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PDF file - 292K, Table S1 shows FACS analysis results of MET, CD133, and CD15 in the freshly isolated patient GBM cells, and Table S2 describes the list of patient-derived GBM cells and their derivatives used in this study. Patient information of 14 matched pre- and postradiated GBM specimens is shown in Table S3

PDF file - 6.4MB, Figure S1: High MET mRNA expression in GBM specimens portends poor patient survival. Figure S2: Dual immunofluorescence of MET and GSC enrichment markers (CD133 and CD15) on the frozen sections of GBM specimens. Figure S3: HGF/MET signaling in GBMs. Figure S4: METhigh cells generated xenograft tumors consisting of both METhigh and METlow/- cells. Figure S5: Immunohistochemical analysis of HGF protein in orthotopic xenograft tumors derived of from GBM 578 (A and C) and 464 (B and D). Figure S6: Effects of MET blockade on cell survival and clonogenicity of GSCs. Figure S7: In vitro migration and invasion assays using GSCs with MET targeting. Figure S8: Immunohistochemical analysis using human nuclei-specific antibody on the sections of GBM xenografts. Figure S9: Limiting dilution sphere forming assay to determine the clonogenic potentials of 822 GSCs after treatments of radiation and SU11274

We report new ruthenium complexes bearing the lipophilic bathophenanthroline (BPhen) ligand and dihydroxybipyridine (dhbp) ligands which differ in the placement of the OH groups ([(BPhen)(2)Ru(n,n′-dhbp)]Cl(2) with n = 6 and 4 in 1(A) and 2(A), respectively). Full characterization data are reported for 1(A) and 2(A) and single crystal X-ray diffraction for 1(A). Both 1(A) and 2(A) are diprotic acids. We have studied 1(A), 1(B), 2(A), and 2(B) (B = deprotonated forms) by UV-vis spectroscopy and 1 photodissociates but 2 is light stable. Luminescence studies reveal that the basic forms have lower energy (3)MLCT states relative to the acidic forms. Complexes 1(A) and 2(A) produce singlet oxygen with quantum yields of 0.05 and 0.68, respectively, in acetonitrile. Complexes 1 and 2 are both photocytotoxic towards breast cancer cells, with complex 2 showing EC(50) light values as low as 0.50 μM with PI values as high as >200 vs. MCF7. Computational studies were used to predict the energies of the (3)MLCT and (3)MC states. An inaccessible (3)MC state for 2(B) suggests a rationale for why photodissociation does not occur with the 4,4′-dhbp ligand. Low dark toxicity combined with an accessible (3)MLCT state for (1)O(2) generation explains the excellent photocytotoxicity of 2.

Ruthenium complexes bearing protic diimine ligands are cytotoxic to certain cancer cells upon irradiation with blue light. Previously reported complexes of the type [(N,N)(2)Ru(6,6’-dhbp)]Cl(2) with 6,6’-dhbp = 6,6’-dihydroxybipyridine and N,N = 2,2’-bipyridine (bipy) (1(A)), 1,10-phenanthroline (phen) (2(A)), and 2,3-dihydro-[1,4]dioxino[2,3-f][1,10]phenanthroline (dop) (3(A)) show EC(50) values as low as 4 μM (for 3(A)) vs. breast cancer cells upon blue light irradiation (Inorg. Chem. 2017, 56, 7519). Herein, subscript A denotes the acidic form of the complex bearing OH groups, and B denotes the basic form bearing O(−) groups. This photocytotoxicity was originally attributed to photodissociation, but recent results suggest that singlet oxygen formation is a more plausible cause of photocytotoxicity. In particular, bulky methoxy substituents enhance photodissociation but these complexes are non-toxic (Dalton Trans. 2018, 47, 15685). Cellular studies are presented herein that show the formation of reactive oxygen species (ROS) and apoptosis indicators upon treatment of cells with complex 3(A) and blue light. Singlet oxygen sensor green (SOSG) shows the formation of (1)O(2) in cell culture for cells treated with 3(A) and blue light. At physiological pH, complexes 1(A)-3(A) are deprotonated to form 1(B)-3(B) in situ. Quantum yields for (1)O(2) (ɸ(Δ) ) are 0.87 and 0.48 for 2(B) and 3(B) respectively, and these are an order of magnitude higher than the quantum yields for 2(A) and 3(A). The values for ɸ(Δ) show an increase with 6,6’-dhbp derived substituents as follows: OMe < OH < O(−). TD-DFT studies show that the presence of a low lying triplet metal-centered ((3)MC) state favors photodissociation and disfavors (1)O(2) formation for 2(A) and 3(A) (OH groups). However, upon deprotonation (O(−) groups), the (3)MLCT state is accessible and can readily lead to (1)O(2) formation, but the dissociative (3)MC state is energetically inaccessible. The changes to the energy of the (3)MLCT state upon deprotonation have been confirmed by steady state luminescence experiments on 1(A)-3(A) and their basic analogs, 1(B)-3(B). This energy landscape favors (1)O(2) formation for 2(B) and 3(B) and leads to enhanced toxicity for these complexes under physiological conditions. The ability to convert readily from OH to O(−) groups allowed us to investigate an electronic change that is not accompanied by steric changes in this fundamental study.

