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Aromia bungii (Faldermann) (Coleoptera: Cerambycidae) is an invasive pest, damaging Rosaceae trees (particularly Prunus ) in Japan and Europe. The establishment of this beetle in Japan was first detected in 2012, and subsequently, it has rapidly expanded its distribution. Currently, Japanese populations of A. bungii are widely distributed in six non-contiguous regions. In this study, we compared the nucleotide sequences of mitochondrial cytochrome oxidase subunit 1 of the populations in these six regions in Japan to examine whether multiple introductions or human-mediated long-distance dispersal have contributed to the non-contiguous distribution of A. bungii . Seven haplotypes were detected from Japanese populations, and one of these was identical to a sequence deposited from China. One to two haplotypes were detected in each region, suggesting a genetic bottleneck. Detected haplotypes differed between introduced regions, although two regions shared a single haplotype. These results suggest that multiple independent introductions of A. bungii have contributed to its non-contiguous distribution in Japan. Quarantine measures for wood-packing materials in trade need to be strengthened to prevent the establishment of further populations of A. bungii ., Competing Interests: The authors declare no conflict of interest.

We investigated the ecological traits of emerging adults and the boring density in Aromia bungii -infested flowering cherry ( Cerasus × yedoensis 'Somei-yoshino') and peach ( Prunus persica ) trees to evaluate their suitability as food resources for A. bungii , and their vulnerability to infestation. The number of adults per m 3 that emerged from P. persica was 10-times larger than from C . × yedoensis , and the numbers of emergence holes, entrance holes, and pupal chambers were also larger in P. persica logs. The lifetime fecundity of adults that emerged from P. persica was also higher. Elytral length, sex ratios, and adult lifespans did not differ between the two host trees. Our results indicate that peach trees provide more suitable conditions than do flowering cherry trees for A. bungii larvae. Although flowering cherry trees, primarily C . × yedoensis , which are currently grown as street or ornamental trees in Japan, have been more severely affected by A. bungii to date, the greater risk in the long term is to P. persica , an agricultural species in the main producing areas surrounding the Kanto region.

In recent years, insecticide trunk injection was put into practical use for controlling wood boring pests. However, few studies have investigated the dose-response relationships between insecticides and wood-boring pests in detail. This study used two commercial formulations of the neonicotinoid insecticides thiamethoxam and dinotefuran and investigated their dose-response relationships with invasive wood borer Aromia bungii (Coleoptera: Cerambycidae) larvae. Neonates and late instar larvae were reared with an artificial diet containing different insecticide concentrations (0.01-100 ppm) in the laboratory, and their diet excavation activity, survival rate, and weight change were recorded. Diet excavation immediately dropped in larvae exposed to high concentrations of thiamethoxam or dinotefuran (≥1 ppm in neonates and ≥10 ppm in late instar larvae). The weight and survival rate gradually declined over 12 weeks in late instar larvae. These results suggest that the two neonicotinoids intoxicate and debilitate A. bungii larvae gradually to death. In practical use, rapid suppression of A. bungii wood boring damage can be expected by trunk injection of neonicotinoid insecticides. However, a relatively long-term retention of the insecticides may be required to kill large larvae. Neonates may be controlled with lower insecticide dosage and shorter exposure than larger larvae.

Pine wilt disease (PWD) invaded southern Japan in the early 1900s and has gradually expanded its range to northern Honshu (Japanese mainland). The disease is caused by a pathogenic North American nematode, which is transmitted by native pine sawyer beetles. Recently, the disease has invaded other portions of East Asia and Europe where extensive mortality of host pines is anticipated to resemble historical patterns seen in Japan. There is a critical need to identify the main drivers of PWD invasion spread so as to predict the future spread and evaluate containment strategies in newly invaded world regions. But the coupling of pathogen and vector population dynamics introduces considerable complexity that is important for understanding this and other plant disease invasions. In this study, we analysed historical (1980-2011) records of PWD infection and vector abundance, which were spatially extensive but recorded at coarse categorical levels (none, low and high) across 403 municipalities in northern Honshu. We employed a multistate occupancy model that accounted both for demographic stochasticity and observation errors in categorical data. Analysis revealed that sparse sawyer populations had lower probabilities of transition to high abundance than did more abundant populations even when regional abundance stayed the same, suggesting the existence of positive density dependence, that is an Allee effect, in sawyer dynamics. Climatic conditions (average accumulated degree days) substantially limited invasion spread in northern regions, but this climatic influence on sawyer dynamics was generally weaker than the Allee effect. Our results suggest that tactics (eg sanitation logging of infected pines) which strengthen Allee effects in sawyer dynamics may be effective strategies for slowing the spread of PWD., (© 2018 The Authors. Journal of Animal Ecology published by John Wiley & Sons Ltd on behalf of British Ecological Society.)

