Preliminary Studies on Hepatic Carnitine Palmitoyltransferase in Dairy Cattle with or without Fatty Liver

The hepatic mitochondrial carnitine palmitoyltransferase (CPT) activity was measured by fluorimetric assay in dairy cows with or without fatty liver. CPT activities in 13 lactating cattle and in 6 non-lactating cows were 304.4±86.6 mol CoA/min per g protein and 169.3±84.8 mol CoA/min per g protein, respectively. This difference was significant (<0.05). CPT activities in early lactation (0–110 days after calving), mid-lactation (111–220 days after calving) and late lactation (over 220 days after calving) were 278.9±68.0, 312.4±124.1 and 320±59.3 mol CoA/min per g protein, respectively. There was no significant difference between the values at different stages of lactation. The CPT activity in 10 lactating cows with fatty liver unrelated to calving was 201.3±80.0 mol CoA/min per g protein. CPT activity in 10 cattle with fatty liver was significantly lower than that in normal lactating cattle. Based on these findings, clinical fatty liver unrelated to calving appears to be associated with a decrease in hepatic CPT activity.     

Synergistic effects of thiols and amines on antiradical efficiency of protocatechuic acid

DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity of protocatechuic acid and its structural analogues (methyl protocatechuate, 3',4'-dihydroxyacetophenone, 3,4-dihydroxybenzaldehyde, and 3,4-dihydroxybenzonitrile) were examined in aprotic and protic solvents. In aprotic acetonitrile, all test compounds scavenged two radicals. In protic methanol, however, these compounds rapidly scavenged five radicals except for protocatechuic acid, which consumed only two radicals. The result indicated that higher radical scavenging activity in methanol than in acetonitrile was due to a nucleophilic addition of the methanol molecule on the oxidized quinones, which led to a regeneration of catechol structures. To investigate the importance of the nucleophilic addition on the quinones for the high radical scavenging activity, DPPH radical scavenging activity of protocatechuic acid and its analogues was examined in the presence of a variety of nucleophiles. The addition of a strong nucleophile such as a cysteine derivative significantly increased the radical scavenging equivalence. Furthermore, thiol adducts at C-2 and C-2,5 of protocatechuic acid and its analogues were isolated from the reaction mixtures. These results strongly suggest that the quinone of protocatechuic acid and its analogues undergo a nucleophilic attack at C-2 to yield 2-substituted-3,4-diols. Then, a regenerated catechol moiety of adducts scavenge two additional radicals by reoxidation into quinones, which undergo the second nucleophilic attack at the C-5. This mechanism demonstrates a possibility of synergistic effects of various nucleophiles on the radical scavenging ability of plant polyphenols containing a 3,4-dihydroxy substructure like protocatechuic acid and its analogues.     

Flavonol Caffeoylglycosides as alpha-Glucosidase Inhibitors from Spiraea cantoniensis Flower

In the screening experiments for rat intestinal alpha-glucosidase inhibitors in 218 plants cultivated in the Japanese temperate region, potent maltase-inhibiting activity was found in the extract of flowers of Spiraea cantoniensis. The enzyme assay guided fractionation of the extract led to the isolation of three flavonol caffeoylglycosides, quercetin 3- O-(6- O-caffeoyl)-beta-galactoside ( 1), kaempferol 3- O-(6- O-caffeoyl)-beta-galactoside ( 2), and kaempferol 3- O-(6- O-caffeoyl)-beta-glucoside ( 3), as rat intestinal maltase inhibitors. This is the first report on the alpha-glucosidase-inhibitory activity of those flavonol caffeoylglycosides. Comparison in the activity of the isolates indicated the importance of caffeoyl substructures in the molecule for the alpha-glucosidase-inhibiting activity. The relatively high contents of the active isolates in the plant suggest that S. cantoniensis could be physiologically useful for treatment of diabetes.     

