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Introduction: Apocynin (Apo), an NADPH oxidase (NOX) inhibitor, has been widely used to treat various inflammatory diseases. However, the therapeutic effects of Apo on benign prostatic hyperplasia (BPH), a multifactorial disease associated with chronic inflammation and hormone imbalance, remain unknown. Objectives: The link between androgen signaling, reactive oxygen species (ROS), and prostate cell proliferation may contribute to the pathogenesis of BPH; therefore, the aim of this study was to identify the specific signaling pathway involved and to demonstrate whether the anti-oxidant Apo plays a role in the prevention and treatment of BPH. Methods: Ingenuity pathway analysis and si-RNA transfection were conducted to demonstrate the androgen receptor (AR) and NOX4 linkage in BPH. Pathological markers of BPH were measured by H&E staining, immunoblotting, ELISA, qRT-PCR, and immunofluorescence to examine the effect of Apo. Rats stimulated with testosterone and BPH-1 cells were used as BPH models. Results: AR and NOX4 network-mediated oxidative stress was upregulated in the BPH model. Next, we examined the effects of Apo on oxidative stress and chronic prostatic inflammation in BPH mouse models. In a testosterone-induced BPH rat model, Apo alleviated pathological prostate enlargement and suppressed androgen/AR signaling. Apo suppressed the upregulation of proinflammatory markers and promoted the expression of anti-oxidant factors. Furthermore, Apo regulated the TGF-β/Glut9/activin pathway and macrophage programming. In BPH-1 cells, Apo suppressed AR-mediated proliferation and upregulation of TGFB and NOX4 expression by alleviating oxidative stress. Apo activated anti-oxidant and anti-inflammatory systems and regulated macrophage polarization in BPH-1 cells. AR knockdown partially abolished the beneficial effects of Apo in prostate cells, indicating AR-dependent effects of Apo. Conclusion: In contrast with existing BPH therapies, Apo may provide a new application for prostatic disease treatment, especially for BPH, by targeting the AR/TGF-β/NOX4 signaling pathway.

Work-related musculoskeletal disorders represent a major occupational disability issue, and 53.4% of these disorders occur in the back or shoulders. Various types of passive shoulder exoskeletons have been introduced to support the weight of the upper arm and work tools during overhead work, thereby preventing injuries and improving the work environment. The general passive shoulder exoskeleton is constructed with rigid links and joints to implement shoulder rotation, but there exists a challenge to align with the flexible joint movements of the human shoulder. Also, a force-generating part using mechanical springs require additional mechanical components to generate torque similar to the shoulder joint, resulting in increased overall volume and inertia to the upper arm. In this study, we propose a new type of passive shoulder exoskeleton that uses magnetic spring joint and link chain. The redundant degrees of freedom in the link chains enables to follow the shoulder joint movement in the horizontal direction, and the magnetic spring joint generates torque without additional parts in a compact form. Conventional exoskeletons experience a loss in the assisting torque when the center of shoulder rotation changed during arm elevation. Our exoskeleton minimizes the torque loss by customizing the installation height and initial angle of the magnetic spring joint. The performances of the proposed exoskeleton were verified by an electromyographic evaluation of shoulder-related muscles in overhead work and box lifting task.

C-Phycocyanin (C-PC) is a photosynthetic pigment found in cyanobacteria, notably in Arthrospira species. The extraction of phycocyanobilin (PCB), the chromophore of C-PC, is a common approach to address the instability of C-PC under light, heat, and acidic conditions. Methanol is typically used for PCB extraction. However, its use poses challenges for industrial applications owing to the need for solvent removal, extensive purification, and safety validation. Therefore, this study proposes ethanol as an alternative to methanol, optimizing the ethanol extraction conditions through response surface methodology (RSM) using a central composite design (CCD) and techno-economic analysis. The parameters evaluated were the extraction temperature, time, and C-PC/solvent ratio. Optimal conditions—68.81 °C, 14.91 h, and a C-PC to solvent ratio of 1:95 (w/v)— yielded a predicted PCB yield of 29.18%, closely aligning with the actual value of 29.67 ± 1.33%. A techno-economic analysis for pilot-scale PCB production showed that optimized ethanol extraction could yield 147.13 kg/year with 506 batches, compared with 84.31 kg/year standard methanol extraction with 317 batches. Furthermore, it was evaluated to have a unit production cost of USD 1,413,588/kg, an internal rate of return (IRR) of 53.36%, and a payback time of 1.6 years with increased yields and reduced toxic solvent disposal costs. This study supports scalable PCB production with a natural blue pigment suitable for the food, beverage, and cosmetics industries.

Despite the many beneficial effects of phycocyanobilin (PCB) on human skin, its cosmetic applications have not been extensively investigated owing to its light and temperature sensitivity. This is the first report of PCB extract (SP) derived from marine Arthrospira maxima having skin anti-wrinkling effects associated with antioxidant efficacy and reduction of intracellular reactive oxygen species (ROS) production. We obtained 46.63 ± 1.72 mg PCB/g dry weight of A. maxima in SP through an ethanol extraction process. PCB extracts showed strong effects in increasing collagen synthesis and decreasing matrix metalloproteinases (MMP-1) production. Interestingly, skin anti-wrinkling effects of the PCB extracts were significantly increased by the addition of wheat bran extracts (WB), up to 20–30% of the effects of PCB at all concentrations, possibly due to the synergistic effects of soluble globulins and other active substances in WB. Moreover, the mixture of SP and WB (SPWB) greatly reduced cell cytotoxicity to approximately 15% of that of PCB. SPWB upregulated and downregulated the expression of collagen type I α1 (Col1A1) and MMP-1, respectively, although the downregulation of MMP-1 was higher than that of Col1A1. The optimal SPWB concentration for maintaining the highest skin anti-wrinkling effects was 0.5 mg/mL. We show that SPWB holds promise as a vegan cosmaceutical.

