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In this study, a mixture of Platycodon grandiflorum , Pyrus serotina , Chaenomeles sinensis , and Raphanus sativus (PPCRE) was investigated for their immuno-enhancing effects, as well as the molecular mechanism of PPCRE in RAW264.7 cells. PPCRE dramatically increased nitric oxide (NO) and prostaglandin E 2 (PGE 2 ) generation depending on the concentration while exhibiting no cytotoxicity. PPCRE markedly upregulated the mRNA and protein expression of immune-related cytotoxic factors such as cyclooxygenase (COX)-1, COX-2, and inducible nitric oxide synthase (iNOS) and pro-inflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor alpha (TNF-α), as well as the mRNA level of IL-4 . PPCRE increased the mitogen-activated protein kinase (MAPK) signaling pathway by upregulating the phosphorylation of extracellular signal-regulated kinase (ERK), stress-activated protein kinase/Jun N-terminal-kinase (SAPK/JNK), and p38. Furthermore, PPCRE considerably activated the nuclear factor kappa B (NF-κB) signaling pathway by increasing phosphorylation of NF-κB-p65. PPCRE-stimulated RAW264.7 cells increased macrophage phagocytic capacity. In conclusion, our study found that PPCRE improved immune function by modulating inflammatory mediators and regulating the MAPK and NF-κB pathway of signaling in macrophages.

The immune-enhancing activity of the combination of Platycodon grandiflorum and Salvia plebeian extracts (PGSP) was evaluated through macrophage activation using RAW264.7 cells. PGSP (250-1000 μg/mL) showed a higher release of NO in a dose-dependent manner. The results showed that PGSP could significantly stimulate the production of PGE2, COX-2, TNF-α, IL-1β, and IL-6 in RAW264.7 cells and promote iNOS, COX-2, TNF-α, IL-1β, IL-4, and IL-6 mRNA expression. Western blot analysis demonstrated that the protein expression of iNOS and COX-2 and the phosphorylation of ERK, JNK, p38, and NF-κB p65 were greatly increased in PGSP-treated cells. PGSP also promoted the phagocytic activity of RAW264.7 cells. All these results indicated that PGSP might activate macrophages through MAPK and NF-κB signaling pathways. Taken together, PGSP may be considered to have immune-enhancing activity and might be used as a potential immune-enhancing agent., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Jang et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Open Reading Frame 6 (ORF6) proteins, which are unique to severe acute respiratory syndrome-related (SARS) coronavirus, inhibit the classical nuclear import pathway to antagonize host antiviral responses. Several alternative models were proposed to explain the inhibitory function of ORF6 [H. Xia et al ., Cell Rep. 33 , 108234 (2020); L. Miorin et al ., Proc. Natl. Acad. Sci. U.S.A. 117 , 28344-28354 (2020); and M. Frieman et al ., J. Virol. 81 , 9812-9824 (2007)]. To distinguish these models and build quantitative understanding of ORF6 function, we developed a method for scoring both ORF6 concentration and functional effect in single living cells. We combined quantification of untagged ORF6 expression level in single cells with optogenetics-based measurement of nuclear transport kinetics, using methods that could be adapted to measure concentration-dependent effects of any untagged protein. We found that SARS-CoV-2 ORF6 is ~15 times more potent than SARS-CoV-1 ORF6 in inhibiting nuclear import and export, due to differences in the C-terminal region that is required for the NUP98-RAE1 binding. The N-terminal region was required for transport inhibition. This region binds membranes but could be replaced by synthetic constructs which forced oligomerization in solution, suggesting its primary function is oligomerization. We propose that the hydrophobic N-terminal region drives oligomerization of ORF6 to multivalently cross-link the NUP98-RAE1 complexes at the nuclear pore complex, and this multivalent binding inhibits bidirectional transport., Competing Interests: Competing interests statement:The authors declare no competing interest.

All exogenous nanomaterials undergo rapid biotransformation once injected into the body and fall short of executing the intended purpose. Here, it is reported that copper-deposited ceria nanoparticles (CuCe NPs) exhibit enhanced antioxidant effects over pristine ceria nanoparticles, as the released copper buffers the depletion of glutathione while providing the bioavailable copper as a cofactor for the antioxidant enzyme, superoxide dismutase 1. The upregulated intracellular antioxidants along with the ceria nanoparticles synergistically scavenge reactive oxygen species and promote anti-inflammation and M2 polarization of macrophages by modulating signal transducer and activator of transcription 1 and 6 (STAT1 and STAT6). The therapeutic effect of CuCe NPs is demonstrated in ischemic vascular diseases (i.e., murine models of hindlimb ischemia and myocardial infarction) in which the copper-deposition affords increased perfusion and alleviation in tissue damage. The results provide rationale that metal oxide nanomaterials can be designed in a way to induce the upregulation of specific biological factors for optimal therapeutic performance., (© 2023 Wiley-VCH GmbH.)