Underground community assemblies have not been studied well compared with aboveground communities, despite their importance for our understanding of whole ecosystems. To investigate underground community assembly over evolutionary timescales, we examined terrestrial earthworm communities (Oligochaeta: Haplotaxida) in conserved mountainous primary forests in Japan as a model system. We collected 553 earthworms mostly from two dominant families, the Megascolecidae and the Lumbricidae, from 12 sites. We constructed a molecular taxonomic unit tree based on the analysis of three genes to examine the effects of a biogeographic factor (dispersal ability) and an evolutionary factor (habitat adaptation) on the earthworm community assembly process. The phylogenetic distance of the earthworm communities among sites was positively correlated with geographic distance when intraspecific variation was included, indicating that the divergence within species was affected by biogeographic factors. The community assembly process in the Megascolecidae has also been affected by environmental conditions in relation to an evolutionary relationship between habitat environment and intestinal cecum type, a trait closely related to habitat depth and diet, whereas that in the Lumbricidae has not been affected as such. Intestinal cecum type showed a pattern of niche conservatism in the Megascolecidae lineage. Our results suggest that investigating the evolution of a key trait related to life history can lead to the clear description of community assembly process over a long timescale and that the community assembly process can differ greatly among related lineages even though they live sympatrically.

The recent development of screening strategies based on the generation and display of large libraries of antibody fragments has allowed considerable advances for the in vitro isolation of monoclonal antibodies (mAbs). We previously developed a technology referred to as the 'ADLib (Autonomously Diversifying Library) system', which allows the rapid screening and isolation in vitro of antigen-specific monoclonal antibodies (mAbs) from libraries of immunoglobulin M (IgM) displayed by the chicken B-cell line DT40. Here, we report a novel application of the ADLib system to the production of chimeric human mAbs. We have designed gene knock-in constructs to generate DT40 strains that coexpress chimeric human IgG and chicken IgM via B-cell-specific RNA alternative splicing. We demonstrate that the application of the ADLib system to these strains allows the one-step selection of antigen-specific human chimeric IgG. In addition, the production of chimeric IgG can be selectively increased when we modulate RNA processing by overexpressing the polyadenylation factor CstF-64. This method provides a new way to efficiently design mAbs suitable for a wide range of purposes including antibody therapy.

Background: The ambrosia beetle, Platypus quercivorus, is the vector of oak wilt, one of the most serious forest diseases in Japan. Population genetics approaches have made great progress toward studying the population dynamics of pests, especially for estimating dispersal. Knowledge of the genetic structuring of the beetle populations should reveal their population history. Using five highly polymorphic microsatellite loci, 605 individuals from 14 sampling sites were assessed to infer the ongoing gene flow among populations as well as the processes of expansion of damaged areas., Results: Population differentiation (FST = 0.047, G'ST = 0.167) was moderate and two major clusters were detected by several methods, dividing the samples into north-eastern and south-western populations, a similar genetic divergence was reported in host oak trees. Within the north-eastern populations, the subgroups mostly corresponded to differences in the collection period. The genetic characteristics of the population might have changed after 2 years due to the mixing of individuals between populations with enhanced migration related to population outbreaks. Because isolation by distance was detected for whole populations and also within the north-eastern populations, migration was considered to be limited between neighbouring populations, and most populations were suggested to be in genetic equilibrium of genetic drift and gene flow. Recent bottlenecks were found in some populations with no geographical bias; however, they were all from newly emerged oak wilt forests. The emergence of oak wilt should have induced intense fluctuations in the beetle population size., Conclusions: Because the genetic boundaries coincide, we suggest that the geographical structuring of the beetle was formed by co-evolution with the host species. Our findings indicate the oak wilt expansion process.