Ulva pertusa Kjellman (U. pertusa) is a seaweed indigenous to the intertidal zone of the Korean coastline. U. pertusa exhibits immune-enhancing and antitumor activities, and its effects on intestinal health have gained attention. However, the mechanisms underlying its beneficial effects on intestinal physiology remain elusive. Here, the effect of U. pertusa intake in ameliorating loperamide-induced constipation in male mice was evaluated. Additionally, cellular levels of proinflammatory cytokines, including nuclear factor-kB and interleukin-1β, were assessed to decipher the intricate interplay between inflammation and improvements in bowel movement. U. pertusa intake increased fecal weight and water content and improved the intestinal transit rate. Moreover, it reduced the levels of proinflammatory cytokines, possibly via short-chain fatty acids implicated in modulating intestinal motility and mucosal inflammation. These findings underscore the efficacy of U. pertusa in improving bowel motility and intestinal functionality, and its potential in ameliorating constipation.

Abstract Despite intensive clinical and scientific efforts, the mortality rate of sepsis remains high due to the lack of precise biomarkers for patient stratification and therapeutic guidance. Secreted human tryptophanyl-tRNA synthetase 1 (WARS1), an endogenous ligand for Toll-like receptor (TLR) 2 and TLR4 against infection, activates the genes that signify the hyperinflammatory sepsis phenotype. High plasma WARS1 levels stratified the early death of critically ill patients with sepsis, along with elevated levels of cytokines, chemokines, and lactate, as well as increased numbers of absolute neutrophils and monocytes, and higher Sequential Organ Failure Assessment (SOFA) scores. These symptoms were recapitulated in severely ill septic mice with hypercytokinemia. Further, injection of WARS1 into mildly septic mice worsened morbidity and mortality. We created an anti-human WARS1-neutralizing antibody that suppresses proinflammatory cytokine expression in marmosets with endotoxemia. Administration of this antibody into severe septic mice attenuated cytokine storm, organ failure, and early mortality. With antibiotics, the antibody almost completely prevented fatalities. These data imply that blood-circulating WARS1-guided anti-WARS1 therapy may provide a novel theranostic strategy for life-threatening systemic hyperinflammatory sepsis.

Recent advancements in the large-scale cultivation of Tetraselmis sp. in Korea have enabled year-round production of this marine microalgae. This study explores the potential industrial applications of Tetraselmis sp. biomass by investigating the antiviral properties of its extracts and primary components. The antiviral effects of Tetraselmis sp. extracts were evaluated in Zika virus (ZIKV)-infected cells. Following extensive isolation and purification, the main compounds were characterized using liquid chromatography–mass spectrometry (LC-MS) and nuclear magnetic resonance (NMR) analyses. Their antiviral activities were confirmed using in vitro and in silico tests. Tetraselmis sp. extracts reduced infectious viral particles and non-structural protein 1 messenger RNA levels in ZIKV-infected cells without inducing cytotoxicity. Additionally, they modulated the interferon-mediated immune system responses. Tetraselmis sp. extracts are composed of four main chlorophylls: chlorophyll a, chlorin e6-131-152-dimethyl-173-phytyl ester, hydroxychlorophyll a, and hydroxypheophytin a. Among them, chlorophyll a, chlorin e6-131-152-dimethyl-173-phytyl ester, and hydroxypheophytin showed the antiviral activities in ZIKV-infected cells and molecular docking simulations predicted interactions between these chlorophylls and ZIKV. Our findings suggest that Tetraselmis sp. chlorophyll extracts exert antiviral effects against ZIKV and could serve as potential therapeutic candidates against ZIKV infection.

Fibroblast growth factor 2 (FGF2) is an attractive biomaterial for pharmaceuticals and functional cosmetics. To improve the thermo-stability of FGF2, we designed two mutants harboring four-point mutations: FGF2-M1 (D28E/C78L/C96I/S137P) and FGF2-M2 (D28E/C78I/C96I/S137P) through bioinformatics, molecular thermodynamics, and molecular modeling. The D28E mutation reduced fragmentation of the FGF2 wild type during preparation, and the substitution of a whale-specific amino acid, S137P, enhanced the thermal stability of FGF2. Surface-exposed cysteines that participate in oligomerization through intermolecular disulfide bond formation were substituted with hydrophobic residues (C78L/C78I and C96I) using the in silico method. High-resolution crystal structures revealed at the atomic level that the introduction of mutations stabilizes each local region by forming more favorable interactions with neighboring residues. In particular, P137 forms CH-π interactions with the side chain indole ring of W123, which seems to stabilize a β-hairpin structure, containing a heparin-binding site of FGF2. Compared to the wild type, both FGF2-M1 and FGF2-M2 maintained greater solubility after a week at 45 °C, with their Tm values rising by ~ 5 °C. Furthermore, the duration for FGF2-M1 and FGF2-M2 to reach 50% residual activity at 45 °C extended to 8.8- and 8.2-fold longer, respectively, than that of the wild type. Interestingly, the hydrophobic substitution of surface-exposed cysteine in both FGF2 mutants makes them more resistant to proteolytic cleavage by trypsin, subtilisin, proteinase K, and actinase than the wild type and the Cys → Ser substitution. The hydrophobic replacements can influence protease resistance as well as oligomerization and thermal stability. It is notable that hydrophobic substitutions of surface-exposed cysteines, as well as D28E and S137P of the FGF2 mutants, were designed through various approaches with structural implications. Therefore, the engineering strategies and structural insights adopted in this study could be applied to improve the stability of other proteins.

