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Key Message: A novel fluorescent i-motif DNA silver nanoclusters system has been developed for visualization of reactive oxygen species in plants, enabling the detection of intracellular signaling in plant cells. Reactive oxygen species (ROS) are crucial in plant growth, defense, and stress responses, making them vital for improving crop resilience. Various ROS sensing methods for plants have been developed to detect ROS in vitro and in vivo. However, each method comes its own advantages and disadvantages, leading to an increasing demand for a simple and effective sensory system for ROS detection in plants. Here, we introduce novel DNA silver nanoclusters (DNA/AgNCs) sensors for visualizing ROS in plants. Two sensors, C 20 /AgNCs and FAM-C 20 /AgNCs-Cy5, detect intracellular ROS signaling in response to stimuli, such as abscisic acid, salicylic acid, ethylene, and bacterial peptide elicitor flg22. Notably, FAM-C 20 /AgNCs-Cy5 exceeds the sensing capabilities of HyPer7, a widely recognized ROS sensor. Taken together, we suggest that fluorescent i-motif DNA/AgNCs system is an effective tool for visualizing ROS signals in plant cells. This advancement is important to advancing our understanding of ROS-mediated processes in plant biology., Competing Interests: Declarations. Conflict of interest: No conflict of interest was declared., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

A low-cost and indium-tin-oxide (ITO)-free electrode-based flexible perovskite solar cell (PSC) that can be fabricated by roll-to-roll processing shall be developed for successful commercialization. High processing temperatures present a challenge for the PSC fabrication on flexible substrates. The most efficient planar n-i-p PSC structures, which utilize a metal oxide as an electron transport layer (ETL), necessitate high annealing temperatures. In addition, the device performance deteriorates owing to the migration of halogen ions, which causes the oxidation of the metal electrodes. These drawbacks conflict with the development of highly efficient flexible PSCs fabricated on ITO-free transparent electrodes. Herein, an efficient ETL material that enables low-temperature processing is presented. Tin dioxide (SnO 2 ) is modified by (sulfobetaine-N,N-dimethylamino)propyl naphthalene diimide (NDI-B) and used as an ETL. The NDI-B effectively reduces the interfacial nonradiative recombination between the ETL and perovskite and suppresses the ion migration by passivating oxygen-vacancy defects in SnO 2 and strongly interacting with halogen ions, respectively. Based on the NDI-B-blended SnO 2 ETL, a record PCE of 17.48% is achieved in the ITO-free flexible PSC fabricated at low temperature., (© 2024 Wiley‐VCH GmbH.)

Metallic nanogaps have emerged as a versatile platform for realizing ultrastrong coupling in terahertz frequencies. Increasing the coupling strength generally involved reducing the gap width to minimize the mode volume, which presents challenges in fabrication and efficient material coupling. Here, we propose employing terahertz nanoslots, which can efficiently squeeze the mode volume in an extra dimension alongside the gap width. Our experiments using 500 nm wide nanoslots integrated with an organic-inorganic hybrid perovskite demonstrate ultrastrong phonon-photon coupling with a record-high Rabi splitting of 48% of the original resonance (Ω = 0.48ω 0 ), despite having a gap width 5 times larger than previously reported structures with Ω = 0.45ω 0 . Mechanisms underlying this effective light--matter coupling are investigated with simulations using coupled mode theory. Moreover, bulk polariton analyses reveal that our results account for 68% of the theoretical maximum Rabi splitting, with the potential to reach 82% through further optimization of the nanoslots.

Interface properties between charge transport and perovskite light-absorbing layers have a significant impact on the power conversion efficiency (PCE) of perovskite solar cells (PSCs). Poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS) is a polyelectrolyte composite that is widely used as a hole transport layer (HTL) to facilitate hole transport from a perovskite layer to an anode. However, PEDOT:PSS must be modified using a functional additive because PSCs with a pristine PEDOT:PSS HTL do not exhibit a high PCE. Herein, we demonstrate an increase in the PCE of PSCs with a polyethylene glycol hexadecyl ether (Brij C10)-mixed PEDOT:PSS HTL. Photoelectron spectroscopy results show that the Brij C10 content becomes significantly high in the HTL surface composition with an increase in the Brij C10 concentration (0-5 wt%). The enhanced PSC performance, e.g., a PCE increase from 8.05 to 11.40%, is attributed to the reduction in non-radiative recombination at the interface between PEDOT:PSS and perovskite by the insulating Brij C10. These results indicate that the suppression of interface recombination is essential for attaining a high PCE for PSCs.

Importance: Moral injury and distress (MID), which occurs when individuals have significant dissonance with their belief system and overwhelming feelings of being powerless to do what is believed to be right, has not been explored in the unique population of military surgeons deployed far forward in active combat settings. Deployed military surgeons provide care to both injured soldiers and civilians under command-driven medical rules of engagement (MROE) in variably resourced settings. This practice setting has no civilian corollary for comparison or current specific tool for measurement., Objective: To characterize MID among military surgeons deployed during periods of high casualty volumes through a mixed-methods approach., Design, Setting, and Participants: This qualitative study using convergent mixed methods was performed from May 2020 to October 2020. Participants included US military surgeons who had combat deployments to a far-forward role 2 treatment facility during predefined peak casualty periods in Iraq (2003-2008) and Afghanistan (2009-2012), as identified by purposeful snowball sampling. Data analysis was performed from October 2020 to May 2021., Main Outcomes and Measures: Measure of Moral Distress for Healthcare Professionals (MMD-HP) survey and individual, semistructured interviews were conducted to thematic saturation., Results: The total cohort included 20 surgeons (mean [SD] age, 38.1 [5.2] years); 16 (80%) were male, and 16 (80%) had 0 or 1 prior deployment. Deployment locations were Afghanistan (11 surgeons [55%]), Iraq (9 surgeons [45%]), or both locations (3 surgeons [15%]). The mean (SD) MMD-HP score for the surgeons was 104.1 (39.3). The primary thematic domains for MID were distressing outcomes (DO) and MROE. The major subdomains of DO were guilt related to witnessing horrific injuries; treating pregnant women, children, and US soldiers; and second-guessing decisions. The major subdomains for MROE were forced transfer of civilian patients, limited capabilities and resources, inexperience in specialty surgical procedures, and communication with command. Postdeployment manifestations of MID were common and affected sleep, medical practice, and interpersonal relationships., Conclusions and Relevance: In this qualitative study, MID was ubiquitous in deployed military surgeons. Thematic observations about MID, specifically concerning the domains of DO and MROE, may represent targets for further study to develop an evaluation tool of MID in this population and inform possible programs for identification and mitigation of MID.

We report the improvement in photoelectrochemical water splitting (PEC-WS) by controlling migration kinetics of photo-generated carriers using InGaN/GaN hetero-structure nanowires (HSNWs) as a photocathode (PC) material. The InGaN/GaN HSNWs were formed by first growing GaN nanowires (NWs) on an Si substrate and then forming InGaN NWs thereon. The InGaN/GaN HSNWs can cause the accumulation of photo-generated carriers in InGaN due to the potential barrier formed at the hetero-interface between InGaN and GaN, to increase directional migration towards electrolyte rather than the Si substrate, and consequently to contribute more to the PEC-WS reaction with electrolyte. The PEC-WS using the InGaN/GaN-HSNW PC shows the current density of 12.6 mA/cm 2 at -1 V versus reversible hydrogen electrode (RHE) and applied-bias photon-to-current conversion efficiency of 3.3% at -0.9 V versus RHE. The high-performance PEC-WS using the InGaN/GaN HSNWs can be explained by the increase in the reaction probability of carriers at the interface between InGaN NWs and electrolyte, which was analyzed by electrical resistance and capacitance values defined therein.

