Leptospiral lipopolysaccharide stimulates the expression of toll‐like receptor 2 and cytokines in pig fibroblasts

Pigs throughout the world are afflicted with leptospirosis, causing serious economic losses and potential hazards to human health. Although it has been known that leptospiral lipopolysaccharide (L‐LPS) is involved in an immunological reaction between an antigen and a host cell, little is known about how the immune system of pigs can respond to L‐LPS. Here, we stimulated pig fibroblasts by L‐LPS and then quantitatively measured gene and protein expression levels of two toll‐like receptors (TLRs), TLR2 and TLR4, by real‐time PCR and Western blotting. As a result, expression of TLR2 was found to be significantly up‐regulated within 24 h after L‐LPS stimulation whereas induction of TLR4 expression was relatively weak. We also revealed that of myeloid differentiation primary response gene 88 (MyD88), interleukin 6 (IL‐6) and IL‐8 gene expressions were markedly up‐regulated by L‐LPS stimulation. These results may suggest that the pig cell can activate TLR2 rather than TLR4 by L‐LPS stimulation, thereby inducing expression of cytokines.     

Quantification of the effect of rice paddy area changes on recharging groundwater

This study quantifies the effects of paddy irrigation water on groundwater recharge. A numerical model of groundwater flow was conducted using MODFLOW in a 600 ha study site in an alluvial plain along the Chikugo River, located in southwestern Japan. To specify the surface boundary condition, data on the land use condition stored in the GIS database were transferred into a numerical model of groundwater flow. The simulated results were consistent with the observed yearly changes of groundwater level. Thus, it was appropriate to use the model to simulate the effects of paddy irrigation on groundwater. To quantify these effects, the groundwater level was simulated during the irrigation period when all farmlands in the study site were ponded. In this situation, the groundwater level was 0.5 to 1.0 m higher, the ground water storage 20% larger, and the return flow of the groundwater to the river 50% larger than in the present land use condition.     

Chemical forms and stability of selenium in soil

The chemical forms of selenium (Se) in soils derived from different parent rocks, and the stability of Se species in soils were investigated. Around 40% of the total Se occurred as organic Se on the average. Total selenite content was higher than that of total selenate. The content of organic Se decreased with heating of the soils at high temperatures, and most of it disappeared at heating temperatures above 500°C. However, the results of incubation experiments indicated that Se species were stable in soil, and that their chemical forms did not change appreciably with the variation of soil conditions. In soluble Se, organic Se accounted for about 50%, and in contrast to the total Se, the selenate content was higher than that of selenite.     

Functional Analysis of Lysosomes During Mouse Preimplantation Embryo Development

Lysosomes are acidic and highly dynamic organelles that are essential for macromolecule degradation and many other cellular functions. However, little is known about lysosomal function during early embryogenesis. Here, we found that the number of lysosomes increased after fertilization. Lysosomes were abundant during mouse preimplantation development until the morula stage, but their numbers decreased slightly in blastocysts. Consistently, the protein expression level of mature cathepsins B and D was high from the one-cell to morula stages but low in the blastocyst stage. One-cell embryos injected with siRNAs targeted to both lysosome-associated membrane protein 1 and 2 (LAMP1 and LAMP2) were developmentally arrested at the two-cell stage. Pharmacological inhibition of lysosomes also caused developmental retardation, resulting in accumulation of lipofuscin. Our findings highlight the functional changes in lysosomes in mouse preimplantation embryos.     

Genetic succession and spatial genetic structure in a natural old growth Cryptomeria japonica forest revealed by nuclear and chloroplast microsatellite markers

Spatial genetic structure and diversity of Cryptomeria japonica trees in old growth forest but selectively logged in approximately 300 years ago were analyzed using seven nuclear microsatellite markers and one chloroplast microsatellite marker. The individuals were sampled from a 4-ha natural forest plot (200 m × 200 m), which are consisted of three size classes including newly regenerated individuals such as regenerated individuals (REG class; stem diameter at breast height (DBH), 5–110 cm), remaining individuals (REM class; DBH ≥ 110 cm), and old stumps. Genetic diversity was similar between the three groups. These results suggest that genes from the old growth forest may have been passed on to the next generation (i.e., the current forest). DNA extraction and analysis from old stumps was partly successful and we could compare the genetic diversity between old stumps and living trees using four microsatellite loci. Patterns of spatial genetic structure detected by analyses of the nuclear and chloroplast markers differed substantially; all nuclear markers showed significant positive autocorrelation (P < 0.05) over short distances, but the chloroplast marker showed no significant autocorrelation over any distance. We concluded that Moran's I spatial autocorrelation pattern for the chloroplast microsatellite may have been influenced by the extensive pollen flow characteristic of this species, because chloroplast DNA of C. japonica is paternally inherited through pollen and is a wind-pollinated species that produces abundant pollen every few years. In contrast, Moran's I spatial autocorrelation pattern for nuclear microsatellites may reflect limited seed dispersal in gaps where the forest has regenerated and low levels of seed shadow overlap due to past logging. The average seed dispersal distance was 86.0 m at the study site and about 34.0% of the seeds were from the outside of the site, which revealed by parentage analysis. The individuals remaining after logging, and the extensive pollen flow from outside our study plot, may play important roles in maintaining the forests on this island. Gaps created by the selective logging activities appear to have been important for the regeneration of this species in the past but in the present time, natural disturbances such as typhoon and landslide are also important for gap creation.     

