Ligand-binding characteristics of feline insulin-binding immunoglobulin G

Polyclonal immunoglobulin (Ig) G autoantibodies against insulin have been identified in sera of healthy cats. We purified and fractionated insulin-binding IgGs from cat sera by affinity chromatography and analyzed affinity of insulin-binding IgGs for insulin and their epitopes. Following the passing of fraction A, which did not bind to insulin, insulin-binding IgGs were eluted into two fractions, B and C, by affinity chromatography using a column fixed with bovine insulin. Dissociation constant (KD) values between insulin-binding IgGs and insulin, determined by surface plasmon resonance analysis (Biacore™system), were 1.64e⁻⁴ M for fraction B (low affinity IgGs) and 2e⁻⁵ M for fraction C (high affinity IgGs). Epitope analysis was conducted using 16 peptide fragments synthesized in concord with the amino acid sequence of feline insulin by an enzyme-linked immunosorbent assay. Fractions B and C showed higher absorbance (affinity) of the peptide fragment of 10 amino acid residues at the carboxyl-terminal of the B chain (peptide No. 19), followed by peptide fragments of 6 to 15 amino acid residues of the B chain (peptide No. 8). Fraction C showed a higher absorbance to 7 to 16 amino acid residues of the B chain (peptide No. 5) compared with the absorbance of fraction B. Polyclonal insulin-binding IgGs may form a macromolecule complex with insulin through the multiple affinity sites of IgG molecules. Feline insulin-binding IgGs are multifocal and may be composed of multiple IgG components and insulin.     

Metabolic characterization of newly isolated Pseudomonas nitroreducens Jin1 growing on eugenol and isoeugenol

Newly isolated soil bacterium strain Jin1 was able to grow on both eugenol and isoeugenol each as sole source of carbon and energy. Based on bacterial 16S rDNA analysis, Jin1 belongs to Pseudomonas nitroreducens with a similarity of 98.92% (14/1297). P. nitroreducens Jin1 was found to biotransform eugenol and isoeugenol to vanillin by different pathways. Eugenol was biotransformed to vanillin through coniferyl alcohol and ferulic acid similarly to the pathway shown previously by Pseudomonassp. HR199 and vanillin produced from eugenol was rapidly metabolized to vanillic acid. Contrastively, Pseudomonas nitroreducens Jin1 did not appear to produce metabolic intermediates during the biotransformation of isoeugenol to vanillin which was finally biotransformed to vanillic acid with much slower rate. These results indicate that there seems to be different metabolic regulation systems for the biotransformation of eugenol and isoeugenol by this bacterium. Herein, we report on Pseudomonas nitroreducens Jin1, a novel bacterium that produces vanillin from eugenol and isoeugenol by two different metabolic pathways.     

Changes in density of under-yearling masu salmon <Emphasis Type="Italic">Oncorhynchus masou</Emphasis> in a natural stream after release

ABSTRACT:   The relation between the density of fish and parameters such as body weight, condition factor and the hepatic triglyceride (TG) content of masu salmon Oncorhynchus masou juveniles after release into a natural stream were studied. Hatchery-reared juveniles of different sizes and numbers were released in May 1999, 2000 and 2001, and investigations were carried out from July to September each year. The density of fish was relatively high in 1999, when juveniles of the largest size and number were released, but then continued to decrease during the investigation. The hepatic TG content remained considerably low in 1999, and there was a significant negative correlation between the density of fish and the hepatic TG content of the juveniles caught over three years. The initial fish size and number at release as well as the nutritional condition of the juveniles may affect the density of fish after release.     

Diet shift of larval and juvenile landlocked Ryukyu-ayu <Emphasis Type="Italic">Plecoglossus altivelis ryukyuensis</Emphasis> in the Fukuji Reservoir,Okinawa Island,Japan

ABSTRACT:   The diet shift of larval and juvenile Ryukyu-ayu Plecoglossus altivelis ryukyuensis was investigated for the landlocked population stocked in the Fukuji Reservoir, Okinawa Island. Gut contents of 274 specimens (5.1–31.4 mm in body length) including 31 yolk-sac larvae (5.7–8.0 mm) were examined. The diet of larval and juvenile landlocked P. a. ryukyuensis was mainly composed of zooplankton such as dinoflagellates, rotifers, copepods, and cladocerans. Feeding incidence was 48% in total and it gradually increased with growth. Diet analysis clearly demonstrated a diet shift during the larval and juvenile periods. For yolk-sac larvae and 5.0–19.9 mm fish, the diet was dominated by dinoflagellates Gymnodinium sp., and Peridinium sp., both numerically and by frequency, followed by rotifers that mainly consisted of Polyarthra spp. Diet breadth (Levin's standardized index B_A) diversified with growth, and feeding on copepods and cladocerans gradually increased in 10.0–14.9 mm fish. Diet overlap (Morisita's index: C) showed that diet drastically changed in 20.0–24.9 mm fish with a specialization on cladocerans Bosminopsis deitersi and Diaphanosoma brachyurum . This diet shift probably corresponded with an increase of feeding ability resulting from an enlargement of the mouth, increased swimming ability, and the development of sense organs.     

Comparison of Quality‐Related Alleles Among Australian and North American Wheat Classes Exported to Japan

Grain hardness, amylose content, and glutenin subunit composition are critical determinants for end‐use properties of wheat. To improve the end‐use properties of domestic wheats, we studied these traits between the Australian and North American wheat classes exported to Japan in 2009 and 2011 by analyzing the corresponding alleles. Most hard classes had Pina‐D1b or Pinb‐D1b. A partial waxy allele (Wx‐B1b) was found in all Australian Standard White (ASW) seeds in 2009 and two‐thirds of ASW seeds in 2011. All or most American hard wheat seeds had Glu‐D1d. Most U.S. Western White (WW) seeds had a null allele (Glu‐A1c) or alleles that lacked one of the two Glu‐B1 subunits. Most hard red winter (HRW) seeds had Glu‐B3b or Glu‐B3g. Quality characteristics of these classes seemed to be consistent with these results. In addition, we also found new Glu‐1 and Glu‐3 alleles in HRW and WW. These results suggested that although there are variations in its allelic composition from year to year, each class has unique quality‐related alleles corresponding to its end use. We proposed two matrices for classification of starch properties on the basis of Pin and Wx allelic combinations and for classification of gluten strength on the basis of glutenin allelic combinations.