Spontaneous Accumulation of Globotriaosylceramide (Gb3) in Proximal Renal Tubules in an ICR Mouse

This report describes spontaneous cytoplasmic vacuolation in the proximal renal tubules of a 7-week-old male ICR [Crlj:CD1(ICR)] mouse. The contents of vacuoles were positively stained with periodic acid-Schiff (PAS) and Sudan black, and the membranes were positive on immunohistochemical staining for lysosomal-associated membrane protein-2 (LAMP-2), a marker of lysosomal membrane. Electron microscopy revealed electron-dense lamellar bodies in the proximal tubular epithelial cells. These histopathological features are similar to those in α-galactosidase A-deficient mice, in which globotriaosylceramide (Gb3), a glycosphingolipid, accumulates in lysosomes. When we performed immunohistochemical staining for Gb3, the contents of vacuoles were positively stained. From these results, spontaneous cytoplasmic vacuolation in the proximal renal tubules in the mouse was identified as lysosomal accumulation of Gb3.     

Effect of thinning on cultivar differences of green stem disorder in soybean

ABSTRACT

Green stem disorder (GSD) in soybean (Glycine max (L.) Merrill) negatively affects harvest efficiency and seed appearances. Breeding GSD-insensitive cultivars is expected to be an effective countermeasure to GSD. However, it is difficult to stably detect cultivar differences in GSD under conventional field conditions because the occurrences of GSD largely vary by location and year. The thinning effect, which had been reported to promote GSD, may help accurate phenotyping for occurrences of GSD in breeding. To verify this possibility, the thinning treatment was applied to four cultivars, the GSD severity values of which were evaluated in an independent study by another group. As a result, the cultivar differences in GSD severity were generally comparable between the present and previous studies. However, the difference was more evident, with the thinning treatment exhibiting the GSD score of 2.8 of ‘Hatsusayaka’ compared with the GSD score of 3.6 of ‘Sachiyutaka’, while the scores of those cultivars were similar without the thinning treatment. A positive correlation between GSD severity and N concentration in the main stem could be seen but the increasing rate of GSD score with the N concentration in the main stem differed between cultivars. Thus, although more cultivars need to be tested to prove, the thinning treatment could be useful as a phenotyping technique in the breeding of GSD-insensitive cultivars.     

Bean husks as a supplemental fiber for ruminants: Potential use for activation of fibrolytic rumen bacteria to improve main forage digestion

This study evaluated the suitability of easily digested fiber sources as a supplemental fiber to improve overall fiber digestion in ruminants. First, the degradation of five fibrous feedstuffs and the stimulatory effects on rumen bacteria were examined in situ. Chickpea and lablab bean husks were selected for their potential use due to their large degradable fraction (> 94%), which had a stimulatory effect on fibrolytic rumen bacteria such as Fibrobacter succinogenes. Second, a possible improvement in the digestibility of rice straw diet by husk supplementation was monitored in vivo. Four dietary treatments comprising RS (rice straw and concentrate), CHM (RS supplemented with Myanmar chickpea husk), CHE (RS with Egyptian chickpea husk) and LH (RS with lablab bean husk) were allocated to four wethers. The digestibility of acid detergent fiber was 3.1–5.5% greater in CHM and LH than RS. Total volatile fatty acid concentration was higher in LH than other treatments. Acetate proportion was higher in LH than RS. Ruminal abundance of F. succinogenes was 1.3–1.5 times greater in CHM and LH than RS. These results suggest that bean husk supplementation, especially lablab bean husk, might improve the nutritive value of rice straw diet by stimulating fibrolytic bacteria.     

Growth pattern and seasonal weight changes of the feral raccoon (Procyon lotor) in Hokkaido,Japan

The growth pattern and seasonal weight fluctuations of feral raccoons in Hokkaido were evaluated between 1999 and 2001. The growth rates inbody length and body weight were described for juveniles (young of the year) and yearlings (animals born in the previous season) using the Gompertz growth model. The asymptotic body sizes for males were greater than those for females. Young raccoons born during spring in the study area could potentially grow up to their asymptotic size at the beginning of their first winter, but they would not reach their full adult sizes until at least their second fall. Adult raccoons (animals > or = 2 -year-old) had seasonal weight fluctuations with annual weight loss of 25% to 28% of mean maximum weights in west-central Hokkaido, but these result would be an underestimate of the degree of annual weight fluctuations. Juvenile raccoons can be distinguished from the older animals by measuring body length or body weight during fall in Hokkaido.     

Partial characterization of structure and function of a xylanase gene from the rumen hemicellulolytic bacterium Eubacterium ruminantium

A gene encoding for xylanase activity in the rumen hemicellulolytic bacterium Eubacterium ruminantium was cloned into pBR322 in Escherichia coli (E. coli ). The primary clone had a 5.7 kb insert produced by Eco RI partial digestion. Subcloning followed by sequencing allowed for the discovery that this enzyme has a glycosyl‐hydrolase family 10 catalytic domain with a family 9 carbohydrate binding module at C‐terminus and a region partially homologous to a family 22 carbohydrate binding module at N‐terminus. Cloned xylanase is specifically active against xylan and oligoxyloside to produce xylobiose and xylotriose, showing optimal pH and temperature at 7.0 and 50°C, respectively. Molecular size of the xylanase (91 kDa) was confirmed by zymogram analysis of the E. coli clone, which agreed with the predicted size from the DNA sequence. Functions of the two modules at C‐ and N‐termini were evaluated by using xylanase variants with and without the respective module and the C‐terminal module was found to be functional in binding to acid‐swollen cellulose and insoluble oat‐spelt xylan, whereas the N‐terminal module was inactive for binding them.     

