Diffusible Cytotoxic Substances and Cell-Mediated Resistance to Syngeneic Tumors: In vitro Demonstration

The interaction between target cells (mouse sarcoma cells) and syngeneic immunocytes (peritoneal cells) in close proximity but without direct contact was studied. The two cell types were separated by Millipore membranes, which allow diffusion of substances of large molecular size, in an assembly which permits cultivation of target cells on one side of the membrane and immunocytes on the other side. When brought into proximity in this manner for 48 to 72 hours, immunocytes from donors which had been immunized against syngeneic tumors caused destruction of the target cells. Since serum from immunized donors had no effect, it appears that the immunocytes produced a diffusible cytotoxic substance (or substances) which may be different from typical antibody.     

Walnut polyphenols prevent liver damage induced by carbon tetrachloride and d-galactosamine: hepatoprotective hydrolyzable tannins in the kernel pellicles of walnut

The polyphenol-rich fraction (WP, 45% polyphenol) prepared from the kernel pellicles of walnuts was assessed for its hepatoprotective effect in mice. A single oral administration of WP (200 mg/kg) significantly suppressed serum glutamic oxaloacetic transaminase (GOT) and glutamic pyruvic transaminase (GPT) elevation in liver injury induced by carbon tetrachloride (CCl 4), while it did not suppress d-galactosamine (GalN)-induced liver injury. In order to identify the active principles in WP, we examined individual constituents for the protective effect on cell damage induced by CCl 4 and d-GalN in primary cultured rat hepatocytes. WP was effective against both CCl 4- and d-GalN-induced hepatocyte damages. Among the constituents, only ellagitannins with a galloylated glucopyranose core, such as tellimagrandins I, II, and rugosin C, suppressed CCl 4-induced hepatocyte damage significantly. Most of the ellagitannins including tellimagrandin I and 2,3- O-hexahydroxydiphenoylglucose exhibited remarkable inhibitory effect against d-GalN-induced damage. Telliamgrandin I especially completely suppressed both CCl 4- and d-GalN-induced cell damage, and thus is likely the principal constituent for the hepatoprotective effect of WP.     

Alpha-glucosidase inhibitory and antihyperglycemic effects of polyphenols in the fruit of Viburnum dilatatum Thunb

Small crimson fruit of Viburnum dilatatum Thunb. (gamazumi), a wild deciduous low tree belonging to a family of Caprifoliaceae, has strong antioxidant activity, and cyanidin 3-sambubioside (C3S) and 5-caffeoyl quinic acid (5-CQA) are identified as active compounds. The freeze-dried powder of V. dilatatum fruit juice (CEV) was orally administered to streptozotocin-induced diabetic rats for 4 weeks repeatedly. Consequently, the elevation of plasma glucose level after oral administration of 2 g/kg glucose was suppressed by the repeated administration of CEV. The action was dependent on the dose of CEV, and plasma glucose level in rats administered 500 mg/kg of CEV was decreased significantly from that in rats without CEV. Increase of insulin secretion was not found in rats with or without administration of CEV. It was expected that CEV had some effects on glucose uptake. In five compounds identified from V. dilatatum fruit, C3S and 5-CQA showed inhibitory activity on sucrase and maltase. Inhibitory activity of cyanidin 3-glucoside and cyanidin aglycon (Cy) was not found markedly, and so it was thought that the activity was a characteristic property in Cy diglycosides. Moreover, 5-CQA and C3S were main polyphenol in the fruit of V. dilatatum. These results suggest that V. dilatatum fruit has the alpha-glucosidase inhibitory activities and the antihyperglycemic action.     

Pharmacokinetics of Eugenol in Japanese Flounder,Paralichthys olivaceus

Eugenol, which has been used as an anesthetic for fish, was administered to Japanese flounder, Paralichthys olivaceus (127 ± 50.8 g), by bath treatment at concentrations of 0.025, 0.125, and 0.25 mL/L seawater for 10 min and intramuscular injection at 40 μL/fish to investigate the pharmacokinetic characteristics of eugenol in the plasma of Japanese flounder. In the bath treatment test, plasma eugenol concentration increased with increasing eugenol concentration up to 0.125 mL/L and reached steady state within 5 min. After a 10‐min bath treatment in 0.25 mL/L eugenol, plasma eugenol concentration was about 58.4 µg/mL. After transfer into running seawater, plasma eugenol concentration decreased biphasically with half‐lives of 0.0296 h (α‐phase) and 0.289 h (β‐phase). The AUC_0000→0800 was about 16.5 µg h/mL. In administration by intramuscular injection, plasma eugenol concentration increased rapidly after administration and decreased biphasically with half‐lives of 0.0329 h (α‐phase) and 8.08 h (β‐phase). The AUC_0000→0800 was about 52.5 µg h/mL. In both methods of administration, Japanese flounder with average weight of 127 g were effectively anesthetized when plasma eugenol concentrations were between 2.19 and 4.88 µg/mL.     

