Quiz-style online training tool helps to learn birdsong identification and support citizen science.

Citizen science is an important approach to monitoring for biodiversity conservation because it allows for data acquisition or analysis on a scale that is not possible for researchers alone. In citizen science projects, the use of online training is increasing to improve such skills. However, the effectiveness of quiz-style online training, assumed to be efficient to enhance participants' skills, has not been evaluated adequately on species identification for citizen science biodiversity monitoring projects. Memory mechanisms in adaptive learning were hypothesized to guide the development of quiz-based online training tools for learning birdsong identification and for improving interest in birds and natural environments. To examine the hypothesis, we developed a quiz-style online training tool called TORI-TORE. We experimentally applied TORI-TORE in Fukushima, Japan, and examined its effectiveness for bird identification training using test scores and questionnaires to determine participants' attitudes in a randomized control trial. We obtained the following key results: (1) TORI-TORE had positive effects on test scores and trainees' attitudes toward birds. (2) Adaptive training, in which questions focused preferentially on unmastered bird species based on the answer history of individual trainees inspired by adaptive learning, unexpectedly led to lower scores and satisfaction in TORI-TORE. (3) Focusing on species that are relatively easy to remember, short lag times between training and testing, and long question intervals positively affected scores. While there is room for improvement, we expect TORI-TORE to contribute to online capacity building and to increase interest in natural environments.

Identification of a novel nonsense NOG mutation in a patient with stapes ankylosis and symphalangism spectrum disorder.

Multiple bone disorders due to mutations in the human noggin (NOG) causes a variety of phenotypes. Hearing impairment due to stapes ankylosis secondary to bony degeneration is also a feature of these syndromes. We describe the case of an individual in a Japanese family with conductive hearing loss due to stapes ankylosis and hyperopia and dactylosymphysis. We revealed a novel NOG mutation, NM_005450.6:c.222 C > A / p.Tyr74*, and confirmed genetic significance.

Pneumococcal conjugate vaccines reduce myringotomy with tympanostomy tube insertion in young children in Japan.

OBJECTIVE: Pneumococcal conjugate vaccines (PCVs) have been reported to reduce the incidence of myringotomy with tympanostomy tube insertion (MTTI) in children. However, little information is available focusing specific ages. We examined the prophylactic efficacy of PCVs on the onset of complex otitis media (ComOM) that requires MTTI. METHOD: From 2011, the public support for PCV7 started with the usual four-dose schedule and an emergency schedule for 2- to 4-year-old children in Japan. PCV7 was replaced with PCV13 in 2013. We reviewed the nationwide database obtained from the JMDC Claims Database (https://www.jmdc.co.jp/en/) to examine the MTTI incidence during the era before and after PCV introduction (from 2008 to 2010 and from 2011 to 2017, respectively). Subjects were analyzed by stratified age groups (from 0 to 8 years old) and in subdivided groups of 6 months (from 0 to 35 months old). We compared the MTTI incidence between the groups for each age as well as between those for each calendar year. RESULTS: A significant reduction in the MTTI incidence was detected in the 1-year-old children of the PCV era compared to those of the pre-PCV era. The reduction rates were more prominent in the 12-17 months group as compared to the 18-23 months group (PCV7 p = .005 and PCV13 p = .011, PCV7 p = .014 and PCV13 p = .153, respectively). The significant difference in the 1-year-old children continued in six of seven calendar years from 2011 to 2017, whereas no significant reduction was detected in children >3 years old. CONCLUSIONS: The introduction of both PCV7 and PCV13 reduced MTTI incidences in children around 1 year old, and the effects were more prominent during the early half-periods. Our results support etiological evidence that pneumococcal infection in children aged 1 year and younger might play roles in the pathogenesis of ComOM that requires MTTI.

Identification of a Novel Copy Number Variation of EYA4 Causing Autosomal Dominant Non-syndromic Hearing Loss.