Diagnosing salivary gland tumors (SGTs) through fine-needle aspiration (FNA) biopsies is challenging due to the overlapping cytomorphologic features between benign and malignant tumors. The authors developed an innovative, multiplexed cycling technology for the rapid analyses of single cells obtained from FNA that can facilitate the molecular analyses and diagnosis of SGTs. Antibodies against 29 protein markers associated with 7 SGT subtypes were validated and chemically modified via custom linker-bio-orthogonal probes (FAST). Single-cell homogenates and FNA samples were profiled by FAST cyclic imaging and computational analysis. A prediction model was generated using a training set of 151,926 cells from primary SGTs (N = 26) and validated on a separate cohort (N = 30). Companion biomarker testing, such as neurotrophic tyrosine receptor kinase (NTRK), was also assessed with the FAST technology. The FAST molecular diagnostic assay was able to distinguish between benign and malignant SGTs with an accuracy of 0.86 for single-cell homogenate samples and 0.88 for FNA samples. Profiling of multiple markers as compared to a single marker increased the diagnostic accuracy (0.82 as compared to 0.65-0.74, respectively), independent of the cell number sampled. NTRK expression was also assessed by the FAST assay, highlighting the potential therapeutic application of this technology. Application of the novel multiplexed single-cell technology facilitates rapid biomarker testing from FNA samples at low cost. The customizable and modular FAST-FNA approach has relevance to multiple pathologies and organ systems where cytologic samples are often scarce and/or indeterminate resulting in improved diagnostic workflows and timely therapeutic clinical decision-making., (© 2022 The Authors. Cancer Cytopathology published by Wiley Periodicals LLC on behalf of American Cancer Society.)

Several studies have examined the effects of micro- and nanoplastics on microbes, cells, and the environment. However, only a few studies have examined their effects-especially, those of their reduced cohesiveness-on cell viability and physiology. We synthesized surfactant-free amine-functionalized polystyrene (PS) nanoparticles (NPs) and PS-NPs with decreased crosslinking density (DPS-NPs) without changing other factors, such as size, shape, and zeta potential and examined their effects on cell viability and physiology. PS- and DPS-NPs exhibited reactive oxygen species (ROS) scavenging activity by upregulating GPX3 expression and downregulating HSP70 (ROS-related gene) and XBP1 (endoplasmic reticulum stress-related gene) expression in human bone marrow-derived mesenchymal stem cells (hBM-MSCs). Additionally, they led to upregulation of MFN2 (mitochondrial fusion related gene) expression and downregulation of FIS1 (mitochondrial fission related gene) expression, indicating enhanced mitochondrial fusion in hBM-MSCs. Cell-cycle analysis revealed that PS- and DPS-NPs increased the proportion of cells in the S phase, indicating that they promoted cell proliferation and, specifically, the adipogenic differentiation of hBM-MSCs. However, the cytotoxicity of DPS-NPs against hBM-MSCs was higher than that of PS-NPs after long-term treatment under adipogenic conditions., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Suppression of the host innate immune response is a critical aspect of viral replication. Upon infection, viruses may introduce one or more proteins that inhibit key immune pathways, such as the type I interferon pathway. However, the ability to predict and evaluate viral protein bioactivity on targeted pathways remains challenging and is typically done on a single-virus or -gene basis. Here, we present a medium-throughput high-content cell-based assay to reveal the immunosuppressive effects of viral proteins. To test the predictive power of our approach, we developed a library of 800 genes encoding known, predicted, and uncharacterized human virus genes. We found that previously known immune suppressors from numerous viral families such as Picornaviridae and Flaviviridae recorded positive responses. These include a number of viral proteases for which we further confirmed that innate immune suppression depends on protease activity. A class of predicted inhibitors encoded by Rhabdoviridae viruses was demonstrated to block nuclear transport, and several previously uncharacterized proteins from uncultivated viruses were shown to inhibit nuclear transport of the transcription factors NF-κB and interferon regulatory factor 3 (IRF3). We propose that this pathway-based assay, together with early sequencing, gene synthesis, and viral infection studies, could partly serve as the basis for rapid in vitro characterization of novel viral proteins. IMPORTANCE Infectious diseases caused by viral pathogens exacerbate health care and economic burdens. Numerous viral biomolecules suppress the human innate immune system, enabling viruses to evade an immune response from the host. Despite our current understanding of viral replications and immune evasion, new viral proteins, including those encoded by uncultivated viruses or emerging viruses, are being unearthed at a rapid pace from large-scale sequencing and surveillance projects. The use of medium- and high-throughput functional assays to characterize immunosuppressive functions of viral proteins can advance our understanding of viral replication and possibly treatment of infections. In this study, we assembled a large viral-gene library from diverse viral families and developed a high-content assay to test for inhibition of innate immunity pathways. Our work expands the tools that can rapidly link sequence and protein function, representing a practical step toward early-stage evaluation of emerging and understudied viruses.