Substrate specificities of microbial transglutaminase for primary amines

Transglutaminase (epsilon-glutaminyl-peptide:amine gamma-glutaminyl-transferase, EC 2.3.2.13) (TGase) is an enzyme that catalyzes acyl transfer reactions between primary amines and Gln residues in proteins and peptides. The substrate specificity of TGase for primary amines was investigated to incorporate various functional groups into proteins and peptides. In this study, microbial transglutaminase and guinea pig liver transglutaminase were used. For the primary amines to be incorporated into benzyloxycarbonyl-L-Gln-Gly (Z-Gln-Gly), they were required to have more than four carbon chains without side chains between the functional groups. These results suggest that with appropriate primary amines as spacers, various functional groups, carboxyl groups, phosphate groups, saccharides, and so on, can be incorporated into proteins by using TGase.     

Isolation and identification of indigestible pyroglutamyl peptides in an enzymatic hydrolysate of wheat gluten prepared on an industrial scale

An enzymatic hydrolysate of wheat gluten was further digested in vitro with porcine pepsin and pancreatin to obtain an indigestible peptide. Indigestible pyroglutamyl peptide was isolated from the digest by strong cation-exchange, size-exclusion, and reversed-phase chromatographies. The pyroglutamyl peptide was digested with pyroglutamate aminopeptidase, and the digest was reacted with phenyl isothiocyanate. The resultant phenylthiocarbamyl (PTC) peptides were purified by reversed-phase HPLC by using binary gradient elution with ammonium acetate buffer, pH 6.0, and acetonitrile. The PTC peptides were analyzed with an automatic peptide sequencer on the basis of the Edman degradation method with a modified program. Some pyroglutamyl peptides were also analyzed by fast-atom bombardment ionization mass spectrometry without the pyroglutamate amino peptidase digestion. Consequently, pyroGlu-Asn-Pro-Gln, pyroGlu-Gln-Gln-Pro-Gln, pyroGlu-Gln-Pro-Gln, pyroGlu-Gln-Pro-Gly-Gln-Gly-Gln, pyroGlu-Gln, pyroGlu-Gln-Pro, pyroGlu-Ile-Pro-Gln, pyroGlu-Ile-Pro, pyroGlu-Gln-Pro-Leu, pyroGlu-Gln-Phe-Pro-Gln, pyroGlu-Ser-Phe-Pro-Gln, pyroGlu-Phe-Pro-Gln, and pyroGlu-Gln-Pro-Pro-Phe-Ser were identified.     

Asphericity in supernova explosions from late-time spectroscopy

Core-collapse supernovae (CC-SNe) are the explosions that announce the death of massive stars. Some CC-SNe are linked to long-duration gamma-ray bursts (GRBs) and are highly aspherical. One important question is to what extent asphericity is common to all CC-SNe. Here we present late-time spectra for a number of CC-SNe from stripped-envelope stars and use them to explore any asphericity generated in the inner part of the exploding star, near the site of collapse. A range of oxygen emission-line profiles is observed, including a high incidence of double-peaked profiles, a distinct signature of an aspherical explosion. Our results suggest that all CC-SNe from stripped-envelope stars are aspherical explosions and that SNe accompanied by GRBs exhibit the highest degree of asphericity.     

Orbiter cloud photopolarimeter investigation

The first polarization measurements of the orbiter cloud photopolarimeter have detected a planet-wide layer of submicrometer aerosols of substantial visible optical thickness, of the order of 0.05 to 0.1, in the lower stratosphere well above the main visible sulfuric acid cloud layer. Early images show a number of features observed by Mariner 10 in 1974, including planetary scale markings that propagate around the planet in the retrograde sense at roughly 100 meters per second and bright- and dark-rimmed cells suggesting convective activity at low latitudes. The polar regions are covered by bright clouds down to latitudes aproximately 50 degrees, with both caps significantly brighter (relative to low latitudes) than the south polar cloud observed by Mariner 10. The cellular features, often organized into clusters with large horizontal scale, exist also at mid-latitudes, and include at least one case in which a cell cuts across the edge of the bright polar cloud of the northern hemisphere.