Ruminal distribution of the cellulolytic bacterium Fibrobacter succinogenes in relation to its phylogenetic grouping

To investigate the ecological importance of the cellulolytic bacterium Fibrobacter succinogenes in fiber digestion, ruminal distribution of F. succinogenes was determined in relation to its phylogenetic grouping. Rumen digesta from wethers and steers fed orchardgrass hay, rice straw or fresh orchardgrass were employed as the materials for the analyses. Orchardgrass hay stem incubated in the rumen was also used. By using total DNA extracted from these materials, population sizes of total F. succinogenes and of four different phylogenetic groups of this species were quantitated through competitive polymerase chain reaction (PCR), and restriction fragment length polymorphism analysis of PCR products targeted the bacterial 16S rDNA. Rumen digesta and ruminally incubated hay stems had a reasonably high population size of F. succinogenes (×10^7?8/g) that was composed of strains belonging to the phylogenetic groups 1 and 3. The relative abundance of each group was different among the samples; group 1 dominated on the ruminally incubated hay stem and in the rumen of wethers fed fresh orchardgrass, while group 3 was major in the rumen of wethers and steers on hay diet. These results suggest that there could be phenotypic differences among the phylogenetic groups of F. succinogenes, and group 1 dominating on hay stem might contribute to rumen fiber digestion more than the other groups.     

Roles of central histaminergic system in glucose metabolisms and food intake in sheep

The neurotransmitter histamine is thought to be involved in the homeostatic regulation of energy balance. The present study was conducted to examine whether the central histaminergic system is involved in the regulation glucose metabolisms and food intake in ruminants. Thioperamide, a presynaptic H3‐receptor antagonist, was intracerebroventricularly infused at a rate of 68 μg/min for 3 h, beginning at 11.00 hours, into castrated Suffolk rams (48–55 kg bodyweight) accustomed to being fed alfalfa hay cubes (2% of body weight) at 12.00 hours. The food intake was measured every 60 min for 2 h after the commencement of feeding (12.00 hours). Blood samples were taken through a jugular vein catheter every 10 or 20 min during the experiment. Plasma glucose levels were significantly (P < 0.05) greater in the thioperamide‐treated group than in the control. The post‐prandial insulin increment was significantly (P < 0.05) greater in the thioperamide‐treated group than in the saline‐treated control. The food intake of the thioperamide‐treated group was significantly (P < 0.05) less than that of the control in the period from 60 to 120 min after feeding. In conclusion, the central histaminergic system is involved in the regulation of glucose metabolisms and food intake in ruminants.     

Dynamic changes in subcellular localization of cattle XLF during cell cycle, and focus formation of cattle XLF at DNA damage sites immediately after irradiation

Clinically, many chemotherapeutics and ionizing radiation (IR) have been applied for the treatment of various types of human and animal malignancies. These treatments kill tumor cells by causing DNA double-strand breaks (DSBs). Core factors of classical nonhomologous DNA-end joining (C-NHEJ) play a vital role in DSB repair. Thus, it is indispensable to clarify the mechanisms of C-NHEJ in order to develop next-generation chemotherapeutics for cancer. The XRCC4-like factor (XLF; also called Cernunnos or NHEJ1) is the lastly identified core NHEJ factor. The localization of core NHEJ factors might play a critical role in regulating NHEJ activity. The localization and function of XLF have not been elucidated in animal species other than mice and humans. Domestic cattle (Bos taurus) are the most common and vital domestic animals in many countries. Here, we show that the localization of cattle XLF changes dynamically during the cell cycle. Furthermore, EYFP-cattle XLF accumulates quickly at microirradiated sites and colocalizes with the DSB marker γH2AX. Moreover, nuclear localization and accumulation of cattle XLF at DSB sites are dependent on 12 amino acids (288–299) of the C-terminal region of XLF (XLF CTR). Furthermore, basic amino acids on the XLF CTR are highly conserved among domestic animals including cattle, goat and horses, suggesting that the CTR is essential for the function of XLF in domestic animals. These findings might be useful to develop the molecular-targeting therapeutic drug taking XLF as a target molecule for human and domestic animals.     

An annual faecal 16S amplicon sequencing of individual sea cucumber (Apostichopus japonicus) demonstrates the feeding behaviours against eukaryotes in natural environments

To fill in the gaps in knowledge as to how individual sea cucumber Apostichopus japonicus behave consuming eukaryotic food sources in natural environments, eukaryotic communities in the faeces of sea cucumbers and sediments were analysed through one whole year based on 16S rRNA gene sequencing of the organelle genomes. A total of 390 eukaryotic features were obtained, and 99.7% of the features were assigned to chloroplasts. The eukaryotic communities in faeces and sediments showed seasonal fluctuations through one whole year based on Bray‐Curtis distance and community composition. Comparison of eukaryotic communities between faeces and sediments showed that 12 families including Chaetocerotaceae and Laminariaceae were more abundant in faeces than in sediments, suggesting that sea cucumbers may choose sediment containing these algal taxa more often compared with others in natural environments. All features of Laminariaceae were assigned to Saccharina japonica, which is consistent with the fact that this alga is one of the most suitable diets in the aquaculture of A. japonicus. Assessments of individual 16S amplicon sequences of both faecal and sediment samples could be an alternative tool to help us understand dynamic feeding behaviours of sea cucumber populations in contributing to bioresource conservation and development of a superior approach to aquaculture.