An urgent need in chemodynamic therapy (CDT) is to achieve high Fenton catalytic efficiency at small doses of CDT agents. However, simple general promotion of the Fenton reaction increases the risk of damaging normal cells along with the cancer cells. Therefore, a tailored strategy to selectively enhance the Fenton reactivity in tumors, for example, by taking advantage of the characteristics of the tumor microenvironment (TME), is in high demand. Herein, a heterogeneous CDT system based on copper-iron peroxide nanoparticles (CFp NPs) is designed for TME-mediated synergistic therapy. CFp NPs degrade under the mildly acidic conditions of TME, self-supply H 2 O 2 , and the released Cu and Fe ions, with their larger portions at lower oxidation states, cooperatively facilitate hydroxyl radical production through a highly efficient catalytic loop to achieve an excellent tumor therapeutic efficacy. This is distinct from previous heterogeneous CDT systems in that the synergism is closely coupled with the Cu + -assisted conversion of Fe 3+ to Fe 2+ rather than their independent actions. As a result, almost complete ablation of tumors at a minimal treatment dose is demonstrated without the aid of any other therapeutic modality. Furthermore, CFp NPs generate O 2 during the catalysis and exhibit a TME-responsive T 1 magnetic resonance imaging contrast enhancement, which are useful for alleviating hypoxia and in vivo monitoring of tumors, respectively.

Noble metal-based multimetallic nanoparticles (NMMNs) have attracted great attention for their multifunctional and synergistic effects, which offer numerous catalytic applications. Combined experimental and theoretical studies have enabled formulation of various design principles for tuning the electrocatalytic performance through controlling size, composition, morphology, and crystal structure of the nanoparticles. Despite significant advancements in the field, the chemical synthesis of NMMNs with ideal characteristics for catalysis, including high activity, stability, product-selectivity, and scalability is still challenging. This review provides an overview on structure-based classification and the general synthesis of NMMN electrocatalysts. Furthermore, postsynthetic treatments, such as the removal of surfactants to optimize the activity, and utilization of NMMNs onto suitable support for practical electrocatalytic applications are highlighted. In the end, future direction and challenges associated with the electrocatalysis of NMMNs are covered., (© 2021 The Authors. Advanced Science published by Wiley-VCH GmbH.)

Macromolecular transport across the nuclear envelope depends on facilitated diffusion through nuclear pore complexes (NPCs). The interior of NPCs contains a permeability barrier made of phenylalanine-glycine (FG) repeat domains that selectively facilitates the permeation of cargoes bound to nuclear transport receptors (NTRs). FG-repeat domains in NPCs are a major site of O-linked N-acetylglucosamine (O-GlcNAc) modification, but the functional role of this modification in nucleocytoplasmic transport is unclear. We developed high-throughput assays based on optogenetic probes to quantify the kinetics of nuclear import and export in living human cells. We found that increasing O-GlcNAc modification of the NPC accelerated NTR-facilitated transport of proteins in both directions, and decreasing modification slowed transport. Superresolution imaging revealed strong enrichment of O-GlcNAc at the FG-repeat barrier. O-GlcNAc modification also accelerated passive permeation of a small, inert protein through NPCs. We conclude that O-GlcNAc modification accelerates nucleocytoplasmic transport by enhancing the nonspecific permeability of the FG-repeat barrier, perhaps by steric inhibition of interactions between FG repeats., (© 2021 Yoo and Mitchison.)

Compared to nanostructured platinum (Pt) catalysts, ordered Pt-based intermetallic nanoparticles supported on a carbon substrate exhibit much enhanced catalytic performance, especially in fuel cell electrocatalysis. However, direct synthesis of homogeneous intermetallic alloy nanocatalysts on carbonaceous supports with high loading is still challenging. Herein, we report a novel synthetic strategy to directly produce highly dispersed MPt alloy nanoparticles (M = Fe, Co, or Ni) on various carbon supports with high catalyst loading. Importantly, a unique bimetallic compound, composed of [M(bpy) 3 ] 2+ cation (bpy = 2,2'-bipyridine) and [PtCl 6 ] 2- anion, evenly decomposes on carbon surface and forms uniformly sized intermetallic nanoparticles with a nitrogen-doped carbon protection layer. The excellent oxygen reduction reaction (ORR) activity and stability of the representative reduced graphene oxide (rGO)-supported L1 0 -FePt catalyst (37 wt %-FePt/rGO), exhibiting 18.8 times higher specific activity than commercial Pt/C catalyst without degradation over 20 000 cycles, well demonstrate the effectiveness of our synthetic approach toward uniformly alloyed nanoparticles with high homogeneity.

Integrated with heat-generating devices, a Li-ion battery (LIB) often operates at 20-40 °C higher than the ordinary working temperature. Although macroscopic investigation of the thermal contribution has shown a significant reduction in the LIB performance, the molecular level structural and chemical origin of battery aging in a mild thermal environment has not been elucidated. On the basis of the combined experiments of the electrochemical measurements, Cs-corrected electron microscopy, and in situ analyses, we herein provide operando structural and chemical insights on how a mild thermal environment affects the overall battery performance using anatase TiO 2 as a model intercalation compound. Interestingly, a mild thermal condition induces excess lithium intercalation even at near-ambient temperature (45 °C), which does not occur at the ordinary working temperature. The anomalous intercalation enables excess lithium storage in the first few cycles but exerts severe intracrystal stress, consequently cracking the crystal that leads to battery aging. Importantly, this mild thermal effect is accumulated upon cycling, resulting in irreversible capacity loss even after the thermal condition is removed. Battery aging at a high working temperature is universal in nearly all intercalation compounds, and therefore, it is significant to understand how the thermal condition contributes to battery aging for designing intercalation compounds for advanced battery electrode materials.

