Phylogeography of the true freshwater crab, Geothelphusa dehaani: Detected dual dispersal routes via land and sea.

Dispersal is an important factor that determines the potential for colonization to pioneer sites. Although most decapods employ seaward migration for reproduction with a planktonic larval phase, true freshwater crabs spend their entire life cycle in freshwater. Therefore, it is expected that genetic regionality can be easily detected. In this study, we focused on true freshwater crabs, Geothelphusa Stimpson, 1858. Herein, we reveal the evolutionary history and dispersal patterns of freshwater crustaceans. We collected and genetically analyzed 283 specimens at 138 localities across the Japanese Islands. Phylogenetic analyses were conducted on the combined dataset (mtDNA COI, 16S, and nDNA ITS1, histone H3 regions) and the data set based on the mtDNA COI region. The phylogenetic relationships detected 10 clades that were highly monophyletic. The highlights of this study were the discovery of several cryptic species or undescribed species, and the completely different heterogeneous dual dispersal pathways within a single species; i.e., both land and ocean routes. Although it was concluded that Japanese crabs are basically genetically divided by straits, strong evidence for dispersion via ocean currents was also detected (i.e., a "sweepstake"). It was also confirmed that Geothelphusa dehaani (White, 1847) could survive in seawater.

A major flood caused by a typhoon did not affect the population genetic structure of a river mayfly metapopulation.

Floods affect the population structure of organisms that inhabit streams. In recent decades, the scale of floods has become larger due to climate change. Under these circumstances, on 12 October 2019, the largest typhoon in the history of observation in Japan struck the Japanese Archipelago. This typhoon caused heavy rainfall in various places, and the Chikuma-Shinano River System (Japan's largest) suffered great damage. Eight years before the large-scale disturbance in the river system, the population structure of the mayfly Isonychia japonica was studied in detail using quantitative sampling (population numbers and biomass) and by sequencing the mtDNA cytochrome c oxidase subunit I. To understand the impact of the flood on the population and genetic structures, we repeated the same research approximately 1 year after the flood. Direct comparison of sites before and after the flood revealed no significant changes between pre- and post-flood population genetic structure. This indicates high in situ resistance and/or resilience recovery of the populations to the disturbance. We hypothesize that this high resistance/resilience to flood disturbance is a result of strong selection for such traits in the rivers of the Japanese Archipelago, which are short, steep, flow rapidly and violently, and are strongly affected by floods.

Genome-wide molecular phylogenetic analyses and mating experiments which reveal the evolutionary history and an intermediate stage of speciation of a giant water bug.

The intermediate stages of speciation are important for understanding the processes involved in the creation of biodiversity, and also comprise a number of interesting phenomena. However, difficulties are associated with dividing clear speciation stages because speciation is a continuous process. Therefore, the elucidation of speciation is an interesting and important task in evolutionary biology. We herein present an example of a species in an intermediate stage of speciation using the giant water bug Appasus japonicus (Heteroptera, Belostomatidae) that was investigated using mating experiments and phylogenetic analyses of the mtDNA COI (658 bp) and 16S rRNA (435 bp) regions, and nDNA SSR (13 loci) and its genome-wide SNPs (11,241 SNPs). The results of our phylogenetic analyses based on their mtDNA data set and the genome-wide SNPs data set strongly supported the paraphyly of the Japanese populations. Therefore, it is suggested that their ancestral lineage which being distributed in the Japanese Archipelago subsequently migrated to the Eurasian Continent (i.e., back-dispersal occurred). Furthermore, the results of the mating experiments suggested that among A. japonicus, even between closely related lineages, premating reproductive isolation has been established by the differentiation of copulatory organ morphologies. In contrast, premating reproductive isolation is not established in the absence of the differentiation of copulatory organ morphologies, even if genetic differentiation is prominent. These results suggested that their phylogenetic distance does not predict premating reproductive isolation. Furthermore, in the present study, we present a clear example of premating reproductive isolation driving speciation between closely related lineages.

Possibility of Undeveloped Egg Absorption during Embryogenesis: A Unique Phenomenon Observed in the Ovoviviparous Mayfly Cloeon dipterum.