Objective: This study is an investigator-initiated, prospective, randomized, controlled study to evaluate the efficacy and safety of the combined use of recombinant human BMP-2 (rhBMP-2) and a hydroxyapatite (HA) carrier in multilevel fusion in patients with adult spinal deformity (ASD)., Methods: Thirty patients underwent posterolateral fusion for lumbar spinal deformities at 3 to 5 segments between L1 and S1. The patients received rhBMP-2+HA or HA on the left or right side of the transverse processes. They were followed up regularly at 1, 3, 6, and 12 months postoperatively. Fusion was defined according to the bone bridging on computed tomography scans. The fusion rate per segment was subanalyzed. Function and quality of life as well as pain in the lower back and lower extremities were evaluated., Results: The union rate for the rhBMP-2+HA group was 100% at 6 and 12 months. The union rate for the HA group was 77.8% (21 of 27) at 6 months and 88.0% (22 of 25) at 12 months (p = 0.014 at 6 months; not significant at 12 months). All segments were fused at 6 and 12 months in the rhBMP-2+HA group (p < 0.001). In the HA group, 108 of 115 segments (93.5%) were fused at 6 months and 105 of 109 segments (96.3%) at 12 months. Other clinical parameters (visual analogue scale, 36-item Short Form Health Survey, and Scoliosis Research Society-22 scores) improved compared to baseline., Conclusion: Combining rhBMP-2 and an HA carrier is a safe and effective method to achieve multilevel fusion in patients with ASD.

We analyzed the changes in patients’ clinical characteristics and transport refusal pre- and post-COVID-19 and identified the reasons for transport refusal using emergency medical services run sheet data from pre-COVID-19 (April−December 2019) and post-COVID-19 (April−December 2020) in Gyeonggi Province, South Korea. We included patients aged ≥18 years. Univariate and multivariate logistic regression analyses were performed to identify the relationship between patients’ personal factors and clinical characteristics and emergency transport refusal. During the control and study periods, 612,681 cases were reported; the transport refusal rates during the control and study periods were 6.7% and 8.2%, respectively. Emergency transport refusal was associated with younger age, the male sex, a normal mental status, a shock index < 1, and trauma in both the pre- and post-COVID-19 periods. Although fever prevented transport refusal during the pre-COVID-19 period (aOR, 0.620; 95% CI, 0.567−0.679), it became a significant risk factor for transport refusal during the post-COVID-19 period (aOR, 1.619; 95% CI, 1.534−1.709). The most common reason for transport refusal by critically ill patients was “because it was not accepted within the jurisdiction and remote transport was required.” It is necessary to expand the response capacity of patients with fever in the community to reduce the refusal of transport by critically ill patients.

The core plant microprocessor consists of DICER-LIKE 1 (DCL1), SERRATE (SE), and HYPONASTIC LEAVES 1 (HYL1) and plays a pivotal role in microRNA (miRNA) biogenesis. However, the proteolytic regulation of each component remains elusive. Here, we show that HYL1-CLEAVAGE SUBTILASE 1 (HCS1) is a cytoplasmic protease for HYL1-destabilization. HCS1-excessiveness reduces HYL1 that disrupts miRNA biogenesis, while HCS1-deficiency accumulates HYL1. Consistently, we identified the HYL1 K154A mutant that is insensitive to the proteolytic activity of HCS1, confirming the importance of HCS1 in HYL1 proteostasis. Moreover, HCS1-activity is regulated by light/dark transition. Under light, cytoplasmic CONSTITUTIVE PHOTOMORPHOGENIC 1 (COP1) E3 ligase suppresses HCS1-activity. COP1 sterically inhibits HCS1 by obstructing HYL1 access into the catalytic sites of HCS1. In contrast, darkness unshackles HCS1-activity for HYL1-destabilization due to nuclear COP1 relocation. Overall, the COP1-HYL1-HCS1 network may integrate two essential cellular pathways: the miRNA-biogenetic pathway and light signaling pathway., Competing Interests: The authors declare no competing interest., (Copyright © 2022 the Author(s). Published by PNAS.)

The optical properties of InGaN/GaN green light-emitting diodes (LEDs) with an undoped graded short-period superlattice (GSL) and a Si-doped GSL (SiGSL) were investigated using photoluminescence (PL) and time-resolved PL spectroscopies. For comparison, an InGaN/GaN conventional LED (CLED) without the GSL structure was also grown. The SiGSL sample showed the strongest PL intensity and the largest PL peak energy because of band-filling effect and weakened quantum- confined stark effect (QCSE). PL decay time of SiGSL sample at 10 K was shorter than those of the CLED and GSL samples. This finding was attributed to the oscillator strength enhancement by the reduced QCSE due to the Coulomb screening by Si donors. In addition, the SiGSL sample exhibited the longest decay time at 300 K, which was ascribed to the reduced defect and dislocation density. These results indicate that insertion of the Si-doped GSL structure is an effective strategy for improving the optical properties in InGaN/GaN green LEDs.

Potential strategies such as surface passivation and perovskite material halide mixing may protect material surfaces, improve luminescence, and reduce charge traps for device stability. In this study, we used deep level transient spectroscopy to investigate the effect of CdSe/ZnS core-shell quantum dots (QDs) on defect states and carrier transport in methylammonium (MA) lead halide perovskites (CH 3 NH 3 PbX 3 where X  = I, Br). In MAPbI 3 and MAPbI 2 Br films with CdSe/ZnS QDs, the density of hole traps located at E v  + 0.37 eV and E v  + 0.56 eV was reduced dramatically. Deep traps at E v  + 0.78 eV and E v  + 1.08 eV were removed, and one broad electron trap signal dominated. Film photoresponsivity under 600-nm wavelength light and a bias voltage of -0.7 V was 10 and 18 mA/W, which is 100 and 27 times larger than the 0.1 and 0.67 mA/W of bare perovskites (PS), respectively. This demonstrates that carrier transport was enhanced due to defect suppression. Our findings on defect suppression and photoresponsivity enhancement provide an important direction for optimizing high-performance PS device fabrication., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2021 Elsevier Inc. All rights reserved.)

The shift of dark-grown seedlings into light causes enormous transcriptome changes followed by a dramatic developmental transition. Here, we show that microRNA (miRNA) biogenesis also undergoes regulatory changes during de-etiolation. Etiolated seedlings maintain low levels of primary miRNAs (pri-miRNAs) and miRNA processing core proteins, such as Dicer-like 1, SERRATE, and HYPONASTIC LEAVES 1, whereas during de-etiolation both pri-miRNAs and the processing components accumulate to high levels. However, the levels of most miRNAs do not notably increase in response to light. To reconcile this inconsistency, we demonstrated that an unknown suppressor decreases miRNA-processing activity and light-induced SMALL RNA DEGRADING NUCLEASE 1 shortens the half-life of several miRNAs in de-etiolated seedlings. Taken together, these data suggest a novel mechanism, miRNA-biogenetic inconsistency, which accounts for the intricacy of miRNA biogenesis during de-etiolation. This mechanism is essential for the survival of de-etiolated seedlings after long-term skotomorphogenesis and their optimal adaptation to ever-changing light conditions., (Copyright © 2019 The Author. Published by Elsevier Inc. All rights reserved.)