Increase in the nitrogen (N)-use efficiency and optimization of N response in crop species are urgently needed. Although transcription factor-based genetic engineering is a promising approach for achieving these goals, transcription factors that play key roles in the response to N deficiency have not been studied extensively. Here, we performed RNA-seq analysis of root samples of 20 Asian rice (Oryza sativa) accessions with differential nutrient uptake. Data obtained from plants exposed to N-replete and N-deficient conditions were subjected to coexpression analysis and machine learning-based pathway inference to dissect the gene regulatory network required for the response to N deficiency. Four transcription factors, including members of the G2-like and bZIP families, were predicted to function as key regulators of gene transcription within the network in response to N deficiency. Cotransfection assays validated inferred novel regulatory pathways, and further analyses using genome-edited knockout lines suggested that these transcription factors are important for N-deficiency responses in planta. Many of the N deficiency-responsive genes, including those encoding key regulators within the network, were coordinately regulated by transcription factors belonging to different families. Transcription factors identified in this study could be valuable for the modification of N response and metabolism.

Supporting multi-language linking such as linking C and handwritten assembly modules in the verified compiler CompCert requires a more compositional verification technique than that used in CompCert just supporting separate compilation. The two extensions, CompCertX and Compositional CompCert, supporting multi-language linking take different approaches. The former simplifies the problem by imposing restrictions that the source modules should have no mutual dependence and be verified against certain well-behaved specifications. On the other hand, the latter develops a new verification technique that directly solves the problem but at the expense of significantly increasing the verification cost. In this paper, we develop a novel lightweight verification technique, called RUSC (Refinement Under Self-related Contexts), and demonstrate how RUSC can solve the problem without any restrictions but still with low verification overhead. For this, we develop CompCertM, a full extension of the latest version of CompCert supporting multi-language linking. Moreover, we demonstrate the power of RUSC as a program verification technique by modularly verifying interesting programs consisting of C and handwritten assembly against their mathematical specifications.

Glioblastoma multiforme (GBM) is the deadliest form of primary brain tumor. GBM tumors are highly heterogeneous, being composed of tumor cells as well as glioblastoma stem cells (GSCs) that contribute to drug resistance and tumor recurrence following treatment. To develop therapeutic strategies, an improved understanding of GSC behavior in their microenvironment is critical. Herein, we have employed three-dimensional (3D) hyaluronic acid (HA) hydrogels that allow the incorporation of brain microenvironmental cues to investigate GSC behavior. U87 cell line and patient-derived D456 cells were cultured as suspension cultures (serum-free) and adherently (in the presence of serum) and were then encapsulated in HA hydrogels. We observed that all the seeded single cells expanded and formed spheres, and the size of the spheres increased with time. Increasing the initial cell seeding density of cells influenced the sphere size distribution. Interestingly, clonal expansion of serum-free grown tumor cells in HA hydrogels was observed. Also, stemness marker expression of serum and/or serum-free grown cells was altered when cultured in HA hydrogels. Finally, we demonstrated that HA hydrogels can support long-term GSC culture (up to 60 days) with retention of stemness markers. Overall, such biomimetic culture systems could further our understanding of the microenvironmental regulation of GSC phenotypes.

