Retrospective diagnosis of feline GM2 gangliosidosis variant 0 (Sandhoff-like disease) in Japan: possible spread of the mutant allele in the Japanese domestic cat population

GM2 gangliosidosis variant 0 (human Sandhoff disease) is a lysosomal storage disease caused by simultaneous deficiencies of acid beta-hexosaminidase (Hex) A and Hex B due to an abnormality of beta-subunit, a common component in these enzyme molecules, which is coded by the HEXB gene. In the present study, a retrospective diagnosis was performed in 2 previous suspected cases of feline Sandhoff-like disease using a DNA test to detect the causative mutation identified previously in 4 cats in 2 other families of Japanese domestic cats. Enzymic analysis was also performed using stored leukocytes and plasma collected from the subject families in order to investigate the usefulness of enzymic diagnosis and genotyping of carriers. The DNA test suggested that the 2 cases were homozygous recessive for the mutation. Consequently, 6 cats homozygous for the same mutation have been found in 4 separate locations of Japan, suggesting that this mutant allele may be spread widely in the Japanese domestic cat populations. In enzymic analysis, Hex A and Hex B activities in leukocytes and plasma measured using 4-methylumbelliferyl N-acetyl-beta-D-glucosaminide as a substrate were negligible in affected cats, compared with those in normal and carrier cats. However, there was a wide overlap in enzyme activity between normal and carrier cats. Therefore, it was concluded that enzymic analysis is useful for diagnosis of affected cats, but is not acceptable for genotyping of carriers.     

Generation of monoclonal antibody against canine neural-cell adhesion molecule

A monoclonal antibody, K9BYU, was generated using Escherichia coli recombinant extracellular domain of canine neural-cell adhesion molecule (N-CAM) as an antigen. Immunoreactivity of K9BYU to insect cell recombinant canine N-CAM was demonstrated by Western blotting using Sf9 insect cells transfected with the canine N-CAM gene. In Western blotting against canine brain tissue, K9BYU detected three isoforms of N-CAM that correspond to three major isoforms of human and mouse N-CAM (N-CAM-120, -140, and -180). From these results, K9BYU was considered to be a useful tool for research of canine N-CAM.     

Circadian variations in salivary chromogranin a concentrations during a 24-hour period in dogs

The purpose of this study was to determine if salivary chromogranin a secretion in dogs exhibits a circadian rhythm. Saliva sampling was performed during three different sessions occurring in three nonconsecutive 24-h periods. Sixteen healthy adult beagle dogs (8 males and 8 females) were moved to a sampling room and housed individually in cages. Saliva samples were obtained every 4 h from 12:00 p.m. to 12:00 p.m. the following day. In the interest of habituation, saliva was obtained hourly from each dog 3 h before the experiment was started. Salivary chromogranin A concentrations were measured using an enzyme-linked immunosorbent assay. No circadian rhythm was detected for salivary chromogranin A secretion, and no differences in salivary chromogranin A concentrations measured every 4 h were demonstrated during the 24-h cycle in dogs.     

Isolation and structural characterization of rhamnogalacturonan II-borate complex fromPinus densiflora

A boron-containing pectic polysaccharide was isolated from Driselase digests of akamatsu (Pinus densiflora) cell wall. The boron-containing polysaccharide was purified by successive column chromatography of Cosmosil C₁₈ OPN, DEAE Sepharose FF, and Superdex 75. The complex had 0.12% (w/w) boron (B) and¹¹B-NMR spectroscopy showed the complex to be the tetrahedral diester form of boron. The sugar composition and glycosyl linkage analyses showed that the sugar portion of the complex was rhamnogalacturonan-II (RG-11). The boron-attached glycosyl residue was identified to be 2-O-Me-xylose-containing apiosyl side chain in RG-II. These results established that the location of borate in the cell wall is conserved among dicots, gramineous monocots, and coniferous gymnosperm.Part of this study was presented at the 8th Cell Wall Symposium, Norwich, September 3, 1998     

Development of a docosahexaenoic acid (DHA)-rich yellowtail <Emphasis Type="Italic">Seriola quinqueradiata</Emphasis> using tuna by-product oil

Because a diet rich in docosahexaenoic acid (DHA) is beneficial to human health, it is desirable for yellowtail Seriola quinqueradiata intended for human consumption to contain a high amount of DHA. We investigated the effects on the growth and fatty acid composition of yellowtail of replacing some of the pollack liver oil (PO) in the yellowtail diet with a DHA-rich tuna oil (TO). Experimental diets were prepared in which PO was replaced with 0, 15, 30, and 60% TO. Replacement of PO with TO did not negatively affect growth in a 16-week trial. In contrast, the feed efficiency of the group that received 60% TO was significantly higher than that of the control group after 6 weeks. The DHA level in the whole fish body increased with increasing dietary TO level. The groups that received 30 and 60% TO replacement diets showed significantly higher DHA levels than those of the control group. DHA level in the fillet was significantly higher in the group that received 60% TO than in the control group. DHA retention tended to increase with increasing TO level. These results show that utilizing TO as a fat source can increase DHA accumulation without incurring any adverse effects on the growth of yellowtail.     