Surface passivation is an effective method to protect the surfaces and improve the luminescence properties of perovskite (PS) films. CdSe/ZnS core-shell quantum dots (QDs) have been employed for surface passivation of PS films because of their size-dependent tunable bandgaps. Herein, the energy transfer (ET) behavior of CH 3 NH 3 PbI 2 Br PS films covered with CdSe/ZnS QDs (QD/PS hybrid structures) is characterized by using photoluminescence (PL) and time-resolved PL spectroscopy. The PL decay time and the integrated PL intensity of the QD/PS hybrid structure increase compared with those of the bare PS films, owing to ET from the QDs to the PS and reduced charge traps. The ET efficiency increases from ~7% to 63% for the QD/PS hybrid structure when the core diameter of the QDs decreases from 6.5 to 2.7 nm, respectively. This can be explained by the charge transfer rate enhancement due to the control of energy level alignment of QDs. These results allow us to understand fundamental mechanisms such as ET from QDs to PS films as a function of the size of the QD.

Dengue virus (DENV) nonstructural 1 (NS1) protein is a specific and sensitive biomarker for the diagnosis of dengue. In this study, an efficient electrochemical biosensor that uses chemically modified affinity peptides was developed for the detection of dengue virus NS1. A series of amino acid-substituted synthetic peptides was rationally designed, chemically synthesized and covalently immobilized to a gold sensor surface. The sensor performance was monitored via square wave voltammetry (SWV) and electrochemical impedance spectroscopy (EIS). Potential affinity peptides specific for NS1 were chosen according to the dynamic current decrease in SWV experiments. Using circular dichroism, the molar ellipticity of peptides (DGV BP1-BP5) was determined, indicating that they had a mostly similar in random coil structure, not totally identical. Using SWV, DGV BP1 was selected as a promising recognition peptide and limit of detection for NS1 was found to be 1.49 μg/mL by the 3-sigma rule. DGV BP1 showed good specificity and stability for NS1, with low signal interference. The validation of the sensor to detect NS1 proteins was confirmed with four dengue virus culture broth (from serotype 1 to 4) as proof-of-concept. The detection performance of our sensor incorporating DGV BP1 peptides showed a statistically significant difference. These results indicate that this strategy can potentially be used to detect the dengue virus antigen, NS1, and to diagnosis dengue fever within a miniaturized portable device in point-of-care testing., Competing Interests: The authors have declared that no competing interests exist.

The plant protein production system is a platform that can not only reduce production costs but also produce monoclonal antibodies that do not have the risk of residual proteins from the host. However, due to the difference between post-translational processes in plants and animals, there may be a modification in the Fab region of the monoclonal antibody produced in the plant; thus, it is necessary to compare the antigen affinity of this antibody with that of the prototype. In this study, ofatumumab, a fully human anti-CD20 IgG1κ monoclonal antibody used for its non-cross resistance to rituximab, was expressed in Nicotiana benthamiana, and its affinities and efficacies were compared with those of native ofatumumab produced from CHO cells. Two forms of plant ofatumumab (with or without HDEL-tag) were generated and their production yields were compared. The HDEL-tagged ofatumumab was more expressed in plants than the form without HDEL-tag. The specificity of the target recognition of plant-derived ofatumumab was confirmed by mCherry-CD20-expressing HEK cells via immuno-staining, and the capping of CD20 after ofatumumab binding was also confirmed using Ramos B cells. In the functional equivalence tests, the binding affinities and complement-dependent cell cytotoxicity efficacy of plant-ofatumumab-HDEL and plant-ofatumumab without HDEL were significantly reduced compared to those of CHO-derived ofatumumab. Therefore, we suggest that although ofatumumab is not a good candidate as a template for plant-derived monoclonal antibodies because of its decreased affinity when produced in plants, it is an interesting target to study the differences between post-translational modifications in mammals and plants., (Copyright © 2019 Elsevier Inc. All rights reserved.)

A spoof fingerprint was fabricated on paper and applied for a spoofing attack to unlock a smartphone on which a capacitive array of sensors had been embedded with a fingerprint recognition algorithm. Using an inkjet printer with an ink made of carbon nanotubes (CNTs), we printed a spoof fingerprint having an electrical and geometric pattern of ridges and furrows comparable to that of the real fingerprint. With this printed spoof fingerprint, we were able to unlock a smartphone successfully; this was due to the good quality of the printed CNT material, which provided electrical conductivities and structural patterns similar to those of the real fingerprint. This result confirms that inkjet-printing CNTs to fabricate a spoof fingerprint on paper is an easy, simple spoofing route from the real fingerprint and suggests a new method for outputting the physical ridges and furrows on a two-dimensional plane., Competing Interests: The authors declare no competing financial interest.

Here, a method to synthesize cellulose nanofiber biotemplated palladium composite aerogels is presented. Noble metal aerogel synthesis methods often result in fragile aerogels with poor shape control. The use of carboxymethylated cellulose nanofibers (CNFs) to form a covalently bonded hydrogel allows for the reduction of metal ions such as palladium on the CNFs with control over both nanostructure and macroscopic aerogel monolith shape after supercritical drying. Crosslinking the carboxymethylated cellulose nanofibers is achieved using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) in the presence of ethylenediamine. The CNF hydrogels maintain their shape throughout synthesis steps including covalent crosslinking, equilibration with precursor ions, metal reduction with high concentration reducing agent, rinsing in water, ethanol solvent exchange, and CO2 supercritical drying. Varying the precursor palladium ion concentration allows for control over the metal content in the final aerogel composite through a direct ion chemical reduction rather than relying on the relatively slow coalescence of pre-formed nanoparticles used in other sol-gel techniques. With diffusion as the basis to introduce and remove chemical species into and out of the hydrogel, this method is suitable for smaller bulk geometries and thin films. Characterization of the cellulose nanofiber-palladium composite aerogels with scanning electron microscopy, X-ray diffractometry, thermal gravimetric analysis, nitrogen gas adsorption, electrochemical impedance spectroscopy, and cyclic voltammetry indicates a high surface area, metallized palladium porous structure.

The interfacial properties of organolead halide perovskite solar cells (PSCs) affect the exciton and charge-transport dynamics significantly. Thus, proper modification of the interfaces between perovskite and charge-transport layers is an efficient method to increase the power conversion efficiency (PCE) of PSCs. In this work, we explore the effect of a nonionic surfactant, that is, Triton X-100 (TX) additive, in the poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT:PSS) hole-transport layer. The electronic structure of TX-modified PEDOT:PSS is investigated with ultraviolet/X-ray photoelectron spectroscopy and X-ray absorption spectroscopy with various TX concentrations. The surface of the TX-modified PEDOT:PSS layer showed high TX content, and thus the semimetallic properties of PEDOT:PSS were suppressed conspicuously by its insulating nature. With the TX-modified PEDOT:PSS, the PCE of methylammonium lead iodide (MAPbI 3 ) PSCs increased significantly. To elucidate the origin of the improved device performance, the electrical properties and photoluminescence were investigated comprehensively. Consequently, it was found that the TX additive inhibits interface recombination between PEDOT:PSS and MAPbI 3 , which is caused by the suppression of semimetallic properties of the PEDOT:PSS surface. Hence, we fabricated flexible PSCs successfully using a graphene electrode and TX-modified PEDOT:PSS.