Oviparous, ovoviviparous and viviparous reproduction are interesting subjects for understanding animals' evolutionary pathways and adaptation to their life history and habitat conditions. In this study, we examined the reproductive mode of the ovoviviparous mayfly Cloeon dipterum, particularly comparing embryogenesis between hand-pairing and unmated females' common oviduct. Our study suggested that the high developmental rate of C. dipterum observed in a recent study could be ascribed to their absorption of unfertilized eggs. The developmental rates of hand-paired females were almost 100%, while their egg-bearing numbers were lower than those of virgin females. Thus, such reduced egg numbers suggest the maternal absorption of unfertilized eggs. This trait is thought to have evolved with the ovoviviparous characteristics of C. dipterum. We identified the basis of the irregularity of this species exhibiting such a high (i.e., 100%) developmental rate in our previous recent study.

Development of microsatellite markers for a giant water bug, Appasus japonicus, distributed in East Asia.

We developed microsatellite markers for Appasus japonicus (Heteroptera: Belostomatidae). This belostomatid bug is distributed in East Asia (Japanese Archipelago, Korean Peninsula and mainland China) and often listed as an endangered species in the Red List or the Red Data Book at the national and local level in Japan. Here, we describe twenty novel polymorphic microsatellite loci developed for A. japonicus, and marker suitability was evaluated using 56 individuals from four A. japonicus populations (Nagano, Hiroshima and Yamaguchi prefectures, Japan, and Chungcheongnam-do, Korea). The number of alleles per locus ranged from 1 to 12 (mean = 2.5), and the average observed and expected heterozygosity and fixation index per locus were 0.270, 0.323 and 0.153, respectively. In addition, a population structure analysis was conducted using the software STRUCTURE, and its result suggested that the 20 markers described here will be useful for investigating the genetic structure of A. japonicus populations, which should contribute to population genetics studies of this species.

Impact of Photo-Excitation on Leakage Current and Negative Bias Instability in InSnZnO Thickness-Varied Thin-Film Transistors.

InSnZnO thin-film transistors (ITZO TFTs), having high carrier mobility, guarantee the benefits of potential applications in the next generation of super-high-definition flat-panel displays. However, the impact of photo-excitation on the leakage current and negative bias stress (NBIS) of ITZO TFTs must be further explored. In this study, the ITZO thickness (TITZO) is designed to tailor the initial performance of devices, especially for the 100 nm TITZO TFT, producing excellent electrical properties of 44.26 cm2V-1s-1 mobility, 92 mV/dec. subthreshold swing (SS), 0.04 V hysteresis, and 3.93 × 1010 ON/OFF ratio, which are superior to those of the reported ITZO TFTs. In addition, incident light coupled with tunable photon energy is introduced to monitor the leakage current of various TITZO devices. The OFF-current results demonstrate that under the identical photon energy, many more electrons are photo-excited for the thicker TITZO TFTs. NBIS-induced Vth shift and SS deterioration in all TFTs are traced and analyzed in real time. As the TITZO thickens to near Debye length, the degree of degradation is exacerbated. When the thickness further increases, the notorious instability caused by NBIS is effectively suppressed. This study provides an important research basis for the application of ITZO-based TFTs in future displays.

Community Structure of Aquatic Insects Adapted to Lentic Water Environments, and Fine-Scale Analyses of Local Population Structures and the Genetic Structures of an Endangered Giant Water Bug Appasus japonicus.