To study the dispersal process of the pine sawyer Monochamus alternatus (Hope) in frontier populations, a microsatellite marker-based genetic analysis was performed on expanding populations at the northern limit of its range in Japan. In Asian countries, M. alternatus is the main vector of pine wilt disease, the most serious forest disease in Japan. Sawyers were collected from nine sites near the frontier of the pine wilt disease damage area. A mountain range divides the population into western and eastern sides. Five microsatellite loci were examined and a total of 188 individuals was genotyped from each locus with the number of alleles ranged from two to nine. The mean observed heterozygosity for all loci varied from 0.282 to 0.480 in the nine sites, with an overall mean of 0.364. None of the populations have experienced a significant bottleneck. Significant differentiation was found across the mountain range, but the genetic composition was similar amongst populations of each side. It is believed that the mountain range acts as a geographical barrier to dispersal and that gene flow without a geographical barrier is high. On the west side of the mountain range, a pattern of isolation by distance was detected. This was likely to be caused by secondary contact of different colonizing routes on a small spatial scale. Based on these data, a process linking genetic structure at local (kilometres) and regional spatial scales (hundreds of kilometres) was proposed.

Background/aims: To examine whether chronic intake of vegetable protein added to animal protein diet affects renal hemodynamics or not, we studied effects of three kinds of diets containing various amounts of animal and vegetable protein with 1-week dietary program in each on renal hemodynamics., Methods: The crossover design of different amounts of vegetable protein added to the constant amount of animal protein was applied to two groups of 7 healthy individuals after the control dietary program. Renal function and 24 hours' urinary albumin excretion rate (AER) were examined on every 7th day of three consecutive 1-week dietary programs., Results: Glomerular filtration rate (GFR; sodium thiosulphate clearance) and renal plasma flow (RPF) significantly decreased after decreasing the intake of animal protein by one third with keeping the amount of vegetable protein constant. The results when substituting vegetable protein for some of the animal protein in the diet without changing the total amount of protein were identical. The filtration fraction and AER did not change over the study periods regardless of dietary composition., Conclusion: The lack of an effect a 1-week intake of vegetable protein added to animal protein on GFR and RPF suggests that vegetable protein may be excluded from lists of restriction in low protein diet therapy in patients with renal insufficiency., (Copyright 2002 S. Karger AG, Basel)

To examine whether urinary excretions of plasma proteins with molecular radii of 45-55 A and different isoelectric points such as IgG (pI = 7.4) and ceruloplasmin (pI = 4.4) increase selectively in normoalbuminuric type 2 diabetic patients, urinary albumin excretion rate (AER), renal clearances of IgG, ceruloplasmin and alpha2-macroglobulin, and creatinine clearance (Ccr) were studied in timed overnight urine samples of 36 diabetic outpatients and 16 control subjects. Furthermore, to examine effect of glycemic control on these urinary protein excretions, the same analysis was performed before and after glycemic control in 17 diabetic inpatients admitted for glycemic control. Renal clearances of IgG and ceruloplasmin were significantly higher in diabetic outpatients than in the control group, whereas AER and renal clearance of alpha2-macroglobulin did not differ. Glycemic control caused significant decreases in renal clearances of IgG and ceruloplasmin, accompanied with tendency for Ccr to decrease (p = 0.055). The present results, together with our previous finding of selectively increased urinary excretions of 45-55 A sized plasma proteins in parallel with enhanced glomerular filtration rate after acute protein loading, led us to conclude that enhanced intraglomerular hydraulic pressure may cause increases in clearances of IgG and ceruloplasmin, and that this change can be reversed by strict glycemic control in normoalbuminuric diabetic patients.

Oxidative stress possibly contributes to the development of diabetic nephropathy. Therefore, the levels of endogenous antioxidants may be one of determinants of the susceptibility to diabetic nephropathy. Glutathione S-transferases (GSTs) can work as one of endogenous antioxidants to protect cells from oxidative stress. The M1 member of GST mu class (GSTM1) is polymorphic and only expressed in 55-60% of Caucasians because of the homozygous deletion of the gene (null genotype). Recent studies have provided evidence that the GSTM1 null genotype, i.e. lack of the GSTM1 activity, is associated with an increased susceptibility to lung cancer and colorectal cancer. The present study was conducted to determine whether the genetic polymorphism influences the development of diabetic nephropathy. We examined 105 patients with diabetic nephropathy and 69 patients without diabetic nephropathy in Japanese type 2 diabetic patients with proliferative diabetic retinopathy. GSTM1 genotyping was performed by polymerase chain reaction. The two patient groups were well matched with regard to age, body mass index and HbAlc. GSTM1 null genotype was observed in 48.6% of patients with nephropathy versus 55.1% of patients without nephropathy. The frequency of GSTM1 null genotype was not significantly higher in the patient group with nephropathy than in the patient group without nephropathy. This study is the first to investigate the association of GSTM1 gene polymorphism with the development of diabetic nephropathy. The present results suggest that GSTM1 null genotype does not contribute to the development of diabetic nephropathy in Japanese type 2 diabetic patients.