Background: Controlled turnover of proteins as mediated by the ubiquitin proteasome system (UPS) is an important element in plant defense against environmental and pathogen stresses. E3 ligases play a central role in subjecting proteins to hydrolysis by the UPS. Recently, it has been demonstrated that a specific class of E3 ligases termed the U-box ligases are directly associated with the defense mechanisms against abiotic and biotic stresses in several plants. However, no studies on U-box E3 ligases have been performed in one of the important staple crops, barley., Results: In this study, we identified 67 putative U-box E3 ligases from the barley genome and expressed sequence tags (ESTs). Similar to Arabidopsis and rice U-box E3 ligases, most of barley U-box E3 ligases possess evolutionary well-conserved domain organizations. Based on the domain compositions and arrangements, the barley U-box proteins were classified into eight different classes. Along with this new classification, we refined the previously reported classifications of U-box E3 ligase genes in Arabidopsis and rice. Furthermore, we investigated the expression profile of 67 U-box E3 ligase genes in response to drought stress and pathogen infection. We observed that many U-box E3 ligase genes were specifically up-and-down regulated by drought stress or by fungal infection, implying their possible roles of some U-box E3 ligase genes in the stress responses., Conclusion: This study reports the classification of U-box E3 ligases in barley and their expression profiles against drought stress and pathogen infection. Therefore, the classification and expression profiling of barley U-box genes can be used as a platform to functionally define the stress-related E3 ligases in barley.

Nobel metal composite aerogel fibers made from flexible and porous biopolymers offer a wide range of applications, such as in catalysis and sensing, by functionalizing the nanostructure. However, producing these composite aerogels in a defined shape is challenging for many protein-based biopolymers, especially ones that are not fibrous proteins. Here, we present the synthesis of silk fibroin composite aerogel fibers up to 2 cm in length and a diameter of ~300 μm decorated with noble metal nanoparticles. Lyophilized silk fibroin dissolved in hexafluoro-2-propanol (HFIP) was cast in silicon tubes and physically crosslinked with ethanol to produce porous silk gels. Composite silk aerogel fibers with noble metals were created by equilibrating the gels in noble metal salt solutions reduced with sodium borohydride, followed by supercritical drying. These porous aerogel fibers provide a platform for incorporating noble metals into silk fibroin materials, while also providing a new method to produce porous silk fibers. Noble metal silk aerogel fibers can be used for biological sensing and energy storage applications.

In this study, we report highly efficient and flexible photosensors with GaN nanowires (NWs) horizontally embedded in a graphene sandwich structure fabricated on polyethylene terephthalate. GaN NWs and the graphene sandwich structure are used as light-absorbing media and the channel for carrier movement, respectively. To form uniform high-quality crystalline GaN NWs on Si(111) substrates, the initial nucleation behavior of the NWs was manipulated by applying the new growth technique of Ga predeposition. High-resolution transmission electron microscopic images obtained along the vertical direction of GaN NWs showed that stacking faults, typically observed in Si-based (In,Ga)As NWs, were rare. Consequently, narrow and strong optical emission was observed from the GaN NWs at wavelengths of 365.12 nm at 300 K. The photocurrent and photoresponsivity of the flexible photosensor with 802 nm long GaN NWs horizontally embedded in the graphene sandwich channel were measured as 9.17 mA and 91.70 A/W, respectively, at the light intensity of 100 mW/cm 2 , which are much higher than those previously reported. The high optical-to-electrical conversion characteristics of our flexible photosensors are attributed to the increase in the effective interface between the light-absorbing media and the carrier channel by the horizontal distribution of the GaN NWs within the graphene sandwich structure. After 200 cyclic-bending test of the GaN NW photosensor at the strain of 3%, the photoresponsivity under strain was measured as 89.04 A/W at 100 mW/cm 2 , corresponding to 97.1% of the photoresponsivity obtained before bending. The photosensor proposed in this study is relatively simple in device design and fabrication, and it requires no sophisticated nanostructural design to minimize the resistance to metal contacts.

Dengue virus type 2 NS1 (DENV2 NS1) is a specific and sensitive protein biomarker for dengue fever diagnosis. In this study we used polyvalent phage display to identify unique affinity peptides that can bind NS1 protein. Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) were used to investigate the binding interactions. The potential affinity peptide-displayed phage from these methods was selected; its sequence was EHDRMHAYYLTR (R3#10). Amino acid sequence analysis showed that the peptide was rich in basic residues (two His and Arg). Among all the peptides tested, R3#10 showed the greatest decrease in current in CV and increase in impedance in EIS upon binding to NS1 proteins. EIS revealed that R3#10 phage clones were more specific towards NS1 proteins, as compared to bovine serum albumin or the M13 wild type used as control. Detection of NS1 proteins is in accordance with the electron-transfer resistance (R ct ) value of the sensor layer, which is confirmed by EIS, and the K d value of the R3#10 peptide while binding to the phage particles was measured. To the best of our knowledge, this is the first example of identification and characterization of NS1 binding affinity peptides using phage display technology and electrochemical methods. We concluded that these new peptide-displayed phages or free peptides from phages may have potential applications in dengue diagnosis., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Light is the most influential environmental stimulus for plant growth. In response to deficient light, plants reprogram their development to adjust their growth in search for a light source. A fine reprogramming of gene expression orchestrates this adaptive trait. Here we show that plants alter microRNA (miRNA) biogenesis in response to light transition. When plants suffer an unusual extended period of light deprivation, the miRNA biogenesis factor HYPONASTIC LEAVES 1 (HYL1) is degraded but an inactive pool of phosphorylated protein remains stable inside the nucleus. Degradation of HYL1 leads to the release of gene silencing, triggering a proper response to dark and shade. Upon light restoration, a quick dephosphorylation of HYL1 leads to the reactivation of miRNA biogenesis and a switch toward a developmental program that maximizes the light uptake. Our findings define a unique and fast regulatory mechanism controlling the plant silencing machinery during plant light response., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Here, a method to synthesize gold, palladium, and platinum aerogels via a rapid, direct solution-based reduction is presented. The combination of various precursor noble metal ions with reducing agents in a 1:1 (v/v) ratio results in the formation of metal gels within seconds to minutes compared to much longer synthesis times for other techniques such as sol-gel. Conducting the reduction step in a microcentrifuge tube or small volume conical tube facilitates a proposed nucleation, growth, densification, fusion, equilibration model for gel formation, with final gel geometry smaller than the initial reaction volume. This method takes advantage of the vigorous hydrogen gas evolution as a by-product of the reduction step, and as a consequence of reagent concentrations. The solvent accessible specific surface area is determined with both electrochemical impedance spectroscopy and cyclic voltammetry. After rinsing and freeze drying, the resulting aerogel structure is examined with scanning electron microscopy, X-ray diffractometry, and nitrogen gas adsorption. The synthesis method and characterization techniques result in a close correspondence of aerogel ligament sizes. This synthesis method for noble metal aerogels demonstrates that high specific surface area monoliths may be achieved with a rapid and direct reduction approach.