Leptolyngbya sp. KIOST-1 has been proposed as a candidate species for use as a protein supplement due to its high protein content and absence of cytotoxicity. The species has also garnered attention due to the photosynthetic pigments it possesses. However, limited information is available on its cultivation. Therefore, this study was conducted to identify the optimal culture medium and fundamental physiological properties of Leptolyngbya sp. KIOST-1 under various culture conditions. In this study, SOT (Society of Toxicology) medium was confirmed as the optimal culture medium for Leptolyngbya sp. KIOST-1 growth. The biomass production, protein content, and photosynthetic pigment content of Leptolyngbya sp. KIOST-1 were significantly higher in SOT medium. The use of this medium allowed for scaling up from laboratory (10 mL) to pilot (200 L) conditions and industrial-scale outdoor conditions (10,000 L), with the biomass containing over 66% protein. The phytochemical composition of Leptolyngbya sp. KIOST-1 cultured at laboratory and industrial-scales was discovered in this study. Furthermore, we observed that reducing the carbon and nitrogen sources to 1/5 of those supplied by the optimal medium did not significantly affect biomass production, and Leptolyngbya sp. KIOST-1 demonstrated favorable growth capabilities in a salinity range of 10–50 psu and at pH levels of 8.3 to 10.3. The results of this study demonstrate the suitability of Leptolyngbya sp. KIOST-1 for various industrial applications and its adaptability to large-scale cultivation.

Microalgae have gained attention as a promising source of chlorophylls and carotenoids in various industries. However, scaling up of conventional bubble columns presents challenges related to cell sedimentation and the presence of non-photosynthetic cells due to non-circulating zones and decreased light accessibility, respectively. Therefore, this study aimed to evaluate the newly developed continuously circulated bioreactor ROSEMAX at both laboratory and pilot scales, compared to a conventional bubble column. There was no significant difference in the biomass production and photosynthetic pigment content of Tetraselmis sp. cultivated at the laboratory scale (p > 0.05). However, at the pilot scale, the biomass cultured in ROSEMAX showed significantly high biomass (1.69 ± 0.11 g/L, dry weight, DW), chlorophyll-a (14.60 ± 0.76 mg/g, DW), and total carotene (5.64 ± 0.81 mg/g, DW) concentrations compared to the conventional bubble column (1.17 ± 0.11 g/L, DW, 10.67 ± 0.72 mg/g, DW, 3.21 ± 0.56 mg/g, DW, respectively) (p ≤ 0.05). Flow cytometric analyses confirmed that the proportion of Tetraselmis sp. live cells in the culture medium of ROSEMAX was 32.90% higher than that in the conventional bubble column, with a photosynthetic efficiency 1.14 times higher. These results support suggestions to use ROSEMAX as a bioreactor for industrial-scale applications.

Osteoarthritis (OA) is the most common joint disease that causes local inflammation and pain, significantly reducing the quality of life and normal social activities of patients. Currently, there are no disease-modifying OA drugs (DMOADs) available, and treatment relies on pain relief agents or arthroplasty. To address this significant unmet medical need, we aimed to develop monoclonal antibodies that can block the osteoclast-associated receptor (OSCAR). Our recent study has revealed the importance of OSCAR in OA pathogenesis as a novel catabolic regulator that induces chondrocyte apoptosis and accelerates articular cartilage destruction. It was also shown that blocking OSCAR with a soluble OSCAR decoy receptor ameliorated OA in animal models. In this study, OSCAR-neutralizing monoclonal antibodies were isolated and optimized by phage display. These antibodies bind to and directly neutralize OSCAR, unlike the decoy receptor, which binds to the ubiquitously expressed collagen and may result in reduced efficacy or deleterious off-target effects. The DMOAD potential of the anti-OSCAR antibodies was assessed with in vitro cell-based assays and an in vivo OA model. The results demonstrated that the anti-OSCAR antibodies significantly reduced cartilage destruction and other OA signs, such as subchondral bone plate sclerosis and loss of hyaline cartilage. Hence, blocking OSCAR with a monoclonal antibody could be a promising treatment strategy for OA.

