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MicroRNA (miRNA) maturation is critically dependent on structural features of primary transcripts (pri-miRNAs). However, the scarcity of determined pri-miRNA structures has limited our understanding of miRNA maturation. Here, we employed selective 2'-hydroxyl acylation analyzed by primer extension and mutational profiling (SHAPE-MaP), a high-throughput RNA structure probing method, to unravel the secondary structures of 476 high-confidence human pri-miRNAs. Our SHAPE-based structures diverge substantially from those inferred solely from computation, particularly in the apical loop and basal segments, underlining the need for experimental data in RNA structure prediction. By comparing the structures with high-throughput processing data, we determined the optimal structural features of pri-miRNAs. The sequence determinants are influenced substantially by their structural contexts. Moreover, we identified an element termed the bulged GWG motif (bGWG) with a 3' bulge in the lower stem, which promotes processing. Our structure-function mapping better annotates the determinants of pri-miRNA processing and offers practical implications for designing small hairpin RNAs and predicting the impacts of miRNA mutations., Competing Interests: Declaration of interests V.N.K. is a member of the Molecular Cell advisory board., (Copyright © 2024 Elsevier Inc. All rights reserved.)

When individuals listen to speech, their neural activity phase-locks to the slow temporal rhythm, which is commonly referred to as "neural tracking". The neural tracking mechanism allows for the detection of an attended sound source in a multi-talker situation by decoding neural signals obtained by electroencephalography (EEG), known as auditory attention decoding (AAD). Neural tracking with AAD can be utilized as an objective measurement tool for diverse clinical contexts, and it has potential to be applied to neuro-steered hearing devices. To effectively utilize this technology, it is essential to enhance the accessibility of EEG experimental setup and analysis. The aim of the study was to develop a cost-efficient neural tracking system and validate the feasibility of neural tracking measurement by conducting an AAD task using an offline and real-time decoder model outside the soundproof environment. We devised a neural tracking system capable of conducting AAD experiments using an OpenBCI and Arduino board. Nine participants were recruited to assess the performance of the AAD using the developed system, which involved presenting competing speech signals in an experiment setting without soundproofing. As a result, the offline decoder model demonstrated an average performance of 90%, and real-time decoder model exhibited a performance of 78%. The present study demonstrates the feasibility of implementing neural tracking and AAD using cost-effective devices in a practical environment., (© 2023. The Author(s).)

The detection of asbestos roof slate by drone is necessary to avoid the safety risks and costs associated with visual inspection. Moreover, the use of deep-learning models increases the speed as well as reduces the cost of analyzing the images provided by the drone. In this study, we developed a comprehensive learning model using supervised and unsupervised classification techniques for the accurate classification of roof slate. We ensured the accuracy of our model using a low altitude of 100 m, which led to a ground sampling distance of 3 cm/pixel. Furthermore, we ensured that the model was comprehensive by including images captured under a variety of light and meteorological conditions and from a variety of angles. After applying the two classification methods to develop the learning dataset and employing the as-developed model for classification, 12 images were misclassified out of 475. Visual inspection and an adjustment of the classification system were performed, and the model was updated to precisely classify all 475 images. These results show that supervised and unsupervised classification can be used together to improve the accuracy of a deep-learning model for the detection of asbestos roof slate.

Selective auditory attention has been shown to modulate the cortical representation of speech. This effect has been well documented in acoustically more challenging environments. However, the influence of top-down factors, in particular topic familiarity, on this process remains unclear, despite evidence that semantic information can promote speech-in-noise perception. Apart from individual features forming a static listening condition, dynamic and irregular changes of auditory scenes-volatile listening environments-have been less studied. To address these gaps, we explored the influence of topic familiarity and volatile listening on the selective auditory attention process during dichotic listening using electroencephalography. When stories with unfamiliar topics were presented, participants' comprehension was severely degraded. However, their cortical activity selectively tracked the speech of the target story well. This implies that topic familiarity hardly influences the speech tracking neural index, possibly when the bottom-up information is sufficient. However, when the listening environment was volatile and the listeners had to re-engage in new speech whenever auditory scenes altered, the neural correlates of the attended speech were degraded. In particular, the cortical response to the attended speech and the spatial asymmetry of the response to the left and right attention were significantly attenuated around 100-200 ms after the speech onset. These findings suggest that volatile listening environments could adversely affect the modulation effect of selective attention, possibly by hampering proper attention due to increased perceptual load., Competing Interests: Declaration of Competing Interest The authors declare no competing financial interests., (Copyright © 2023. Published by Elsevier B.V.)

Physical unclonable function (PUFs) utilize inherent random physical variations of solid-state devices and are a core ingredient of hardware security primitives. PUFs promise more robust information security than that provided by the conventional software-based approaches. While silicon- and memristor-based PUFs are advancing, their reliability and scalability require further improvements. These are currently limited by output fluctuations and associated additional peripherals. Here, highly reliable spintronic PUFs that exploit field-free spin-orbit-torque switching in IrMn/CoFeB/Ta/CoFeB structures are demonstrated. It is shown that the stochastic switching polarity of the perpendicular magnetization of the top CoFeB can be achieved by manipulating the exchange bias directions of the bottom IrMn/CoFeB. This serves as an entropy source for the spintronic PUF, which is characterized by high entropy, uniqueness, reconfigurability, and digital output. Furthermore, the device ensures a zero bit-error-rate under repetitive operations and robustness against external magnetic fields, and offers scalable and energy-efficient device implementations., (© 2022 The Authors. Advanced Materials published by Wiley-VCH GmbH.)

Active research on crack detection technology for structures based on unmanned aerial vehicles (UAVs) has attracted considerable attention. Most of the existing research on localization of cracks using UAVs mounted the Global Positioning System (GPS)/Inertial Measurement Unit (IMU) on the UAVs to obtain location information. When such absolute position information is used, several studies confirmed that positioning errors of the UAVs were reflected and were in the order of a few meters. To address these limitations, in this study, without using the absolute position information, localization of cracks was defined using relative position between objects in UAV-captured images to significantly reduce the error level. Through aerial photography, a total of 97 images were acquired. Using the point cloud technique, image stitching, and homography matrix algorithm, 5 cracks and 3 reference objects were defined. Importantly, the comparative analysis of estimated relative position values and ground truth values through field measurement revealed that errors in the range 24-84 mm and 8-48 mm were obtained on the x- and y-directions, respectively. Also, RMSE errors of 37.95-91.24 mm were confirmed. In the future, the proposed methodology can be utilized for supplementing and improving the conventional methods for visual inspection of infrastructures and facilities.

Protein tyrosine phosphatases (PTPs), along with protein tyrosine kinases, control signaling pathways involved in cell growth, metabolism, differentiation, proliferation, and survival. Several PTPs, such as PTPN1, PTPN2, PTPN9, PTPN11, PTPRS, and DUSP9, disrupt insulin signaling and trigger type 2 diabetes, indicating that PTPs are promising drug targets for the treatment or prevention of type 2 diabetes. As part of an ongoing study on the discovery of pharmacologically active bioactive natural products, we conducted a phytochemical investigation of African mango ( Irvingia gabonensis ) using liquid chromatography-mass spectrometry (LC/MS)-based analysis, which led to the isolation of terminalin as a major component from the extract of the seeds of I. gabonensis . The structure of terminalin was characterized by spectroscopic methods, including one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) and high-resolution (HR) electrospray ionization (ESI) mass spectroscopy. Moreover, terminalin was evaluated for its antidiabetic property; terminalin inhibited the catalytic activity of PTPN1, PTPN9, PTPN11, and PTPRS in vitro and led to a significant increase in glucose uptake in differentiated C2C12 muscle cells, indicating that terminalin exhibits antidiabetic effect through the PTP inhibitory mechanism. These findings suggest that terminalin derived from African mango could be used as a functional food ingredient or pharmaceutical supplement for the prevention of type 2 diabetes.