Objective: The Discrimination and Stigma Scale (DISC 12), which assesses behavioral and experienced stigma, has not been translated into Korean. We developed and standardized the Korean version of the DISC 12 (DISC 12-K) in patients with depressive disorders., Methods: The study included 230 patients with depressive disorders who were assessed on the four subscales of the DISC 12-K: Unfair Treatment, Stopping Self, Overcoming Stigma, and Positive Treatment. Additionally, stigma was assessed using the Internalized Stigma of Mental Illness scale, depressive symptoms using the Hamilton Depression Rating Scale and Beck Depression Inventory, level of functioning using the Social and Occupational Functioning Assessment Scale, self-esteem using the Rosenberg Self-Esteem Scale, and quality of life was assessed using the EuroQol-5D. The reliability of DISC 12 was assessed by internal consistency using Cronbach's alpha coefficient and estimating the intercorrelation of items and corrected item-total correlations; interrater reliability and test-retest reliability were assessed using intraclass correlation coefficients at the item and subscale levels; and the concurrent validity of the DISC 12-K relative to the other assessment scales was assessed using Spearman's correlation coefficient., Results: All of the DISC 12-K subscales had high reliability. The validity was good for the Unfair Treatment and Stopping Self subscales, but only fair for the Overcoming Stigma and Positive Treatment subscales., Conclusion: The Unfair Treatment and Stopping Self subscales of the new DISC 12-K are reliable and valid measures of stigma in patients with depressive disorders. Future studies are needed to test the validity of this scale in other mental disorders.

Mechanisms involved in an electrochemically assisted oxidation process using persulfate and nanosized zero-valent iron (NZVI) were elucidated. Initially, Fe 0 acted as a source of Fe 2+ to activate the persulfate, then Fe 2+ /Fe 3+ redox mediation between cathode and persulfate played a decisive role in persulfate activation at a current density low enough not to inhibit Fe 0 corrosion. An excessive current density which resulted in a low cathodic potential limited Fe 0 corrosion and therefore limited the supply of dissolved Fe to activate the persulfate. Direct oxidation of phenol at the anode therefore became more important under the excessive current density than oxidation by sulfate radicals. At a low current density, Fe 0 in the NZVI particles was completely transformed into iron (oxyhydr)oxides such as ferrihydrite, lepidocrocite, and magnetite. Fe 0 was transformed into Fe 2+ little when the current density was high. Increasing the current density increased the energy cost by increasing the amount of electrical energy dissipated in side reactions that decreased sulfate radical formation. The results indicated that a low current density can generally be used to give a high reaction rate and a high energy efficiency and that a high current density can be used when the NZVI particles need to be preserved., (Copyright © 2019 Elsevier B.V. All rights reserved.)

A carbonation/granulation process for treating mine tailings using a MgO/ground-granule blast-furnace-slag (GGBS) binder was developed. The materials were mixed and granules produced using a granulator, then the granules were cured in a CO 2 atmosphere. The optimum granulator rotation speed and retention time were 60 rpm and 7 min, respectively. The binder composition MgO 0.5 GGBS 0.5 and binder: mine tailings ratio 3:10 gave the strongest granules. Carbonation generally increased the granule strength, but different CO 2 concentrations, between 0.04% and 100%, changed the granule strength to different degrees. Granules cured in 20% CO 2 for 28 d had a strength of 4.71 MPa, which was higher than the strengths of granules cured in other CO 2 concentrations and of granules produced using Portland cement. The granules had relatively high CO 2 storage capacities of 0.157-0.167 kg CO 2 /kg binder and good acid-neutralizing capacities (higher than the acid-neutralizing capacity of granules produced using Portland cement). The strength of the granules cured in 20% CO 2 for 28 d was probably mainly attributed to the formation of hydromagnesite during carbonation. The hydromagnesite contributed dense and connected structures within the granules. The granules produced show great potential for use as aggregates for reclamation work and backfilling in mining areas., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Development of quantitative, safe and rapid techniques for assessing embryo quality provides significant advances in Assisted Reproductive Technologies (ART). Instead of assessing the embryo quality by the standard morphologic evaluation, we apply the phasor-FLIM (Fluorescence Lifetime Imaging Microscopy) method to capture endogenous fluorescent biomarkers of pre-implantation embryos as a non-morphological caliber for embryo quality. Here, we identify, under hypoxic and non-hypoxic conditions, the unique spectroscopic trajectories at different stages of mouse pre-implantation development, which is referred to as the developmental, or "D-trajectory", that consists of fluorescence lifetime from different stages of mouse pre-implantation embryos. The D-trajectory correlates with intrinsic fluorescent species from a distinctive energy metabolism and oxidized lipids, as seen with Third Harmonic Generation (THG) that changes over time. In addition, we have defined a non-morphological Embryo Viability Index (EVI) to distinguish pre-implantation embryo quality using the Distance Analysis (DA), a machine learning algorithm to process the fluorescence lifetime distribution patterns. We show, under our experimental conditions, that the phasor-FLIM approach provides a much-needed non-invasive quantitative technology for identifying healthy embryos at the early compaction stage with 86% accuracy. The DA and phasor-FLIM method may provide the opportunity to improve implantation success rates for in vitro fertilization clinics.

