Neospora caninum is an Apicomplexan parasite that causes repeated abortion and stillbirth in cattle. The aim of this study was to clone the gene encoding the N. caninum orthologue (NcBAG1) of the ...Toxoplasma gondii bradyzoite-specific protein TgBAG1 and characterize its expression pattern in the parasite. Isolation of the full-length 684-bp gene revealed that it shared 78.3% sequence similarity with TgBAG1. NcBAG1 encodes a predicted protein of 227 amino acids with 80.3% similarity to TgBAG1. A putative signal peptide sequence and an invariant GVL motif characteristic of small heat-shock proteins were identified in the predicted N. caninum amino acid sequence. We expressed the NcBAG1 gene as a recombinant glutathione S-transferase fusion protein (rNcBAG1) in Escherichia coli and used the purified 60 kDa protein to obtain a monoclonal antibody (Mab). rNcBAG1 reacted to Mabs specific for NcBAG1 and TgBAG1. No reaction between the NcBAG1 Mab and N. caninum tachyzoites was observed. Although the predicted molecular mass of NcBAG1 is 25 kDa, Western blot analysis of parasite lysates using the NcBAG1 Mab revealed a cross-reactive protein of approximately 30 kDa. Additionally, immunofluorescence assays using the tachyzoite-specific Mab for NcSAG1 and the bradyzoite-specific Mab for TgBAG1 or NcSAG4 revealed NcBAG1-specific expression in bradyzoites in cultures exposed to sodium nitroprusside, a reagent that increases the frequency of bradyzoites. Interestingly, the NcBAG1 protein was identified in the cytoplasm of the bradyzoite-stage parasites. This preliminary analysis of the NcBAG1 gene will assist investigations into the role of this protein in N. caninum.
A number of US overseas bases were deployed around the world to protect allies and maintain regional peace. Some bases have been stationed in the partner countries for the long term, whereas others ...were withdrawn from their partners' territories in the face of strong local opposition. Understanding local support for US overseas bases is indispensable for managing alliance politics and pursuing US grand strategy. This article addresses the 1972–2006 Okinawa gubernatorial elections where the US base issue had been chronically politicized and locals supported pro-base candidates six out of ten times contrary to their anti-base preferences. This article addresses external threats as a determinant of vote choice. We analyze the gubernatorial elections as the opportunities for Okinawans to convey their support for or opposition to the current national security policy since US bases in Okinawa are critical to Japan's security. We find that external threats do encourage Okinawans to support pro-base candidates, but the effect of perceived security-related risks is moderate. Moreover, physical and psychological costs such as airplane crashes, environmental and noise pollution, and rape incidents have larger influence on the election outcomes rather than material benefits such as the fiscal transfers and base-related subsidies, which is contrary to the conventional view.
Effects of soil freeze-thaw cycles on soil microbial biomass were examined using 8 soil samples collected from various locations, including 4 arable land sites and 2 forest sites in temperate regions ...and 2 arable land sites in tropical regions. The amounts of soil microbial biomass C and N. determined by the chloroform fumigation and extraction method, significantly decreased by 6 to 40% following four successive soil freeze-thaw cycles ( - 13 and 4 deg C at 12 h-intervals) compared with the unfrozen control (kept at 4 deg C during the same period of time as that of the freeze-thaw cycles). In other words, it was suggested that 60 to 94% of the soil microorganisms might survive following the successive freeze-thaw cycles. Canonical correlation analysis revealed a significantly positive correlation between the rate of microbial survival and organic matter content of soil (r = 0.948*). Correlation analysis showed that the microbial survival rate was also positively correlated with the porespace whose size ranged from 9.5 to 6.0 micro m (capillary-equivalent-diameter; r = 0.995**), pH(KCl) values (r = 0.925**), EC values (r = 0.855*), and pH(H,O) values (r = 0.778*), respectively. These results suggested that the soil physicochemical properties regulating the amount of unfrozen water in soil may affect the rate of microbial survival following the soil freeze-thaw cycles. The potential of organic matter decomposition of the soils was examined to estimate the effects of the soil freeze-thaw cycles on the soil processes associated with the soil microbial communities. The soil freeze-thaw cycles led to significant 6% increase in chitin decomposition and 7% decrease in rice straw decomposition (p < 0.05), suggesting that the partial sterilization associated with the soil freeze-thaw cycles might disturb the soil microbial functions.
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