Maturation of canonical microRNA (miRNA) is initiated by DROSHA that cleaves the primary transcript (pri-miRNA). More than 1,800 miRNA loci are annotated in humans, but it remains largely unknown whether and at which sites pri-miRNAs are cleaved by DROSHA. Here, we performed in vitro processing on a full set of human pri-miRNAs (miRBase version 21) followed by sequencing. This comprehensive profiling enabled us to classify miRNAs on the basis of DROSHA dependence and map their cleavage sites with respective processing efficiency measures. Only 758 pri-miRNAs are confidently processed by DROSHA, while the majority may be non-canonical or false entries. Analyses of the DROSHA-dependent pri-miRNAs show key cis-elements for processing. We observe widespread alternative processing and unproductive cleavage events such as "nick" or "inverse" processing. SRSF3 is a broad-acting auxiliary factor modulating alternative processing and suppressing unproductive processing. The profiling data and methods developed in this study will allow systematic analyses of miRNA regulation., Competing Interests: Declaration of interests V.N.K. is a member of the Molecular Cell Advisory Board. The authors declare no other competing interests., (Copyright © 2021 Elsevier Inc. All rights reserved.)

L1 retrotransposons can pose a threat to genome integrity. The host has evolved to restrict L1 replication. However, mechanisms underlying L1 propagation out of the host surveillance remains unclear. Here, we propose an evolutionary survival strategy of L1, which exploits RNA m 6 A modification. We discover that m 6 A 'writer' METTL3 facilitates L1 retrotransposition, whereas m 6 A 'eraser' ALKBH5 suppresses it. The essential m 6 A cluster that is located on L1 5' UTR serves as a docking site for eukaryotic initiation factor 3 (eIF3), enhances translational efficiency and promotes the formation of L1 ribonucleoprotein. Furthermore, through the comparative analysis of human- and primate-specific L1 lineages, we find that the most functional m 6 A motif-containing L1s have been positively selected and became a distinctive feature of evolutionarily young L1s. Thus, our findings demonstrate that L1 retrotransposons hijack the RNA m 6 A modification system for their successful replication.

As part of our systematic study on Korean toxic mushrooms, bioactivity-guided fractionation of the MeOH extract of Amanita spissacea (Amanitaceae) fruiting bodies and chemical investigation of its cytotoxic fractions led to the isolation of (9 E )-8-oxo-9-octadecenoic acid ( 1 ), (10 E )-9-oxo-10-octadecenoic acid ( 2 ), (9 E )-8-oxo-9-octadecenoate methyl ester ( 3 ), (9 Z )-9-octadecenoate-(2' S )-2',3'-dihydroxypropyl ester ( 4 ), (9 Z )-9-octadecenoic acid ( 5 ), and palmitic acid ( 6 ). The structures of the isolates were elucidated by NMR spectroscopic analysis and LC/MS analysis. Among the isolated compounds, compounds 1 and 2 exhibited the most potent cytotoxic activity in all human lung cancer cell lines examined, with IC 50 values ranging from 255.7 to 321.0 μM and 250.2 to 322.5 μM, respectively. The cytotoxicity of these compounds was also found to be mediated by apoptosis associated with caspase-3 activation. These findings provide experimental evidence suggesting the potential of A. spissacea as a promising natural source for the discovery of novel anticancer drug candidates.

Safflower ( Carthamus tinctorius ) is an annual herb belonging to the Compositae family; it has a history of use as a food colorant, dye, and medicine in oriental countries. LC-MS-UV-based chemical analysis of extract of the florets of C. tinctorius led to the isolation of two new C 10 -polyacetylene glycosides, (8 Z )-decaene-4,6-diyne-1,10-diol-1- O - β -d-glucopyranoside ( 1 ) and (8 S )-deca-4,6-diyne-1,8-diol-1- O - β -d-glucopyranoside ( 2 ), together with five known analogs ( 3 - 7 ). The structures of the new compounds were determined by using 1D and 2D NMR spectroscopic data and HR-MS data, as well as chemical transformations. Of compounds 1 - 7 , compounds 2 , 3 , and 4 inhibited the adipogenesis of 3T3-L1 preadipocytes, whereas compounds 1 and 6 promoted adipogenesis. Compounds 2 , 3 , and 4 also prevented lipid accumulation through the suppression of the expression of lipogenic genes and the increase of the expression of lipolytic genes. Moreover, compounds 3 and 4 activated AMPK, which is known to facilitate lipid metabolism. Our findings provide a mechanistic rationale for the use of safflower-derived polyacetylene glycosides as potential therapeutic agents against obesity.

Auditory attention detection (AAD) is the tracking of a sound source to which a listener is attending based on neural signals. Despite expectation for the applicability of AAD in real-life, most AAD research has been conducted on recorded electroencephalograms (EEGs), which is far from online implementation. In the present study, we attempted to propose an online AAD model and to implement it on a streaming EEG. The proposed model was devised by introducing a sliding window into the linear decoder model and was simulated using two datasets obtained from separate experiments to evaluate the feasibility. After simulation, the online model was constructed and evaluated based on the streaming EEG of an individual, acquired during a dichotomous listening experiment. Our model was able to detect the transient direction of a participant's attention on the order of one second during the experiment and showed up to 70% average detection accuracy. We expect that the proposed online model could be applied to develop adaptive hearing aids or neurofeedback training for auditory attention and speech perception.

The microprocessor complex cleaves the primary transcript of microRNA (pri-miRNA) to initiate miRNA maturation. Microprocessor is known to consist of RNase III DROSHA and dsRNA-binding DGCR8. Here, we identify Enhancer of Rudimentary Homolog (ERH) as a new component of Microprocessor. Through a crystal structure and biochemical experiments, we reveal that ERH uses its hydrophobic groove to bind to a conserved region in the N-terminus of DGCR8, in a 2:2 stoichiometry. Knock-down of ERH or deletion of the DGCR8 N-terminus results in a reduced processing of suboptimal pri-miRNAs in polycistronic miRNA clusters. ERH increases the processing of suboptimal pri-miR-451 in a manner dependent on its neighboring pri-miR-144. Thus, the ERH dimer may mediate 'cluster assistance' in which Microprocessor is loaded onto a poor substrate with help from a high-affinity substrate in the same cluster. Our study reveals a role of ERH in the miRNA biogenesis pathway., (© The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Bioassay-guided fractionation of a 90% ethanol extract of Periostracum Cicadae led to the isolation of two new N-acetyldopamine dimers (1a/1b) along with six known dimers (2a/2b, 3a/3b, and 4a/4b) and two monomers (5a/5b); compounds 2a/2b, 4a/4b and 5a/5b were newly isolated from this material. All compounds were isolated as enantiomeric mixtures and each enantiomer was successfully separated by chiral-phase HPLC. The structures including absolute configurations were confirmed by high-resolution electrospray ionization mass spectrometry (HR-ESIMS), 1D/2D nuclear magnetic resonance (NMR) spectroscopy, 1 H iterative Full Spin Analysis (HiFSA), and electronic circular dichroism (ECD) spectroscopy. Subsequently, the bioactivities of these isolates were evaluated via CD4 + T cell differentiations, which are critical for immune responses and inflammation. The results revealed that compound 5b was observed to enhance the IFN-γ + Th1 differentiation, which may have a potential for cancer immunotherapy., 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 © 2020 Elsevier Inc. All rights reserved.)

The antidepressant activities of hispidol and decursin (both potent monoamine oxidase A (MAO-A) inhibitors) were evaluated using the forced swimming test (FST) and the tail suspension test (TST) in mice, and thereafter, levels of neurotransmitter monoamines and metabolites in brain tissues were analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Hispidol (15 mg/kg) caused less or comparable immobility than fluoxetine (15 mg/kg; the positive control) in immobility time, as determined by FST (9.6 vs 32.0 s) and TST (53.1 vs 48.7 s), respectively, and its effects were dose-dependent and significant. Decursin (15 mg/kg) also produced immobility comparable to that of fluoxetine as determined by FST (47.0 vs 43.4 s) and TST (55.6 vs 63.4 s), and its effects were also dose-dependent and significant. LC-MS/MS analysis after FST showed that hispidol (15 mg/kg) greatly increased dopamine (DA) and serotonin levels dose-dependently in brain tissues as compared with the positive control. Decursin (15 mg/kg) dose-dependently increased DA level after TST. Slight changes in norepinephrine and 3,4-dihydroxyphenylacetic acid levels were observed after FST and TST in hispidol- or decursin-treated animals. It was observed that hispidol and decursin were effective and comparable to fluoxetine in immobility tests. These immobility and monoamine level results suggest that hispidol and decursin are potential antidepressant agents for the treatment of depression, and that they act mainly through serotonergic and/or dopaminergic systems.