Spindle microtubules, whose dynamics vary over time and at different locations, cooperatively drive chromosome segregation. Measurements of microtubule dynamics and spindle ultrastructure can provide insight into the behaviors of microtubules, helping elucidate the mechanism of chromosome segregation. Much work has focused on the dynamics and organization of kinetochore microtubules, that is, on the region between chromosomes and poles. In comparison, microtubules in the central-spindle region, between segregating chromosomes, have been less thoroughly characterized. Here, we report measurements of the movement of central-spindle microtubules during chromosome segregation in human mitotic spindles and Caenorhabditis elegans mitotic and female meiotic spindles. We found that these central-spindle microtubules slide apart at the same speed as chromosomes, even as chromosomes move toward spindle poles. In these systems, damaging central-spindle microtubules by laser ablation caused an immediate and complete cessation of chromosome motion, suggesting a strong coupling between central-spindle microtubules and chromosomes. Electron tomographic reconstruction revealed that the analyzed anaphase spindles all contain microtubules with both ends between segregating chromosomes. Our results provide new dynamical, functional, and ultrastructural characterizations of central-spindle microtubules during chromosome segregation in diverse spindles and suggest that central-spindle microtubules and chromosomes are strongly coupled in anaphase.

We previously described a MgO-based binder for treating fine sediment and simultaneously store CO 2 . Here, we describe a study of the physical/mechanical characteristics and carbonation reactions of the MgO-based binder used to solidify/stabilize fine sediment in atmospheres containing different CO 2 concentrations. Carbonation of the sediment treated with the MgO-based binder at the atmospheric CO 2 concentration markedly improved the compressive strength of the product. The compressive strength was 4.78 MPa after 365 days of curing, 1.3 times higher than the compressive strength of sediment treated with portland cement. This improvement was caused by the formation of carbonation products, such as hydromagnesite, nesquehonite, and lansfordite, and the constant high pH (~ 12) of the specimen, which favored the growth of hydration products such as calcium silicate hydrates and portlandite. Very low compressive strengths were found when 50 and 100% CO 2 atmospheres were used because of excessive formation of carbonation products, which occupied 78% of the specimen depth. Abundant carbonation products increased the specimen volume and decreased the pH to 10.2, slowing the growth of hydration products. The absence of brucite in specimens produced in a 100% CO 2 atmosphere indicated that MgO carbonation is favored over hydration at high CO 2 concentrations.

Proper kinetochore-microtubule attachments, mediated by the NDC80 complex, are required for error-free chromosome segregation. Erroneous attachments are corrected by the tension dependence of kinetochore-microtubule interactions. Here, we present a method, based on fluorescence lifetime imaging microscopy and Förster resonance energy transfer, to quantitatively measure the fraction of NDC80 complexes bound to microtubules at individual kinetochores in living human cells. We found that NDC80 binding is modulated in a chromosome autonomous fashion over prometaphase and metaphase, and is predominantly regulated by centromere tension. We show that this tension dependency requires phosphorylation of the N-terminal tail of Hec1, a component of the NDC80 complex, and the proper localization of Aurora B kinase, which modulates NDC80 binding. Our results lead to a mathematical model of the molecular basis of tension-dependent NDC80 binding to kinetochore microtubules in vivo., Competing Interests: TY, JC, WC, CY, RP, DN No competing interests declared, (© 2018, Yoo et al.)

The mechanisms involved in the activation of persulfate by nanosized zero-valent iron (NZVI) were elucidated and the NZVI transformation products identified. Two distinct reaction stages, in terms of the kinetics and radical formation mechanism, were found when phenol was oxidized by the persulfate/NZVI system. In the initial stage, lasting 10 min, Fe 0 (s) was consumed rapidly and sulfate radicals were produced through activation by aqueous Fe 2+ . The second stage was governed by Fe catalyzed activation in the presence of aqueous Fe 3+ and iron (oxyhydr)oxides in the NZVI shells. The second stage was 3 orders of magnitude slower than the initial stage. An electron balance showed that the sulfate radical yield per mole of persulfate was more than two times higher in the persulfate/NZVI system than in the persulfate/Fe 2+ system. Radicals were believed to be produced more efficiently in the persulfate/NZVI system because aqueous Fe 2+ was supplied slowly, preventing sulfate radicals being scavenged by excess aqueous Fe 2+ . In the second stage, the multilayered shell conducted electrons, and magnetite in the shell provided electrons for the activation of persulfate. Iron speciation analysis (including X-ray absorption spectroscopy) results indicated that a shrinking core/growing shell model explained NZVI transformation during the persulfate/NZVI process.