Genomic evaluation using SNP‐ and haplotype‐based genomic relationship matrices in a closed line of Duroc pigs

A simulation analysis and real phenotype analysis were performed to evaluate the impact of three different relationship matrices on heritability estimation and prediction accuracy in a closed‐line breeding of Duroc pigs. The numerator relationship matrix (NRM), single nucleotide polymorphism (SNP)‐based genomic relationship matrix (GRM) (G_S), and haplotype‐based GRM (G_H) were applied in this study. We used PorcineSNP60 genotype array data (38 114 SNPs) of 831 Duroc pigs with four selection traits. In both heritability estimation and prediction accuracy, the accuracy depended on the number of animals with records. For heritability estimation, a large difference in the results among three relationship matrices was not shown, but the trend of the estimated heritabilities between GRMs, that is G_S < G_H, was shown in this population. For the accuracy of prediction values in test animals, the accuracies of prediction values obtained by two GRMs were higher than that by the NRM in this population. The accuracies obtained by GRMs using animals with no records were lower than that by the NRM using animals with their performance records, but were close to that by the NRM using animals with full‐sib testing records.     

Capsule types of Klebsiella pneumoniae isolated from the genital tract of mares with metritis, extra-genital sites of healthy mares and the genital tract of stallions

A survey of K. pneumoniae was performed on cervical swabs, feces and nasal swabs of mares and on samples from the genital tract of stallions from 1980 to 1986 in south-western Hokkaido, Japan. K1 was the predominant type (79 of 88, 89.8%) in the metritis cases due to K. pneumoniae in mares of racing breeds. The same type was isolated from semen and swabs of the fossa glandis of 6 of 20 (30.0%) of the stallions of racing breeds. Heavily encapsulated and less heavily encapsulated K1 strains were isolated from the stallions. Mares bred to stallions carrying heavily encapsulated strains developed metritis, while those bred to stallions carrying less heavily encapsulated strains did not. K39 was isolated from cervical swabs solely from metritis-infected mares of draft breeds and not from any mares of the racing breeds examined. Untypable strains were isolated from cervical swabs in 7 of 88 (8.0%) metritis cases of mares of racing breeds and from semen in 7 of 19 (36.8%) stallions of racing breeds and they were predominant in feces (19 of 21, 90.5%) and nasal swabs (3 of 4, 75.0%) of healthy mares of racing breeds.     

Magnitude of mitral valve closure plays a pivotal role in enhancing the forward blood flow during cardiac massage in dogs with ventricular fibrillation

Motion of mitral valve during cardiac massage was examined using beagle dogs with ventricular fibrillation (n=4). Active compression-decompression cardiac massage (ACD-CM) exhibited greater peak aortic pressure than standard cardiac massage (S-CM), reverse of which was true for peak pulmonary capillary wedge pressure in each animal. Accordingly, peak aortic pressure was greater than peak pulmonary capillary wedge pressure with ACD-CM, whereas its reverse was true with S-CM. Transesophageal echocardiography revealed that mitral valve was incompletely closed with S-CM with showing regurgitation. The valve was more effectively closed during ACD-CM. These results indicate that effective closure of mitral valve during cardiac massage may increase forward blood flow, supporting “cardiac pump theory” rather than “thoracic pump theory” as a principle in dogs.     

Allelopathy of buckwheat: Assessment of allelopathic potential of extract of aerial parts of buckwheat and identification of fagomine and other related alkaloids as allelochemicals

The allelopathic potential of buckwheat ( Fagopyrum esculentum ) was investigated. Field study showed that living buckwheat reduced weed biomass compared with plots without buckwheat. Laboratory study revealed that root exudates suppressed root and shoot growth of weeds and reduced weed dry weight. Sequential partitioning of the aqueous ethanolic extract of the aerial parts of buckwheat showed that the chloroform extract caused an 80% reduction in radicle elongation of lettuce seedlings at a concentration of less than 100 p.p.m. Fagomine, 4-piperidone and 2-piperidinemethanol were isolated and identified by ¹H and ¹³C nuclear magnetic resonance from the chloroform extract. These compounds caused a 50% inhibition of radicle elongation in lettuce seedlings at concentrations of less than 100 p.p.m. These results suggest that compounds 1, 2, and 3 might act as allelochemicals and affect the growth or germination of different plant species.