Carthamus tinctorius L. (Compositae; safflower or Hong Hua) has been used in Korean traditional medicine for maintaining the homeostasis of body circulation. Phytochemical investigation was performed on the florets of C. tinctorius by liquid chromatography-mass spectrometry (LC/MS), which afforded two dihydrophaseic acid glucosides ( 1 and 2 ). Isolated compounds were structurally confirmed using a combination of spectroscopic methods including 1D and 2D nuclear magnetic resonance and high-resolution electrospray ionization mass spectroscopy. Their absolute configurations were established by quantum chemical electronic circular dichroism calculations and enzymatic hydrolysis. The anti-adipogenesis activity of the isolated compounds was evaluated using 3T3-L1 preadipocytes. Treatment with the dihydrophaseic acid glucoside ( 1 ) during adipocyte differentiation prevented the accumulation of lipid droplets and reduced the expression of adipogenic genes, Fabp4 and Adipsin . However, compound 2 did not affect adipogenesis. Our study yielded a dihydrophaseic acid glucoside derived from C. tinctorius, which has potential advantages for treating obesity.

Nineteen compounds were isolated from the stems of Maackia amurensis by activity-guided screening for new human monoamine oxidase-B (hMAO-B) inhibitors. Among the compounds isolated, flavonoids calycosin (5) and 8-O-methylretusin (6) were found to potently and selectively inhibit hMAO-B (IC 50  = 0.24 and 0.23 μM, respectively) but not hMAO-A with high selectivity index (SI) values (SI = 293.8 and 81.3, respectively). In addition, 5 and 6 reversibly and competitively inhibited hMAO-B with K i values of 0.057 and 0.054 μM, respectively. A pterocarpan (-)-medicarpin (18) was also observed to strongly inhibit hMAO-B (IC 50  = 0.30 μM). Most of the compounds weakly inhibited AChE, except isolupalbigenin (13) (IC 50  = 20.6 μM), which suggested 13 be considered a potential dual function inhibitor of MAO-B and AChE. Molecular docking simulation revealed that the binding affinities of 5 and 6 for hMAO-B (both -9.3 kcal/mol) were higher than those for hMAO-A (-7.4 and -7.2 kcal/mol, respectively). Compound 5 was found to interact by hydrogen bonding with hMAO-B at Cys172 residue (distance: 3.250 Å); no hydrogen bonding was predicted between 5 and hMAO-A. These findings suggest that compounds 5 and 6 be considered novel potent, selective, and reversible hMAO-B inhibitors and candidates for the treatment of neurological disorders., Competing Interests: Declaration of competing interest The authors have no conflict of interest to declare., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Nuclear processing of most miRNAs is mediated by Microprocessor, comprised of RNase III enzyme Drosha and its cofactor DGCR8. Here, we uncover a hidden layer of Microprocessor regulation via studies of Dicer-independent mir-451, which is clustered with canonical mir-144. Although mir-451 is fully dependent on Drosha/DGCR8, its short stem and small terminal loop render it an intrinsically weak Microprocessor substrate. Thus, it must reside within a cluster for normal biogenesis, although the identity and orientation of its neighbor are flexible. We use DGCR8 tethering assays and operon structure-function assays to demonstrate that local recruitment and transfer of Microprocessor enhances suboptimal substrate processing. This principle applies more broadly since genomic analysis indicates suboptimal canonical miRNAs are enriched in operons, and we validate several of these experimentally. Proximity-based enhancement of suboptimal hairpin processing provides a rationale for genomic retention of certain miRNA operons and may explain preferential evolutionary emergence of miRNA operons., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Opuntia humifusa , known as the eastern prickly pear cactus and locally called "Cheonnyuncho" in Korea, is cultivated widely on Jeju Island, Korea. Phytochemical analysis of the methanolic extract of the cladodes of O. humifusa , for which previous research is relatively limited, was performed under the guidance of LC/MS-based analysis. As a result, one new megastigmane ( 1 ) and four new megastigmane glucosides ( 2 - 5 ) were isolated along with 18 known compounds ( 6 - 23 ). The structures of the new compounds were established by 1D and 2D NMR and HRESIMS, and their absolute configurations were established by chemical reactions, quantum chemical electronic circular dichroism calculations, and DP4+ analysis using the gauge-including atomic orbital NMR chemical shift calculations as well as the application of Snatzke's method. The isolated compounds ( 1 - 23 ) were tested for NO production inhibition in lipopolysaccharide (LPS)-induced RAW 264.7 cells to investigate their anti-inflammatory effects. Compounds 10 and 11 exhibited significant inhibitory effects on LPS-induced NO production in a dose-dependent manner. The potential mechanistic pathway of 10 and 11 was also investigated using Western blotting, indicating that compounds 10 and 11 inhibit NO through iNOS expression.

Hippophae rhamnoides L. (Elaeagnaceae; commonly known as "sea buckthorn" and "vitamin tree"), is a spiny deciduous shrub whose fruit is used in foods and traditional medicines. The H. rhamnoides fruit (berry) is rich in vitamin C, with a level exceeding that found in lemons and oranges. H. rhamnoides berries are usually washed and pressed to create pomace and juice. Today, the powder of the aqueous extract of H. rhamnoides berries are sold as a functional food in many countries. As part of our ongoing effort to identify bioactive constituents from natural resources, we aimed to isolate and identify those from the fruits of H. rhamnoides . Phytochemical analysis of the extract of H. rhamnoides fruits led to the isolation and identification of six compounds, namely, a citric acid derivative ( 1 ), a phenolic ( 2 ), flavonoids ( 3 and 4 ), and megastigmane compounds ( 5 and 6 ). Treatment with compounds 1 - 6 did not have any impact on the cell viability of RAW 264.7 mouse macrophages. However, pretreatment with these compounds suppressed lipopolysaccharide (LPS)-induced NO production in RAW 264.7 mouse macrophages in a concentration-dependent manner. Among the isolated compounds, compound 1 was identified as the most active, with an IC 50 of 39.76 ± 0.16 μM. This value was comparable to that of the N G -methyl-L-arginine acetate salt, a nitric oxide synthase inhibitor with an IC 50 of 28.48 ± 0.05 μM. Western blot analysis demonstrated that compound 1 inhibited the LPS-induced expression of IKKα/β (IκB kinase alpha/beta), I-κBα (inhibitor of kappa B alpha), nuclear factor kappa-B (NF-κB) p65, iNOS (inducible nitric oxide synthase), and COX-2 (cyclooxygenase-2) in RAW 264.7 cells. Furthermore, LPS-stimulated cytokine production was detected using a sandwich enzyme-linked immunosorbent assay. Compound 1 decreased interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) production in LPS-stimulated RAW 264.7 cells. In summary, the mechanism of action of 1 included the suppression of LPS-induced NO production in RAW 264.7 cells by inhibiting IKKα/β, I-κBα, NF-κB p65, iNOS, and COX-2, and the activities of IL-6 and TNF-α.

Garlic ( Allium sativum L.) is utilized worldwide for culinary and medicinal use and has diverse health benefits. As part of our ongoing research to identify bioactive components from natural resources, phytochemical analysis of the methanolic extract of garlic led to the isolation and characterization of six compounds: Three eugenol diglycosides ( 1 - 3 ) and three β-carboline alkaloids ( 4 - 6 ). In particular, the absolute configurations of β-carboline alkaloids ( 5 and 6 ) were established by gauge-including atomic orbital nuclear magnetic resonance chemical shift calculations, followed by DP4+ analysis. Here, we evaluated the effects of compounds 1 - 6 on 3T3-L1 preadipocyte adipogenesis and lipid metabolism. 3T3-L1 adipocyte differentiation was evaluated using Oil Red O staining; the expression of adipogenic genes was detected using RT-qPCR. Among compounds 1 - 6 , (1 R, 3 S )-1-methyl-1,2,3,4-tetrahydro- β- carboline-3-carboxylic acid ( 6 ) inhibited 3T3-L1 preadipocyte adipogenesis and reduced the expression of adipogenic genes ( Fabp4 , PPARγ , C/EBPβ , Adipsin , and Adipoq ). Moreover, it markedly decreased the actylation of α-tubulin, which is crucial for cytoskeletal remodeling during adipogenesis. Anti-adipogenic effects were observed upon treatment with compound 6, not only during the entire process, but also on the first two days of adipogenesis. Additionally, treatment with compound 6 regulated the expression of genes involved in adipocyte lipid metabolism, decreasing the lipogenic gene ( SREBP1 ) and increasing lipolytic genes ( ATGL and HSL ). We provide experimental evidence of the health benefits of using (1 R, 3 S )-1-methyl-1,2,3,4-tetrahydro- β- carboline-3-carboxylic acid obtained from garlic to prevent excessive adipogenesis in obesity.