OBJECTIVES: The F1Fo ATP synthase is a multienzyme complex that produces mitochondrial ATP. Aberrant expression or assembly of F1Fo ATP synthase subunits leads to alterations in energy metabolism. We recently found that breast cancer cells exposed to fluid shear stress (FSS) have significantly enhanced metastatic behavior including chemoresistance and cell proliferation. Chemoresistance depends upon active transport systems, and cell division and growth require ATP. Therefore, we hypothesized that circulating breast cancer cells undergo altered energy metabolism via FSS-induced changes in F1Fo ATP synthase subunits and subsequent mitochondrial remodeling. METHODS: Non-metastatic MCF7 and metastatic MDA-MB-231 human breast cancer cells were treated with or without FSS and cultured. Cellular proliferation was assayed by measuring cell number and gap distance. Metabolic profile including intracellular ATP and oxygen consumption rate were analyzed. We also quantified abundance of F1Fo ATP synthase subunits using immunoblotting. RESULTS: Treatment with FSS significantly increased proliferation of both MCF7 and MDA-MB-231 human breast cancer cells. FSS significantly increased intracellular ATP in MDA-MB-231 breast cancer cells while ATP levels in MCF7 were not significantly changed. MDA-MB-231 cells retained increased ATP after treatment with the uncoupler FCCP, indicating remodeling and decreased reliance on mitochondrial energy metabolism. Interestingly, oxygen consumption rate was significantly increased in both MCF7 and MDA-MB-231 by FSS. We further quantified the abundance of F1Fo ATP synthase subunits in both cell lines. The β- and c-subunits of the F1Fo ATP synthase were significantly depleted in both lines of FSS-treated breast cancer cells. CONCLUSIONS: Our data show that FSS alters abundance of the F1Fo ATP synthase subunits leading to metabolic remodeling. We suggest that FSS may influence non-metastatic (MCF7) and metastatic cancer cells (MDA-MB-231) differently. Underlying changes in mitochondrial and cytoplasmic ATP production in these cells is still under investigation. However, it is possible that reactive oxygen species generated during FSS may signal a switch to cytoplasmic intracellular energy metabolism. FUNDING SOURCES: Alabama Life Research Institute Pilot Project (University of Alabama)

Even with rigorous treatments, glioblastoma multiforme (GBM) has an abysmal median survival rate, greatly due to the drug-resistant glioblastoma stem cell (GSC) population. GSCs are known to remodel their microenvironment, but the precise role of extracellular matrix components hyaluronic acid (HA) and hyaluronidases (HAases) on the GSC population is still largely unknown. Our objective was to determine how HAase can sensitize GSCs to chemotherapy drugs by disrupting the HA-CD44 signaling. GBM cell line U87-MG and patient-derived D456 cells were grown in GSC-enriching media and treated with HA or HAase. Expressions of GSC markers, HA-related genes, and drug resistance genes were measured via flow cytometry, confocal microscopy, and qRT-PCR. Proliferation after combined HAase and temozolomide (TMZ) treatment was measured via WST-8. HA supplementation promoted the expression of GSC markers and CD44 in GBM cells cultured in serum-free media. Conversely, HAase addition inhibited GSC gene expression while promoting CD44 expression. Finally, HAase sensitized GBM cells to TMZ. We propose a combined treatment of HAase and chemotherapy drugs by disrupting the stemness-promoting HA to target GSCs. This combination therapy shows promise even when temozolomide treatment alone causes resistance. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1007/s12307-019-00224-2) contains supplementary material, which is available to authorized users.

