Characterization of 38 kDa and 42 kDa chitinase isozymes from the liver of Japanese common squid Todarodes pacificus

ABSTRACT: Characterization was investigated on the 38 kDa and 42 kDa chitinase (EC3.2.1.14) isozymes from the liver of Japanese common squid Todarodes pacificus . Optimum pH toward colloidal chitin was observed at pH 3.0 for the 38 kDa chitinase, and pH 3.0 and 9.0 for the 42 kDa chitinase. K _m and k _cat of the 38 kDa and 42 kDa chitinases toward a longer substrate, glycol chitin, were 0.071 mg/mL and 1.22/s, and 0.074 mg/mL and 0.196/s, respectively. Alternatively, strong substrate inhibition of both chitinases were observed toward a short substrate, N -acetylchitopentaose (GlcNAc₅). Both chitinases decomposed not only chitin but also chitosan (D. A. 95%). The cleavage pattern and reaction rate were investigated using N -acetylchitooligosaccharides (GlcNAc_n, n  = 2–6). Both chitinases hydrolyzed GlcNAc_n ( n  = 4,5, and 6). The release of GlcNAc was not observed. The speed of the reaction was observed to be in the following order: GlcNAc₄ > GlcNAc₅ > GlcNAc₆ for the 38 kDa chitinase, and GlcNAc₆ > GlcNAc₅ > GlcNAc₄ for the 42 kDa chitinase. Both the chitinases released p -nitrophenol from p -nitrophenyl GlcNAc_n ( n  = 2, 3, and 4). N-terminal amino acid sequences of the 38 kDa and 42 kDa chitinases were YLLSXYFTNWSQYRPGAGKYFPQNI and EYRKVXYYTNWSQYREVPAKFFPEN, respectively.     

The Arterial Supply of the Female Reproductive System in the Hamster

The arterial supply to the pelvis and female reproductive system was studied in 54 hamsters of the CBN, APG and ACN strains, 3 rats of the Wistar strain and 3 mice of the ICR strain. All animals were prepared by intravascular injection of a mixture of barium and red latex for gross dissection and radiographic examination. The vascular patterns of the three strains of hamsters were compared with each other and with those of rats and mice. The possible reasons for variations within and between species are discussed.     

Regulation of secondary follicle growth by theca cells and insulin-like growth factor 1

Ovaries contain follicles at various stages of development, including primordial, primary, secondary, antral and Graafian follicles. Although the growth of these follicles is controlled to maintain regular ovulation, the mechanism through which this occurs remains unclear. In our study, we found that the growth rate of cultured secondary follicles separated from mice ovaries differed between follicles. After 4 days of culture, the size of some secondary follicles was markedly increased, while that of others had either slightly increased, remained unchanged or shrunk. We compared the expression levels of growth factors between these secondary follicles and found that the growth rate of cultured secondary follicles correlated with the expression level of insulin-like growth factor 1 (Igf1) mRNA. Igf1 mRNA expression level in secondary follicles containing theca cells was higher than that in secondary follicles without theca cells, and the granulosa cell proliferation around follicles containing theca cells was increased. Furthermore, an IGF1 inhibitor also inhibited the granulosa cell proliferation, and administration of IGF1 to secondary follicles without growth promoted granulosa cell proliferation. These results indicated that the theca cells of secondary follicles induced the expression of IGF1 and promoted the follicle growth.     

Blood biochemical values in Japanese Black breeding cows in Kagoshima Prefecture,Japan

To obtain blood biochemical basic data of Japanese Black breeding cows in Kagoshima Prefecture, Japan, blood samples were obtained from 857 clinically healthy multiparous cows on 71 farms. Cows were divided into three stages: the prepartum stage (between 20 and 80 days before parturition, n=290), lactation stage (between 20 and 80 days after parturition, n=283) and maintenance stage (between 120 and 200 days after parturition, n=284). The mean concentration of total protein and total cholesterol, and the mean activities of aspartate aminotransferase in the lactation stage tended to be higher than those in the prepartum and maintenance stages. The mean concentration of glucose in the prepartum stage tended to be lower than that in the lactation and the maintenance stages. The mean concentration of nonesterified fatty acids and beta-hydroxybutyrate in the prepartum stage tended to be higher than those in the lactation and maintenance stages. The mean concentration of serum retinol was approximately 30 µg/dl in all stages. These results suggest that the blood biochemical values of multiparous Japanese Black cows vary with breeding stages, and it is considered that the blood parameters obtained in this study are useful as indices for health management of Japanese Black breeding cows.     