The present study documents the synthesis of oxygenated chalcone (O1-O26) derivatives and their abilities to inhibit monoamine oxidases. All 26 derivatives examined showed potent inhibitory activity against MAO-B. Compound O23 showed the greatest inhibitory activity against MAO-B with an IC 50 value of 0.0021 µM, followed by compounds O10 and O17 (IC 50  = 0.0030 and 0.0034 µM, respectively). In addition, most of the derivatives potently inhibited MAO-A and O6 was the most potent inhibitor with an IC 50 value of 0.029 µM, followed by O3, O4, O9, and O2 (IC 50  = 0.035, 0.053, 0.072, and 0.082 µM, respectively). O23 had a high selectivity index (SI) value for MAO-B of 138.1, and O20 (IC 50 value for MAO-B = 0.010 µM) had an extremely high SI of >4000. In dialysis experiments, inhibitions of MAO-A and MAO-B by O6 and O23, respectively, were recovered to their respective reversible reference levels, demonstrating both are reversible inhibitors. Kinetic studies revealed that O6 and O23 competitively inhibited MAO-A and MAO-B, respectively, with respective K i values of 0.016 ± 0.0007 and 0.00050 ± 0.00003 µM. Lead compound are also non-toxic at 200 µg/mL in normal rat spleen cells. Molecular docking simulations and subsequent Molecular Mechanics/Generalized Born Surface Area calculations provided a rationale that explained experimental data., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Withania somnifera (L.) Dunal (Solanaceae), known as Indian ginseng or ashwagandha, has been used in Indian Ayurveda for the treatment of a variety of disorders, such as diabetes and reproductive and nervous system disorders. It is particularly used as a general health tonic, analgesic, and sedative. As part of continuing projects to discover unique bioactive natural products from medicinal plants, phytochemical investigation of the roots of W. somnifera combined with a liquid chromatography-mass spectrometry (LC/MS)-based analysis has led to the isolation of two novel phenylpropanoid esters, Withaninsams A (1) and B (2), as an inseparable mixture, along with three known phenolic compounds (3, 4, and 6) and a pyrazole alkaloid (5). The structures of the new compounds were elucidated using a combination of spectroscopic methods, including one-dimensional (1D) and two-dimensional (2D) nuclear magnetic resonance (NMR) and high-resolution electrospray ionization mass spectroscopy (HR-ESIMS). Withaninsams A (1) and B (2) are phenylpropanoid esters that contain a side chain, 4-methyl-1,4-pentanediol unit. To the best of our knowledge, the present study is the first to report on phenylpropanoid esters with 4-methyl-1,4-pentanediol unit. The anti-inflammatory activity of the isolated compounds (1-6) was evaluated by determining their inhibitory effects on nitric oxide (NO) production in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages, where compound 3 inhibited LPS-induced NO production (IC 50 = 33.3 μM) and TNF-α production, a pro-inflammatory cytokine (IC 50 = 40.9 μM). The anti-inflammatory mechanism through the inhibition of transcriptional iNOS protein expression was confirmed by western blotting experiments for the active compound 3 , which showed decreased iNOS protein expression.

To discover new pharmacologically active natural products, here, we performed the phytochemical analysis of a Korean medicinal plant. Euonymus alatus (Thunb.) Sieb. is a traditional medicinal plant that has been used as a remedy for various diseases in Asian countries. In particular, the cork cambium on the twigs of E. alatus has been used to treat dysmenorrhea, tumors, diabetes, and wound. Phytochemical analysis of the methanolic extract of E. alatus twigs led to the isolation of a sterol, which was identified as (3 β ,16 α )-3,16-dihydroxypregn-5-en-20-one ( 1 ) by 1D and 2D nuclear magnetic resonance (NMR) spectroscopy and high-resolution electrospray ionization mass spectrometry. The stereochemistry of 1 was established with nuclear Overhauser effect spectroscopy (NOESY) analysis and comparison of electronic circular dichroism (ECD) data. To the best of our knowledge, the isolation of compound 1 from nature is first reported here, as well as the complete and revised NMR data assignment of 1 . In lipopolysaccharide (LPS)-stimulated RAW-264.7 macrophages, compound 1 significantly inhibited nitric oxide (NO) production at an IC 50 value of 12.54 ± 0.05 μM as well as the protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). Moreover, the pre-treatment with compound 1 attenuated the LPS-induced phosphorylation of nuclear factor kappa B (NF-κB) p65 through the inhibition of the phosphorylation of IκB kinase alpha (IKKα), IKKβ, and inhibitor of kappa B alpha (IκBα). Compound 1 also inhibited the LPS-induced phosphorylation of p38, c-Jun NH 2 -terminal kinase (JNK), and extracellular signal-regulated kinase (ERK). Taken together, compound 1 may serve as an anti-inflammatory constituent of E. alatus twigs and its anti-inflammatory property is thought to be associated with the inhibition of NO production via suppression of iNOS and COX-2 expression through inhibition of IKKα/β, I-κBα and NF-κB p65 activation and downregulation of p38, JNK, and ERK mitogen-activated protein kinase signal pathways in RAW 264.7 macrophages. These findings also provide experimental evidence that compound 1 identified from E. alatus twigs could be a candidate for an anti-inflammatory agent.

Six synthetic (1-6) and six natural (7-12) chalcones were tested for human monoamine oxidases (hMAOs) and acetylcholinesterase (AChE) inhibitory activities. Compounds 4-dimethylaminochalcone (2), 4'-chloro-4-dimethylaminochalcone (5), and 2,4'-dichloro-4-dimethylaminochalcone (1) potently inhibited hMAO-B with IC 50 values of 0.029, 0.061, and 0.075 μM, respectively. 4-Nitrochalcone (4) and 4-chlorochalcone (3) also potently inhibited hMAO-B with IC 50 values of 0.066 and 0.082 μM, respectively (2.3- and 2.6-fold less than compound 2). Compound 2 had a high selectivity index (113.1) for hMAO-B over hMAO-A (IC 50  = 3.28 μM). Compounds 1 and 2,2'-dihydroxy-4',6'-dimethoxychalcone (12) potently inhibited hMAO-A with IC 50 values of 0.18 and 0.39 μM, respectively. In addition, compounds 4 and 2 also effectively inhibited AChE with IC 50 values of 1.25 and 6.07 μM, respectively, and thus, exhibited dual-targeting. Compound 2 reversibly and competitively inhibited hMAO-B with a K i value of 0.0066 μM. Docking simulations showed binding affinities of compounds 1 to 5 for hMAO-B were higher than those for hMAO-A or AChE and suggested these five chalcones form hydrogen bonds with MAO-B at Cys172 but that they do not form hydrogen bonds with hMAO-A or AChE. These findings suggest compound 2 be considered a promising and dual-targeting lead compound for the treatment of Alzheimer's disease., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Pinus eldarica (Pinaceae), an evergreen plant, is distributed across the warm and dry climates of western Asia, including Asia Minor, the Middle East, and land surrounding the Caspian Sea. Essential oils (EOs) from different aerial parts of this tree have been used in traditional medicine. We aimed to investigate the chemical profile and antimicrobial activity of the EO from P. eldarica grown in northwestern Iran. EO from the needles, bark, and pollen were extracted with boiling water using a Clevenger apparatus at yield of 0.7-1.2 cm 3 /100 g of dry plant material. The main chemical components of the EO from the needles were D-germacrene (18.17%), caryophyllene (15.42%), γ-terpinene (12.96%), and β-pinene (10.62%); those from the bark were limonene (16.99%), caryophyllene oxide (13.22%), and drimenol (13.2%); and those from the pollen were α-pinene (25.64%) and limonene (19.94%). In total, 83 constituents were characterized in the EOs, using gas chromatography mass spectrometry analysis; mainly, sesquiterpene hydrocarbons in needle EO and monoterpene hydrocarbons in pollen and bark EOs. β-Pinene, β-myrcene, limonene, and caryophyllene were identified in the EOs from all three plant parts. The antibacterial and antifungal properties of the EOs were examined: pollen EO exhibited antibacterial activity against Escherichia coli ; bark EO inhibited the growth of Candida albicans and Staphylococcus aureus ; and the needle EO inhibited the growth of S. aureus . Thus, the EOs from aerial parts of P. eldarica can benefit the EO industry and antibiotic development.

