Distribution of glycoproteins on feline testicular sperm, epididymal sperm and ejaculated sperm

Glycoproteins (GPs) are known to be involved in the phenomenon of sperm maturation and capacitation. In the present study, we investigated the attachment of GPs on sperm cell membrane during the process of feline sperm maturation from testicular sperm to ejaculated sperm by using 8 FITC-labeled lectins. The results showed that 3 types of GPs were presented on testicular sperm and 7 on caput epididymal sperm. Corpus and cauda epididymal sperm and ejaculated sperm had GPs detected by 8 FITC-labeled lectins used in the present study. This study demonstrates the part of the characteristic of GPs that are present on the feline sperm cell membrane during the process of sperm maturation.     

1NM-PP1 treatment of mice infected with Toxoplasma gondii

Bumped kinase inhibitors (BKIs) target analog-sensitive kinases, which the genomes of mammals rarely encode. Previously, we demonstrated that a BKI effectively suppressed the in vitro replication of Toxoplasma gondii, the causative pathogen of toxoplasmosis, by targeting T. gondii calcium-dependent protein kinase 1 (TgCDPK1) (Eukaryotic Cell, 9: 667-670). Here, we examined whether the BKI 1NM-PP1 reduced parasite replication in vivo. A high dose of 1NM-PP1, by intraperitoneal injection, just before the parasite inoculation effectively reduced the parasite load in the brains, livers, and lungs of T. gondii-infected mice, however, a low dose of 1NM-PP1 with oral administration didn't change the survival rates of infected mice.     

Developmental ability of vitrified mouse oocytes expressing water channels

Previously, we showed that the exogenous expression of aquaporin 3 (AQP3), an aquaglyceroporin, improved the tolerance of mouse oocytes to vitrification with a glycerol-based solution. In the present study, we examined conditions suitable for the expression of AQP3 and the ability of vitrified oocytes to develop in vitro and in vivo after fertilization. After only partial remove of cumulus cells, immature mouse oocytes (germinal vesicle stage) were injected with 5, 10 or 20 pg of AQP3 cRNA and cultured for 12 h for maturation. When matured oocytes were vitrified with a glycerol-based solution, 57-61% survived regardless of the amount of cRNA injected (5-20 pg). By contrast, no oocytes injected with water (control) survived. When the zona pellucida was removed from the vitrified oocytes and the oocytes were then fertilized in vitro and cultured, the proportions that were fertilized and developed into blastocysts were higher when the amount of cRNA injected was 5 pg than 10-20 pg. When 16 blastocysts were transferred to a pseudopregnant mouse, 5 developed to term, demonstrating that oocytes vitrified after injection of AQP3 cRNA retained the ability to develop to term. The water-permeability of cRNA-injected oocytes was higher than that of control oocytes from the maturing stage to the 1-cell zygote stage, whereas glycerol-permeability was higher only at metaphase II. This indicates that AQP3 was expressed for a relatively short period of time. These results suggest that the transient expression of water/cryoprotectant channels is effective for cryopreserving cells that have low membrane-permeability, such as mammalian oocytes.     

Kudzu (Pueraria lobata) vine ethanol extracts improve ovariectomy-induced bone loss in female mice

Bone-loss-improving action of kudzu vine ethanol extracts (PVEE) was clarified. PVEE was composed roughly of 80% fiber, 10% puerarin, 3.6% daidzin, 2.5% 6″-O-malonyldaidzin, and the other minor isoflavones. Ten-week-old ovariectomized (OVX) mice were fed diets containing PVEE (20 mg/kg body weight/day) for 8 weeks. The bone resorption markers (urinary deoxypyridinoline and tartrate-resistant acid phosphatase activity) was elevated in OVX mice and was significantly decreased in OVX mice that consumed PVEE for 8 weeks. Consistent with the decrease in the markers, the number of matured osteoclasts in the distal femur was diminished in OVX mice fed PVEE diets. PVEE diets also suppressed the decrease in femoral bone mineral density (BMD) by OVX. PVEE showed the affinity for estrogen receptor α and β nearly 1/10000 weaker than 17β-estradiol. No hypertrophy in the uterus by the PVEE diet was observed. These results suggest that PVEE could be a promising resource for a functional food that improves osteoporosis.     

Conversion of sulfuric acid lignin generated during bioethanol production from lignocellulosic materials into polyesters with <Emphasis Type="Italic">ɛ</Emphasis>-caprolactone

In this study, polyesters were prepared from sulfuric acid lignin (SAL) that was generated as a by-product during the production of bioethanol from lignocellulosic materials using ɛ-caprolactone (CL). Carboxylic groups were introduced into SAL by a hydrothermal reaction under alkali conditions. The acid form (-COOH) of the obtained polymer (HSAL) was dissolved into ɛ-caprolactone and the mixture was heated at 150°C for 12 h with ZnCl₂ as a catalyst. The structure of the obtained polymer (PCL-HSAL) was proposed based on results of differential scanning calorimetric analysis, swelling tests, and dynamic mechanical thermal analysis. The hydroxyl group in HSAL participated in the ring-opening polymerization of CL. PCL-HSAL with a high HSAL content formed a three-dimensional network that did not dissolve in organic solvents and did not melt at high temperature. PCL-HSAL showed a high swelling property for a suitable ratio of CL to HSAL.     