We propose a novel distance-based regularization method for deep metric learning called Multi-level Distance Regularization (MDR). MDR explicitly disturbs a learning procedure by regularizing pairwise distances between embedding vectors into multiple levels that represents a degree of similarity between a pair. In the training stage, the model is trained with both MDR and an existing loss function of deep metric learning, simultaneously; the two losses interfere with the objective of each other, and it makes the learning process difficult. Moreover, MDR prevents some examples from being ignored or overly influenced in the learning process. These allow the parameters of the embedding network to be settle on a local optima with better generalization. Without bells and whistles, MDR with simple Triplet loss achieves the-state-of-the-art performance in various benchmark datasets: CUB-200-2011, Cars-196, Stanford Online Products, and In-Shop Clothes Retrieval. We extensively perform ablation studies on its behaviors to show the effectiveness of MDR. By easily adopting our MDR, the previous approaches can be improved in performance and generalization ability., Accepted to AAAI 2021

The dominant underwater noise source of a ship is known to be propeller cavitation. Recently, attempts have been made to quantify the source strength using on-board pressure sensors near the propeller, as this has advantages over conventional noise measurement. In this study, a beamforming method was used to estimate the source strength of a cavitating propeller. The method was validated against a model-scale measurement in a cavitation tunnel, which showed good agreement between the measured and estimated source levels. The method was also applied to a full-scale measurement, in which the source level was measured using an external hydrophone array. The estimated source level using the hull pressure sensors showed good agreement with the measured one above 400 Hz, which shows potential for noise monitoring using on-board sensors. A parametric study was carried out to check the practicality of the method. From the results, it was shown that a sufficient recording time is required to obtain a consistent level at high frequencies. Changing the frequency resolution had little effect on the result, as long as enough data were provided for the one-third octave band conversion. The number of sensors affected the mid- to low-frequency data.

INTRODUCTION: Glioblastoma multiforme (GBM), the most common primary brain tumor in adults, is extremely malignant and lethal. GBM tumors are highly heterogenous, being comprised of cellular and matrix components, which contribute to tumor cell invasion, cancer stem cell maintenance, and drug resistance. Here, we developed a heterotypic 3D spheroid model integrating GBM cells with astrocytes and endothelial cells (ECs) to better simulate the cellular components of the tumor microenvironment and investigate their impact on the stemness marker expression of GBM cells, which has not been previously investigated. METHODS: We used U87 GBM cells, C8-D1A mouse astrocytes, and human umbilical vein ECs to construct co- and tri-culture spheroid models in low-attachment U-well plates. We characterized the expression of known stemness markers NESTIN, SOX2, CD133, NANOG, and OCT4 in these models and compared it to respective mixed monoculture spheroids (control) using qRT-PCR and immunostaining. RESULTS: We incorporated GBM cells and astrocytes/ECs in 1:1, 1:2, 1:4, and 1:9 ratio and observed spontaneous self-assembled spheroids in all coculture conditions. We observed changing spheroid size dynamics over 7 days and an increased expression in stemness markers in GBM-astrocyte and GBM-EC coculture spheroids in 1:4 and 1:9 coculture conditions, respectively. In a triculture model employing GBM cells, astrocytes, and ECs in a 1:4:9 ratio, we found an increased expression of all the stemness markers. CONCLUSIONS: We elucidated the impact of astrocytes and ECs on GBM stemness marker expression. This multicellular spheroid model may provide an important tool for investigating the crosstalk between cell types in GBM. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12195-021-00691-y.