Summary: While cytoplasmic tryptophanyl-tRNA synthetase (WARS1) ligates tryptophan (Trp) to its cognate tRNAs for protein synthesis, it also plays a role as an innate immune activator in extracellular space. However, its secretion mechanism remains elusive. Here, we report that in response to stimuli, WARS1 can be secreted via two distinct pathways: via Trp-dependent secretion of naked protein and via Trp-independent plasma-membrane-derived vesicles (PMVs). In the direct pathway, Trp binding to WARS1 induces a “closed” conformation, generating a hydrophobic surface and basic pocket. The Trp-bound WARS1 then binds stable phosphatidylinositol (4,5)-biphosphate and inner plasma membrane leaflet, passing across the membrane. In the PMV-mediated secretion, WARS1 recruits calpain 2, which is activated by calcium. WARS1 released from PMVs induces inflammatory responses in vivo. These results provide insights into the secretion mechanisms of WARS1 and improve our understanding of how WARS1 is involved in the control of local and systemic inflammation upon infection.

Abstract Serum is a stable medium supplement for in vitro cell culture. Live cells are used in stem cell research, drug toxicity and safety testing, disease diagnosis and prevention, and development of antibiotics, drugs, and vaccines. However, use of serum in culture involves concerns such as an ethical debate regarding the collection process, lack of standardized ingredients, and high cost. Herein, therefore, we evaluated the possibility of using edible cyanobacterium (Spirulina maxima), which is a nutrient-rich, sustainable, and ethically acceptable source, as a novel substitute for fetal bovine serum (FBS). H460 cells were cultured to the 10th generation by adding a mixture of spirulina animal cell culture solution (SACCS) and FBS to the culture medium. Cell morphology and viability, cell cycle, apoptosis, proteomes, and transcriptomes were assessed. We observed that SACCS had better growth-promoting capabilities than FBS. Cell proliferation was promoted even when FBS was replaced by 50–70% SACCS; there was no significant difference in cell shape or viability. There were only slight differences in the cell cycle, apoptosis, proteomes, and transcriptomes of the cells grown in presence of SACCS. Therefore, SACCS has the potential to be an effective, low-cost, and eco-friendly alternative to FBS in in vitro culture.

Protein dimerization or oligomerization resulting from swapping part of the protein between neighboring polypeptide chains is known to play a key role in the regulation of protein function and in the formation of protein aggregates. Glutaredoxin-1 from Clostridium oremlandii (cGrx1) was used as a model to explore the formation of multiple domain-swapped conformations, which were made possible by modulating several hinge-loop residues that can form a pivot for domain swapping. Specifically, two alternative domain-swapped structures were generated and analyzed using nuclear magnetic resonance (NMR), X-ray crystallography, circular-dichroism spectroscopy and hydrogen/deuterium-exchange (HDX) mass spectrometry. The first domain-swapped structure (β3-swap) was formed by the hexameric cGrx1–cMsrA complex. The second domain-swapped structure (β1-swap) was formed by monothiol cGrx1 (C16S) alone. In summary, the first domain-swapped structure of an oxidoreductase in a hetero-oligomeric complex is presented. In particular, a single point mutation of a key cysteine residue to serine led to the formation of an intramolecular disulfide bond, as opposed to an intermolecular disulfide bond, and resulted in modulation of the underlying free-energy landscape of protein oligomerization.

Indoleamine 2,3-dioxygenase 1 (IDO1), a tryptophan catabolising enzyme, is known as a tumour cell survival factor that causes immune escape in several types of cancer. Flavonoids of Sophora flavescens have a variety of biological benefits for humans; however, cancer immunotherapy effect has not been fully investigated. The flavonoids (1–6) isolated from S. flavescens showed IDO1 inhibitory activities (IC50 4.3–31.4 µM). The representative flavonoids (4–6) of S. flavescens were determined to be non-competitive inhibitors of IDO1 by kinetic analyses. Their binding affinity to IDO1 was confirmed using thermal stability and surface plasmon resonance (SPR) assays. The molecular docking analysis and mutagenesis assay revealed the structural details of the interactions between the flavonoids (1–6) and IDO1. These results suggest that the flavonoids (1–6) of S. flavescens, especially kushenol E (6), as IDO1 inhibitors might be useful in the development of immunotherapeutic agents against cancers.

Abstract G-protein coupled receptors (GPCRs) play indispensable physiological roles in cell proliferation, differentiation, and migration; therefore, identifying the mechanisms by which ligands bind to GPCRs is crucial for developing GPCR-targeting pharmaceutics and for understanding critical biological functions. Although some structural information is available regarding the interactions between GPCRs and their small molecule ligands, knowledge of how GPCRs interact with their corresponding macromolecule ligands, such as peptides and proteins, remains elusive. In this study, we have developed a novel strategy to investigate the precise ligand recognition mechanisms involved in the interaction of endothelin receptor type A (ETA) with its ligand, endothelin-1 (ET-1); we call this method “directed degeneration” method. Through flow cytometric screening of a randomized ETA library, statistical analysis of the identified sequences, and biochemical studies, the ligand interaction map was successfully obtained.

