Preparation of phytate-removed deamidated soybean globulins by ion exchangers and characterization of their calcium-binding ability.

Phytate-removed deamidated soybean globulins were prepared using ion-exchange resins to provide them with a functional property to enhance calcium absorption in the body. The phosphorus level was reduced from 45 to 20 micromol/g using 0.05 g/mL of AE-4, an anion-exchange resin with a (2-hydroxyethyl)dimethylammonium group, in 0.2% soybean globulin solution for 1 h at 4 degrees C, and 90-92% of the phosphorus in defatted soybeans could be removed. As for deamidation, CE-4, a cation-exchange resin of the carboxylate type, showed a much higher deamidation activity than CE-1 and CE-2, cation-exchange resins of the sulfonate type. No peptide bond hydrolysis was observed for any cation-exchange resin treated at 4 degrees C. There was no significant difference in the amount of acid amide deamidated at temperatures between 4 and 50 degrees C. The deamidation level was able to increase to 73% using 0.10 g/mL of CE-4 in a 0.2% soybean globulin solution for 6 h at 4 degrees C. The amount of calcium bound to the soybean globulins decreased with removal of the phytate but increased with deamidation.     

Effect of 3-O-octanoyl-(+)-catechin on the responses of GABA(A) receptors and Na+/glucose cotransporters expressed in xenopus oocytes and on the oocyte membrane potential

Recently, 3-O-octanoyl-(+)-catechin (OC) was synthesized from (+)-catechin (C) by incorporation of an octanoyl chain into C in the light of (-)-epicatechin gallate (ECg) and (-)-epigallocatechin gallate (EGCg), which are the major polyphenols found in green tea and have strong physiological activities. OC was found to inhibit the response of ionotropic gamma-aminobutyric acid (GABA) receptors (GABA(A) receptors) and Na+/glucose cotransporters expressed in Xenopus oocytes in a noncompetitive manner more strongly than does C. OC also induced a nonspecific membrane current and decreased the membrane potential of the oocyte, and thus death of the oocyte occurred even at lower concentrations than that induced by C or EGCg. Although EGCg produced H2O2 in aqueous solution, OC did not. This newly synthesized catechin derivative OC possibly binds to the lipid membrane more strongly than does C, Ecg, or EGCg and as a result perturbs the membrane structure.     

Comparison of Ni/Zn Accumulation Ability of Thlaspi japonicum from Three Different Areas in Hokkaido

Thlaspi japonicum is a Japanese Ni-hyperaccumulator plant growing in areas with serpentine soil containing a high level of Ni. In the present study, we investigated the ability of wild Thlaspi japonicum plants inhabiting non-ultramafic soil areas of Hokkaido to accumulate Ni/Zn. Wild T. japonicum plants growing on Rebun Island (Rebun-T) accumulated small amounts of Ni, while the plants cultivated on ultramafic rock soil accumulated large amounts of Ni, like T. japonicum growing on Mt. Yubari (Yubari-T), and this change was also observed for T. japonicum growing on Mt. Tengu (Tengu-T). All the T. japonicum plants transplanted on both ultramaflc soil or Andosol accumulated over 100 mg kg⁻¹ Zn, especially the Ni/Zn content of Tengu-T plants tended to be higher than that of the plants growing in the other two areas. In contrast, transplanting of Yubari-T plants from ultramafic soil with a high level of Ni to soil with a lower-Ni content decrease of the Ni concentration in the leaves. Wild Rebun-T plants showed a smaller biomass than the Yubari-T and Tengu-T plants, and some individuals of the Rebun-T population were smaller than those of the other populations, with tiny radical leaves. Zinc content of the small leaves was not significantly different from that in large leaves. These results suggested that T. japonicum plants are able to accumulate high Ni/Zn accumulation, regardless ecotype, habitat and leaf size.     

Analysis of properties of monoclonal antibodies to transmissible gastroenteritis virus using TO-163 strain

Nineteen monoclonal antibodies (MAbs), reactive in enzyme-linked immunosorbent assay (ELISA), with porcine transmissible gastroenteritis (TGE) virus TO-163 were obtained. Of these MAbs, 5 showed neutralizing (NT) activity (x 3,200 to 25,600) against TO-163. One of the MAbs which had NT activity showed hemagglutination inhibition activity (x 5,120) too. 14 hybridomas of polypeptide specificity against TO-163 strain were developed from which 11, 2, and 1 were specific for protein E2, N, and E1, respectively. Immunofluorescence staining patterns in TGE virus-infected cells reacted with MAbs were divided into three groups (types I, II and III). The fluorescence staining of E2 specific MAbs having NT activity were limited to the perinuclear area. All MAbs having NT activity showed the same fluorescence staining pattern.     

