tasks that require nematode extraction and microscopic examination. To develop a more efficient detection method for Bursaphelenchus xylophilus, we first generated monoclonal antibodies (MAbs) specific to B. xylophilus. Among 2,304 hybridoma fusions screened, a hybridoma clone named 3-2A7-2H5 recognized a single protein from B. xylophilus specifically. We finally selected the MAb clone 3-2A7-2H5-D9-F10 (D9-F10) for further studies. To identify the antigenic target of MAb-D9-F10, we analyzed proteins in spots, fractions or bands via nano liquid chromatography electrospray ionization quadrupole ion trap mass spectrometry (nano-LC-ESI-Q-IT-MS). Peptides of galactose-binding lectin-1 of B. xylophilus (Bx-LEC-1) were commonly detected in several proteomic analyses, demonstrating that this LEC-1 is the antigenic target of MAb-D9-F10. The localization of MAb-D9-F10 immunoreactivities at the area of the median bulb and esophageal glands suggested that the Bx-LEC-1 may be involved in food perception and digestion. The Bx-LEC-1 has two non-identical galactose-binding lectin domains important for carbohydrate binding. The affinity of the Bx-LEC-1 to D-(+)-raffinose and N-acetyllactosamine were much higher than that to L-(+)-rhamnose. Based on this combination of evidences, MAb-D9-F10 is the first identified molecular biomarker specific to the Bx-LEC-1.

Three acetylcholinesterases (AChEs) were identified from the pinewood nematode, Bursaphelenchus xylophilus. Sequence comparison with known AChEs in conjunction with three-dimensional structure analysis suggested that all BxAChEs share typical characteristics of AChE at the major catalytic structures. BgAChE3 was most predominantly transcribed and then followed by AChE1 and AChE2. Immunohistochemistry using anti-BxAChEs antibodies revealed that BxAChE1 is most widely distributed whereas BxAChE2 exhibits more localized distribution in neuronal tissues. BxAChE3 was detected from entire body together with some limited tissues, including mouth parts and alimentary lining, and determined to be the only soluble AChE, suggesting its localization in hemolymph or/and extracellular space. Kinetic analysis of in vitro expressed BxAChEs revealed that BxAChE1 has the highest substrate specificity whereas BxAChE2 has the highest catalytic efficiency with BxAChE3 having the lowest catalytic efficiency. Interestingly, presence of BxAChE3 in the pool of BxAChEs significantly reduced the inhibition of BxAChE1 and BxAChE2 by inhibitors. Knockout of BxAChE3 by RNAi significantly increased the toxicity of nematicides, suggesting the protective role of BxAChE3 against these toxicants. Based on several features, including tissue distribution, expression level, substrate kinetics and inhibition property, it appeared that BxAChE1 is the major AChE with the function of postsynaptic transmission whereas BxAChE3 has been evolved to acquire the function of chemical defense, perhaps intrinsically against secondary toxic compounds from host pine trees, such as α-pinene and limonene. BxAChE2 appears to play a role in post-synaptic transmission in specialized neurons but its detailed physiological function still remains to be elucidated.

Seeking to improve the weak resolution of deeper divergences in an initial study based on five nuclear genes (6.6kb total) in 123 exemplars, we nearly tripled the total sequence (to 26 genes, 18.4 kb total) in one third (41) of the taxa. The expanded, deliberately incomplete data matrix consistently increased bootstrap support for previously-identified groupings, while introducing no contradictory groupings of the kind that missing data have been predicted to produce. To test the relative effectiveness of “more genes” versus “more taxa” sind that we compared two largely complete matrices, the initial 5 gene × 123 taxon and the 26 gene × 41 taxon data sets, that contain roughly equal amounts of sequence. The “more genes” data set yielded consistently, sometimes dramatically higher bootstrap support that is generally not attributable to taxon number alone. We also found that a gene-rich taxon subset provides reassuring evidence of strong underlying signal that is not obvious in subsequent larger analyses, helping to encourage and guide the search for deep relationships amid the noise of expanded taxon sampling.

In this paper, Fe-TiO2 and Fe-fullerene/TiO2 composite photocatalysts were prepared with titanium (IV) n-butoxide (TNB) by a sol-gel method. TiO2, Fe-TiO2 and Fe-fullerene/TiO2 were characterized by scanning electron microscopy (SEM), Transmission electron microscope (TEM), specific surface area (BET), X-ray diffraction analysis (XRD) and energy dispersive X-ray spectroscopy (EDX). The photocatalytic activities were evaluated by the photocatalytic oxidation of methylene blue (MB) solution. XRD patterns of the composites showed that the photocatalyst composite contained a typical single and clear anatase phase. The surface properties shown by SEM presented a characterization of the texture on Fe-fullerene/TiO2 composites and showed a homogenous composition in the particles for the titanium sources used. The EDX spectra for the elemental identification showed the presence of O, C and Ti elements. Moreover, peaks of the Fe element were observed in the Fe-TiO2 and Fe-fullerene/TiO2 composites. The degradation of MB solution by UV-light irradiation in the presence of photocatalyst compounds was investigated in complete darkness. The degradation of MB concentration in aqueous solution occurred via three kinds of physical phenomena: quantum efficiency of the fullerene; organo-metallic reaction of the Fe compound; and decomposition of TiO2. The degradation rate of the methylene blue solution increased when using Fe-fullerene/TiO2 compounds.

We report relative proper motion measurements of H2O masers in massive star-forming region W51 Main, based on data sets of VLBI observations for H2O masers at 22 GHz with Japanese VERA telescopes from 2003 to 2006. Data reductions and single-beam imaging analysis are to measure internal kinematics of maser spots and eventually to estimate the three-dimensional kinematics of H2O masers in W51 Main. Average space motions and proper motion measurements of H2O masers are given both graphical and in table formats. We find in this study that W51 Main appears to be associated with hyper-compact H II region with multiple massive proto-stars whose spectral types are of late O.