There is increasing demand for essential fatty acids (EFAs) from non-fish sources such as microalgae, which are considered a renewable and sustainable biomass. The open raceway system (ORS) is an affordable system for microalgae biomass cultivation for industrial applications. However, seasonal variations in weather can affect biomass productivity and the quality of microalgal biomass. The aim of this study was to determine the feasibility of year-round Tetraselmis sp. cultivation in a semi-ORS in Korea for biomass and bioactive lipid production. To maximize biomass productivity of Tetraselmis sp., f medium was selected because it resulted in a significantly higher biomass productivity (1.64 ± 0.03 g/L) and lower omega-6/omega-3 ratio (0.52/1) under laboratory conditions than f/2 medium (0.70/1). Then, we used climatic data-based building information modeling technology to construct a pilot plant of six semi-ORSs for controlling culture conditions, each with a culture volume of 40,000 L. Over 1 year, there were no significant variations in monthly biomass productivity, fatty acid composition, or the omega-6/omega-3 ratio; however, the lipid content correlated significantly with photosynthetic photon flux density. During year-round cultivation from November 2014 to October 2017, areal productivity was gradually increased by increasing medium salinity and injecting CO2 gas into the culture medium. Productivity peaked at 44.01 g/m2/d in October 2017. Throughout the trials, there were no significant differences in average lipid content, which was 14.88 ± 1.26%, 14.73 ± 2.44%, 12.81 ± 2.82%, and 13.63 ± 3.42% in 2014, 2015, 2016, and 2017, respectively. Our results demonstrated that high biomass productivity and constant lipid content can be sustainably maintained under Korean climate conditions.

Tryptophanyl-tRNA synthetase 1 (WARS1) is an endogenous ligand of mammalian Toll-like receptors (TLR) 2 and TLR4. Microarray data, using mRNA from WARS1-treated human peripheral blood mononuclear cells (PBMCs), had indicated WARS1 to mainly activate innate inflammatory responses. However, exact molecular mechanism remains to be understood. The triggering receptor expressed on myeloid cells (TREM)-1 is an amplifier of pro-inflammatory processes. We found WARS1 to significantly activate TREM-1 at both mRNA and protein levels, along with its cell surface expression and secretion in macrophages. WARS1 stimulated TREM-1 production via TLR2 and TLR4, mediated by both MyD88 and TRIF, since targeted deletion of TLR4, TLR2, MyD88, and TRIF mostly abrogated TREM-1 activation. Furthermore, WARS1 promoted TREM-1 downstream phosphorylation of DAP12, Syk, and AKT. Knockdown of TREM-1 and inhibition of Syk kinase significantly suppressed the activation of inflammatory signaling loop from MyD88 and TRIF, leading to p38 MAPK, ERK, and NF-κB inactivation. Finally, MyD88, TRIF, and TREM-1 signaling pathways were shown to be cooperatively involved in WARS1-triggered massive production of IL-6, TNF-α, IFN-β, MIP-1α, MCP-1, and CXCL2, where activation of Syk kinase was crucial. Taken together, our data provided a new insight into WARS1′s strategy to amplify innate inflammatory responses via TREM-1.

We analyze both the s- and p -wave collision induced frequency shifts and propose an over- π pulse scheme to cancel the shifts in optical lattice clocks interrogated by a Rabi pulse. The collisional frequency shifts are analytically solved as a function of the pulse area and the inhomogeneity of the Rabi frequencies. Experimentally measured collisional frequency shifts in an Yb optical lattice clock are in good agreement with the analytical calculations. Based on our analysis, the over- π pulse combined with a small inhomogeneity below 0.1 allows a fractional uncertainty on a level of 10 ^−18 in both Sr and Yb optical lattice clocks by canceling the collisional frequency shift.

Recently, extracellular vesicles (EVs) have been developed as therapeutic targets for various diseases. Biodistribution is crucial for EVs intended for therapeutic purposes because it can determine the degree of on‐ and off‐target effects. This study aimed to explore techniques to evaluate the biodistribution of unmodified EVs. We devised a novel quantitative polymerase chain reaction (qPCR)‐based assay to detect unmodified EVs by targeting mitochondrial deoxyribonucleic acid (mtDNA), a constituent of EVs. We focused on specific mtDNA regions that exhibited homologous variations distinct from their rodent mtDNA counterparts to establish this analytical approach. Herein, we successfully designed primers and probes targeting human and rodent mtDNA sequences and developed a highly specific and sensitive qPCR method. Furthermore, the quantification range of EVs isolated from various cells differed based on the manufacturer and cell source. IRDye 800CW‐labelled Expi293F EV mimetics were administered to the animals via the tail vein to compare the imaging test and mtDNA‐qPCR results. The results obtained from imaging tests and mtDNA‐qPCR to investigate EV biodistribution patterns revealed differences. The results revealed that our newly developed method effectively determined the biodistribution of unmodified EVs with high sensitivity and reproducibility. [ABSTRACT FROM AUTHOR]

Thermodynamic analysis for inclusion complexation between molecular tube (MT) and poly(ethylene oxide)-block-poly(tetrahydrofuran)-block-poly(ethyleneoxide) triblock copolymer was carried out in terms of isothermal titration calorimetry (ITC). The thermodynamic parameters suggest that van der Waals interaction, hydrophobic interaction, and hydrogen bond interaction participate in inclusion complexation.

