Development and characterization of monoclonal antibodies against canine trypsin

Canine cationic trypsin was purified by salting-out, gel filtration and affinity chromatography. Purity was confirmed by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The molecular weight was ca. 28 kDa by SDS-PAGE.

Thirty hybridomas were obtained which produced mAb to canine cationic trypsin by the cell fusion technique. Twenty-two of these recognized cationic trypsin only, while eight hybridomas recognized both cationic and anionic trypsin. Several of the anti-canine cationic trypsin mAb were purified by salting-out and DEAE ion-change chromatography using ascites fluid of immunized BALB/c mice. The mAb proved to have very high specificity to canine cationic trypsin as shown by immunoblotting and it may be possible to use them to develop clinical assays.     

Evaluation of Cold Tolerance in NERICAs Compared with Japanese Standard Rice Varieties at the Reproductive Stage

New Rice for Africa (NERICA) is a general name for interspecific rice varieties derived from a cross between the high‐yielding Asian rice (Oryza sativa L.) between locally adapted African rice (Oryza glaberrima Steud.). Eight NERICAs were evaluated for cold tolerance (CT) at the reproductive stage and compared with their O. sativa parents and three Japanese standard rice varieties over 3 years. Cold tolerance was evaluated based on the filled grain ratio (FGR) after cold water irrigation. The FGR was greatly reduced by cold water irrigation. NERICA 1, 2 and 7 had higher FGR (51.9–57.9 %), while NERICA 6, 15 and 16 had lower FGR (6.2–14.5 %). NERICA 1, 2 and 7 were less affected by cold stress, with a 31 % mean reduction in FGR, while NERICA 6, 15 and 16 were greatly affected, with their FGRs being reduced by more than 80 %. NERICA 3 and 4 were moderately affected by cold stress, with about 45 % reduction rate in FGR. FGR significantly influenced the grain weights of the varieties with strong positive correlations (r = 0.83–0.91; P < 0.001), and thus, similar trends in grain weights were observed. Grain weights were reduced by 61.7–96.4 % under cold stress. NERICA 1, 2 and 7 showed significantly better performance than NERICA 3 and 4, while NERICA 6, 15 and 16 performed poorly under cold water irrigation. The Japanese varieties Koshihikari (very tolerant) and Ozora (moderately tolerant) were more affected by cold water irrigation than NERICA 1, 2 and 7. On the basis of the mean reduction rate (%) in FGR under cold stress, the varieties were classified as follows: NERICA 1, 2 and 7 as tolerant; NERICA 3 and 4 as moderately tolerant; and NERICA 6, 15 and 16 as susceptible to cold stress. However, NERICA 7 grain yields were lower under cold stress due to both greatly reduced number of panicles per plant and number of spikelets per panicle. Therefore, NERICA 1 and 2 are suitable candidates for production in the highland regions of East Africa and should be promoted for production.     

Utilization of digital differential display to identify differentially expressed genes related to rumen development

This study aimed to identify the genes associated with the development of the rumen epithelium by screening for candidate genes by digital differential display (DDD) in silico. Using DDD in NCBI's UniGene database, expressed sequence tag (EST)‐based gene expression profiles were analyzed in rumen, reticulum, omasum, abomasum and other tissues in cattle. One hundred and ten candidate genes with high expression in the rumen were derived from a library of all tissues. The expression levels of 11 genes in all candidate genes were analyzed in the rumen, reticulum, omasum and abomasum of nine Japanese Black male calves (5‐week‐old pre‐weaning: n = 3; 15‐week‐old weaned calves: n = 6). Among the 11 genes, only 3‐hydroxy‐3‐methylglutaryl‐CoA synthase 2 (HMGCS2), aldo‐keto reductase family 1, member C1‐like (AKR1C1), and fatty acid binding protein 3 (FABP3) showed significant changes in the levels of gene expression in the rumen between the pre‐ and post‐weaning of calves. These results indicate that DDD analysis in silico can be useful for screening candidate genes related to rumen development, and that the changes in expression levels of three genes in the rumen may have been caused by weaning, aging or both. © 2015 Japanese Society of Animal Science     

Changes in intestinal villi, cell area and intracellular autophagic vacuoles related to intestinal function in chickens

1. Morphological changes in the intestinal villi, cell area and cell mitosis number in the duodenal epithelial cells were compared in cockerels fasted for 1, 2 and 3 d, and also when refed for 1 and 2 d after 3 d of fasting, to demonstrate whether these morphological changes are related to intestinal function. Alterations in the fine structure of vacuoles in epithelial cells were also examined in each group to investigate whether the vacuolar changes are associated with these morphological changes, and to obtain an index for judging the nutritional condition of the chicken intestine. 2. Fasting induced decreases in villus height, cell area and cell mitosis number, which recovered rapidly after refeeding, suggesting that these parameters are related to changes in intestinal function and may be useful for assessing intestinal function. 3. At 1 d of fasting, small electron-dense bodies appeared in the absorptive epithelial cells, some of them fusing with each other. As the fasting period increased, these small bodies developed to moderate-sized nascent autophagic vacuoles containing various kinds of electron-dense contents and finally became large autophagic vacuoles with electron-lucent contents. Some vacuoles showed positive acid phosphatase reactions, which indicated that they were lysosomal autophagic vacuoles containing hydrolytic enzymes. 4. After 1 d of refeeding the large autophagic vacuoles seen after 3 d fasting rapidly decreased to the small electron-dense bodies seen after 1 d of fasting. 5. These findings suggest that intestinal epithelial cells have the ability to digest their own cell components to supply nutrients during fasting by means of lysosomal active autophagic transport mechanisms: after refeeding, the epithelial cells return to the absorption of nutrients. 6.The present results demonstrate that the autophagic vacuolar changes are correlated with changes in intestinal villus height, cell area and cell mitosis number induced by fasting and refeeding; this indicates that autophagic vacuoles are a useful index of the nutritional condition of chicken intestine. The greater the number of electron-lucent vacuoles there are in the duodenal absorptive cells, the lower the nutritional condition of the chicken intestine.     