Noble metal aerogels offer a wide range of catalytic applications due to their high surface area and tunable porosity. Control over monolith shape, pore size, and nanofiber diameter is desired in order to optimize electronic conductivity and mechanical integrity for device applications. However, common aerogel synthesis techniques such as solvent mediated aggregation, linker molecules, sol⁻gel, hydrothermal, and carbothermal reduction are limited when using noble metal salts. Here, we present the synthesis of palladium aerogels using carboxymethyl cellulose nanofiber (CNF) biotemplates that provide control over aerogel shape, pore size, and conductivity. Biotemplate hydrogels were formed via covalent cross linking using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) with a diamine linker between carboxymethylated cellulose nanofibers. Biotemplate CNF hydrogels were equilibrated in precursor palladium salt solutions, reduced with sodium borohydride, and rinsed with water followed by ethanol dehydration, and supercritical drying to produce freestanding aerogels. Scanning electron microscopy indicated three-dimensional nanowire structures, and X-ray diffractometry confirmed palladium and palladium hydride phases. Gas adsorption, impedance spectroscopy, and cyclic voltammetry were correlated to determine aerogel surface area. These self-supporting CNF-palladium aerogels demonstrate a simple synthesis scheme to control porosity, electrical conductivity, and mechanical robustness for catalytic, sensing, and energy applications.

The practical applicability of electronic devices is largely determined by the reliability of field effect transistors (FETs), necessitating constant searches for new and better-performing semiconductors. We investigated the stress-induced degradation of MoS 2 multilayer FETs, revealing a steady decrease of drain current by 56% from the initial value after 30 min. The drain current recovers to the initial state when the transistor is completely turned off, indicating the roles of soft-traps in the apparent degradation. The noise current power spectrum follows the model of carrier number fluctuation-correlated mobility fluctuation (CNF-CMF) regardless of stress time. However, the reduction of the drain current was well fitted to the increase of the trap density based on the CNF-CMF model, attributing the presence of the soft-type traps of dielectric oxides to the degradation of the MoS 2 FETs.

Cholera toxin is a major virulent agent of Vibrio cholerae, and it can rapidly lead to severe dehydration, shock, causing death within hours without appropriate clinical treatments. In this study, we present a method wherein unique and short peptides that bind to cholera toxin subunit B (CTX-B) were selected through M13 phage display. Biopanning over recombinant CTX-B led to rapid screening of a unique peptide with an amino acid sequence of VQCRLGPPWCAK, and the phage-displayed peptides analyzed using ELISA, were found to show specific affinities towards CTX-B. To address the use of affinity peptides in development of the biosensor, sequences of newly selected peptides were modified and chemically synthesized to create a series of affinity peptides. Performance of the biosensor was studied using plasmonic-based optical techniques: localized surface plasmon resonance (LSPR) and surface-enhanced Raman scattering (SERS). The limit of detection (LOD) obtained by LSPR with 3σ-rule was 1.89ng/mL, while SERS had a LOD of 3.51pg/mL. In both cases, the sensitivity was much higher than the previously reported values, and our sensor system was specific towards actual CTX-B secreted from V. cholera, but not for CTX-AB 5 ., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths. Therefore, more sensitive and early diagnostic methods for CRC are urgently needed. In this study, an efficient electrochemical biosensor for early diagnosis of adenoma-to-carcinoma progression that employs a series of chemically modified affinity peptides was developed. A series of amino acid-substituted and cysteine-incorporated synthetic peptides with flexible linkers was chemically synthesized and immobilized to a gold sensor layer; performance of the sensor was monitored using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Potential affinity peptides (LRG1 BP1-BP4) specific for the LRG1 biomarker as a target protein were chosen according to a quantitative current decrease and dynamic impedance increase by CV and EIS, respectively. Using EIS, the K d value of the LRG1 BP3 peptide was found to be 8.3 ± 2.7nM. The applicability of the sensor to detect LRG1 proteins was confirmed in human plasma from colorectal adenomas and carcinomas (n = 20 in each group). The detection of LRG1 in accordance with the ΔR ct value (electron-transfer resistance at the electrode surface) of the sensor layer incorporating LRG1 BP3 peptides showed a statistically significant difference (p < 0.001) between adenomas and carcinomas, indicating that the potential use of this biosensing platform for detecting the CRC biomarker, as well as for monitoring the colorectal adenoma-to-carcinoma transition in an electrochemically miniaturized biosensor (e-chem biosensor) in point-of-care testing, is possible., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Ubiquitin E3 ligases are crucial for eliminating misfolded proteins before they form cytotoxic aggregates that threaten cell fitness and survival. However, it remains unclear how emerging misfolded proteins in the cytoplasm can be selectively recognized and eliminated by E3 ligases in plants. We found that Misfolded Protein Sensing RING E3 ligase 1 (MPSR1) is an indispensable E3 ligase required for plant survival after protein-damaging stress. Under no stress, MPSR1 is prone to rapid degradation by the 26S proteasome, concealing its protein quality control (PQC) E3 ligase activity. Upon proteotoxic stress, MPSR1 directly senses incipient misfolded proteins and tethers ubiquitins for subsequent degradation. Furthermore, MPSR1 sustains the structural integrity of the proteasome complex at the initial stage of proteotoxic stress. Here, we suggest that the MPSR1 pathway is a constitutive mechanism for proteostasis under protein-damaging stress, as a front-line surveillance system in the cytoplasm., Competing Interests: The authors declare no conflict of interest., (Copyright © 2017 the Author(s). Published by PNAS.)

Plants are constantly exposed to a variety of abiotic stresses, such as drought, heat, cold, flood, and salinity. To survive under such unfavorable conditions, plants have evolutionarily developed their own resistant-mechanisms. For several decades, many studies have clarified specific stress response pathways of plants through various molecular and genetic studies. In particular, it was recently discovered that ubiquitin proteasome system (UPS), a regulatory mechanism for protein turn over, is greatly involved in the stress responsive pathways. In the UPS, many E3 ligases play key roles in recognizing and tethering poly-ubiquitins on target proteins for subsequent degradation by the 26S proteasome. Here we discuss the roles of RING ligases that have been defined in related to abiotic stress responses in plants. [BMB Reports 2017; 50(8): 393-400].

AtAIRP2 is a cytosolic RING-type E3 ubiquitin ligase that positively regulates an abscisic acid (ABA) response in Arabidopsis ( Arabidopsis thaliana ). Yeast two-hybrid screening using AtAIRP2 as bait identified ATP1 (AtAIRP2 Target Protein1) as a substrate of AtAIRP2. ATP1 was found to be identical to SDIRIP1, which was reported recently to be a negative factor in ABA signaling and a target protein of the RING E3 ligase SDIR1. Accordingly, ATP1 was renamed ATP1/SDIRIP1. A specific interaction between AtAIRP2 and ATP1/SDIRIP1 and ubiquitination of ATP1/SDIRIP1 by AtAIRP2 were demonstrated in vitro and in planta. The turnover of ATP1/SDIRIP1 was regulated by AtAIRP2 in cell-free degradation and protoplast cotransfection assays. The ABA-mediated germination assay of 35S : ATP1/SDIRIP1-RNAi/atairp2 double mutant progeny revealed that ATP1/SDIRIP1 acts downstream of AtAIRP2. AtAIRP2 and SDIR1 reciprocally complemented the ABA- and salt-insensitive germination phenotypes of sdir1 and atairp2 mutants, respectively, indicating their combinatory roles in seed germination. Subcellular localization and bimolecular fluorescence complementation experiments in the presence of MG132, a 26S proteasome inhibitor, showed that AtAIRP2 and ATP1/SDIRIP1 were colocalized to the cytosolic spherical body, which lies in close proximity to the nucleus, in tobacco ( Nicotiana benthamiana ) leaf cells. The 26S proteasome subunits RPN12a and RPT1 and the molecular chaperones HSP70 and HSP101 were colocalized to these discrete punctae-like structures. These results raised the possibility that AtAIRP2 and ATP1/SDIRIP1 interact in the cytosolic spherical compartment. Collectively, our data suggest that the down-regulation of ATP1/SDIRIP1 by AtAIRP2 and SDIR1 RING E3 ubiquitin ligases is critical for ABA and high-salinity responses during germination in Arabidopsis., (© 2017 American Society of Plant Biologists. All Rights Reserved.)