Locomotion of an organism interacting with an environment is the consequence of a symmetry-breaking action in space-time. Here we show a minimal instantiation of this principle using a thin circular sheet, actuated symmetrically by a pneumatic source, using pressure to change shape nonlinearly via a spontaneous buckling instability. This leads to a polarized, bilaterally symmetric cone that can walk on land and swim in water. In either mode of locomotion, the emergence of shape asymmetry in the sheet leads to an asymmetric interaction with the environment that generates movement--via anisotropic friction on land, and via directed inertial forces in water. Scaling laws for the speed of the sheet of the actuator as a function of its size, shape, and the frequency of actuation are consistent with our observations. The presence of easily controllable reversible modes of buckling deformation further allows for a change in the direction of locomotion in open arenas and the ability to squeeze through confined environments--both of which we demonstrate using simple experiments. Our simple approach of harnessing elastic instabilities in soft structures to drive locomotion enables the design of novel shape-changing robots and other bioinspired machines at multiple scales. [ABSTRACT FROM AUTHOR]

Toxoplasma gondii, a ubiquitous, intracellular parasite of the phylum Apicomplexa, infects an estimated onethird of the human population as well as a broad range of warm-blooded animals. We have observed that some tyrosine kinase inhibitors suppressed the growth of T. gondii within host ARPE-10 cells. Among them, afatinib, human epithermal growth factor receptor 2 and 4 (HER2/4) inhibitor, may be used as a therapeutic agent for inhibiting parasite growth with minimal adverse effects on host. In this report, we conducted a proteomic analysis to observe changes in host proteins that were altered via infection with T. gondii and the treatment of HER2/4 inhibitors. Secreting proteins were subjected to a procedure of micor basic reverse phase liquid chromatography, nano-liquid chromatography-mass spectrometry, and ingenuity pathway analysis serially. As a result, the expression level of heterogeneous nuclear ribonucleoprotein K, semaphorin 7A, a GPI membrane anchor, serine/threonine-protein phosphatase 2A, and calpain small subunit 1 proteins were significantly changed, and which were confirmed further by western blot analysis. Changes in various proteins, including these 4 proteins, can be used as a basis for explaining the effects of T. gondii infections and HER2/4 inhibitors. [ABSTRACT FROM AUTHOR]

The characteristics of the zinc oxide (ZnO) inorganic film deposited by radio-frequency magnetron sputtering as an alternative alignment layer for liquid crystal display (LCD) applications are described. Ion beam (IB) bombardment at various incident angles is used for inducing liquid crystal (LC) alignment and has caused the measured pretilt angle on ZnO films to assume a triangular contour.

The effects of tyrosine kinase inhibitors (TKIs) were evaluated on growth inhibition of intracellular Toxoplasma gondii in host ARPE-19 cells. The number of tachyzoites per parasitophorous vacuolar membrane (PVM) was counted after treatment with TKIs. T. gondii protein expression was assessed by western blot. Immunofluorescence assay was performed using Programmed Cell Death 4 (PDCD4) and T. gondii GRA3 antibodies. The TKIs were divided into 3 groups; non-epidermal growth factor receptor (non-EGFR), anti-human EGFR 2 (anti-HER2), and anti-HER2/4 TKIs, respectively. Group I TKIs (nintedanib, AZD9291, and sunitinib) were unable to inhibit proliferation without destroying host cells. Group II TKIs (lapatinib, gefitinib, erlotinib, and AG1478) inhibited proliferation up to 98% equivalent to control pyrimethamine (5 μM) at 20 μM and higher, without affecting host cells. Group III TKIs (neratinib, dacomitinib, afatinib, and pelitinib) inhibited proliferation up to 98% equivalent to pyrimethamine at 1-5 μM, but host cells were destroyed at 10-20 μM. In Group I, TgHSP90 and SAG1 inhibitions were weak, and GRA3 expression was moderately inhibited. In Group II, TgHSP90 and SAG1 expressions seemed to be slightly enhanced, while GRA3 showed none to mild inhibition; however, AG1478 inhibited all proteins moderately. Protein expression was blocked in Group III, comparable to pyrimethamine. PDCD4 and GRA3 were well localized inside the nuclei in Group I, mildly disrupted in Group II, and were completely disrupted in Group III. This study suggests the possibility of a vital T. gondii TK having potential HER2/4 properties, thus anti-HER2/4 TKIs may inhibit intracellular parasite proliferation with minimal adverse effects on host cells. [ABSTRACT FROM AUTHOR]

Nanostructured surfaces with quasi-random geometries can manipulate light over broadband wavelengths and wide ranges of angles. Optimization and realization of stochastic patterns have typically relied on serial, direct-write fabrication methods combined with real-space design. However, this approach is not suitable for customizable features or scalable nanomanufacturing. Moreover, trial-and-error processing cannot guarantee fabrication feasibility because processing–structure relations are not included in conventional designs. Here, we report wrinkle lithography integrated with concurrent design to produce quasi-random nanostructures in amorphous silicon at wafer scales that achieved over 160% light absorption enhancement from 800 to 1,200 nm. The quasi-periodicity of patterns, materials filling ratio, and feature depths could be independently controlled. We statistically represented the quasi-random patterns by Fourier spectral density functions (SDFs) that could bridge the processing–structure and structure–performance relations. Iterative search of the optimal structure via the SDF representation enabled concurrent design of nanostructures and processing. [ABSTRACT FROM AUTHOR]