Developmental changes in pituitary–thyroid axis, and formation of gonads in leptocephali and glass eels of Anguilla spp.

SUMMARY: Pituitary, thyroid gland and gonads in leptocephali of Japanese eel Anguilla japonica (19.8–32.6 mm in total length), A. obscura (45.0 mm), and A. bicolor pacifica (49.5 mm) and those in glass eels of the Japanese eel were histologically and immunohistochemically examined in order to observe the developmental changes of these endocrine organs in the Anguillidae. The pituitary, consisting of adenohypophysis and neurohypophysis in Japanese eel leptocephali over 22.5 mm, did not contain thyroid stimulating hormone (TSH) immunoreactive cells. Such cells were, however, detectable in the more developed pituitaries of leptocephali of A. obscura and A. bicolor pacifica and in those of glass eels. Conversely, thyroxine (T₄)-immunoreactive thyroid follicles could be detected in all specimens, both leptocephalic and glass eel. Only in glass eels, gonads were found in the body cavity, and these gonads harbored one or two primordial germ cells (PGC) per cross-section. Our results indicate that thyroid hormones (TH) production started prior to TSH production, and that TSH and TH are both secreted during the metamorphosis from leptocephalus to glass eel. Therefore, it is plausible that the TSH–TH axis is involved in the metamorphosis from leptocephalus to glass eel, but not in the early growth from preleptocephalus to leptocephalus.     

Distribution of boron in wood treated with aqueous and methanolic boric acid solutions

The distributions of boron in Japanese cedar (Cryptomeria japonica D. Don) sapwood blocks treated with aqueous or methanolic boric acid [B(OH)₃] solutions were explored through Raman spectroscopy and prompt gamma-ray analysis (PGA). B(OH)₃ was the sole boron species observed in Raman spectra of the wood blocks treated with either solution. Plots of weight gain of the treated wood blocks versus boron concentration in treatment solutions were found to be linear. The results indicated that the methanolic solution makes it possible to impregnate wood with much larger amounts of boron than the aqueous solution. PGA confirmed that B(OH)₃ was highly enriched near the end grains of the treated wood blocks. Raman measurements suggested that boron content in the bulk of the wood block is not as large as expected from the weight gain of the treated wood blocks when an ordinary air-drying method is used. It was concluded that the aqueous solution impregnates the cell walls of wood with boron more easily than the methanolic solution. Part of this report was presented at the 55th (Kyoto, March 2005) and 56th (Akita, August 2006) Annual Meetings of the Japan Wood Research Society. This article follows the previous rapid communication “Analysis of boron in wood treated with boric acid solutions using Doppler broadening method of prompt gamma-rays.” J Wood Sci (2006) 52:279–281     

Design and pilot production of a “spiral-winder” for the manufacture of cylindrical laminated veneer lumber

A new spiral-winder was developed for continuous manufacturing of cylindrical laminated veneer lumber (LVL), and a suitable resin adhesive for this cylindrical LVL manufacturing system was investigated. This phase was followed by trial manufacturing and evaluation of cylindrical LVL with the optimum resin adhesive identified. The results are summarized as follows. (1) The shortest gelation time was recorded with a mixture of two commercial resorcinol based resins (DF-1000 and D-33) at a weight ratio of 2575. (2) Bath temperature had a remarkable effect on the gelation time of the adhesive mix. (3) High bonding strength was recorded by 2575 DF-1000/D-33 adhesive mix at a high press temperature despite a short pressing duration. Based on the results of items (1) to (3), 2575 DF-1000/ D-33 is recommended for use in the new spiral-winder. (4) The mechanical properties of cylindrical LVL could be improved by using 2575 DF-1000/D-33 with wider veneer width and longer pressing time. (5) The mechanical properties, especially the modulus of rupture, of the cylindrical LVL manufactured require further improvement for practical structural application.     

Bending characteristics of bamboo (<Emphasis Type="Italic">Phyllostachys pubescens</Emphasis>) with respect to its fiber–foam composite structure

The bending properties of split bamboo culm were compared with those of spruce and beech wood specimens. The bamboo allowed large flexural deformation since its outer layer retains the tensile stress while the softer inner layer undergoes large compressive deformation. The results suggested that the combination of the fiber-rich outer part and the compressible inner part was responsible for the flexural ductility of split bamboo. To clarify the compressible nature of the inner part of bamboo, the longitudinal surfaces of the bamboo and wood specimens were microscopically observed before and after a large longitudinal compression. Although the wood specimens showed serious and localized buckling, the inner part of the bamboo specimens showed no such visible buckling. In the latter case, the foam-like parenchyma cells absorbed the large compressive deformation by their microscopic buckling and simultaneously, the alignment of sclerenchyma fibers was maintained by the surrounding parenchyma matrix. The flexural elasticity of the bamboo was compared to that of the wood in respect of remaining strain during cyclic bending tests. No clear difference was recognized between their remaining strains. This fact indicated that the bamboo was not so flexible elastically, although its fiber–foam combination and intelligent fiber distribution improve flexural ductility.