Circulating tumor cells (CTCs) are cancer cells that detach from the primary site and travel in the blood stream. A higher number of CTCs increases the risk of breast cancer metastasis, and it is inversely associated with the survival rates of patients with breast cancer. Although the numbers of CTCs are generally low and the majority of CTCs die in circulation, the survival of a few CTCs can seed the development of a tumor at a secondary location. An increasing number of studies demonstrate that CTCs undergo modification in response to the dynamic biophysical environment in the blood due in part to fluid shear stress. Fluid shear stress generates reactive oxygen species (ROS), triggers redox-sensitive cell signaling, and alters the function of intracellular organelles. In particular, the mitochondrion is an important target organelle in determining the metastatic phenotype of CTCs. In healthy cells, mitochondria produce adenosine triphosphate (ATP) via oxidative phosphorylation in the electron transport chain, and during oxidative phosphorylation, they produce physiological levels of ROS. Mitochondria also govern death mechanisms such as apoptosis and mitochondrial permeability transition pore opening to, in order eliminate unwanted or damaged cells. However, in cancer cells, mitochondria are dysregulated, causing aberrant energy metabolism, redox homeostasis, and cell death pathways that may favor cancer invasiveness. In this review, we discuss the influence of fluid shear stress on CTCs with an emphasis on breast cancer pathology, then discuss alterations of cellular mechanisms that may increase the metastatic potentials of CTCs.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

In the field of face recognition, a model learns to distinguish millions of face images with fewer dimensional embedding features, and such vast information may not be properly encoded in the conventional model with a single branch. We propose a novel face-recognition-specialized architecture called GroupFace that utilizes multiple group-aware representations, simultaneously, to improve the quality of the embedding feature. The proposed method provides self-distributed labels that balance the number of samples belonging to each group without additional human annotations, and learns the group-aware representations that can narrow down the search space of the target identity. We prove the effectiveness of the proposed method by showing extensive ablation studies and visualizations. All the components of the proposed method can be trained in an end-to-end manner with a marginal increase of computational complexity. Finally, the proposed method achieves the state-of-the-art results with significant improvements in 1:1 face verification and 1:N face identification tasks on the following public datasets: LFW, YTF, CALFW, CPLFW, CFP, AgeDB-30, MegaFace, IJB-B and IJB-C., Comment: Accepted to CVPR 2020

As oxygen may occupy a major volume of oxides, a densification of amorphous oxides under extreme compression is dominated by reorganization of oxygen during compression. X-ray Raman scattering (XRS) spectra for SiO_{2} glass up to 1.6 Mbar reveal the evolution of heavily contracted oxygen environments characterized by a decrease in average O-O distance and the potential emergence of quadruply coordinated oxygen (oxygen quadcluster). Our results also reveal that the edge energies at the centers of gravity of the XRS features increase linearly with bulk density, yielding the first predictive relationship between the density and partial density of state of oxides above megabar pressures. The extreme densification paths with densified oxygen in amorphous oxides shed light upon the possible existence of stable melts in the planetary interiors.

The purpose of dissolved gas analysis (DGA) in utilities is to help detect the presence of abnormality within transformer. When faults occur in transformers, transformer engineers need to determine the location and risk of the faults. Ultimately, they have to decide, based on DGA, whether to continue operating or not, perform internal inspection, or dispose the transformer. In this study, the fault and failure types in the transformer are suggested to determine the location and risk of the faults. In particular, turn-to-turn insulation faults are classified as degradation and breakdown. These faults are difficult to identify during internal inspection, and have a high possibility of failure. Urgent decision and action are thus required to avoid failure. In degradation of turn-to-turn insulation faults, failures may occur by generating thermal gases in paper during a long period of time. In breakdown of turn-to-turn insulation faults, thermal gases are not generated in paper, and failures are rather due to sudden breakdown of insulation. This study also presents a typical example of a turn-to-turn insulation fault. This example shows the progress of the fault from thermal to discharge, which is common phenomena in winding fault. Based on the findings of this work, transformer engineers can determine by DGA if transformers can be operated with or without internal inspection, or disposed when the fault has not been identified during internal inspection.