Six hundred forty natural compounds were tested for acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activities. Of those, sargachromanol I (SCI) and G (SCG) isolated from the brown alga Sargassum siliquastrum, dihydroberberine (DB) isolated from Coptis chinensis, and macelignan (ML) isolated from Myristica fragrans, potently and effectively inhibited AChE with IC 50 values of 0.79, 1.81, 1.18, and 4.16 µM, respectively. SCI, DB, and ML reversibly inhibited AChE and showed mixed, competitive, and noncompetitive inhibition, respectively, with K i values of 0.63, 0.77, and 4.46 µM, respectively. Broussonin A most potently inhibited BChE (IC 50  = 4.16 µM), followed by ML, SCG, and SCI (9.69, 10.79, and 13.69 µM, respectively). In dual-targeting experiments, ML effectively inhibited monoamine oxidase B with the greatest potency (IC 50  = 7.42 µM). Molecular docking simulation suggested the binding affinity of SCI (-8.6 kcal/mol) with AChE was greater than those of SCG (-7.9 kcal/mol) and DB (-8.2 kcal/mol). Docking simulation indicated SCI interacts with AChE at Trp81, and that SCG interacts at Ser119. No hydrogen bond was predicted for the interaction between AChE and DB. This study suggests SCI, SCG, DB, and ML be viewed as new reversible AChE inhibitors and useful lead compounds for the development for the treatment of Alzheimer's disease., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Two series of fluorinated chalcones containing morpholine and imidazole-based compounds (f1-f8) were synthesized and evaluated for recombinant human monoamine oxidase (MAO)-A and -B as well as acetylcholinesterase inhibitory activities. Our results indicate that morpholine containing chalcones are highly selective MAO-B inhibitors having reversibility properties. All the imidazole-based fluorinated chalcones showed weak MAO inhibitions in both isoforms. Among the tested compounds, (2E)-3-(3-fluorophenyl)-1-[4-(morpholin-4-yl)phenyl]prop-2-en-1-one (f2) showed potent inhibitory activity for recombinant human MAO-B (IC 50  = 0.087 μM) with a high selectivity index (SI) of 517.2. In the recovery experiments using dialysis, the residual activity of MAO-B inhibited by f2 was close to that with the reversible reference inhibitor. Inhibition assays revealed that the K i values of f1 and f2 for MAO-B were 0.027 and 0.020 μM, respectively, with competitive patterns. All the morpholine-based compounds (f1-f4) showed moderate inhibition toward acetylcholinesterase with IC 50 values ranging between 24 and 54 μM. All morpholine-containing compounds exhibit good blood-brain barrier permeation in the PAMPA method. The rational approach regarding the highly selective MAO-B inhibitor f2 was further ascertained by induced fit docking and molecular dynamics simulation studies., (© 2019 Deutsche Pharmazeutische Gesellschaft.)

Osthenol (6), a prenylated coumarin isolated from the dried roots of Angelica pubescens, potently and selectively inhibited recombinant human monoamine oxidase-A (hMAO-A) with an IC 50 value of 0.74 µM and showed a high selectivity index (SI > 81.1) for hMAO-A versus hMAO-B. Compound 6 was a reversible competitive hMAO-A inhibitor (K i  = 0.26 µM) with a potency greater than toloxatone (IC 50  = 0.93 µM), a marketed drug. Isopsoralen (3) and bakuchicin (1), furanocoumarin derivatives isolated from Psoralea corylifolia L., showed slightly higher IC 50 values (0.88 and 1.78 µM, respectively) for hMAO-A than 6, but had low SI values (3.1 for both). Other coumarins tested did not effectively inhibit hMAO-A or hMAO-B. A structural comparison suggested that the 8-(3,3-dimethylallyl) group of 6 increased its inhibitory activity against hMAO-A compared with the 6-methoxy group of scopoletin (4). Molecular docking simulations revealed that the binding affinity of 6 for hMAO-A (-8.5 kcal/mol) was greater than that for hMAO-B (-5.6 kcal/mol) and that of 4 for hMAO-A (-7.3 kcal/mol). Docking simulations also implied that 6 interacted with hMAO-A at Phe208 and with hMAO-B at Ile199 by carbon hydrogen bondings. Our findings suggest that osthenol, derived from natural products, is a selective and potent reversible inhibitor of MAO-A, and can be regarded a potential lead compound for the design of novel reversible MAO-A inhibitors., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Three flavanones and two flavones were isolated from the leaves of Prunus padus var. seoulensis by the activity-guided screening for new monoamine oxidase (MAO) inhibitors. Among the compounds isolated, rhamnocitrin (5) was found to potently and selectively inhibit human MAO-A (hMAO-A, IC 50  = 0.051 µM) and effectively inhibit hMAO-B (IC 50  = 2.97 µM). The IC 50 value of 5 for hMAO-A was the lowest amongst all natural flavonoids reported to date, and the potency was 20.2 times higher than that of toloxatone (1.03 µM), a marketed drug. In addition, 5 reversibly and competitively inhibited hMAO-A and hMAO-B with K i values of 0.030 and 0.91 µM, respectively. Genkwanin (4) was also observed to strongly inhibit hMAO-A and hMAO-B (IC 50  = 0.14 and 0.35 µM, respectively), and competitively inhibit hMAO-A and hMAO-B (K i  = 0.097 and 0.12 µM, respectively). Molecular docking simulation reveals that the binding affinity of 5 with hMAO-A (-18.49 kcal/mol) is higher than that observed with hMAO-B (0.19 kcal/mol). Compound 5 interacts with hMAO-A at four possible residues (Asn181, Gln215, Thr336, and Tyr444), while hMAO-B forms a single hydrogen bond at Glu84. These findings suggest that compound 5 as well as 4 can be considered as novel potent and reversible hMAO-A and/or hMAO-B inhibitors or useful lead compounds for future development of hMAO inhibitors in neurological disorder therapies., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Microprocessor, composed of DROSHA and its cofactor DGCR8, initiates microRNA (miRNA) biogenesis by processing the primary transcripts of miRNA (pri-miRNAs). Here we investigate the mechanism by which Microprocessor selects the cleavage site with single-nucleotide precision, which is crucial for the specificity and functionality of miRNAs. By testing ∼40,000 pri-miRNA variants, we find that for some pri-miRNAs the cleavage site is dictated mainly by the mGHG motif embedded in the lower stem region of pri-miRNA. Structural modeling and deep-sequencing-based complementation experiments show that the double-stranded RNA-binding domain (dsRBD) of DROSHA recognizes mGHG to place the catalytic center in the appropriate position. The mGHG motif as well as the mGHG-recognizing residues in DROSHA dsRBD are conserved across eumetazoans, suggesting that this mechanism emerged in an early ancestor of the animal lineage. Our findings provide a basis for the understanding of miRNA biogenesis and rational design of accurate small-RNA-based gene silencing., (Copyright © 2018 Elsevier Inc. All rights reserved.)