A flag-type atom shutter based on a rotating lever that is driven by a bender piezoelectric actuator was developed to manipulate atomic beams. The shutter flag was displaced by ~10 mm to open and close a 5-mm-diameter aperture with a shutter time of 13 ms that produced small mechanical vibrations. The short-term shutter time stability for each cycle was 0.03 ms and the long-term stability over an average of 20 000 cycles was 0.02 ms. The operational cycle number (lifetime) of the shutter reached 2.0 × 106 cycles after an intermittent operation over a period of eight months in an ultra-high vacuum chamber, and another shutter in an atmospheric environment swung for 2.6 × 107 cycles of continuous operation at 5 Hz for a period of 60 days without major problems. The shutter was shown to be compatible with the operation in an ultra-high vacuum at a low 10-7 Pa level after a gentle baking treatment. [ABSTRACT FROM AUTHOR]

The Center for Time and Frequency at the Korea Research Institute of Standards and Science (KRISS) is responsible for the maintenance, improvement, and dissemination of time and frequency standards in Korea. In this paper, we will introduce current research activities regarding the development of precision atomic clocks, atomic gravimeters, miniature atomic clocks, and Bloch oscillation interferometers. [ABSTRACT FROM AUTHOR]

Plasmonic nanostructures with enhanced localized optical fields as well as narrow linewidths have driven advances in numerous applications. However, the active engineering of ultranarrow resonances across the visible regime—and within a single system—has not yet been demonstrated. This paper describes how aluminum nanoparticle arrays embedded in an elastomeric slab may exhibit high-quality resonances with linewidths as narrow as 3 nm at wavelengths not accessible by conventional plasmonic materials. We exploited stretching to improve and tune simultaneously the optical response of as-fabricated nanoparticle arrays by shifting the diffraction mode relative to single-particle dipolar or quadrupolar resonances. This dynamic modulation of particle–particle spacing enabled either dipolar or quadrupolar lattice modes to be selectively accessed and individually optimized. Programmable plasmon modes offer a robust way to achieve real-time tunable materials for plasmon-enhanced molecular sensing and plasmonic nanolasers and opens new possibilities for integrating with flexible electronics. [ABSTRACT FROM AUTHOR]

This paper describes how delamination-free, hierarchical patterning of graphene can be achieved on prestrained thermoplastic sheets by surface wrinkling. Conformal contact between graphene and the substrate during strain relief was maintained by the presence of a soft skin layer, resulting in the uniform patterning of three-dimensional wrinkles over large areas (>cm²). The graphene wrinkle wavelength was tuned from the microscale to the nanoscale by controlling the thickness of the skin layer with 1 nm accuracy to realize a degree of control not possible by crumpling, which relies on delamination. Hierarchical patterning of the skin layers with varying thicknesses enabled multiscale graphene wrinkles with predetermined orientations to be formed. Significantly, hierarchical graphene wrinkles exhibited tunable mechanical stiffness at the nanoscale without compromising the macroscale electrical conductivity. [ABSTRACT FROM AUTHOR]

We report the design of three-dimensional (3D) hierarchical wrinkle substrates that can maintain their superhydrophobicity even after being repeatedly stretched. Monolithic poly(dimethysiloxane) with multiscale features showed wetting properties characteristic of static superhydrophobicity with water contact angles (<160°) and very low contact angle hysteresis (>5°). To examine how superhydrophobicity was maintained as the substrate was stretched, we investigated the dynamic wetting behavior of bouncing and splashing upon droplet impact with the surface. On hierarchical wrinkles consisting of three different length scales, superhydrophobic bouncing was observed. The substrate remained superhydrophobic up to 100% stretching with no structural defects after 1000 cycles of stretching and releasing. Stretchable superhydrophobicity was possible because of the monolithic nature of the hierarchical wrinkles as well as partial preservation of nanoscale structures under stretching. [ABSTRACT FROM AUTHOR]

Intercellular bridges are a conserved feature of spermatogenesis in mammalian germ cells and derive from arresting cell abscission at the final stage of cytokinesis. However, it remains to be fully understood how germ cell abscission is arrested in the presence of general cytokinesis components. The TEX14 (testis-expressed gene 14) protein is recruited to the midbody and plays a key role in the inactivation of germ cell abscission. To gain insights into the structural organization of TEX14 at the midbody, we have determined the crystal structures of the EABR [endosomal sorting complex required for transport (ESCRT) and ALIX-binding region] of CEP55 bound to the TEX14 peptide (or its chimeric peptides) and performed functional characterization of the CEP55-TEX14 interaction by multiexperiment analyses. We show that TEX14 interacts with CEP55-EABR via its AxGPPx3Y (Ala793, Gly795, Pro796, Pro797, and Tyr801) and PP (Pro803 and Pro804) sequences, which together form the AxGPPx3YxPP motif. TEX14 competitively binds to CEP55-EABR to prevent the recruitment of ALIX, which is a component of the ESCRT machinery with the AxGPPx3Y motif. We also demonstrate that a high affinity and a low dissociation rate of TEX14 to CEP55, and an increase in the local concentration of TEX14, cooperatively prevent ALIX from recruiting ESCRT complexes to the midbody. The action mechanism of TEX14 suggests a scheme of how to inactivate the abscission of abnormal cells, including cancer cells. [ABSTRACT FROM AUTHOR]

