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.     

Absorption of dissolved and dispersed nutrients from sea-water by Panulirus japonicus phyllosoma larvae

This study examined the capacity of Japanese spiny lobster Panulirus japonicus phyllosoma larvae to absorb nutrients directly from the surrounding water. For this purpose, 48-, 81-, 124-, 160-, and 264-day-old intermoult larvae were starved for 48 h, held for 30 min to 4 h in 0.05 g L^–1 solutions or dispersions of cholesterol, peanut oil, saccharose and horseradish peroxidase in seawater, and subsequently examined by histological and ultrastructural methods. Lipids, carbohydrate, and protein were detected in either the lumen of the midgut gland tubules, the midgut gland cells, the haemolymph or in the cells of the epidermis of all larvae examined after each experimental period, with general improvement of the cell and tissue characteristics over unfed controls. The lipids were also found in the cuticle. Thus, the results provide evidence of nutrient absorption via the digestive tract, and possibly also via the integument. Structural adaptations that may be involved in the process of absorption were tentatively identified. Our observations suggest the possibility that particulate organic matter (POM) and dissolved organic matter (DOM) can be utilized by the P. japonicus phyllosoma larvae, perhaps as supplementary sources of nutrients to macroscopic diets.     

Quantitative assessment of muscle mass and gene expression analysis in dogs with glucocorticoid-induced muscle atrophy

The present study aimed to quantitatively evaluate muscle mass and gene expression in dogs with glucocorticoid-induced muscle atrophy. Five healthy beagles received oral prednisolone for 4 weeks (1 mg/kg/day), and muscle mass was then evaluated via computed tomography. Histological and gene expression analyses were performed using biopsy samples from the biceps femoris before and after prednisolone administration. The cross-sectional area of the third lumbar paraspinal and mid-femoral muscles significantly decreased after glucocorticoid administration (from 27.5 ± 1.9 to 22.6 ± 2.0 cm² and from 55.1 ± 4.7 to 50.7 ± 4.1 cm², respectively; P<0.01). The fast- and slow-twitch muscle fibers were both atrophied (from 2,779 ± 369 to 1,581 ± 207 μm² and from 2,871 ± 211 to 1,971 ± 169 μm², respectively; P<0.05). The expression of the growth factor receptor-bound protein 10 (GRB10) significantly increased after prednisolone administration (P<0.05). Because GRB10 suppresses insulin signaling and the subsequent mammalian target of rapamycin complex 1 activity, increased expression of GRB10 may have resulted in a decrease in protein anabolism. Taken together, 1 mg/kg/day oral prednisolone for 4 weeks induced significant muscle atrophy in dogs, and GRB10 might participate in the pathology of glucocorticoid-induced muscle atrophy in canines.     

Clinical evaluation of urinary liver-type fatty acid-binding protein for the diagnosis of renal diseases in dogs

Liver-type fatty acid-binding protein (L-FABP) is a biomarker for the early detection of renal diseases in humans. L-FABP is a cytotoxic oxidation product secreted from the proximal tubules under ischemic and oxidative stress conditions. First, L-FABP gene expression in the kidney and liver was evaluated. Next, the urinary L-FABP concentrations in dogs with or without renal diseases were measured using a novel enzyme-linked immunosorbent assay kit. Urinary L-FABP was normalized relative to urinary creatinine (uCre) concentrations (µg/g uCre). Finally, the relationships between urinary L-FABP and renal biomarkers used in canine medicine or serum alanine transaminase (ALT) as an indicator of liver damage were examined. Serum and urine samples from 94 client-owned dogs including 23 dogs with renal diseases and 71 dogs without renal diseases were used for analysis. Relative L-FABP gene expression was confirmed both in the liver and kidney. Dogs with renal diseases had a significantly higher urinary L-FABP than those without, and its predictive cutoff value was 26 µg/g uCre. Urinary L-FABP was significantly correlated with serum creatinine (r=0.4674, P<0.01), urea nitrogen (r=0.4907, P<0.01), urine specific gravity (r=−0.5100, P<0.01), and urine protein/creatinine ratio (r=0.7216, P<0.01), but not with serum ALT. Hence, dogs with a high urinary L-FABP value were more likely to have renal diseases.     

The first detection of Dicrocoelium chinensis sporocysts from the land snail Aegista vulgivaga in Gifu Prefecture, Japan

Trematodes of the genus Dicrocoelium are one of the most common parasites in ruminant animals; however, their life cycles in Japan are unclear. To find the sporocysts of D. chinensis in the natural field, we sampled 269 land snails (14 species) at a location with high level infection of sika deer in Gifu Prefecture, Honshu Island, Japan in autumn between 2017 and 2019. During the sampling period, we found mother sporocysts in the hepatopancreas of Aegista vulgivaga and Cyclophorus herklotsi. DNA barcoding based on the sequences of cytochrome c oxidase subunit 1 showed that the sporocysts from A. vulgivaga belonged to D. chinensis, indicating that this snail has potential as the first intermediate host of D. chinensis at this location.     

Usefulness of histological criteria for assessing the adequacy of diets for Panulirus japonicus phyllosoma larvae

Mass seed production depends a great deal on the development of adequate artificial diets, which in turn depends on the development of methods to speedily and reliably assess the nutritional value of experimental diets. This study examined the sensitivity of light histological and ultrastructural criteria to detect differences in the nutritional value of diets for the Japanese spiny lobster, Panulirus japonicus (V. Siebold), phyllosoma larvae. The acceptability of diets, the apparent absorption of lipids and carbohydrates by the midgut gland cells, and the characteristics of various cell types were determined in 43-, 106- and 143-day-old intermoult phyllosoma fed diets of Mytilus edulis (L.) ovary alone (MO) or in combination with Anemia (MO+AR), which so far have given the best results in terms of growth and percentage of metamorphosis into puerulus, and two other diets considered less efficient, viz. Mytilus edulis testis (MT) and Artemia (AR). Acceptability of all diets attained 100% within 10 min of feeding as revealed by observation through the semitransparent cuticle of the larvae. Histological examination of larvae that ingested comparable amounts of food revealed pronounced variation in the absorption of lipids and carbohydrates by the midgut gland cells (AR相似文献   

A serological survey of porcine reproductive and respiratory syndrome virus in wild boar in Gifu Prefecture,Japan

Porcine reproductive and respiratory syndrome (PRRS) is an infectious swine disease caused by the PRRS virus (PRRSV) that results in economic loss to the pig-rearing industry. To study PRRSV infection in wild boars and pigs, we conducted a serological survey in Gifu Prefecture, Japan, from 2020 to 2021. Three out of 453 (0.7%) wild boar sera were positive for PRRSV antibodies in a commercial ELISA. However, given that PRRSV RNA was not detected in these three wild boars and the specificity and sensitivity of the test kit, these are considered as false positives. Although seropositive pigs were found in multiple pig farms in the study area, the role of wild boars as a source of PRRS to pig farms appeared to be minimal.