Background: Chalcones are considered as the selective scaffold for the inhibition of MAO-B., Objectives: A previously synthesized ethyl acetohydroxamate-chalcones (L1-L22) were studied for their inhibitory activities against human recombinant monoamine oxidase A and B (hMAO-A and hMAO-B, respectively) and acetylcholinesterase (AChE) as multi-target directed ligands for the treatment of Alzheimer's Disease (AD)., Methods: Enzyme inhibition studies of MAO-A, MAO-B and AChE is carried out. Computational studies such as Molecular docking, Molecular Mechanics/Generalized Born Surface Area calculations, ADMET prediction, and protein target prediction are also performed., Results: Among the screened compounds, compound L3 has most potent hMAO-B inhibition with an IC50 value of 0.028 ± 0.0016 µM, and other compounds, L1, L2, L4, L8, L12, and L21 showed significant potent hMAO-B inhibition with IC50 values of 0.051 ± 0.0014, 0.086 ± 0.0035, 0.036 ± 0.0011, 0.096 ± 0.0061, 0.083 ± 0.0016, and 0.038 ± 0.0021 µM, respectively. On the other hand, among the tested compounds, compound L13 showed highest hMAO-A inhibition with an IC50 value of 0.51± 0.051 µM and L9 has a significant value of 1.85 ± 0.045 µM. However, the compounds L3 and L4 only showed high selectivities for hMAO-B with Selectivity Index (SI) values of 621.4 and 416.7, respectively. Among the substituents in ring A of ethyl acetohydroxamate-chalcones (L1-L9), F atom at p-position (L3) showed highest inhibitory effect against hMAO-B. This result supports the uniqness and bizarre behavior of fluorine. Moreover, chalcones L3, L4, L9, L11, and L12 showed potential AChE inhibitory effect with IC50 values of 0.67, 0.85, 0.39, 0.30, and 0.45 µM, respectively. Inhibitions of hMAO-B by L3 or L4 were recovered to the level of the reversible reference (lazabemide), and were competitive with Ki values of 0.0030 ± 0.0002 and 0.0046 ± 0.0005 µM, respectively. Inhibitions of AChE by L3 and L11 were of the competitive and mixed types with Ki values of 0.30 ± 0.044 and 0.14 ± 0.0054 µM, respectively., Conclusion: The studies indicated that L3 and L4 are considered to be promising multitarget drug molecules with potent, selective, and reversible competitive inhibitors of hMAO-B and with highly potent AChE inhibitory effect., (Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.)

LC/MS-based phytochemical analysis of an EtOH extract of the roots of rice (Oryza sativa; Gramineae), which takes a crucial role in the stable crop population in Asia, resulted in the isolation of a new lignan, oryzativol C (1), as a minor component. The chemical structure of compound 1 was unambiguously confirmed using spectroscopic evidence (including 1D- and 2D-NMR data), HR-ESI-MS, and CD data analysis. Considering the traditional medicinal efficacy of O. sativa and its importance as a food crop, compound 1 was evaluated for effects on breast cancer cell lines (MDA-MB-231) and on glucose-stimulated insulin secretion in an INS-1 pancreatic β-cell line. Compound 1 showed mild cytotoxicity toward the MDA-MB-231. Furthermore, compound 1 stimulated insulin secretion in INS-1 pancreatic β-cells without inducing cytotoxicity. These results indicate that compound 1 is an active ingredient of O. sativa that offers health benefits including inhibition of breast cancer cell proliferation and hyperglycemia control., (© 2018 Wiley-VHCA AG, Zurich, Switzerland.)

In the present study, series of eleven (2E)-1-[4-(1H-imidazol-1-yl)substituted phenyl]-3-phenylprop-2-en-1-one (IM1-IM11) derivatives were synthesized and evaluated as inhibitors of recombinant human monoamine oxidase (MAO) A and B. The results indicate that (2E)-3-[4-(dimethylamino) phenyl]-1-[4-(1H-imidazol-1-yl) phenyl] prop-2-en-1-one (IM5) is a nonselective and reversible competitive inhibitor of MAO-A and MAO-B with IC 50 values of 0.30 ± 0.010 and 0.40 ± 0.017 μM, respectively ; those of (2E)-1-[4-(1H-imidazol-1-yl) phenyl]-3-(4-methylphenyl) prop-2-en-1-one (IM4) were 1.06 ± 0.090 and 0.32 ± 0.021 μM, respectively. Kinetic studies document that both IM5 and IM4 are competitive inhibitors of MAO-A and MAO-B with Ki value of 0.11 ± 0.0085 and 0.085 ± 0.0064 μM, respectively. Molecular docking studies of lead compounds further explained the binding modes in the inhibitor binding cavity of both MAO-A and MAO-B., (Copyright © 2018 Elsevier Masson SAS. All rights reserved.)

A series of 13 phenyl substituted thiosemicarbazones ( SB1-SB13 ) were synthesized and evaluated for their inhibitory potential towards human recombinant monoamine oxidase A and B (MAO-A and MAO-B, respectively) and acetylcholinesterase. The solid state structure of SB4 was ascertained by the single X-ray diffraction technique. Compounds SB5 and SB11 were potent for MAO-A (IC 50 1.82 ± 0.14) and MAO-B (IC 50 0.27 ± 0.015 μM), respectively. Furthermore, SB11 showed a high selectivity index (SI > 37.0) for MAO-B. The effects of fluorine orientation revealed that SB11 ( m -fluorine) showed 28.2 times higher inhibitory activity than SB12 ( o -fluorine) against MAO-B. Furthermore, inhibitions by SB5 and SB11 against MAO-A and MAO-B, respectively, were recovered to near reference levels in reversibility experiments. Both SB5 and SB11 showed competitive inhibition modes, with K i values of 0.97 ± 0.042 and 0.12 ± 0.006 μM, respectively. These results indicate that SB5 and SB11 are selective, reversible and competitive inhibitors of MAO-A and MAO-B, respectively. Compounds SB5 , SB7 and SB11 showed moderate inhibition against acetylcholinesterase with IC 50 values of 35.35 ± 0.47, 15.61 ± 0.057 and 26.61 ± 0.338 μM, respectively. Blood-brain barrier (BBB) permeation was studied using the parallel artificial membrane permeation assay (PAMPA) method. Molecular docking studies were carried out using AutoDock 4.2.

Chelerythrine, an isoquinoline alkaloid isolated from the herbaceous perennial Chelidonium majus, was found to potently and selectively inhibit an isoform of recombinant human monoamine oxidase-A (MAO-A) with an IC 50 value of 0.55 µM. Chelerythrine was a reversible competitive MAO-A inhibitor (K i  = 0.22 µM) with a potency much greater than toloxatone (IC 50  = 1.10 µM), a marketed drug. Other isoquinoline alkaloids tested did not effectively inhibit MAO-A or MAO-B. A structural comparison with corynoline suggested the 1- and/or 2-methoxy groups of chelerythrine increase its inhibitory activity against MAO-A. Molecular docking simulations revealed that the binding affinity of chelerythrine for MAO-A (-9.7 kcal/mol) was greater than that for MAO-B (-4.6 kcal/mol). Docking simulation implied that Cys323 and Tyr444 of MAO-A are key residues for hydrogen-bond interaction with chelerythrine. Our findings suggest chelerythrine is one of the most reversible selective and potent natural inhibitor of MAO-A, and that it be regarded a potential lead compound for the design of novel reversible MAO-A inhibitors., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Bridge inspection using unmanned aerial vehicles (UAV) with high performance vision sensors has received considerable attention due to its safety and reliability. As bridges become obsolete, the number of bridges that need to be inspected increases, and they require much maintenance cost. Therefore, a bridge inspection method based on UAV with vision sensors is proposed as one of the promising strategies to maintain bridges. In this paper, a crack identification method by using a commercial UAV with a high resolution vision sensor is investigated in an aging concrete bridge. First, a point cloud-based background model is generated in the preliminary flight. Then, cracks on the structural surface are detected with the deep learning algorithm, and their thickness and length are calculated. In the deep learning method, region with convolutional neural networks (R-CNN)-based transfer learning is applied. As a result, a new network for the 384 collected crack images of 256 × 256 pixel resolution is generated from the pre-trained network. A field test is conducted to verify the proposed approach, and the experimental results proved that the UAV-based bridge inspection is effective at identifying and quantifying the cracks on the structures.

