Method for Estimating the Content of the Chloroform-Extractable Green Fraction (CEGF) in HCl-DMSO Extract of Soils

Chloroform-extractable green fraction (CEGF) was detected in the supernatant obtained by alkali precipitation from the HCI-dimethylsulfoxide (DMSO) extract of Pg-rich soil. In the alkaline solution, the color of CEGF was green and CEGF showed strong Pg-like absorption bands. Ultraviolet and visible (UV-VIS) spectral analysis and gel chromatography on Sephadex G-50 were performed to compare several properties between CEGF and Pg. CEGF, which was purified by gel chromatography on Bio-Beads SX-1, displayed strong absorption bands at 609, 562, 445 and 280 inn in the alkaline solution. These absorption bands were almost similar to those of Pg. Furthermore, the UV-VIS spectrum of CEGF in the organic solvents showed a similar characteristic pattern of 4,9-dihydroxyperylene-3,10-quinone (DHPQ), which was considered to be a chromophore of Pg. Based on the results of gel chromatography on Sephadex G-50, CEGF mainly consisted of two fractions, corresponding to the G₂ and G₃ fractions of Pg. These results indicated that the method for extracting CEGF from Pg-rich soil in the present study was easy and selective and that CEGF was one of the components of, or a closely related substance to Pg. A colorimetric method for the estimation of the CEGF content in soils was developed. The calibration curve of CEGF was linear over a wide range of contents from 2.75 to 220 mg L⁻¹. The CEGF content in twelve samples of various soils was examined. CEGF was detected in all the soil sampled (5 orders) including three samples (3 orders) where Pg was not detected, and the content ranged from 0.07 to 1.66 g kg⁻¹ (dry soil). Therefore, the method for estimating the CEGF content in soils developed in the present study was found to be suitable for various soil orders and it was assumed that CEGF occurred in various soil orders.     

Enantiomeric purity and odor characteristics of 2- and 3-acetoxy-1, 8-cineoles in the rhizomes of Alpinia galanga willd

(S)-(+)-O-methylmandelate esters of trans- and cis-1,3, 3-trimethyl-2-oxabicyclo[2.2.2]octan-5- and 6-ols (2- and 3-hydroxy-1,8-cineoles) were prepared, and eight diastereomers were separated. The absolute configuration of the asymmetric carbons of the cineole moiety of each diastereomer was determined by (1)H NMR data according to the Mosher theory. Each mandelate was reduced with LiAlH(4) to obtain optically pure hydroxy-1,8-cineoles, this being followed by acetylation to afford optically pure acetoxy-1, 8-cineoles. These acetates were subjected to chiral GC, using a cyclodextrin column, and the enantiomeric purity of trans- and cis-1, 3,3-trimethyl-2-oxabicyclo[2.2.2]octan-5- and 6-yl acetates in the aroma concentrate from the rhizomes of Alpinia galanga was determined as 93.9 (5S), 19.4 (5R), 63.5 (6R), and 100 (6R) % ee, respectively. The aroma character of each enantiomer was also evaluated by GC-sniffing.     

Anti-hyperglycemic effect of diacylated anthocyanin derived from Ipomoea batatas cultivar Ayamurasaki can be achieved through the alpha-glucosidase inhibitory action

To clarify a postprandial glucose suppression effect of diacylated anthocyanin with alpha-glucosidase (AGH) inhibitory activity, a single oral administration study of it in male 8-week-old Sprague-Dawley rats was performed. The diacylated anthocyanin used in this study was peonidin 3-O-[2-O-(6-O-E-feruloyl-beta-D-glucopyranosyl)-6-O-E-caffeoyl-beta-D-glucopyranoside]-5-O-beta-D-glucopyranoside isolated from storage roots of the purple sweet potato (Ipomoea batatas cv. Ayamurasaki), which showed a potent maltase inhibitory activity with an IC(50) value of 200 microM preferable to sucrase inhibition. When the diacylated anthocyanin (100 mg/kg) was administered following maltose (2 g/kg), a maximal blood glucose level (BGL) at 30 min was significantly decreased by 16.5% (P < 0.01) compared to vehicle. A minimum 10 mg/kg dose of the anthocyanin was necessary for the suppression of glycemic rise, and the ED(20) (69 mg/kg) was estimated to be approximately 30-fold lower than that of the therapeutic drug acarbose (ED(20) = 2.2 mg/kg). A reduction of serum insulin secretion was also observed corresponding to the decrease in BGL. No significant change in BGL was observed when sucrose or glucose was ingested, suggesting that the anti-hyperglycemic effect of the anthocyanin was achieved by maltase inhibition, not by sucrase or glucose transport inhibition at the intestinal membrane.     

Determination of spherosomes from lees materials

To evaluate the effectiveness in adsorbing organochlorine compounds such as chloroform, dichloromethane, or benzene by lees materials, the determination of spherosomes from different lees materials was established by using a hemacytometer under an optical microscope. Rice bran, wheat bran, rapeseed, linseed, okara, and sakekasu were used for this investigation, and activated carbon was also used as a standard adsorbent material. The number of spherosomes varied from 1.82 x 10(10) particles/g for sakekasu to 4.95 x 10(10) particles/g for wheat bran. There was a high correlation between the removal efficiency in adsorbing organochlorine compounds such as chloroform, dichloromethane, or benzene by lees materials and the number of spherosomes from different lees materials.     