OBJECTIVE: Eyes absent 4 (EYA4) is the causative gene of autosomal dominant non-syndromic hereditary hearing loss, DFNA10. We aimed to identify a copy number variation of EYA4 in a non-syndromic sensory neural hearing loss pedigree. FAMILY AND CLINICAL EVALUATION: A Japanese family showing late-onset and progressive hearing loss was evaluated. A pattern of autosomal dominant inheritance of hearing loss was recognized in the pedigree. No cardiac disease was observed in any of the individuals. METHODS: Targeted exon sequencing was performed using massively parallel DNA sequencing (MPS) analysis. Scanning of the array comparative genomic hybridization (aCGH) was completed and the copy number variation (CNV) data from the aCGH analysis was confirmed by matching all CNV calls with MPS analysis. Breakpoint detection was performed by whole-genome sequencing and direct sequencing. Sequencing results were examined, and co-segregation analysis of hearing loss was completed. RESULTS: We identified a novel hemizygous indel that showed CNV in the EYA4 gene from the position 133,457,057 to 133,469,892 on chromosome 6 (build GRCh38/hg38) predicted as p.(Val124_Pro323del), and that was segregated with post-lingual and progressive autosomal dominant sensorineural hearing loss by aCGH analysis. CONCLUSION: Based on the theory of genotype-phenotype correlation with EYA4 mutations in terms of hearing loss and comorbid dilated cardiomyopathy, the region of amino acids 124 to 343 is hypothesized not to be the pathogenic region causing dilated cardiomyopathy. Additionally, the theory of genotype-phenotype correlation about the prevalence of dilated cardiomyopathy is thought to be rejected because of no correlation of deleted amino acid region with the prevalence of dilated cardiomyopathy. These results will help expand the research on both the coordination of cochlear transcriptional regulation and normal cardiac gene regulation via EYA4 transcripts and provide information on the genotype-phenotype correlations of DFNA10 hearing loss.

Characterization of the human E2F4 promoter region and its response to 12-O-tetradecanoylphorbol-13-acetate.

The E2F transcription factors (TFs), which control the progression of the cell cycle in response to DNA-damage and various stresses, are known to interact with a tumour suppressor, Retinoblastoma 1 (RB1). We previously showed that the response of the human RB1 promoter to a 12-O-tetradecanoylphorbol-13-acetate (TPA) in HL-60 cells is mediated by a duplicated GGAA motif, which is also present in the 5'-upstream of the E2F family genes. The motifs are especially rich in the 5'-upstream of the E2F4 gene. In the present study, we constructed luciferase (Luc) expression vectors containing a 466 bp of the 5'-upstream of the human E2F4 gene. The transfection of this plasmid and deletion/mutation-introduced derivatives into HL-60 cells and a Luc reporter assay showed that duplicated and triplicated GGAA (TTCC) motifs in the E2F4 promoter respond to TPA. As expected, electrophoretic mobility shift assay indicated that SPI1 (PU.1) binds to the GGAA motif-containing element. A quantitative RT-PCR and western blotting showed that the E2F4 transcripts and its encoding proteins accumulate during the differentiation of HL-60 into macrophage-like cells. In contrast, the expression of the E2F1 gene and the protein, which possibly acts as a cell cycle accelerator, was greatly diminished.

Catalyst-Selective Growth of Single-Orientation Hexagonal Boron Nitride toward High-Performance Atomically Thin Electric Barriers.

The ability to control the crystal orientation of 2D van der Waals (vdW) layered materials grown on large-scale substrates is crucial for tailoring their electrical properties, as well as for integration of functional 2D devices. In general, multiple orientations, i.e., two or four orientations, appear through the crystal rotational symmetry matching between the material and its substrate. Here, it is reported that hexagonal boron nitride (h-BN), an ideal electric barrier in the family of 2D materials, has a single orientation on inclined Cu (1 0 1) surfaces, where the Cu planes are tilted from the (1 0 1) facet around specific in-plane axes. Density functional theory (DFT) calculation indicates that this is a manifestation of only one favored h-BN orientation with the minimum vdW energy on the inclined Cu (1 0 1) surface. Moreover, thanks to the high interfacial strength with the underlying Cu, the single-orientation h-BN is free of thermal wrinkles, and exhibits a spatially homogeneous morphology and tunnel conductance. The findings point to a feasible approach to direct growth of single-orientation, wrinkle-free h-BN thin film for high-performance 2D electrical devices, and will be of benefit for controllable synthesis of other vdW materials.

Self-Folded Three-Dimensional Graphene with a Tunable Shape and Conductivity.

Three-dimensional (3D) graphene architectures are of great interest as applications in flexible electronics and biointerfaces. In this study, we demonstrate the facile formation of predetermined 3D polymeric microstructures simply by transferring monolayer graphene. The graphene adheres to the surface of polymeric films via noncovalent π-π stacking bonding and induces a sloped internal strain, leading to the self-rolling of 3D microscale architectures. Micropatterns and varied thicknesses of the 2D films prior to the self-rolling allows for control over the resulting 3D geometries. The strain then present on the hexagonal unit cell of the graphene produces a nonlinear electrical conductivity across the device. The driving force behind the self-folding process arises from the reconfiguration of the molecules within the crystalline materials. We believe that this effective and versatile way of realizing a 3D graphene structure is potentially applicable to alternative 2D layered materials as well as other flexible polymeric templates.

