Oryzacystatin-II, a cystatin from rice (Oryza sativa L. japonica), is a dimeric protein: possible involvement of the interconversion between dimer and monomer in the regulation of the reactivity of oryzacystatin-II

We examined the biochemical and structural properties of oryzacystatin-II, a phytocystatin in rice (Oryza sativa L. japonica), under heat-stress conditions. The enzyme inhibitory reactivity of oryzacystatin-II was enhanced by heating in a temperature-dependent manner and reached a maximum level by heating at 65 degrees C for 10 min. Size-exclusion chromatography showed that oryzacystatin-II forms a homodimer at ambient temperature and that the enhancement of inhibitory reactivity is due to the conversion of the dimeric to a monomeric form. The monomeric form of oryzacystatin-II reverted to the dimer during storage at 4 degrees C, suggesting that dimerization is an intrinsic property of oryzacystatin-II. The affinity of the monomer for cysteine proteinases was significantly higher than that of the dimer. This is the first paper to describe the noncovalent dimerization for a cystatin under nonstress conditions.     

Annual net production of the five Sargassaceae species in Yoro,western Wakasa Bay,Sea of Japan

ABSTRACT:   Annual net production of the five Sargassaceae species was investigated in Yoro, western Wakasa Bay, Sea of Japan. The Sargassaceae species were collected at a depth of 2.0–2.5 m and subjected to stratified clipping for 13 months. Annual net production was estimated at 2407 g dry wt/m² for Sargassum patens , 2132 g dry wt/m² for Sargassum macrocarpum , 1458 g dry wt/m² for Sargassum siliquastrum , 1197 g dry wt/m² for Myagropsis myagroides , and 1471 g dry wt/m² for Sargassum piluliferum . The ratio of annual net production to maximum biomass (P/B_max) for these species ranged between 1.3 and 1.7, being higher than in previous studies which used the same method. Winter storms with strong wave exposure and relatively calm condition in Yoro were considered to augment P/B_max. The total losses estimated from the different units of stratified clipping were compared. If larger and less segregated units were applied, the total loss would decrease. It was suggested that a 50 cm stratum would be applicable to the larger species (3 m or longer) and a stratum of 20 cm or less would be applicable to mid-sized or smaller species (less than 3 m).     

The effectiveness of a new physical property evaluation method of kamaboko which applied the principal component analysis to the puncture and stress relaxation tests

ABSTRACT:   The physical properties of 49 commercial kamabokos were measured by the puncture and stress relaxation tests. The principal component analysis was applied to the physical parameters of both tests, and their cumulative contribution ratios were over 90% with the first and second principal components, respectively. The comparison among the kamabokos was carried out using the synthetic physical parameters. The kamabokos produced in same area showed the peculiar distribution. The relative positional relation of kamabokos measured by the stress relaxation test was different from that by the puncture test for many of the kamabokos. Physical property evaluation using a principal component analysis is very effective for intensiveness of many measurement parameters. If much more kamabokos were measured by this method, regional characteristics of kamabokos would be clarified.     

Changes in estrogen receptor expression in the chick thymus during late embryonic development

In chickens, although estrogen receptors (ER) are reported to be associated with the immunological processes, detailed information about the differences in ER expression in the tissues related to the development of lymphocytes is not fully known, especially during the developmental stage. To learn more about this immunological relationship, we used semi‐quantitative polymerase chain reaction method to detect the ER expression levels in the thymus tissues of chicks during the developmental stage. Furthermore, ER‐expressing cells were detected by immunohistochemistry. The results of this study show that the expression level of ER increased on embryonic day 16 and decreased on day 20. Furthermore, ER expression was significantly higher in male than in female chickens at day 16. The increased expression on day 16 and decreased level on day 20 were also reproduced in the incidence of immunoreactive cells, although there was a 1‐day delay in the elevated incidence of the cells. This study revealed the changes in ER expression and the incidence of ER‐positive cells in the thymus of chickens during the developmental stage.     

Effect of dietary addition of seaweed and licorice on the immune performance of pigs

In pig production, dietary additive antibiotics are usually used for growth stimulation and disease prevention, although there is public concern about the increased incidence of resistant antibiotics and food safety. It is possible that such antibiotics might be replaced by naturally derived products such as seaweed and licorice. In this study, we evaluated the effect of dietary addition of seaweed and licorice on enhancing the immune function in swine. The animals of each group (eight animals per group) were sensitized at day 42 and 49, and the immunoglobulin production and the expression of cytokines were detected by the ELISA and real‐time PCR. As the results, saliva IgA production of the seaweed‐treated group increased around five times compared to that of control (day 56). Delayed hypersensitivity reaction and IgG production of the seaweed‐treated group increased around 1.8–2.0 times. In addition, enhanced saliva IgA production was detected at day 50 (around two times) and day 51 (around five times) by the licorice treatment, and lower expression level of tumor necrosis factor‐α messenger RNA at day 51 (around 1/25) was observed in the licorice treatment. We conclude that the replacement of antibiotics by naturally derived dietary additives might be feasible for immune system enhancement.     