Chemical doping has been investigated as an alternative method of conventional ion implantation for two-dimensional materials. We herein report chemically doped multilayer molybdenum disulfide (MoS 2 ) field effect transistors (FETs) through n-type channel doping, wherein triethanolamine (TEOA) is used as an n-type dopant. As a result of the TEOA doping process, the electrical performances of multilayer MoS 2 FETs were enhanced at room temperature. Extracted field effect mobility was estimated to be ∼30 cm 2 V -1 s -1 after the surface doping process, which is 10 times higher than that of the pristine device. Subthreshold swing and contact resistance were also improved after the TEOA doping process. The enhancement of the subthreshold swing was demonstrated by using an independent FET model. Furthermore, we found that the doping level can be effectively controlled by the heat treatment method. These results demonstrate a promising material system that is easily controlled with high performance, while elucidating the underlying mechanism of improved electrical properties by the doping effect in a multilayered scheme.

Norovirus is known as the major cause of highly infection for gastrointestinal tracts. In this study, robust and highly sensitive biosensors for detecting human norovirus by employing a recognition affinity peptide-based electrochemical platform were described. A series of amino acid-substituted and cysteine-incorporated recognition peptides isolated from evolutionary phage display technique was chemically synthesized and immobilized to a gold sensor layer, the detection performance of the gold-immobilized synthetic peptide-based sensor system was assessed using QCM, CV and EIS. Using EIS, the limit of detection with Noro-1 as a molecular binder was found to be 99.8nM for recombinant noroviral capsid proteins (rP2) and 7.8copies/mL for human norovirus, thereby demonstrating a high degree of sensitivity for their corresponding targets. These results suggest that a biosensor which consists of affinity peptides as a molecular binder and miniaturized microdevices as diagnostic tool could be served as a new type of biosensing platform for point-of-care testing., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Post-translational modifications (PTMs) of proteins are essential to increase the functional diversity of the proteome. By adding chemical groups to proteins, or degrading entire proteins by phosphorylation, glycosylation, ubiquitination, neddylation, acetylation, lipidation, and proteolysis, the complexity of the proteome increases, and this then influences most biological processes. Although small RNAs are crucial regulatory elements for gene expression in most eukaryotes, PTMs of small RNA microprocessor and RNA silencing components have not been extensively investigated in plants. To date, several studies have shown that the proteolytic regulation of AGOs is important for host-pathogen interactions. DRB4 is regulated by the ubiquitin-proteasome system, and the degradation of HYL1 is modulated by a de-etiolation repressor, COP1, and an unknown cytoplasmic protease. Here, we discuss current findings on the PTMs of microprocessor and RNA silencing components in plants.

The cyanobacterial aldehyde deformylating oxygenase (cADO) is a key enzyme that catalyzes the unusual deformylation of aliphatic aldehydes for alkane biosynthesis and can be applied to the production of biofuel in vitro and in vivo. In this study, we determined crystal structures of two ADOs from Limnothrix sp. KNUA012 (LiADO) and Oscillatoria sp. KNUA011 (OsADO). The structures of LiADO and OsADO resembled those of typical cADOs, consisting of eight α-helices found in ferritin-like di-iron proteins. However, structural comparisons revealed that while the LiADO active site was vacant of iron and substrates, the OsADO active site was fully occupied, containing both a coordinated metal ion and substrate. Previous reports indicated that helix 5 is capable of adopting two distinct conformations depending upon the existence of bound iron. We observed that helix 5 of OsADO with an iron bound in the active site presented as a long helix, whereas helix 5 of LiADO, which lacked iron in the active site, presented two conformations (one long and two short helices), indicating that an equilibrium exists between the two states in solution. Furthermore, acquisition of a structure having a fully occupied active site is unique in the absence of higher iron concentrations as compared with other cADO structures, wherein low affinity for iron complicates the acquisition of crystal structures with bound iron. An in-depth analysis of the ADO apo-enzyme, the enzyme with substrate bound, and the enzyme with both iron and substrate bound provided novel insight into substrate-binding modes in the absence of a coordinated metal ion and suggested a separate two-step binding mechanism for substrate and iron co-factors. Moreover, our results provided a comprehensive structural basis for conformational changes induced by binding of the substrate and co-factor., (Copyright © 2016 Elsevier Inc. All rights reserved.)

MicroRNAs (miRNAs), small non-coding RNA molecules, are important biomarkers for research and medical purposes. Here, we describe the development of a fast and simple method using highly fluorescent oligonucleotide-silver nanocluster probes (DNA/AgNCs) to efficiently detect specific miRNAs. Due to the great sequence diversity of miRNAs in humans and other organisms, a uniform strategy for miRNA detection is attractive. The concept presented is an oligonucleotide-based locking-to-unlocking system that can be endowed with miRNA complementarity while maintaining the same secondary structure. The locking-to-unlocking system is based on fold-back anchored DNA templates that consist of a cytosine-rich loop for AgNCs stabilization, an miRNA recognition site and an overlap region for hairpin stabilization. When an miRNA is recognized, fluorescence in the visible region is specifically extinguished in a concentration-dependent manner. Here, the exact composition of the fold-back anchor for the locking-to-unlocking system has been systematically optimized, balancing propensity for loop-structure formation, encapsulation of emissive AgNCs and target sensitivity. It is demonstrated that the applied strategy successfully can detect a number of cancer related miRNAs in RNA extracts from human cancer cell lines., (© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.)

We report significant improvement in optical and electrical properties of green InGaN/GaN light-emitting diodes (LEDs) by using Si-doped graded short-period InGaN/GaN superlattice (SiGSL) formed by so called indium-conversion technique. For comparison, a conventional LED without the superlattice (C-LED) and a LED with undoped graded superlattice (unGSL-LED) were prepared, respectively. The photoluminescence (PL) intensity of the SiGSL-LED was increased more than 3 times at room temperature (RT) as compared to C-LED. The PL intensity ratios of RT to 10K for the C-LED, unGSL-LED, and SiGSL-LED were measured to be 25, 40.9, and 47.5%, respectively. The difference in carrier lifetimes between 10K and RT for the SiGSL-LED is relatively small compared to that of the C-LED, which is consistent with the variation in PL intensity. The output power of a transistor-outline type SiGSL-LED was increased more than 2 times higher than that of the C-LED.