This paper describes how a memory-based,sequential wrinkling process can transform flat polystyrene sheetsinto multiscale, three-dimensional hierarchical textures. Multiplecycles of plasma-mediated skin growth followed by directional strainrelief of the substrate resulted in hierarchical architectures withcharacteristic generational (G) features. Independent control overwrinkle wavelength and wrinkle orientation for each G was achievedby tuning plasma treatment time and strain-relief direction for eachcycle. Lotus-type superhydrophobicity was demonstrated on three-dimensionalG1–G2–G3 hierarchical wrinkles as well as tunable superhydrophilicityon these same substrates after oxygen plasma. This materials systemprovides a general approach for nanomanufacturing based on bottom-upsequential wrinkling that will benefit a diverse range of applicationsand especially those that require large area (>cm2),multiscale, three-dimensional patterns. [ABSTRACT FROM AUTHOR]

Proteomic analyses of the excretory/secretory proteins from the RH strain of Toxoplasma gondii have been performed to understand their functions in the host-parasite interaction. A total of 34 proteins were identified from LC/MS/MS analysis and their abundance was estimated by spectral counting methods. Among them, 8 species of micronemal proteins (MICs), 2 species of rhoptry proteins (ROPs), and 6 species of dense granular proteins (GRAs) were confirmed. Besides these, 18 species of protein were newly identified, and their cellular functions were estimated from sequence analysis. The three most abundant of the 34 identified extractor/secretory proteins GRA1, GRA7 and GRA2 were confirmed to be highly expressed in T. gondii using the spectral count method. This phenomenon is another demonstration of the importance of GRA proteins for the penetration and survival of T. gondii. [ABSTRACT FROM AUTHOR]

Rhoptry protein 6 (ROP6) from Toxoplasma gondii is a 480-amino acid protein with no homology to any reported excretory or secretory protein. Especially, unlike the many other rhoptry protein types, ROP6 does not have a kinase domain. The biochemical and biophysical properties of ROP6 are unknown. Here, we investigated its structure using an in silico analysis method and overexpression and purification using an Escherichia coli system. The protein was purified to more than 85% homogeneity using immobilized metal affinity chromatography in denaturing conditions. After purification, ROP6 showed slow migration in SDS-PAGE, including fast proteolysis. This implies that ROP6 has a high percentage of flexible regions or extended loop structures. Secondary structure prediction and prediction of intrinsically disordered regions by using various bioinformatics tools, indicated that approximately 60% of ROP6 is predicted to be intrinsically disordered or random coil regions. These observations indicate that ROP6 is an intrinsically disordered protein. [ABSTRACT FROM AUTHOR]

Protein kinase CK2 is a ubiquitous kinase that can phosphorylate hundreds of cellular proteins and plays important roles in cell growth and development. Deregulation of CK2 is related to a variety of human cancers, and CK2 is regarded as a suppressor of apoptosis; therefore, it is a target of anticancer therapy. Nucleolar phosphoprotein 140 (Nopp140), which is an intrinsically disordered protein, interacts with CK2 and inhibits the latter’s catalytic activity in vitro. Interestingly, the catalytic activity of CK2 is recovered in the presence of D-myo-inositol 1,2,3,4,5,6-hexakisphosphate (IP6). IP6 is widely distributed in animal cells, but the molecular mechanisms that govern its cellular functions in animal cells have not been completely elucidated. In this study, the crystal structure of CK2 in complex with IP6 showed that the lysine-rich cluster of CK2 plays an important role in binding to IP6. The biochemical experiments revealed that a Nopp140 fragment (residues 568–596) and IP6 competitively bind to the catalytic subunit of CK2 (CK2α), and phospho-Ser574 of Nopp140 significantly enhances its interaction with CK2α. Substitutions of K74E, K76E, and K77E in CK2α significantly reduced the interactions of CK2α with both IP6 and the Nopp140-derived peptide. Our study gives an insight into the regulation of CK2. In particular, our work suggests that CK2 activity is inhibited by Nopp140 and reactivated by IP6 by competitive binding at the substrate recognition site of CK2. [ABSTRACT FROM AUTHOR]

The aligned liquid crystals (LCs) display on SiNx thin films using ion-beam (IB) irradiation was studied with controllability of pretilt angle depending on incident energies of the IB. The LCs alignment property for the SiNx thin films were observed to verify the practical application potential. A good LCs alignment of vertical alignment LCs cells on SiNx thin film surfaces irradiated with incident IB energy of 1800 eV was achieved. The atomic force microscopy (AFM) images of LCs on SiNx thin film surfaces irradiated with IB energy was used for the surface analysis. [ABSTRACT FROM AUTHOR]

We studied liquid crystal (LCs) alignment effects on homogeneous polyimide (PI) surfaces induced by polarised UV (PUV) exposure and rubbing in different directions. We found that the LC orientation is not affected by PUV exposure times of less than 90 min after rubbing, and that PUV exposure times over 90 min after rubbing resulted in LC orientations that were more closely aligned with the direction of PUV exposure than the direction of rubbing. Short-term PUV exposure affects the variation of the pretilt angle of the PI and LC. In addition, as PUV exposure time increases, it can convert the LC alignment direction from the rubbing direction to the PUV direction. [ABSTRACT FROM AUTHOR]