Magnetic torques generated through spin-orbit coupling 1-8 promise energy-efficient spintronic devices. For applications, it is important that these torques switch films with perpendicular magnetizations without an external magnetic field 9-14 . One suggested approach 15 to enable such switching uses magnetic trilayers in which the torque on the top magnetic layer can be manipulated by changing the magnetization of the bottom layer. Spin currents generated in the bottom magnetic layer or its interfaces transit the spacer layer and exert a torque on the top magnetization. Here we demonstrate field-free switching in such structures and show that its dependence on the bottom-layer magnetization is not consistent with the anticipated bulk effects 15 . We describe a mechanism for spin-current generation 16,17 at the interface between the bottom layer and the spacer layer, which gives torques that are consistent with the measured magnetization dependence. This other-layer-generated spin-orbit torque is relevant to energy-efficient control of spintronic devices.

A cellulolytic fungus, YDJ14, was isolated from compost and identified as an Aspergillus sp. strain. Three extracellular β-glucosidases, BGL-A1, BGL-A2, and BGL-A3, were separated using ultrafiltration, ammonium sulfate fractionation, and High-Q chromatography. The molecular masses of the three enzymes were estimated to be 100, 45, and 40 kDa, respectively, by SDS-PAGE. The optimum pH and temperature of BGL-A3 were 5.0 and 50°C, respectively, whereas the optimum pH and temperature of BGL-A1 and BGL-A2 were identical (4.0 and 60°C, respectively). The half-life of BGL-A3 at 70°C (2.8 min) was shorter than that of BGL-A1 and BGL-A2 (12.1 and 8.8 min, respectively). All three enzymes preferred p -nitrophenyl-β- D -glucopyranoside ( p NPG) and hardly hydrolyzed cellobiose, suggesting that these enzymes were aryl β-glucosidases. The K m of BGL-A3 (1.26 mM) for p NPG was much higher than that of BGL-A1 and BGL-A2 (0.25 and 0.27 mM, respectively). These results suggested that BGL-A1 and BGL-A2 were similar in their enzymatic properties, whereas BGL-A3 differed from the two enzymes. When tilianin (a flavone glycoside of acacetin) was reacted with the three enzymes, the inhibitory activity for monoamine oxidase, a target in the treatment of neurological disorders, was similar to that shown by acacetin. We conclude that these enzymes may be useful in the hydrolysis of flavone glycosides to improve their inhibitory activities.

A cellulolytic fungus YDJ216 was isolated from a compost and identified as an Aspergillus sp. strain. Two extracellular β-glucosidases, BGL1 and BGL2, were purified using ultrafiltration, ammonium sulfate fractionation, and High-Q chromatography. Molecular masses of BGL1 and BGL2 were estimated to be 97 and 45 kDa, respectively, by SDS-PAGE. The two enzymes eluted as one peak at 87 kDa by Sephacryl S-200 chromatography, and located at similar positions in a zymogram after intact gel electrophoresis, suggesting BGL1 and BGL2 might be monomeric and dimeric, respectively. Both enzymes showed similar enzymatic properties; they were optimally active at pH 4.0-4.5 and 60 °C, and had similar half-lives at 70 °C. Two enzymes also preferred p-nitrophenyl glucose (pNPG) with the same K m and hardly hydrolyzed cellobiose, suggesting both enzymes are aryl β-glucosidases. However, V max for pNPG of BGL1 (953.2 U/mg) was much higher than those of BGL2 (66.5U/mg) and other β-glucosidases reported. When tilianin (a flavone glycoside of acacetin) was reacted with both enzymes, inhibitory activity for monoamine oxidase, relating to oxidation of neurotransmitter amines, was increased closely to the degree obtained by acacetin. These results suggest that BGL1 and BGL2 could be used to hydrolyze flavone glycosides to improve their inhibitory activities., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A new intracellular serine protease gene of Bacillus megaterium, ispK, encoding a protein composed of 332 amino acid residues with a predicted pI of 4.7 was cloned into Escherichia coli. The deduced amino acid sequence of IspK showed 49-56% similarity with the other microbial intracellular serine proteases described in the literature. The enzyme was effectively purified by one-step chromatography after heat-treatment, and showed a homogeneous band corresponding to 35kDa by SDS-PAGE analysis. Amino acid analysis showed that 16 amino acids of the N-terminus of IspK were removed by post-translational protease activity. The optimum pH and temperature of IspK were 6.0-7.0 and 50°C, respectively. In the presence of 2mM of Ca 2+ ion, the optimum temperature was increased to 65°C and thermostability (t 1/2 ) increased 32.9-fold from 3.3min to 108.5min at 60°C. The enzyme was activated by Ca 2+ and Mg 2+ , almost completely inhibited by phenylmethanesulfonyl fluoride (PMSF) and EDTA, but tolerant to nonionic surfactants, such as, Triton X-100 or Tween 80. IspK efficiently hydrolyzed natural proteins, such as, casein and hemoglobin, and improved blood stain removal. These results suggest IspK can be used as a useful additive for detergent formulations and for deproteinizations., (Copyright © 2017 Elsevier B.V. All rights reserved.)

A series of twelve furanochalcones (F1-F12) was synthesized and investigated for their human monoamine oxidase inhibitory activities. Among the series, compound (2E, 4E)-1-(furan-2-yl)-5-phenylpenta-2, 4-dien-1-one (F1), which was analyzed by single-crystal X-ray diffraction, showed potent and selective MAO-B inhibitory activity with an inhibition constant (K i ) value of 0.0041 μM and selectivity index of (SI) 172.4, and exhibited competitive inhibition. Introduction of a cinnamyl group to the furanochalcone significantly increased the inhibitory activity. In the dilution-recovery experiments, the residual activities of MAO-A and MAO-B by F1 under the diluted condition fully recovered as compared with the undiluted condition, indicating F1 is a reversible inhibitor. The K i value of F1 is the lowest among the values of chalcone derivatives and furthermore lower than that (0.0079 μM) of the reversible MAO-B inhibitor, lazabemide, a marketed drug. Molecular docking study against hMAO-B provided the binding site interactions of the lead compound, including strong π-π stacking between the phenyl system and FAD nucleus., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Hispidol, an aurone, isolated from Glycine max Merrill, was found to potently and selectively inhibit an isoform of recombinant human monoamine oxidase-A (MAO-A), with an IC 50 value of 0.26 µM, and to inhibit MAO-B, but with lower potency (IC 50  = 2.45 µM). Hispidol reversibly and competitively inhibited MAO-A with a K i value of 0.10 µM with a potency much greater than toloxatone (IC 50  = 1.10 µM), a marketed drug. It also reversibly and competitively inhibited MAO-B (K i  = 0.51 µM). Sulfuretin, an analog of hispidol, effectively inhibited MAO-A (IC 50  = 4.16 µM) but not MAO-B (IC 50  > 80 µM). A comparison of their chemical structures showed that the 3'-hydroxyl group of sulfuretin might reduce its inhibitory activities against MAO-A and MAO-B. Flexible docking simulation revealed that the binding affinity of hispidol for MAO-A (-9.1 kcal/mol) was greater than its affinity for MAO-B (-8.7 kcal/mol). The docking simulation showed hispidol binds to the major pocket of MAO-A or MAO-B. The findings suggest hispidol is a potent, selective, reversible inhibitor of MAO-A, and that it be considered a novel lead compound for development of novel reversible inhibitors of MAO-A., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Five compounds were isolated from the leaves of Agastache rugosa and tested for monoamine oxidase (MAO) inhibitory activity. Acacetin, a flavonoid, potently inhibited recombinant human MAO-A and MAO-B (IC 50 =0.19 and 0.17μM, respectively), and reversibly and competitively inhibited MAO-A and MAO-B (K i =0.045 and 0.037μM, respectively). Acacetin 7-O-(6-O-malonylglucoside) (AMG) was also found to effectively inhibit MAO-A and MAO-B (IC 50 =2.34 and 1.87μM, respectively), and to reversibly and competitively inhibit MAO-A and MAO-B (K i =1.06 and 0.38μM, respectively). Tilianin (a glucoside derivative of acacetin) had little inhibitory activity, but the introduction of a malonyl group at sugar moiety significantly increased inhibitory activity. Molecular docking simulation revealed the binding energy of acacetin for MAO-B (-44.2kcal/mol) was greater than its energy for MAO-A (-27.0kcal/mol), and that the Cys172 residue of MAO-B was important for hydrogen bonding with acacetin. AMG was predicted to bind to MAO-B with an energy of -23.1kcal/mol by possible hydrogen-bond formation between an oxygen atom of Ile477 residue and a hydrogen atom (H17) of AMG. However, the interaction between AMG and MAO-A was not verified by the docking simulation. This study suggests acacetin and AMG be viewed as new reversible MAO inhibitors, and useful lead compounds for the inhibitor development., (Copyright © 2017 Elsevier B.V. All rights reserved.)