The chromosomal passenger complex (CPC) is a conserved, essential regulator of cell division. As such, significant anti-cancer drug development efforts have been focused on targeting it, most notably by inhibiting its AURKB kinase subunit. The CPC is activated by AURKB-catalyzed autophosphorylation on multiple subunits, but how this regulates CPC interactions with other mitotic proteins remains unclear. We investigated the hydrodynamic behavior of the CPC in Xenopus laevis egg cytosol using sucrose gradient sedimentation and in HeLa cells using fluorescence correlation spectroscopy. We found that autophosphorylation of the CPC decreases its sedimentation coefficient in egg cytosol and increases its diffusion coefficient in live cells, indicating a decrease in mass. Using immunoprecipitation coupled with mass spectrometry and immunoblots, we discovered that inactive, unphosphorylated CPC interacts with nucleophosmin/nucleoplasmin proteins, which are known to oligomerize into pentamers and decamers. Autophosphorylation of the CPC causes it to dissociate from nucleophosmin/nucleoplasmin. We propose that nucleophosmin/nucleoplasmin complexes serve as chaperones that negatively regulate the CPC and/or stabilize its inactive form, preventing CPC autophosphorylation and recruitment to chromatin and microtubules in mitosis., (© 2017 Hanley et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).)

Time-resolvable quantitative measurements of polymer concentration are very useful to elucidate protein polymerization pathways. There are numerous techniques to measure polymer concentrations in purified protein solutions, but few are applicable in vivo. Here we develop a methodology combining microscopy and spectroscopy to overcome the limitations of both approaches for measuring polymer concentration in cells and cell extracts. This technique is based on quantifying the relationship between microscopy and spectroscopy measurements at many locations. We apply this methodology to measure microtubule assembly in tissue culture cells and Xenopus egg extracts using two-photon microscopy with FLIM measurements of FRET. We find that the relationship between FRET and two-photon intensity quantitatively agrees with predictions. Furthermore, FRET and intensity measurements change as expected with changes in acquisition time, labeling ratios, and polymer concentration. Taken together, these results demonstrate that this approach can quantitatively measure microtubule assembly in complex environments. This methodology should be broadly useful for studying microtubule nucleation and assembly pathways of other polymers., (© 2017 Kaye et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).)

FRET measurements can provide dynamic spatial information on length scales smaller than the diffraction limit of light. Several methods exist to measure FRET between fluorophores, including Fluorescence Lifetime Imaging Microscopy (FLIM), which relies on the reduction of fluorescence lifetime when a fluorophore is undergoing FRET. FLIM measurements take the form of histograms of photon arrival times, containing contributions from a mixed population of fluorophores both undergoing and not undergoing FRET, with the measured distribution being a mixture of exponentials of different lifetimes. Here, we present an analysis method based on Bayesian inference that rigorously takes into account several experimental complications. We test the precision and accuracy of our analysis on controlled experimental data and verify that we can faithfully extract model parameters, both in the low-photon and low-fraction regimes., Competing Interests: TYY was supported in part by the Samsung scholarship (https://www.ssscholarship.com/community/HomeEn.screen). This does not alter our adherence to PLOS ONE policies on sharing data and materials, and Samsung had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Kinetochores play essential roles in coordinating mitosis, as a mechanical connector between chromosome and microtubule and as a source of numerous biochemical signals. These mechanical and biochemical behaviors of kinetochores change dynamically in cells during mitosis. Therefore, understanding kinetochore function requires an imaging tool that quantifies the protein-protein interactions or biochemical changes with high spatiotemporal resolution. FRET has previously been used in combination with biosensors to probe protein-protein interactions and biochemical activity. In this chapter, we introduce FLIM-FRET, a lifetime-based method that quantifies FRET, and describe the use of FLIM-FRET as a method for studying dynamic kinetochore behavior in cells with high spatiotemporal resolution.

We report a simple approach for the production of copper nanoparticles by a wire explosion process that creates different structures in deionized (DI) water versus isopropyl alcohol (IPA) liquid media. In DI water, copper nanoparticles (CNs) are formed, while multi-layer graphene-synthesized copper nanoparticles (MGCNs) with a high degree of graphitization are formed in the IPA liquid media. The nanoparticles have an average diameter ranging from 10 nm to 300 nm and a quasi-spherical morphology. The morphologies and sizes of nanoparticles formed via this method were characterized by high-resolution transmission electron microscopy (HRTEM), field-emission scattering electron microscopy (FESEM), and analysis of dynamic light scattering (DLS). The microstructures and chemical bonding of the nanoparticles were studied by X-ray diffraction (XRD), Raman spectra measurement, and X-ray photoelectron spectroscopy (XPS). This results show an easily reproducible way to synthesize metal-core nanoparticles with multi-layer graphene shells based onto the liquid media used during synthesis. These materials can be used in the field of energy storage and as additives in the near future.