Atomically Thin Graphene Windows That Enable High Contrast Electron Microscopy without a Specimen Vacuum Chamber.

Scanning electron microscopes (SEMs) require a high vacuum environment to generate and shape an electron beam for imaging; however, the vacuum conditions greatly limit the nature of specimens that can be examined. From a purely scattering physics perspective, it is not necessary to place the specimen inside the vacuum chamber-the mean free paths (MFPs) for electron scattering in air at typical SEM beam voltages are 50-100 µm. This is the idea behind the airSEM, which removes the specimen vacuum chamber from the SEM and places the sample in air. The thickness of the gas layer is less than a MFP from an electron-transparent window to preserve the shape and resolution of the incident beam, resulting in comparable imaging quality to an all-vacuum SEM. Present silicon nitride windows scatter far more strongly than the air gap and are currently the contrast and resolution limiting factor in the airSEM. Graphene windows have been used previously to wrap or seal samples in vacuum for imaging. Here we demonstrate the use of a robust bilayer graphene window for sealing the electron optics from the room environment, providing an electron transparent window with only a 2% drop in contrast. There is a 5-fold-increase in signal/noise ratio for imaging compared to multi-MFP-thick silicon nitride windows, enabling high contrast in backscattered, transmission, and surface imaging modes for the new airSEM geometry.

Inductive Effect of Palmatine on Apoptosis in RAW 264.7 Cells.

Osteoporosis is a serious public health problem characterized by low bone density and deterioration of the bone microarchitecture. Current treatment options target either osteoclast resorption or osteoblast formation. It has been reported that berberine, a close structural analog of palmatine, inhibited bone loss in an osteoporosis model. In this study, osseous metabolism was observed in vitro with osteoclast bone resorbing cells. We proved that mouse preosteoclastic cell line (RAW 264.7) has a higher sensitivity to palmatine than mouse osteoblastic cell line (MC3T3-E1); the cell survival rates significantly decreased at 40 µM palmatine. The NO2 (-) level, a metabolic product of nitric monoxide (NO), and iNOS mRNA expression, an osteoclast with NO induced enzyme, also increased with higher dosage of palmatine. Furthermore, it was recognized that the cell viability decrease from palmatine was caused by apoptosis rather than necrosis. Additionally, osteoclast apoptosis from palmatine did not occur when iNOS was inhibited with N(G)-nitro-L-arginine methyl ester hydrochloride (pan NOS inhibitor). These results indicate that palmatine plays an important role in osteoclast apoptosis via the NOS system. Hence, palmatine could be considered as a viable pharmaceutical candidate for osteoporosis bone resorption inhibitor.

Chiral atomically thin films.

Chiral materials possess left- and right-handed counterparts linked by mirror symmetry. These materials are useful for advanced applications in polarization optics, stereochemistry and spintronics. In particular, the realization of spatially uniform chiral films with atomic-scale control of their handedness could provide a powerful means for developing nanodevices with novel chiral properties. However, previous approaches based on natural or grown films, or arrays of fabricated building blocks, could not offer a direct means to program intrinsic chiral properties of the film on the atomic scale. Here, we report a chiral stacking approach, where two-dimensional materials are positioned layer-by-layer with precise control of the interlayer rotation (θ) and polarity, resulting in tunable chiral properties of the final stack. Using this method, we produce left- and right-handed bilayer graphene, that is, a two-atom-thick chiral film. The film displays one of the highest intrinsic ellipticity values (6.5 deg µm(-1)) ever reported, and a remarkably strong circular dichroism (CD) with the peak energy and sign tuned by θ and polarity. We show that these chiral properties originate from the large in-plane magnetic moment associated with the interlayer optical transition. Furthermore, we show that we can program the chiral properties of atomically thin films layer-by-layer by producing three-layer graphene films with structurally controlled CD spectra.

Influence of Palmatine on Bone Metabolism in Ovariectomized Mice and Cytokine Secretion of Osteoblasts.