Drought stress strongly affects plant growth and development, directly connected with crop yields, accordingly. However, related to the function of U-BOX E3 ligases, the underlying molecular mechanisms of desiccation stress response in plants are still largely unknown. Here we report that PUB22 and PUB23, two U-box E3 ligase homologs, tether ubiquitins to 19S proteasome regulatory particle (RP) subunit RPN6, leading to its degradation. RPN6 was identified as an interacting substrate of PUB22 by yeast two-hybrid screening, and in vitro pull-down assay confirmed that RPN6 interacts not only with PUB22, but also with PUB23. Both PUB22 and PUB23 were able to conjugate ubiquitins on RPN6 in vitro. Furthermore, RPN6 showed a shorter protein half-life in PUB22 overexpressing plants than in wild-type, besides RPN6 was significantly stabilized in pub22pub23 double knockout plants. Taken together, these results solidify a notion that PUB22 and PUB23 can alter the activity of 26S proteasome in response to drought stress., (Copyright © 2015 Elsevier Inc. All rights reserved.)

Resonance energy transfer (RET) has been employed for interpreting the energy interaction of graphene combined with semiconductor materials such as nanoparticles and quantum-well (QW) heterostructures. Especially, for the application of graphene as a transparent electrode for semiconductor light emitting diodes, the mechanism of exciton recombination processes such as RET in graphene-semiconductor QW heterojunctions should be understood clearly. Here, we characterized the temperature-dependent RET behaviors in graphene/semiconductor QW heterostructures. We then observed the tuning of the RET efficiency from 5% to 30% in graphene/QW heterostructures with ∼60 nm dipole-dipole coupled distance at temperatures of 300 to 10 K. This survey allows us to identify the roles of localized and free excitons in the RET process from the QWs to graphene as a function of temperature.

Dental implant insertion on a site with low bone quality or bone defect should be preceded by a bone graft or artificial bone graft insertion to heal the defect. We generated a beta-tricalcium phosphate (β-TCP) and poloxamer 407-based hydrogel composite and penetration of the β-TCP/hydrogel composite into the peri-implant area of bone was evaluated by porous bone block experiments. The maximum penetration depth for porous bone blocks and dense bone blocks were 524 μm and 464 μm, respectively. We report the in-vivo performance of a composite of β-TCP/hydrogel composite as a carrier of recombinant human bone morphogenetic protein (rhBMP-2), implanted into a rabbit tibial defect model. Three holes drilled into each tibia of eight male rabbits were (1) grafted with dental implant fixtures; (2) filled with β-TCP/hydrogel composite (containing 5 μg of rhBMP-2), followed by grafting of the dental implant fixtures. Four weeks later, bone-implant contact ratio and peri-implant bone formation were analyzed by radiography, micro-CT and histology of undecalcified specimens. The micro-CT results showed a significantly higher level of trabecular thickness and new bone and peri-implant new bone formation in the experimental treatment compared to the control treatment. Histomorphometry revealed a significantly higher bone-implant contact ratio and peri-implant bone formation with the experimental treatment. The use of β-TCP/poloxamer 407 hydrogel composite as a carrier of rhBMP-2 significantly promoted new bone formation around the dental implant fixture and it also improved the quality of the new bone formed in the tibial marrow space.

Bone formation in tooth defect areas and the osseointegration of dental implants are very important for successful dental implant surgery. The aim of the present study was to assess the strengthening effect of a β-TCP microsphere-hydrogel composite containing recombinant human bone morphogenetic protein-2 (rhBMP-2) on bone healing and implant osseointegration. The molars and premolars on the left and right sides of the maxilla were extracted from six male minipigs, and dental implants were placed using either the β-TCP microsphere-hydrogel carrier alone or the carrier loaded with rhBMP-2 (500 μg). The animals were kept alive for a further 8 weeks. The molars and premolars from the left and the right sides of the mandibles of another six minipigs were extracted, and the animals were kept alive for 4 weeks. Two 5-mm-diameter bone defects were then made on both sides of the mandible. The defects were filled with saline, β-TCP microsphere-hydrogel carrier, or the carrier loaded with rhBMP-2 (300 μg), and dental implant fixtures were inserted. The animals were kept alive for a further 4 weeks. Bone formation was examined using plane radiographs, micro-CT, and the histology of undecalcified specimens. The group treated with the rhBMP-2-loaded carrier composite showed a significantly higher percentage bone volume and a greater trabecular thickness for the newly formed bone in the tooth defect areas when compared to the group treated with the carrier alone. The rhBMP-2 group had a significantly higher osseointegration, a larger percentage bone volume, greater trabecular thickness in the newly formed bone in tooth defect areas, a larger newly formed bone fraction in the fixture pitch, and a greater number of newly formed trabecular bones when compared to the other groups. We confirmed that the rhBMP-2-loaded carrier composite promotes new bone formation after tooth extraction and strengthens osseointegration of dental fixtures by improving the degree of osseointegration around the dental implant fixture., (© 2014 Wiley Periodicals, Inc. and International Center for Artificial Organs and Transplantation.)

Beta-tricalcium phosphate ( β -TCP) and hydroxyapatite (HA) are widely used as bone graft extenders due to their osteoconductivity and high bioactivity. This study aims to evaluate the possibility of using porous substrate with composite ceramics ( β -TCP: HA = 60% : 40%, 60TCP40HA) as a bone graft extender and comparing it with Bio-Oss. Interconnectivity and macroporosity of β -TCP porous substrate were 99.9% and 83%, respectively, and the macro-porosity of packed granule after crushing was 69%. Calvarial defect model with 8 mm diameter was generated with male Sprague-Dawley rats and 60TCP40HA was implanted. Bio-Oss was implanted for a control group and micro-CT and histology were performed at 4 and 8 weeks after implantation. The 60TCP40HA group showed better new bone formation than the Bio-Oss group and the bone formation at central area of bone defect was increased at 8 weeks in micro-CT and histology. The percent bone volume and trabecular number of the 60TCP40HA group were significantly higher than those of Bio-Oss group. This study confirms the usefulness of the porous 60TCP40HA composite as a bone graft extender by showing increased new bone formation in the calvarial defect model and improved bone formation both quantitatively and qualitatively when compared to Bio-Oss.

Recombinant human bone morphogenetic protein-2 (rhBMP-2) requires carriers for clinical effectiveness. In this study, whether porous beta-tricalcium phosphate (β-TCP)-based ceramics are ideal carriers for rhBMP-2 was investigated. Hydroxyapatite (HA), β-TCP, TCP/HA (80 %/20 %), HA with rhBMP-2, TCP with rhBMP-2, and TCP/HA (80 %/20 %) with rhBMP-2 were manufactured by a sponge method with a pore size of 300 μm or more and macro-porosity of 83 %. The alkaline phosphatase (ALP) activity and ALP expression of the cells with 100 % β-TCP granules were more increased than the those of cells with 100 % HA and TCP/HA (80 %/20 %) at the baseline or when treated with 15 ng/ml of rhBMP-2. In an SD rat calvarial defect model, new bone formation was evidently shown in the TCP 100 %-rhBMP-2 and TCP/HA (80 %/20 %)-rhBMP-2 groups, showing that the most affected area was filled with newly-formed bone, that the percent bone volume and trabecular number were larger when compared to the groups without rhBMP-2 treatment at both 4 and 8 weeks after surgery using micro-CT and histology. Porous TCP-based ceramic granules enhanced the osteoblastic differentiation in the hMSC system when treated with 15 ng/ml of rhBMP-2 and accelerated bone-healing by trabecular number in a rat calvarial defect model. Thus, in this study it was proposed that TCP-based ceramics might be useful carriers of rhBMP-2.