We achieved a method for power enhancement of heavy-duty lithium-ion batteries (LIBs) by synthesizing a graphene interfacial layer onto the anode copper current collector (ACCC). We tested fabricated coin cells, which used either 35-μm-thick rolled pristine copper foil or graphene synthesized onto the pristine copper foil for power output estimation of the LIBs. We observed the copper surface morphology with a scanning electron microscope (SEM). Raman spectroscopy was used to measure the bonding characteristics and estimate the layers of graphene films. In addition, transmittance and electrical resistance were measured by ultra-violet visible near-infrared spectroscopy (UV-Vis IR) and 4 point probe surface resistance measurement. The graphene films on polyethylene terephthalate (PET) substrate obtained a transmittance of 97.5% and sheet resistance of 429 Ω/square. Power enhancement performances was evaluated using LIB coin cells. After 5C current discharge rate of -1.7 A/g reversible capacity of 293 mAh/g and 326 mAh/g were obtained for pristine and synthesized graphene anode current collectors, respectively. The graphene synthesized onto the ACCC showed superior power performance. The results presented herein demonstrate a power enhancement of LIBs by a decrease in electron flow resistivity between active materials and the ACCC and removal of the native oxide layer on the anode copper surface using high quality graphene synthesized onto the ACCC.

Electroless Ni-P films were investigated with the aim of application as barrier and seed layers in 3D interconnect technology. Different shapes of blind-via holes were fabricated with a deep reactive ion etcher and SiO2 formed on these holes as an insulating layer. The surface of the substrate has been made hydrophilic by O2 plasma treatment with 100 W of power for 20 min. Electroless Ni-P films were deposited as both a diffusion barrier and a seed layer for Cu filling process. Prior to plating, substrates were activated in a palladium chloride solution after sensitization in a tin chloride solution with various conditions in order to deposit uniform films in TSV. After the formation of the electroless barrier layer, electro Cu was plated directly on the barrier layer. Ni-P films fabricated in blind-via holes were observed by scanning electron microscope. Energy dispersive spectroscopy line scanning was carried out for evaluating the diffusion barrier properties of the Ni-P films. The electroless Ni-P layer worked well as a Cu diffusion barrier until 300 degrees C. However, Cu ions diffused into barrier layer when the annealing temperature increases over 400 degrees C.

The spatial organization of microtubule polarity, and the interplay between microtubule polarity and protein localization, is thought to be crucial for spindle assembly, anaphase, and cytokinesis, but these phenomena remain poorly understood, in part due to the difficulty of measuring microtubule polarity in spindles. We develop and implement a method to nonperturbatively and quantitatively measure microtubule polarity throughout spindles using a combination of second-harmonic generation and two-photon fluorescence. We validate this method using computer simulations and by comparison to structural data on spindles obtained from electron tomography and laser ablation. This method should provide a powerful tool for studying spindle organization and function, and may be applicable for investigating microtubule polarity in other systems., (Copyright © 2014 Biophysical Society. Published by Elsevier Inc. All rights reserved.)

Progress in understanding protein folding relies heavily upon an interplay between experiment and theory. In particular, readily interpretable experimental data that can be meaningfully compared to simulations are required. According to standard mutational ϕ analysis, the transition state for Protein L contains only a single hairpin. However, we demonstrate here using ψ analysis with engineered metal ion binding sites that the transition state is extensive, containing the entire four-stranded β sheet. Underreporting of the structural content of the transition state by ϕ analysis also occurs for acyl phosphatase [Pandit, A. D., Jha, A., Freed, K. F. & Sosnick, T. R., (2006). Small proteins fold through transition states with native-like topologies. J. Mol. Biol.361, 755-770], ubiquitin [Sosnick, T. R., Dothager, R. S. & Krantz, B. A., (2004). Differences in the folding transition state of ubiquitin indicated by ϕ and ψ analyses. Proc. Natl Acad. Sci. USA 101, 17377-17382] and BdpA [Baxa, M., Freed, K. F. & Sosnick, T. R., (2008). Quantifying the structural requirements of the folding transition state of protein A and other systems. J. Mol. Biol.381, 1362-1381]. The carboxy-terminal hairpin in the transition state of Protein L is found to be nonnative, a significant result that agrees with our Protein Data Bank-based backbone sampling and all-atom simulations. The nonnative character partially explains the failure of accepted experimental and native-centric computational approaches to adequately describe the transition state. Hence, caution is required even when an apparent agreement exists between experiment and theory, thus highlighting the importance of having alternative methods for characterizing transition states., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