Distribution of mineral elements in the outer layer of rice and wheat grains,using electron microprobe X-Ray analysis

Electron microprobe X-ray analysis was used to determine the transversal microdistribution of P, K, Mg, Ca, Fe, and Mn, with special attention to the outer layers of rice and wheat grains. P, K, Mg, Fe, and Mn were concentrated in the aleurone layer in each case. In particular, P, Mg, and K were highly concentrated in the subcellular particles of the aleurone layer, and had very similar distribution patterns in the outer layers of the matured grain of rice and wheat. By contrast, Ca was abundant in the pericarp.     

Localization of MT2a Gene promoter expression is different from the site(s) of copper accumulation in the roots of transgenic Arabidopsis thaliana

Accumulation of mRNA for MT2a, a metallothionein of Arabidopsis thaliana, is known to be upregulated by excess copper (Cu). We compared the localization of the promoter activity of the MT2a gene and the accumulation site(s) of CU in the roots of transgenic A. thaliana. Open reading frame of the β-glucuronidase (GUS) gene was fused with the MT2a gene promoter and introduced into A. thaliana. Strong GUS activity was observed in the region near or within the stele in the presence of CU at a high concentration (high-CU condition). Electron probe X-ray micro-analysis demonstrated that under high-CU conditions, CU accumulated most abundantly in the cortex, where GUS activity was not significantly induced. These results indicate that the sites showing the MT2a promoter activity and CU accumulation did not coincide with each other, suggesting that CU is not a direct activator of the MT2a promoter.     

Effect of the long-term application of anaerobically digested residual slurry on methane emissions in a rice paddy field

ABSTRACT

The anaerobic digestion of livestock manure is an environmentally compatible technology used for the production of renewable energy. Anaerobically digested residual slurry has been used worldwide as a liquid fertilizer in both upland and paddy fields. However, a controversial question remains as to whether the application of slurry to rice paddy fields increases methane emissions; although methane is one of the most prevalent greenhouse gases, little is known about the effects of the long-term application of residual slurry on methane emission. In this study, we repeatedly applied slurry to a paddy field for six years at different application rates (10, 15, and 20 g N m^?2 based on ammonium-nitrogen content). At the fifth and sixth years of application, we evaluated the effect in terms of methane flux and soil total carbon content. The effect of the long-term application of the slurry (10 g N m^?2) on grain yield was equivalent to that of chemical fertilizer (10 g N m^?2). The application of the residual slurry was likely to increase the cumulative methane emissions during rice growing season in both 2006 and 2007. On the other hand, we observed that soil total carbon did not accumulate significantly in the soil. Thus, we cannot rule out the potential risk of additional methane emissions caused by the application of the residuary slurry to paddy fields.     

Food sources for the bivalve Corbicula japonica in the foremost fishing lakes estimated from stable isotope analysis

ABSTRACT:   Carbon and nitrogen stable isotope ratios in tissue of the bivalve corbicula Corbicula japonica and particulate organic matter (POM) were measured in Lake Jusan, Lake Ogawara and Lake Shinji, which are the foremost fishing grounds for the corbicula in Japan, to determine their food sources. The bivalves in Lake Ogawara and Lake Shinji showed enriched isotope composition, while those in Lake Jusan were depleted. In addition, the difference in the isotope ratios between the sampling sites was remarkable in Lake Jusan. Chlorophyll concentrations were significantly higher in Lake Ogawara and Lake Shinji than those in the inflow rivers, although that in Lake Jusan was equivalent to that in the river. Residence time of river water was estimated at 1 day, 455 days and 88 days in Lake Jusan, Lake Ogawara and Lake Shinji, respectively. These values indicate that the bivalves in Lake Ogawara and Lake Shinji assimilate autochthonous phytoplankton, while those in Lake Jusan assimilate terrestrial matter in the upper reaches and marine phytoplankton in the lower reaches because of low production in the lake.     

Effect of stocking density and addition of proteins on larval survival in Japanese flounder, Paralichthys olivaceus

The effects of larval density, rearing media, and protein additions on fish larval survival were tested by rearing larvae in small containers. Yolk-sac larvae of Japanese flounder, Paralichthys olivaceus, were transferred 1 day after hatching to plastic dishes and cell-culture microplates by Pasture pipette at densities ranging from 0.3 to 30 individuals/ml and incubated at 16 °C without food. Dead larvae were removed and counted every 12 h. Three peaks of larval mortality were observed: the first peak appeared just after the transfer, the second corresponded to the completion of yolk absorption, and the third occurred 10–12 days after the transfer. Since about 50% of larvae survived until the third peak, it was clear that mortality at the second peak (completion of yolk absorption; equivalent to the first feeding) was not due to starvation. At the first mortality peak, better survival was unexpectedly recorded at higher larval densities (30 individuals/ml), suggesting that some material beneficial for survival is derived from the larvae. This idea was further supported by the improved survival in media previously used to rear larvae at high densities. The addition of bovine serum albumin (BSA) at 10 μg/ml effectively reduced mortality in microplates and in 30-l tanks. To reduce the larval mortality just after handling, addition of egg white at a dilution rate of 1/30,000 (vol/vol) or less is recommended as a practical alternative to BSA. Since the addition of polyethylene glycol 6000 (1 μg/ml) also effectively reduced mortality, reduction in mortality could not be attributed to a specific mechanism. The precise determination of the magnitude and timing of larval mortality was attained through the use of small containers, and a method to prevent mortality due to handling was identified in this study. These types of tests may allow the logical evaluation and establishment of better rearing procedures for seed production in aquaculture.