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.     

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.     

Improved survival of vitrified in vivo-derived porcine embryos

An efficient cryopreservation protocol for porcine morulae was investigated with three types of vitrification having different cooling rates (Exp. 1). Survival of embryos vitrified after removal of cytoplasmic lipid droplets was also examined by means of the minimum volume cooling (MVC) method (Exp. 2). In Exp. 1, the morula stage embryos were vitrified with a 0.25 ml plastic straw (ST-method), gel loading tip (GLT-method) and the MVC-method, respectively, and stored in liquid nitrogen after which they were warmed in sucrose solutions with cryoprotectants being subsequently removed in a stepwise manner. In Exp. 2, morulae were centrifuged with 7.5 microg/ml cytocharasin B at 12000 x g for 20 min to polarize the cytoplasmic lipid droplets that were then removed from the embryos by micromanipulation (delipation). Both those delipated at the morula stage and the intact embryos at the morula to blastocyst stages were vitrified by the MVC-method. In vitro survival of the vitrified embryos was assessed in both experiments by culturing in NCSU-23 + 10% FCS for 48 h. In vitro developments of vitrified embryos after warming to blastocysts were 20% (6/30) for the ST-method, 39% (18/46) for the GLT-method, and 60% (26/43) for the MVC-method. Embryo survival was further improved by vitrification after delipation (95%, 35/37) compared to intact vitrified morulae (24/42, 57%, P<0.001) and blastocysts (23/31, 74%, P<0.05). Moreover, the number of cells in blastocysts (92 +/- 25) derived from the delipated-vitrified morulae was comparable to those derived from intact control non-vitrified embryos (103 +/- 31). Our results demonstrate that vitrified porcine morulae have the highest survival when using the MVC-method in conjunction with delipation.     

Evaluation of the clinical efficacy of benazepril in the treatment of chronic renal insufficiency in cats

BACKGROUND: Chronic renal insufficiency (CRI) is a common disease in cats. Angiotensin-converting enzyme inhibitors (ACEI) have beneficial effects in humans with CRI by reducing the loss of protein in the urine and increasing life expectancy. HYPOTHESIS: The ACEI benazepril has beneficial effects on survival, clinical variables, or both as compared with placebo in cats with CRI. ANIMALS: 61 cats with naturally occurring CRI. METHODS: The cats were enrolled into a prospective, randomized, double-blind, placebo-controlled clinical trial. Cats received placebo or 0.5-1 mg/kg benazepril once daily for up to 6 months. RESULTS: Urine protein/urine creatinine ratios were significantly (P < .05) lower with benazepril as compared with placebo at days 120 and 180. Three cats with placebo and 1 cat with benazepril were removed prematurely from the study because of deterioration of CRI or death. Cats were classified into 4 stages of CRI according to the International Renal Interest Society (IRIS) classification scheme. Incidence rates of cats with IRIS classification stage 2 or stage 3 that remained in stage 2 or 3 without progressing to stage 4 were higher with benazepril (93 +/- 5%) as compared with placebo (73 +/- 13%). CLINICAL IMPORTANCE: These results suggest a potential for benazepril to delay the progression of disease, extend survival time, or both in cats with CRI.