The results of more than a dozen single-molecule Förster resonance energy transfer (smFRET) experiments suggest that chemically unfolded polypeptides invariably collapse from an expanded random coil to more compact dimensions as the denaturant concentration is reduced. In sharp contrast, small-angle X-ray scattering (SAXS) studies suggest that, at least for single-domain proteins at non-zero denaturant concentrations, such compaction may be rare. Here, we explore this discrepancy by studying protein L, a protein previously studied by SAXS (at 5 °C), which suggested fixed unfolded-state dimensions from 1.4 to 5 M guanidine hydrochloride (GuHCl), and by smFRET (at 25 °C), which suggested that, in contrast, the chain contracts by 15-30% over this same denaturant range. Repeating the earlier SAXS study under the same conditions employed in the smFRET studies, we observe little, if any, evidence that the unfolded state of protein L contracts as the concentration of GuHCl is reduced. For example, scattering profiles (and thus the shape and dimensions) collected within ∼4 ms after dilution to as low as 0.67 M GuHCl are effectively indistinguishable from those observed at equilibrium at higher denaturant. Our results thus argue that the disagreement between SAXS and smFRET is statistically significant and that the experimental evidence in favor of obligate polypeptide collapse at low denaturant cannot be considered conclusive yet., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Primary adenocarcinoma of the nonpigmented ciliary epithelium (NPCE) is a very rare disease and the majority of acquired cases were treated by enucleation. We report the case of a 19-year-old man who had an adenocarcinoma arising from the NPCE. The tumor was found incidentally due to changed pupil shape. An excisional biopsy was performed and histopathological examination showed primary adenocarcinoma of the NPCE with a positive resection margin. Because of the positive resection margin and to save the eye, radiotherapy rather than enucleation was performed, followed by chemotherapy. Two years after the diagnosis, the patient continues to be followed up without evidence of relapse or cataract change. Our case suggests that adjunctive radiotherapy should be considered as a sight-saving treatment modality for primary malignant tumors of the NPCE.

Those who dye their hair frequently manifest allergic contact dermatitis (ACD) from p-phenylenediamine (PPD)-containing hair dye. PPD is known to be the most frequent sensitizer in hair dye, but there has been no documentation of this sensitizer having a role in chronic dermatologic conditions. Our department experienced a case of a 62-year-old woman with lichen simplex chronicus (LSC), who complained of aggravation after hair dyeing and made such an association. We conducted a prospective and retrospective study to further evaluate the clinical associations between the two. In our prospective study, patch testing was performed in selected patients who regularly carried out hair dyeing and also had clinical manifestations of LSC. Also a retrospective examination was conducted in cases where patch testing had been performed with PPD in the past for suspected ACD and further selected cases with concurrent LSC and/or other neurodermatitis. 11 and 14 patients in our prospective and retrospective study, respectively, presented with both LSC and positive findings to PPD. 5 (45.5%) and 4 (28.6%) patients in our prospective and retrospective study showed clinical relevance from clinical improvement after stopping use and rechallenge. We report several cases of patients diagnosed as having LSC and/or prurigo nodularis who showed clinical improvement after discontinuing the use of hair dye. The suggestion can therefore be made that hair dye could be a possible aetiologic agent causing LSC in those using hair dyes.

The current study investigated the impact of requiring respondents to elaborate on their answers to a biodata measure on mean scores, the validity of the biodata item composites, subgroup mean differences, and correlations with social desirability. Results of this study indicate that elaborated responses result in scores that are much lower than nonelaborated responses to the same items by an independent sample. Despite the lower mean score on elaborated items, it does not appear that elaboration affects the size of the correlation between social desirability and responses to biodata items or that it affects criterion-related validity or subgroup mean differences in a practically significant way., (((c) 2003 APA, all rights reserved))

Study Design: A case report and review of the literature are presented., Objective: To present the first case of paraplegia as a complication of percutaneous vertebroplasty with polymethylmethacrylate in osteoporotic compression fracture., Summary of Background Data: Complications of percutaneous vertebroplasty with polymethylmethacrylate (PMMA) for the treatment of osteoporotic compression fracture were found to be rare and minor, except in two cases of major neurologic complication., Methods: The reported case is that of a 66-year-old woman with multilevel vertebral osteopenia and compression fractures. Percutaneous vertebroplasty using polymethylmethacrylate was performed at three vertebral bodies (L2, L1, and T11) using careful techniques including venography, large cannula, proper preparation and amount of polymethylmethacrylate, and continuous visualization with fluoroscopy., Results: Immediately after surgery, the patient had complete motor and sensory deficits at T11. Computed tomography scan showed spinal cord compression caused by venous leakage of polymethylmethacrylate. In anticipation of recovery from paraplegia, posterior decompression was performed from L2 to T10., Conclusions: Percutaneous vertebroplasty with polymethylmethacrylate is not as simple and risk free as advocated in the literature. Careful safeguards and modifications are needed for the procedure, and new and physiologic material could be substituted for polymethylmethacrylate.

This paper deals with the estimation of the shadow prices of pollutants with a nonparametric directional distance function approach, where the inefficiency involved in the production process is taken into account unlike the previous studies. The directional vector, which is critical to the estimation and subject to the criterion for an appropriate efficiency rule proposed here, is calculated by using the annual plans of power plants in terms of production and environment. In the empirical study for Korea's electric power industry during the period of 1990-1995, we find that the average shadow prices of sulfur oxides (SOx), nitrogen oxides (NOx), and total suspended particulates (TSP) are approximately 10% lower than those calculated under the assumption of full efficiency. The methodology we propose and the findings obtained in the empirical study allow us to undertake better decision-making over a broad range of environmental policy issues.