Detection of bovine rhinotracheitis virus antibody by neutralizing test and ELISA in experimentally infected rabbits.

The antibody response of rabbit to infectious bovine rhinotracheitis (IBR) virus was examined. Two rabbits were inoculated with IBR virus strain Los Angeles into the trachea and intravenously, and intravenously two times, respectively. The patterns of antibody titers in the rabbits measured by neutralization test and ELISA were similar to those in the case of bovine. The antibody was detected after the inoculation, and much more antibody was detected after the second inoculation. The fact suggests that rabbits are a very useful laboratory animal for IBR virus infection studies.     

Effects of medium‐chain fatty acid‐cyclodextrin complexes on ruminal methane production in vitro

The purpose of the present study was to evaluate effects of medium‐chain fatty acid‐cyclodextrin (CD) complexes on ruminal methane and volatile fatty acid production, and protozoal activity in vitro. Medium‐chain fatty acid‐CDs used in this study were caprylic acid (C8)‐αCD or ‐βCD, capric acid (C10)‐αCD or ‐βCD, and lauric acid (C12)‐αCD or ‐βCD. A 60‐mL of diluted rumen fluid was incubated anaerobically at 38°C for 6 h with the addition of the complex (10–40 mg as fatty acid). Each of the fatty acid‐CDs reduced the number of protozoa, with the order C10 > C12 > C8, and βCD complexes were more effective than αCD complexes. Molar proportions of acetic acid remained unchanged with the addition of fatty acid‐CD, while that of propionic acid increased, being significant for C8‐αCD and βCD, and C10‐αCD and βCD (P < 0.05). Hydrogen production decreased by about 70% of control with the addition of 40 mg of C8 and C10‐CD, on the other hand, it tended to increase with the addition of C12‐CD in both αCD and βCD. Methane production decreased by about 20% with the addition of 40 mg of complexes, except for C10‐βCD, which significantly reduced methane production by about 60%. In conclusion, the addition of C8 or C10‐CD to ruminant diets may be effective in reducing methane production.     

Safety and efficacy of the cell-associated vaccine prepared by an attenuated infectious laryngotracheitis virus.

The safety and efficacy of the cell-associated (C-A) vaccine prepared by chicken embryo fibroblast (CEF) cells infected with the tissue-culture-modified strain of infectious laryngotracheitis (ILT) virus were studied in chickens. Over seventy percent of chickens inoculated with the C-A vaccine by the subcutaneous (S.C.) or intramuscular (I.M.) route at 1 day of age was protected against challenge with a virulent strain of ILT virus without any clinical signs. Chickens vaccinated with the C-A vaccine at 1 day of age acquired immunity within 6 days after vaccination, and the protection rate maintained more than 60% until 10 weeks post-vaccination. The C-A vaccine was invariably effective for chickens at various age. There was no evidence that the development of immunity was hindered by further vaccination with Newcastle disease and infectious bronchitis combined live vaccine. In addition, the C-A vaccine was safe when chickens were inoculated with 10 doses. In the field trials of the C-A vaccine, no adverse reaction was observed, and over 65% of vaccinated chickens was protected against the challenge of the virulent ILT virus at 8 weeks after vaccination.     

Evaluation of the pathogenicity and immunogenicity of vaccinia virus to piglets

The pathogenicity and immunogenicity of vaccinia virus were examined in order to evaluate the possibility of its application as a recombinant viral vaccine in pigs. Following scarification inoculation with vaccinia virus, a mild reddish papulation developed only on the scarified part of the skin. No symptoms of illness such as fever or stunting were noted. Vaccinia virus was recovered in titers from scarified skin 4 and 7 days after inoculation. Control piglets cohabited with inoculated animals remained normal for the whole 5 week observation period. Hemagglutination inhibition and indirect immunofluorescence tests detected antibodies against vaccinia virus in the inoculated piglets, whereas no anti-vaccinia virus antibodies were detected in the contact control animals. Antigen-induced blastogenic tests of peripheral blood lymphocytes from animals, revealed that lymphocytes obtained from inoculated donors 5 weeks after inoculation, had a higher stimulation index (P less than 0.05) than did those from uninoculated piglets. These results suggested that vaccinia virus would be useful as a recombinant viral vector for pigs.