Differential Expression of Two Cytosolic Ascorbate Peroxidases and Two Superoxide Dismutase Genes in Response to Abiotic Stress in Rice

Superoxide dismutase (SOD) and ascorbate peroxidase (APX) play central roles in the pathway for scavenging reactive oxygen species in plants, thereby contributing to the tolerance against abiotic stress. Here we report the responses of cytosolic SOD (cSOD; sodCc1 and sodCc2) and cytosolic APX (cAPX; OsAPX1 and OsAPX2) genes to oxidative and abiotic stress in rice. RNA blot analyses revealed that methyl viologen treatment caused a more prominent induction of cAPXs compared with cSODs, and hydrogen peroxide t...     

CONTRAST-ENHANCED ULTRASONOGRAPHY FOR CHARACTERIZATION OF CANINE FOCAL LIVER LESIONS

In six normal beagles and 27 dogs with spontaneous focal or multifocal liver lesions, contrast-enhanced ultrasonography using Sonazoid^® was performed. Sonazoid^® is a newly developed second-generation contrast agent with the ability to be used for real-time contrast imaging along with parenchymal imaging. An appropriate protocol for the evaluation of all three phases (arterial, portal, and parenchymal) was established based on the results for normal beagles. By evaluation of the echogenicity of hepatic nodules during the arterial and parenchymal phases it was possible to differentiate malignant tumors from benign nodules with very high accuracy. In 15 of 16 dogs diagnosed as malignant tumors, nodules were clearly hypoechoic to the surrounding normal liver during the parenchymal phase. Additionally, malignant tumors had different echogenicity compared with the surrounding normal liver during the arterial phase in 14 of 15 dogs. In the portal phase, there were no characteristic findings. Contrast-enhanced ultrasonography with Sonazoid^® appears to improve the characterization of canine focal and multifocal hepatic lesions.     

KAv-1 is Better Suited to Chick Fibroblast Culture than DMEM or 199 Media

Cultured cells are a useful resource for poultry scientists, since these cells allow scientists to evaluate biological responses to conditions such as infectious diseases in vitro while mimicking the whole-body response in birds. However avian cell culture requires an optimized basal medium, and there are currently relatively few options for this basal medium (medium 199 and KAv-1). This means that there is still room for the development of an optimal basal medium for avian cell culture. Here we compare KAv-1 medium, Dulbecco''s modified Eagle medium (DMEM) and medium 199 during the culture of chick fibroblasts and determine that KAv-1 remains the optimal medium for these assays. Our results show that DNA damage is reduced in fibroblasts cultured in the KAv-1 medium, when compared to both DMEM and Medium 199 and that these cells also display improved growth dynamics in KAv-1 medium when compared to both DMEM and medium 199. To the best of our knowledge, this is the first study to describe a comparative analysis of culture media for avian cells, which would provide useful information for poultry scientists.     

QUANTITATIVE CONTRAST-ENHANCED ULTRASONOGRAPHY OF CANINE SPLEEN

Contrast-enhanced ultrasonography has an important role in the detection of tumors in humans. The second-generation contrast agent Sonazoid has the ability of real-time contrast imaging along with parenchymal imaging. The purposes of this study were to determine the effect and duration of Sonazoid on the changes in gray-scale enhancement of canine spleen and to establish an appropriate protocol for contrast-enhanced ultrasonography of canine spleen. Six healthy beagles were injected with an intravenous bolus of Sonazoid. In the spleen parenchyma, the enhancement was maintained up to 30 min after injection. Moreover, for 5–22 s after injection, gray-scale enhancement of splenic arteries afforded arterial imaging. Perfusion of the kidney may be investigated from 3.6 s to 3.5 min after injection of Sonazoid. These results suggest that Sonazoid is applicable to canine spleen parenchymal imaging and that the optimal time for the parenchymal imaging is 7–30 min after injection. The findings of this quantitative study should prove useful in the evaluation of diffuse or focal splenic and renal diseases in dogs.