Dietary intake of iodine‐enriched eggs decreases the incidence of mouse mammary tumors caused by the activated ErbB2 oncogene

Human epigenetic studies suggest that consumption of seaweed prevents mammary cancer, which possibly is explained by iodine daily intake. In this study, we evaluated the efficacy of dietary intake of iodine‐enriched eggs on mammary tumor incidence caused by the expression of activated type ErbB2. Female transgenic mice were divided into three groups, and fed a basic diet, a diet supplemented with ordinary eggs, or with iodine‐enriched eggs. The number of mammary tumors greater than 5 mm in diameter was recorded in mice at 6 months of age. We report that the average number of mammary tumors per mouse was significantly lower in the iodine‐enriched egg‐added diet group than in either the basic diet or ordinary egg diet groups. These results indicate that iodine intake through livestock‐derived products can reduce the incidence of mammary cancers caused by the expression of activated type ErbB2.     

Effect of plastic-film mulching on leaching of nitrate nitrogen in an upland field converted from paddy

A lysimeter experiment was conducted to examine the effects of plastic film mulching on the leaching rate of nitrate nitrogen (NO₃-N) from chemical fertilizer that was applied to an upland field that had been converted from paddy rice production. Leaching was monitored in two lysimeters filled with sandy loam soil, which contained low soil organic matter content, under different surface mulch conditions. One was mulched only on the ridge (ridge-mulch treatment) and another one was mulched fully, including the furrow, with black plastic film (full-mulch treatment). Chemical fertilizer was mixed into the top 0.2 m of soil in the two lysimeters before installing the mulch. After transplanting broccoli, the amount of subsurface discharge water and the NO₃-N concentrations in the discharge water were measured every day. Larger NO₃-N discharges occurred in the ridge-mulch treatment for three days after heavy rainfalls in which cumulative precipitation exceeded 10 mm, and the daily NO₃-N load was twice as large as the full-mulch treatment. The differences in the amount of subsurface discharge water and NO₃-N discharged between treatments were not significant when there was no rainfall. Cumulative NO₃-N loads for the ridge- and full-mulch treatment during the last month of the experimental period were 0.246 and 0.195 g m^–2, respectively. The effect of mulching on the reduction of NO₃-N discharge rate was higher for the full-mulch treatment. This result showed that a plastic-film mulching system would be effective as an appropriate fertilizer management to reduce nitrate-leaching losses.     

The relationship between water intake rates,paddy ponding depth,and farmers’ water management techniques

Japanese farmers manage their irrigation water based on their past experiences and preferences, considering such factors as weather and available water (hereafter defined as empirical water management). They elaborately control the intake and drainage rates of their own paddy fields to maintain optimal ponding depths. But these well-managed systems will drastically change because of the decreasing number of farmers. Therefore, it is necessary to clarify if the optimal ponding depth will be maintained within the limits of traditionally-allowed water intake rate from the main river. The first objective of this study was the quantification of actual water use in the paddy fields, resulting from the farmers water management on the basis of their experience. The significance of the present water intake rate under empirical water management was studied for a paddy field command area of about 230 ha. Water intake rates and the water requirements of the whole area were investigated by measuring the flow rate at 17 points of irrigation and drainage canals. Characteristics of the farmers empirical water management were investigated by measuring the hourly changes in inflow and outflow rates for a sub-area using an automatic measurement system, and an inferential method of determining water management patterns for the paddy fields was proposed. The newly-proposed inferential method was introduced in the tank model, which expresses the characteristics of water management in the command area. The Shuffled Complex Evolution Algorithm (SCE-UA) method was used for optimizing the model parameters. It was proven that the model accuracy improved when the farmers empirical water management was taken into account. The optimal amount of water to be applied to the command area was quantified by the simulation. The second objective was to predict the effect of the decreasing number of farmers on future water use conditions. The simulated result indicates the difficulty of maintaining optimal ponding depth for the whole command area when the farmers empirical water management is not maintained. In other words, results indicated that efficient water use requires an automatic water management system or a new pipeline system to replace the farmers present empirical water management.     

Genetic and antigenic characterization of bovine viral diarrhea viruses isolated from cattle in Hokkaido,Japan

In our previous study, we genetically analyzed bovine viral diarrhea viruses (BVDVs) isolated from 2000 to 2006 in Japan and reported that subgenotype 1b viruses were predominant. In the present study, 766 BVDVs isolated from 2006 to 2014 in Hokkaido, Japan, were genetically analyzed to understand recent epidemics. Phylogenetic analysis based on nucleotide sequences of the 5′-untranslated region of viral genome revealed that 766 isolates were classified as genotype 1 (BVDV-1; 544 isolates) and genotype 2 (BVDV-2; 222). BVDV-1 isolates were further divided into BVDV-1a (93), 1b (371) and 1c (80) subgenotypes, and all BVDV-2 isolates were grouped into BVDV-2a subgenotype (222). Further comparative analysis was performed with BVDV-1a, 1b and 2a viruses isolated from 2001 to 2014. Phylogenetic analysis based on nucleotide sequences of the viral glycoprotein E2 gene, a major target of neutralizing antibodies, revealed that BVDV-1a, 1b and 2a isolates were further classified into several clusters. Cross-neutralization tests showed that BVDV-1b isolates were antigenically different from BVDV-1a isolates, and almost BVDV-1a, 1b and 2a isolates were antigenically similar among each subgenotype and each E2 cluster. Taken together, BVDV-1b viruses are still predominant, and BVDV-2a viruses have increased recently in Hokkaido, Japan. Field isolates of BVDV-1a, 1b and 2a show genetic diversity on the E2 gene with antigenic conservation among each subgenotype during the last 14 years.