Environments such as floodplains and the marshlands of rivers, lakes and ponds, are important habitats for aquatic insects adapted to lentic water conditions. In addition, ponds and paddy fields artificially created for agriculture are also important alternative habitats for lotic water-dependent wildlife. In this study, we focused on aquatic insects in ponds in the Matsumoto Basin, located in the center of Japan. Although this is an urbanized area, aquatic animals adapted to floodplains inhabit it at a relatively high density for Japan. We conducted a multifaceted evaluation of the environments of the 33 ponds in this region and conducted a survey of the aquatic insect fauna inhabiting them. In this study, we conducted quantitative sampling, focusing on two insect orders adapted to large-scale lentic water environments (i.e., Heteroptera and Coleoptera), and observed five species of three families and 16 species of five families from the Matsumoto Basin, respectively. Within these species, eight endangered species were included. Furthermore, we carried out a genetic structure analysis for the giant water bug, Appasus japonicus, inhabiting these ponds in high density, and conducted a comparative evaluation of their genetic diversity between these ponds. A total of 530 specimens of A. japonicus were genetically analyzed for the mitochondrial DNA COI region, and 26 haplotypes were observed. The degree of genetic diversity between the ponds was clearly demonstrated. In addition, we discussed the wintering possibilities for the giant water bugs based on their corresponding surrounding environmental factors, and comprehensively discussed their "source-sink" relationships in this region. Therefore, this is a comprehensive study focused on the relevant environmental factors, diversification of their community structures, their population structures, and their genetic structure at a fine scale.

Understanding the Role of Temperature and Drain Current Stress in InSnZnO TFTs with Various Active Layer Thicknesses.

Thin-film transistor (TFT) devices composed of metal oxide semiconductors have attracted tremendous research attention globally in recent years. Owing to their ability to offer mobility, metal oxide semiconductor materials can enable high-performance TFTs for next-generation integrated display devices. Nevertheless, further breakthroughs of metal oxide TFTs are mainly obstructed by their long-term variability, the reason for which is not yet fully understood. Herein, TFTs based on InSnZnO (ITZO) with various thicknesses (TITZO) were prepared and their long-term stabilities under test temperatures and drain current stress were investigated. The results indicate that ITZO TFTs exhibit outstanding electrical properties regardless of the TITZO, including a high saturated mobility of over 35 cm2V-1s-1 and sharp subthreshold swing. Note that the transfer and output characteristic curves of the device with a thick TITZO of 100 nm express an abnormal current surge when high gate and drain voltages are exerted, which is attributed to the floating body effect, caused when the imposed electric field induces impact ionization near the drain side. More interestingly, these drain current stress results further suggest that the abnormal shift behavior of the electrical properties of the ITZO TFTs with a TITZO of greater than 75 nm is observed to deteriorate gradually with increasing temperature and drain current bias. This study addresses that such a degradation effect should be restrained for the operation of high-mobility devices.

Genealogical Position of Japanese Populations of the Globally Distributed Mayfly Cloeon dipterum and Related Species (Ephemeroptera, Baetidae): A Molecular Phylogeographic Analysis.

In the present study, we add genetic data of the mayfly Cloeon dipterum collected from the Japanese Islands to the established molecular phylogenetic knowledge in the mitochondrial COI gene of Cloeon mayflies. Cloeon dipterum is a typical cosmopolitan species that includes six intraspecific haplotype groups. The present phylogenetic analysis revealed that haplotypes of the Japanese C. dipterum constitute a seventh group together with a haplotype from Korea. This East Asian group forms a sister group with previously known European and North American haplotype groups (i.e., the clade CT1 to CT3). The present phylogenetic analysis further revealed the occurrence of two described species (C. dipterum and Cloeon ryogokuensis) and possibly three species (Cloeon sp. 1 to 3) in Japan. Consideration is given to the degree of genetic differentiation, divergence time, and differentiation process among these seven genetic groups.

Construction of sinusoid-scale microvessels in perfusion culture of a decellularized liver.