Numerical simulation on effect of irrigation conditions on water temperature distribution in a paddy field

Water management methods regulate water temperature in paddy fields, which affects rice growth and the environment. To understand the effect of irrigation conditions on water temperature in a paddy field, water temperature distribution under 42 different irrigation models including the use of ICT water management, which enables remote and automatic irrigation, was simulated using a physical model of heat balance. The following results were obtained: (1) Irrigation water temperature had a more significant effect on paddy water temperature close to the inlet. As the distance from the inlet increased, the water temperature converged to an equilibrium, which was determined by meteorological conditions and changes in water depth. (2) Increasing the irrigation rate with higher irrigation water amount increased the extent and magnitude of the effects of the irrigation water temperature. (3) When total irrigation water amount was the same, increasing the irrigation rate decreased the time-averaged temperature gradient effect over time across the paddy field. (4) Irrigation during the lowest and highest paddy water temperatures effectively decreased and increased the equilibrium water temperature, respectively. The results indicate that irrigation management can be used to alter and control water temperature in paddy fields, and showed the potential of ICT water management in enhancing the effect of water management in paddy fields. Our results demonstrated that a numerical simulation using a physical model for water temperature distribution is useful for revealing effective water management techniques under various irrigation methods and meteorological conditions.

     

Physiological difference between free and triglyceride-type conjugated linoleic acid on the immune function of C57BL/6N mice

Previous studies have shown the physiological significance of dietary conjugated linoleic acid (CLA) in various experimental animals and in human beings. One of the important problems to better elucidate is the difference between triglyceride (TG) and free (FFA) dietary CLA. Here, using splenocytes, this study assesses how TG- and FFA-CLA modulate immunoglobulin and various cytokine productions. In this study, C57BL/6N mice were fed an experimental diet containing 0% CLA, 0.1 or 1% FFA-CLA, or 0.1 or 1% TG-CLA for 3 weeks. The production of immunoglobulin tended to be up-regulated by 1% FFA-CLA. As a result of protein array analysis using the supernatant from splenocytes cultured with no CLA, 1% FFA-CLA, and TG-CLA, some cytokine production was shown to be remarkably regulated by dietary FFA- and TG-CLA. A total of 32 cytokines were examined, and 11-14 produced cytokines that were 2-fold up-regulated as compared with control for FFA- or TG-CLA, respectively. Especially, the production of IL-9 and MCP-5 and other cytokines was remarkably up-regulated by both FFA- and TG-CLA. In addition, seven cytokines were 2-fold down-regulated by TG-CLA. These data show that there is a slight but significant difference between the functionalities of FFA- and TG-CLA.     

Post-mortem evaluation of expanded polytetrafluoroethylene (ePTFE) used in mitral valve repair in dogs

Mitral valve repair is one of the treatment options for mitral regurgitation. Expanded polytetrafluoroethylene (ePTFE) is a polymer that has been widely used in cardiovascular surgery. In this case series, we report the autopsy and histological findings in 6 dogs that underwent cardiopulmonary bypass for mitral annuloplasty using ePTFE sheets and chordoplasty using ePTFE sutures.From May 2005 to October 2009, 3 female and 3 male dogs with severe mitral regurgitation underwent mitral valve repair. This case series included 3 Cavalier King Charles spaniels, 2 Maltese, and 1 Shih Tzu. The survival period after surgery was 19–72 (35 ± 19) months.In all the cases, autopsy revealed that the ePTFE sheets and sutures were not damaged and well integrated into the surrounding highly differentiated, connective tissues. Low-power microscopy revealed that in all cases, the tissues surrounding the ePTFE sheet in the mitral valve annulus had almost completely been covered by granulation tissue. No inflammatory infiltrate or thrombogenesis was observed around the ePTFE in any of the cases.There was no evidence of reactive changes in the region surrounding the ePTFE. These results suggest that ePTFE has excellent tissue compatibility and durability and can be effectively used for canine mitral valve repair.     

New localities and awn morphology variation in Miscanthus ×ogiformis

Miscanthus ×ogiformis is a hybrid between Miscanthus sacchariflorus and Miscanthus sinensis and its several synonyms indicate the insufficient understanding of the morphological variation of its awn on the spikelet, which is a key characteristic for identifying Miscanthus species. In the present study, all the Miscanthus specimens in 19 herbaria were examined in order to find new localities for Miscanthus ×ogiformis. Thirty‐nine Miscanthus ×ogiformis specimens appeared to be naturally occurring hybrids, as their callus hair length, awn length, and callus‐to‐spikelet ratio differed from those found in the parental species. Therefore, new Miscanthus ×ogiformis individuals with morphological variations are likely to be discovered and these strains might be useful as new genetic resources for this biomass crop.