β-1,3-1,4-Glucanase (BGlc8H) from Paenibacillus sp. X4 was mutated by error-prone PCR or truncated using termination primers to improve its enzyme properties. The crystal structure of BGlc8H was determined at a resolution of 1.8 Å to study the possible roles of mutated residues and truncated regions of the enzyme. In mutation experiments, three clones of EP 2-6, 2-10, and 5-28 were finally selected that exhibited higher specific activities than the wild type when measured using their crude extracts. Enzyme variants of BG 2-6 , BG 2-10 , and BG 5-28 were mutated at two, two, and six amino acid residues, respectively. These enzymes were purified homogeneously by Hi-Trap Q and CHT-II chromatography. Specific activity of BG 5-28 was 2.11-fold higher than that of wild-type BG wt , whereas those of BG 2-6 and BG 2-10 were 0.93- and 1.19-fold that of the wild type, respectively. The optimum pH values and temperatures of the variants were nearly the same as those of BG wt (pH 5.0 and 40 °C, respectively). However, the half-life of the enzyme activity and catalytic efficiency (k cat /K m ) of BG 5-28 were 1.92- and 2.12-fold greater than those of BG wt at 40 °C, respectively. The catalytic efficiency of BG 5-28 increased to 3.09-fold that of BG wt at 60 °C. These increases in the thermostability and catalytic efficiency of BG 5-28 might be useful for the hydrolysis of β-glucans to produce fermentable sugars. Of the six mutated residues of BG 5-28 , five residues were present in mature BGlc8H protein, and two of them were located in the core scaffold of BGlc8H and the remaining three residues were in the substrate-binding pocket forming loop regions. In truncation experiments, three forms of C-terminal truncated BGlc8H were made, which comprised 360, 286, and 215 amino acid residues instead of the 409 residues of the wild type. No enzyme activity was observed for these truncated enzymes, suggesting the complete scaffold of the α 6 /α 6 -double-barrel structure is essential for enzyme activity.

Background: Current state-of-the-art approaches to biological event extraction train statistical models in a supervised manner on corpora annotated with event triggers and event-argument relations. Inspecting such corpora, we observe that there is ambiguity in the span of event triggers (e.g., "transcriptional activity" vs. 'transcriptional'), leading to inconsistencies across event trigger annotations. Such inconsistencies make it quite likely that similar phrases are annotated with different spans of event triggers, suggesting the possibility that a statistical learning algorithm misses an opportunity for generalizing from such event triggers., Methods: We anticipate that adjustments to the span of event triggers to reduce these inconsistencies would meaningfully improve the present performance of event extraction systems. In this study, we look into this possibility with the corpora provided by the 2009 BioNLP shared task as a proof of concept. We propose an Informed Expectation-Maximization (EM) algorithm, which trains models using the EM algorithm with a posterior regularization technique, which consults the gold-standard event trigger annotations in a form of constraints. We further propose four constraints on the possible event trigger annotations to be explored by the EM algorithm., Results: The algorithm is shown to outperform the state-of-the-art algorithm on the development corpus in a statistically significant manner and on the test corpus by a narrow margin., Conclusions: The analysis of the annotations generated by the algorithm shows that there are various types of ambiguity in event annotations, even though they could be small in number.

The phenomena based on spin-orbit interaction in heavy metal/ferromagnet/oxide structures have been investigated extensively due to their applicability to the manipulation of the magnetization direction via the in-plane current. This implies the existence of an inverse effect, in which the conductivity in such structures should depend on the magnetization orientation. In this work, we report a systematic study of the magnetoresistance (MR) of W/CoFeB/MgO structures and its correlation with the current-induced torque to the magnetization. We observe that the MR is independent of the angle between the magnetization and current direction but is determined by the relative magnetization orientation with respect to the spin direction accumulated by the spin Hall effect, for which the symmetry is identical to that of so-called the spin Hall magnetoresistance. The MR of ~1% in W/CoFeB/MgO samples is considerably larger than those in other structures of Ta/CoFeB/MgO or Pt/Co/AlOx, which indicates a larger spin Hall angle of W. Moreover, the similar W thickness dependence of the MR and the current-induced magnetization switching efficiency demonstrates that MR in a non-magnet/ferromagnet structure can be utilized to understand other closely correlated spin-orbit coupling effects such as the inverse spin Hall effect or the spin-orbit spin transfer torques.

The coarse-grained structural model such as Gaussian network has played a vital role in the normal mode studies for understanding protein dynamics related to biological functions. However, for the large proteins, the Gaussian network model is computationally unfavorable for diagonalization of Hessian (stiffness) matrix for the normal mode studies. In this article, we provide the coarse-graining method, referred to as "dynamic model condensation," which enables the further coarse-graining of protein structures consisting of small number of residues. It is shown that the coarser-grained structures reconstructed by dynamic model condensation exhibit the dynamic characteristics, such as low-frequency normal modes, qualitatively comparable to original structures. This sheds light on that dynamic model condensation and may enable one to study the large protein dynamics for gaining insight into biological functions of proteins.

Mutations of p53 and PTCH gene, two candidate tumor suppressor genes for basal cell carcinoma (BCC), were screened in 15 cases of sporadic BCCs that developed in sun-exposed skin region in a Korean population. p53 and PTCH mutations were detected at a frequency of 33 and 40%, respectively, and the mutations were predominantly UV-signature transition, C-->T transitions at dipyrimidine sites and CC-->TT tandem mutations. In both genes, the most common mutations were missense mutations resulting in amino acid substitution, which is different than the results from Caucasian BCCs where mutations are frequently predicted to make truncated or absent proteins. All mutations, except for one, occurred on the nontranscribed strand where is little efficient removal of UV-induced pyrimidine dimers relative to the transcribed strand. Loss of heterozygocity (LOH) of 9q22 for PTCH loci was found in eight of 15 informative cases of BCCs (53%), but none of the cases were informative for LOH of 17p13 for p53 loci. Not only do our data indicate the key role played by p53 and PTCH in the development of BCCs, these findings also suggest that UVB may significantly contribute to BCC tumorigenesis. Moreover, molecular epidemiology composed of incidence of p53 and PTCH mutations, difference in the type of mutation and repair bias of UV-induced DNA lesions might affect the distinct features of BCCs between different racial population.

The skin offers a tissue site accessible for delivery of gene-based therapeutics. To develop the capability for sustained systemic polypeptide delivery via cutaneous gene transfer, we generated and injected pseudotyped HIV-1 lentiviral vectors intradermally at a range of doses into human skin grafted on immune-deficient mice. Unlike Moloney murine leukemia virus (MLV)-based retrovectors, which failed to achieve detectable cutaneous gene transfer by this approach, lentivectors effectively targeted all major cell types within human skin tissue, including fibroblasts, endothelial cells, keratinocytes, and macrophages. After a single injection, lentivectors encoding human erythropoietin (EPO) produced dose-dependent increases in serum human EPO levels and hematocrit that increased rapidly within one month and remained stable subsequently. Delivered gene expression was confined locally at the injection site. Excision of engineered skin led to rapid and complete loss of human EPO in the bloodstream, confirming that systemic EPO delivery was entirely due to lentiviral targeting of cells within skin rather than via spread of the injected vector to visceral tissues. These findings indicate that the skin can sustain dosed systemic delivery of therapeutic polypeptides via direct lentivector injection and thus provide an accessible and reversible approach for gene-based delivery to the bloodstream.