AtPUB18 and AtPUB19 are homologous U-box E3 ubiquitin ligases in Arabidopsis (Arabidopsis thaliana). AtPUB19 is a negative regulator of abscisic acid (ABA)-mediated drought responses, whereas the role of AtPUB18 in drought responses is unknown. Here, loss-of-function and overexpression tests identified AtPUB18 as a negative regulator in ABA-mediated stomatal closure and water stress responses. The atpub18-2atpub19-3 double mutant line displayed more sensitivity to ABA and enhanced drought tolerance than each single mutant plant; therefore, AtPUB18 and AtPUB19 are agonistic. Stomatal closure of the atpub18-2atpub19-3 mutant was hypersensitive to hydrogen peroxide (H(2)O(2)) but not to calcium, suggesting that AtPUB18 and AtPUB19 exert negative effects on the ABA signaling pathway downstream of H(2)O(2) and upstream of calcium. AtPUB22 and AtPUB23 are other U-box E3 negative regulators of drought responses. Although atpub22atpub23 was more tolerant to drought stress relative to wild-type plants, its ABA-mediated stomatal movements were highly similar to those of wild-type plants. The atpub18-2atpub19-3atpub22atpub23 quadruple mutant exhibited enhanced tolerance to drought stress as compared with each atpub18-2atpub19-3 and atpub22atpub23 double mutant progeny; however, its stomatal behavior was almost identical to the atpub18-2atpub19-3 double mutant in the presence of ABA, H(2)O(2), and calcium. Overexpression of AtPUB18 and AtPUB19 in atpub22atpub23 effectively hindered ABA-dependent stomatal closure, but overexpression of AtPUB22 and AtPUB23 in atpub18-2atpub19-3 did not inhibit ABA-enhanced stomatal closure, highlighting their ABA-independent roles. Overall, these results suggest that AtPUB18 has a linked function with AtPUB19, but is independent from AtPUB22 and AtPUB23, in negative regulation of ABA-mediated drought stress responses.

Among approximately 480 RING domain-containing E3 Ub ligases in Arabidopsis, three, At3g46620, At5g59550, and At2g39720, have a domain-of-unknown-function (DUF) 1117 motif in their C-terminal regions. At3g46620 and At5g59550 were identified as homologous ABA- and drought-induced RING-DUF1117 genes and were designated AtRDUF1 and AtRDUF2, respectively. Single and double knock-out mutations of AtRDUFs resulted in hyposensitive phenotypes toward ABA in terms of germination rate and stomatal closure and markedly reduced tolerance to drought stress relative to wild-type plants. These results are discussed in the context that AtRDUF1 and AtRDUF2 play combinatorial, but still distinguishable, roles in ABA-mediated dehydration stress responses., (Copyright © 2012 Elsevier Inc. All rights reserved.)

The ubiquitin (Ub)-26S proteasome pathway is implicated in various cellular processes in higher plants. AtAIRP1, a C3H2C3-type RING (for Really Interesting New Gene) E3 Ub ligase, is a positive regulator in the Arabidopsis (Arabidopsis thaliana) abscisic acid (ABA)-dependent drought response. Here, the AtAIRP2 (for Arabidopsis ABA-insensitive RING protein 2) gene was identified and characterized. AtAIRP2 encodes a cytosolic C3HC4-type RING E3 Ub ligase whose expression was markedly induced by ABA and dehydration stress. Thus, AtAIRP2 belongs to a different RING subclass than AtAIRP1 with a limited sequence identity. AtAIRP2-overexpressing transgenic (35S:AtAIRP2-sGFP) and atairp2 loss-of-function mutant plants exhibited hypersensitive and hyposensitive phenotypes, respectively, to ABA in terms of seed germination, root growth, and stomatal movement. 35S:AtAIRP2-sGFP plants were highly tolerant to severe drought stress, and atairp2 alleles were more susceptible to water stress than were wild-type plants. Higher levels of drought-induced hydrogen peroxide production were detected in 35S:AtAIRP2-sGFP as compared with atairp2 plants. ABA-inducible drought-related genes were up-regulated in 35S:AtAIRP2-sGFP and down-regulated in atairp2 progeny. The positive effects of AtAIRP2 on ABA-induced stress genes were dependent on SNF1-related protein kinases, key components of the ABA signaling pathway. Therefore, AtAIRP2 is involved in positive regulation of ABA-dependent drought stress responses. To address the functional relationship between AtAIRP1 and AtAIRP2, FLAG-AtAIRP1 and AtAIRP2-sGFP genes were ectopically expressed in atairp2-2 and atairp1 plants, respectively. Constitutive expression of FLAG-AtAIRP1 and AtAIRP2-sGFP in atairp2-2 and atairp1 plants, respectively, reciprocally rescued the loss-of-function ABA-insensitive phenotypes during germination. Additionally, atairp1/35S:AtAIRP2-sGFP and atairp2-2/35S:FLAG-AtAIRP1 complementation lines were more tolerant to dehydration stress relative to atairp1 and atairp2-2 single knockout plants. Overall, these results suggest that AtAIRP2 plays combinatory roles with AtAIRP1 in Arabidopsis ABA-mediated drought stress responses.

Ubiquitination is a eukaryotic posttranslational protein modification that is mediated by the cascade of E1, E2, and E3 ubiquitin (Ub) ligases and is involved in regulating numerous cellular functions. In this study, we obtained 100 different Arabidopsis (Arabidopsis thaliana) T-DNA insertion mutant plants in which RING E3 Ub ligase genes were suppressed and monitored their phenotypes in the presence of exogenous abscisic acid (ABA), a plant stress hormone. One of these loss-of-function mutants displayed ABA-insensitive phenotypes at the germination stage and was named atairp1 (for Arabidopsis ABA-insensitive RING protein 1). AtAIRP1 encodes a cytosolic protein containing a single C3H2C3-type RING motif with in vitro E3 Ub ligase activity. AtAIRP1 was significantly induced by ABA and drought stress. In contrast to atairp1 mutant plants, AtAIRP1-overexpressing transgenic plants (35S:AtAIRP1-sGFP) were hypersensitive to exogenous ABA in terms of radicle emergence, cotyledon development, root elongation, and stomatal closure. Ectopic expression of AtAIRP1-sGFP in atairp1 effectively rescued the loss-of-function ABA-insensitive phenotype. Both 35S:AtAIRP1-sGFP and atairp1/35S:AtAIRP1-sGFP plants accumulated higher amounts of hydrogen peroxide in response to exogenous ABA than did wild-type and atairp1 mutant plants. AtAIRP1 overexpressors were markedly tolerant to severe drought stress, as opposed to atairp1, which was highly susceptible. The levels of drought stress-related genes and basic leucine zipper transcription factor genes were up-regulated in the 35S:AtAIRP1-sGFP lines relative to wild-type and atairp1 mutant plants in response to ABA. Overall, these results suggest that AtAIRP1, a C3H2C3-type RING E3 Ub ligase, is a positive regulator in the Arabidopsis ABA-dependent drought response.