There is a great deal of demand for the construction of transplantable liver grafts. Over the last decade, decellularization techniques have been developed to construct whole liver tissue grafts as potential biomaterials. However, the lack of intact vascular networks, especially sinusoids, in recellularized liver scaffolds leads to hemorrhage and thrombosis after transplantation, which is a major obstacle to the development of transplantable liver grafts. In the present study, we hypothesized that both mechanical (e.g., fluid shear stress) and chemical factors (e.g., fibronectin coating) can enhance the formation of hierarchical vascular networks including sinusoid-scale microvessels. We demonstrated that perfusion culture promoted formation of sinusoid-scale microvessels in recellularized liver scaffolds, which was not observed in static culture. In particular, perfusion culture at 4.7 ml/min promoted the formation of sinusoid-scale microvessels compared to perfusion culture at 2.4 and 9.4 ml/min. In addition, well-aligned endothelium was observed in perfusion culture, suggesting that endothelial cells sensed the flow-induced shear stress. Moreover, fibronectin coating of decellularized liver scaffolds enhanced the formation of sinusoid-scale microvessels in perfusion culture at 4.7 ml/min. This study represents a critical step in the development of functional recellularized liver scaffolds, which can be used not only for transplantation but also for drug screening and disease-modeling studies. STATEMENT OF SIGNIFICANCE: Decellularized liver scaffolds are promising biomaterials that allow production of large-scale tissue-engineered liver grafts. However, it is difficult to maintain recellularized liver grafts after transplantation due to hemorrhage and thrombosis. To overcome this obstacle, construction of an intact vascular network including sinusoid-scale microvessels is essential. In the present study, we succeeded in constructing sinusoid-scale microvessels in decellularized liver scaffolds via a combination of perfusion culture and surface coating. We further confirmed that endothelial cells in decellularized liver scaffolds responded to flow-derived mechanical stress by aligning actin filaments. Our strategy to construct sinusoid-scale microvessels is critical for the development of intact vascular networks, and addresses the limitations of recellularized liver scaffolds after transplantation.

Detection of an endangered aquatic heteropteran using environmental DNA in a wetland ecosystem.

The use of environmental DNA (eDNA) has recently been employed to evaluate the distribution of various aquatic macroorganisms. Although this technique has been applied to a broad range of taxa, from vertebrates to invertebrates, its application is limited for aquatic insects such as aquatic heteropterans. Nepa hoffmanni (Heteroptera: Nepidae) is a small (approx. 23 mm) aquatic heteropteran that inhabits wetlands, can be difficult to capture and is endangered in Japan. The molecular tool eDNA was used to evaluate the species distribution of N. hoffmanni in comparison to that determined using hand-capturing methods in two regions of Japan. The eDNA of N. hoffmanni was detected at nearly all sites (10 eDNA-detected sites out of 14 sites), including sites where N. hoffmanni was not captured by hand (five eDNA-detected sites out of six captured sites). Thus, this species-specific eDNA technique can be applied to detect small, sparsely distributed heteropterans in wetland ecosystems. In conclusion, eDNA could be a valuable technique for the detection of aquatic insects inhabiting wetland habitats, and could make a significant contribution to providing distribution data necessary to species conservation.

Excited state properties of a short π-electron conjugated peridinin analogue.

C29-peridinin is a synthetic analogue of the important, naturally-occurring carotenoid, peridinin, found in several marine algal species. C29-peridinin has five conjugated carbon-carbon double bonds compared to eight possessed by peridinin and also lacks the methyl group functionalities typically present along the polyene chain of carotenoids. These structural modifications lead to unique excited state properties and important insights regarding the factors controlling the photophysics of peridinin and other carbonyl-containing carotenoids, which are critical components of the light-harvesting systems of many photosynthetic organisms.

Superconductivity in transparent zinc-doped In2O3 films having low carrier density.

Thin polycrystalline zinc-doped indium oxide (In2O3-ZnO) films were prepared by post-annealing amorphous films with various weight concentrations x of ZnO in the range 0⩽x ⩽0.06. We have studied the dependences of the resistivity ρ and Hall coefficient on temperature T and magnetic field H in the range 0.5⩽T ⩽300 K, H⩽6 Tfor 350 nm films annealed in air. Films with 0⩽x⩽0.03 show the superconducting resistive transition. The transition temperature Tc is below 3.3 K and the carrier density n is about 1025-1026 m-3. The annealed In2O3-ZnO films were examined by transmission electron microscopy and x-ray diffraction analysis revealing that the crystallinity of the films depends on the annealing time. We studied the upper critical magnetic field Hc2 (T) for the film with x = 0.01. From the slope of dHc2 /dT, we obtain the coherence length ξ (0) ≈ 10 nm at T = 0 K and a coefficient of electronic heat capacity that is small compared with those of other oxide materials.