Their Suitability to the Fluke

Seven laboratory mammal and bird species were orally inoculated with 200–1,000 encysted Metagonimus hakubaensis metacercariae that had been isolated from naturally infected lampreys (Lethenteron reissneri) captured in Aomori Prefecture. At 8 and 15 days post-infection, adult flukes were recovered from all of the laboratory animals tested, and therefore, hamster, rat, mouse, dog, cat, chicken and quail were considered as final hosts of M. hakubaensis. Recovery rates of the fluke were higher in dogs and hamsters than in cats, rats, mice, chickens and quails. The flukes recovered from dogs and hamsters showed increased body length and higher fecundity than those recovered from the other hosts. These results indicate that the suitability of dogs and hamsters for M. hakubaensis infection is higher than that of the other laboratory animals.     

Strain differences in morphometrical characteristics of rat kidneys

To clarify the strain differences in the morphology of the rat kidneys, we investigated the morphometrical characteristics of the kidneys of Slc:Wistar, Slc:SD, and F344/NSlc rats. The diameter of the renal corpuscles in female F344/N rats is smaller than that in female Wistar rats. Although sex differences (males>females) were shown in SD and F344/N rats, no effects of castration were detected in any of the groups. Strain-dependent differences in the percentage of renal corpuscles with a cuboidal parietal layer were found in both male and female groups. The highest percentage of them was noted in male Wistar rats. Effects of castration were observed in female Wistar and male F344/N rats, and the values after castration were significantly higher than those in the intact animals. As for the number of proximal convoluted tubular nuclei, no strain differences were detected in either the male or female groups. Although a sex difference was found in SD rats (female>male), no effects of castration were detected in any of the groups. In female F344/N rats, numerous numbers of PAS-positive granules, which were observed in the proximal convoluted and straight tubular epithelia, were noted. Orchiectomy induced an increase of these granules in male SD and F344/N rats, but ovariectomy showed no effects on them in any strains. This is the first study to clarify the strain differences in the morphological characteristics of the kidneys in ordinary rat strains.     

Morphometrical analysis of the kidney from nonobese diabetic (NOD) mice in the non-diabetic stage

The kidneys of non-diabetic NOD and wild type ICR mice were examined morphometrically at 3 and 6 months of age. Kidney weights and diameter of renal corpuscles of non-diabetic NOD mice were less than those of ICR mice. No lesions were observed in glomeruli or uriniferous tubules. Renin-positive areas were more common in NOD mice than in ICR mice, but no differences were detected in the Western blot analyses.     

Rodent renal structure differs among species

In the present study, we histologically and morphometrically investigated species differences in renal structure using laboratory rodents (mice, gerbils, hamsters, rats, and guinea pigs). Morphometric parameters were as follows, 1) diameter of the cortical renal corpuscles, 2) diameter of the juxtamedullary renal corpuscles, 3) percentage of the renal corpuscles with a cuboidal parietal layer, 4) number of nuclei in proximal convoluted tubules (PCTs) per unit area of cortex, 5) semi-quantitative score of the periodic acid-Schiff (PAS) -positive granules in PCTs, and 6) semi-quantitative score of the PAS-positive granules in proximal straight tubules (PSTs). Significant species differences were detected for each parameter, and particularly severe differences were observed in the PAS-positive granules of PCTs and PSTs. Granular scores varied among species and sexes. Vacuolar structures that did not stain with PAS or hematoxylin-eosin were observed in the renal proximal tubules. The appearance and localization of these vacuolar structures differed remarkably between species and sexes.     

Proposal of screening method for intestinal mucus adhesive lactobacilli using the enzymatic activity of glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH)

Adhesion tests are complex, time‐consuming and expensive, while the most important criterion for a probiotic lactobacilli is the ability to adhere to the human intestine. Thirty lactobacilli isolates from human intestinal tissues were measured for cell surface glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) activity using a microtiter plate screening method. GAPDH activities were detected in 21 out of 30 samples from 12 h cultures and in all samples from 18 h cultures. This suggests GAPDH is universally expressed on the bacterial cell surfaces from many lactobacilli. A statistically significant positive correlation was shown between GAPDH activity and adhesion using the BIACORE adhesion assay (P < 0.01). The new screening method using GAPDH enzymatic activity without an adhesion test may be possible due to the significant positive correlation of GAPDH activity with adhesion of lactobacilli derived from the human intestine.     

Age-related morphological changes in kidneys of SPF C57BL/6Cr mice maintained under controlled conditions

Age-related morphological changes were examined in the kidneys of inbred C57BL/6Cr mice maintained in a controlled environment. The specific pathogen free status of animals used in the present study was confirmed by microbiological monitoring. Kidneys were histologically and histometrically investigated at 3, 5, 12, 15, 24 and 27-months-old. Kidney weights did not change with age. Renal corpuscles increased in number at 24- and 27-months-old, but diameter remained constant. The percentage of renal corpuscles with a cuboidal glomerular capsule decreased at 24- and 27-months-old. Score indicating glomerular damage increased from 5- to 27-months-old. Changes to the proximal convoluted tubules were severe. Vacuolar degeneration was observed from 12-months-old. Tubular atrophy was observed at 24- and 27-months-old, with number of nuclei per unit area increasing at the same ages. Amyloidosis and scar lesions were observed at 27-months-old. Focal cell infiltration around vessels was found at 24- and 27-months-old. Electron microscopy at 27-months-old revealed expansion of the mesangial matrices and fusion of foot processes in the glomeruli. Enlarged lysosomes with lipid content were observed in the proximal convoluted tubules.