BACKGROUND/AIM: Receptor activator of nuclear factor-κB ligand (RANKL) and osteoprotegerin (OPG), regulate the cognate receptor RANK on osteoclast precursor cells. Herein we examined the inhibitory effects of palmatine on bone metabolism using ovariectomized (OVX) mice. MATERIALS AND METHODS: The first experimentaI set was designed to histologically and biochemically examine mice randomly divided into four groups: sham-operated, OVX, and OVX-palmatine intake groups (1 mg/kg and 10 mg/kg). The second experimental set examined the influence of palmatine on osteoblast-like cells in vitro. RESULTS: Palmatine caused significant suppression of osteoclast numbers in tissues. In palmatine-treated mice, RANKL and OPG expression decreased. In the culture supernatant of MC3T3-E1 cells, RANKL and OPG levels were significantly reduced by palmatine addition. CONCLUSION: Palmatine may attenuate osteoclast differentiation through inhibition of RANKL and OPG expression by osteoblasts. Therefore, palmatine might be a candidate anti-resorptive agent for osteoporosis therapy.

Polycrystalline graphene with single crystalline electronic structure.

We report the scalable growth of aligned graphene and hexagonal boron nitride on commercial copper foils, where each film originates from multiple nucleations yet exhibits a single orientation. Thorough characterization of our graphene reveals uniform crystallographic and electronic structures on length scales ranging from nanometers to tens of centimeters. As we demonstrate with artificial twisted graphene bilayers, these inexpensive and versatile films are ideal building blocks for large-scale layered heterostructures with angle-tunable optoelectronic properties.

Structure and transport properties of the interface between CVD-grown graphene domains.

During the chemical vapor deposition (CVD) growth of graphene, graphene domains grown on a Cu surface merge together and form a uniform graphene sheet. For high-performance electronics and other applications, it is important to understand the interfacial structure of the merged domains, as well as their influence on the physical properties of graphene. We synthesized large hexagonal graphene domains with controlled orientations on a heteroepitaxial Cu film and studied the structure and properties of the interfaces between the domains mainly merged with the same angle. Although the merged domains have various interfaces with/without wrinkles and/or increased defect-related Raman D-band intensity, the intra-domain transport showed higher carrier mobility reaching 20,000 cm(2) V(-1) s(-1) on SiO2 at 280 K (the mean value was 7200 cm(2) V(-1) s(-1)) than that measured for inter-domain areas, 6400 cm(2) V(-1) s(-1) (mean value 2000 cm(2) V(-1) s(-1)). The temperature dependence of the mobility suggests that impurity scattering dominates at the interface even for the merged domains with the same orientation. This study highlights the importance of domain interfaces, especially on the carrier transport properties, in CVD-grown graphene.

Lattice-oriented catalytic growth of graphene nanoribbons on heteroepitaxial nickel films.

Graphene nanoribbons (GNRs) are a promising material for electronic applications, because quantum confinement in a one-dimensional nanostructure can potentially open the band gap of graphene. However, it is still a challenge to synthesize high-quality GNRs by a bottom-up approach without relying on lithographic techniques. In this work, we demonstrate lattice-oriented catalytic growth of single-layer GNRs on the surface of a heteroepitaxial Ni film. Catalytic decomposition of a poly(methyl methacrylate) film on the Ni(100) film at 1000 °C gives narrow nanoribbons with widths of 20-30 nm, which are aligned along either [011] or [011] directions of the Ni lattice. Furthermore, low-energy electron microscope (LEEM) analysis reveals that orientation of carbon hexagons in these GNRs is highly controlled by the underlying Ni(100) lattice, leading to the formation of zigzag edges. This heteroepitaxial approach would pave a way to synthesize nanoribbons with controlled orientation for future development of electronic devices based on graphene nanostructures.

Dense arrays of highly aligned graphene nanoribbons produced by substrate-controlled metal-assisted etching of graphene.

Dense arrays of aligned graphene nanoribbons (GNRs) are fabricated by substrate-controlled etching of large-area single-layer graphene. An adequate choice of etching substrate and catalyst deposition method allows densities up to 25 nanoribbons µm(-1) to be obtained with average widths of 19 nm. The efficacy of the method is evidenced by the high on/off ratios of back-gated transistors made with these GNRs, which can go up to 5000.

Catalytic Growth of Graphene: Toward Large-Area Single-Crystalline Graphene.

For electronic applications, synthesis of large-area, single-layer graphene with high crystallinity is required. One of the most promising and widely employed methods is chemical vapor deposition (CVD) using Cu foil/film as the catalyst. However, the CVD graphene is generally polycrystalline and contains a significant amount of domain boundaries that limit intrinsic physical properties of graphene. In this Perspective, we discuss the growth mechanism of graphene on a Cu catalyst and review recent development in the observation and control of the domain structure of graphene. We emphasize the importance of the growth condition and crystallinity of the Cu catalyst for the realization of large-area, single-crystalline graphene.