Polymeric procyanidins as radical-scavenging components in red-hulled rice

The extracts from white-, black-, and red-hulled rice were prepared by sequential extraction with six different polar solvents, and their radical-scavenging activities were measured by methods using 1,1-diphenyl-2-picrylhydrazyl radical (DPPH*) and tert-butyl hydroperoxyl radical (t-BuOO*). The extracts prepared with highly polar solvents, methanol and deionized water, exhibited higher DPPH* and t-BuOO* scavenging activities in all three cultivars. In addition, the acetone extract from red-hulled rice exhibited a high DPPH* and t-BuOO* scavenging activity, while no such activity was detected for the acetone extracts from white- and black-hulled rice. The major components responsible for the radical scavenging in the acetone extract from red-hulled rice were identified as procyanidins by acidic hydrolysis, vanillin assay, and Sephadex LH-20 chromatography. GPC analysis of the acetylated procyanidins revealed that the average molecular weight is about 5000, in a range of about 500-18,000.     

Characterization of Newcastle disease viruses isolated from field cases in Japan

Seven Newcastle disease viruses isolated in Japan from 1930 to 1984 were cloned on chicken embryo fibroblasts (CEFs) and characterized biologically. All seven produced two or more types of plaques on CEFs. The plaques were classified into four types. Plaque cloning was carried out five times, and 22 cloned viruses were established. The biological characters of the cloned viruses suggested that the strains contain different clones and that their clones are different even among close cases, such as G strain and H strain.     

Properties of nine continuous B-cell lines established from enzootic bovine leukosis tumors.

We established 9 cell lines from 63 tumor cases of enzootic bovine leukosis and studied their properties. Cells of all lines formed small clumps and floated in culture medium, indicating growth. Four of the 9 cell lines were surface immunoglobulin (SIg)-positive, but the remaining 5 line cells were negative for SIg or, if SIg was detected, the percentage of SIg-positive cells was very low. Tests for the properties of the cells with monoclonal antibodies to lymphocytes revealed that the established line cells are B-lymphocytes. Morphological observation also revealed that they had the morphology of B-lymphoblastic cell. The results of E and EAC rosette assay were negative, but 6 of 8 cell lines were positive for EA rosetting. All the 9 cell lines reacted with MoAb C-143, which recognizes the tumor-associated antigen (TAA) of the EBL tumor cell. All 9 cell lines produced bovine leukosis virus (BLV). These results suggest that the 9 cell lines are tumor cells derived from B-lymphocytes of EBL.     

Tryptophan availability of some feedstuffs determined by pig growth assay

Three experiments were conducted with young pigs to measure the availability of tryptophan in soybean meal, meat and bone meal, cottonseed meal, corn and sorghum. In Exp. 1, a 17.5% crude protein corn and gelatin-based diet, deficient in tryptophan, was supplemented with graded levels of L-tryptophan to establish the growth response of 10- to 20-kg pigs to graded additions of dietary tryptophan. From these data the requirement for maximal growth was calculated to be .16% of the diet using a broken-line model. In Exp. 2, the effects of excess amino acids in a test feedstuff, i.e., soybean meal, on the growth assay method for estimating tryptophan availability was evaluated. The addition of excess crystalline amino acids to the basal diet in proportions equal to the excesses contributed by a test level of soybean meal resulted in a 17.7 percentage unit reduction in the estimate of tryptophan availability (82.3%). When the standard diet was supplemented with amino acids to provide the pattern of excess amino acids found in the corn-gelatin basal diet with added soybean meal, the availability of tryptophan in soybean meal was estimated to be 95.2%. In Exp. 3, the tryptophan availabilities for meat and bone meal, cottonseed meal, corn and sorghum were estimated to be 82.2%, 80.9%, 94.0% and 86.4%, respectively. The diets used in this experiment were supplemented to contain excesses of individual amino acids in the same proportions as found in the basal diet, with the test feed ingredient added as a source of tryptophan. In these experiments the tryptophan content of the various feedstuffs was measured by ion-exchange liquid chromatography following alkaline hydrolysis.     

Prevalence of circulating antibodies to p10, a non-structural protein of the Borna disease virus in cats with ataxia.

Japanese domestic cats were surveyed for circulating antibodies to the plO and p24 proteins of the Borna disease virus (BDV) by Western blotting. Twenty-four of 52 cats (46.2%) with ataxia and other neurologic symptoms of unknown cause were positive for antibodies to BDV p10 and/or p24. In contrast, cats without neurological symptoms gave a significantly lower prevalence of anti-BDV antibodies to p10 and/or p24 (36 of 152 cats, 23.7%). Thirty specific pathogen-free (SPF) cats tested as controls were uniformly negative to BDV pl0 and p24 antigens. These results suggest that BDV may play a role in ataxia in cats. Additionally, our results suggest that it is necessary to use both p10 and p24 as antigens to detect circulating antibodies to BDV in cats.     

An outbreak of copper poisoning in Mute swans (Cygnus olor).

Eight of 9 Mute swans (Cygnus olor) untied in the river acrossing the central part of Tottori-city died within the period of 40 days of summer in 1989. Seven of 8 Mute swans were pathologically examined. In all swans many yellowish-brown to greenish-brown granules were found in the cytoplasm of hepatocytes. The granules were intensely stained with rhodanine copper stain, schmorl method, and Berlin blue stain. Ultrastructurally, many lysosomes increased in size and density in the cytoplasm of hepatocytes. Other three swans, that died at other places, were served as controls. In control swans, many brown granules intensely stained with schmorl method and Berlin blue stain were also found in hepatocytes, but the number of rhodanine-positive granules were fewer than those of the affected cases. X-ray qualitative analysis showed three peaks corresponding to copper, zinc and sodium in the liver of the affected and control swans. Quantitative analysis demonstrated that mean hepatic copper concentration of the affected group was significantly higher than that of control group (P less than 0.01). From these findings, we concluded that all of 7 Mute swans died of copper poisoning.