Effects of natural feeding stimulants and glutamic acid supplementation on the feed intake,growth performance and digestive enzyme activities of red sea bream (Pagrus major) fed fish meal‐free soy protein concentrate (SPC)‐based diet

Two experiments were conducted for red sea bream (Pagrus major). In experiment 1, the optimum level of glutamic acid and natural feeding stimulants to enhance feed intake were determined and found that glutamic acid level of 0.5% and fish meat hydrolysate (FMH) were effective. In experiment 2, fish were fed with soy protein concentrate (SPC)‐based diet with synthetic feeding stimulants (Basal diet), the Basal diet with FMH (FMH diet), the FMH diet with glutamic acid (FMHG diet) and with fish meal diet (FM diet) as a control until satiation for 8 weeks. Feed intake of FMHG‐fed fish was significantly higher than others (p < 0.05). Specific growth rate and the feed conversion ratio of FMHG were comparable to those of FM‐fed fish (p > 0.05). Relative visceral fat ratio and crude lipid content of any SPC‐based diet‐fed fish tended to be lower than those of FM diet‐fed fish. There were no significant differences in trypsin and lipase activities hepatopancreas among treatments. SPC can be utilized as a sole protein source in a diet for red sea bream. The lower growth performance in SPC‐based diet‐ fed fish was not due to poor digestive enzyme secretion but could be associated with lipid utilization disorder.     

Low-fidelity homing behaviour of Biwa salmon <Emphasis Type="Italic">Oncorhynchus</Emphasis> sp. landlocked in Lake Biwa as inferred from otolith elemental and Sr isotopic compositions

Biwa salmon Oncorhynchus sp. is endemic to Lake Biwa, Japan, where it is an important commercial and recreational fisheries species. However, no information is currently available on its population structure and migration ecology. Therefore, here we evaluated whether otolith Sr/Ca, Ba/Ca and ⁸⁷Sr/⁸⁶Sr ratios can be used as natural signatures in Biwa salmon and then used these to determine the natal origins of lake-migration-phase individuals and spawning adults, and the homing ability of spawning adults in the Lake Biwa water system. Quadratic discriminant function analysis demonstrated that the lake-migration school comprised individuals with multiple origins, including rivers to the east, west and north of Lake Biwa, and that the homing rate of spawning adults was low (18 out of 80 individuals), with ca. 78% of fish straying into non-natal rivers. However, this straying behaviour was not spatially random, with fish tending to migrate upstream in rivers neighbouring their natal rivers. The high rate of straying in spawning adults is considered important for establishing and maintaining this species, which is highly adapted to life in the Lake Biwa water system where environmental disturbances often occur.     

Two β-amylase genes,OsBAM2 and OsBAM3, are involved in starch remobilization in rice leaf sheaths

To identify mechanisms of starch degradation in rice leaf sheaths at the post-heading stage, we investigated the function of OsBAM2 and OsBAM3, which encode plastid-targeted active β-amylase isoforms, in starch remobilization in leaf sheaths. The starch content in the second leaf sheaths below the flag leaf (the third leaf sheaths) peaked at the flag leaf emergence stage and gradually decreased until 15 days after heading. The mRNA levels of OsBAM2 and OsBAM3 in the third leaf sheaths increased from the flag leaf emergence stage to the heading stage when the starch content began to decrease. However, these mRNA levels did not always remain high during post-heading. Overexpression of OsBAM2 or OsBAM3 markedly repressed starch accumulation in the third leaf sheaths, showing that OsBAM2 and OsBAM3 function in starch degradation in rice leaf sheaths. In contrast, no significant differences in starch content in the third leaf sheaths were detected between knockdown plants of OsBAM2 or OsBAM3 and non-transgenic wild-type plants. Our results suggest that reduced expression of the individual genes, OsBAM2 or OsBAM3, does not result in excess accumulation of starch in the leaf sheaths, probably because of the complementary function of another gene or the action of other genes encoding starch-degrading .     

Spatial and interspecific variability in phenological responses to warming temperatures

A comprehensive understanding of species phenological responses to global warming will require observations that are both long-term and spatially extensive. Here we present an analysis of the spring phenological response to climate variation of twelve taxa: six plants, three birds, a frog, and two insects. Phenology was monitored using standardized protocols at 176 meteorological stations in Japan and South Korea from 1953 to 2005, and in some cases even longer. We developed a hierarchical Bayesian model to examine the complex interactions of temperature, site effects, and latitude on phenology. Results show species-specific variation in the magnitude and even in the direction of their responses to increasing temperature, which also differ from site-to-site. At most sites the differences in phenology among species are forecast to become greater with warmer temperatures. Our results challenge the assertion that trends in one geographic region can be extrapolated to others, and emphasize the idiosyncratic nature of the species response to global warming. Field studies are needed to determine how these patterns of variation in species response to climate change affect species interactions and the ability to persist in a changing climate.