Structural transition in amorphous oxides, including glasses, under extreme compression above megabar pressures (>1 million atmospheric pressure, 100 GPa) results in unique densification paths that differ from those in crystals. Experimentally verifying the atomistic origins of such densifications beyond 100 GPa remains unknown. Progress in inelastic X-ray scattering (IXS) provided insights into the pressure-induced bonding changes in oxide glasses; however, IXS has a signal intensity several orders of magnitude smaller than that of elastic X-rays, posing challenges for probing glass structures above 100 GPa near the Earth’s core–mantle boundary. Here, we report megabar IXS spectra for prototypical B2O3 glasses at high pressure up to ∼120 GPa, where it is found that only four-coordinated boron ([4]B) is prevalent. The reduction in the [4]B–O length up to 120 GPa is minor, indicating the extended stability of sp3-bonded [4]B. In contrast, a substantial decrease in the average O–O distance upon compression is revealed, suggesting that the densification in B2O3 glasses is primarily due to O–O distance reduction without the formation of [5]B. Together with earlier results with other archetypal oxide glasses, such as SiO2 and GeO2, the current results confirm that the transition pressure of the formation of highly coordinated framework cations systematically increases with the decreasing atomic radius of the cations. These observations highlight a new opportunity to study the structure of oxide glass above megabar pressures, yielding the atomistic origins of densification in melts at the Earth’s core–mantle boundary.

We report a multi adsorbent-based method using combinations of metal-organic frameworks (MOFs) and a commercial sorbent Tenax-TA for sampling and thermal desorption (TD) gas chromatography-mass spectrometry (GC-MS) quantification of mixtures of six (C1 to C5) aldehydes. The feasibility of this approach was demonstrated along with the optical analytical conditions for maximum recovery. Optimal TD conditions for adsorption and desorption of aldehydes using MOF-5 (Zn-based MOF)+ Tenax-TA were determined as −25 °C and 150 °C, respectively (purge volume: 100 ml). These conditions yielded good linearity (R2 = 0.997), precision, and high sensitivity. Analysis of the aldehyde mixtures yielded slightly smaller R2 values than the analysis of single species. Additionally, the performance of MOF-5+ Tenax-TA was compared with other combinations comprising of Cu-based MOF-199 and Zr-based MOF of UiO-66 topology. The results of the theoretical modelling analyses propose simultaneous interaction of the C=O group of aldehydes with open metal sites of the studied MOFs and van der Waals interaction of hydrocarbon “tail” of aldehydes with linkers of MOFs. The combined interactions significantly increased the enthalpy (eV/molecule) of formaldehyde adsorption on MOF. Our findings unravel a potential way to extend the application of GC-based detection toward concurrent analysis of organic molecules of variable sizes.

Background The primary cilium is an organelle that can act as a master regulator of cellular signaling. Despite the presence of primary cilia in hippocampal neurons, their function is not fully understood. Recent studies have demonstrated that the primary cilium influences interleukin (IL)-1β-induced NF-κB signaling, ultimately mediating the inflammatory response. We, therefore, investigated ciliary function and NF-κB signaling in lipopolysaccharide (LPS)-induced neuroinflammation in conjunction with ciliary length analysis. Methods Since TLR4/NF-κB signaling is a well-known inflammatory pathway, we measured ciliary length and inflammatory mediators in wild type (WT) and TLR4−/− mice injected with LPS. Next, to exclude the effects of microglial TLR4, we examined the ciliary length, ciliary components, inflammatory cytokine, and mediators in HT22 hippocampal neuronal cells. Results Primary ciliary length decreased in hippocampal pyramidal neurons after intracerebroventricular injection of LPS in WT mice, whereas it increased in TLR4−/− mice. LPS treatment decreased primary ciliary length, activated NF-κB signaling, and increased Cox2 and iNOS levels in HT22 hippocampal neurons. In contrast, silencing Kif3a, a key protein component of cilia, increased ARL13B ciliary protein levels and suppressed NF-κB signaling and expression of inflammatory mediators. Conclusions These data suggest that LPS-induced NF-κB signaling and inflammatory mediator expression are modulated by cilia and that the blockade of primary cilium formation by Kif3a siRNA regulates TLR4-induced NF-κB signaling. We propose that primary cilia are critical for regulating NF-κB signaling events in neuroinflammation and in the innate immune response. Electronic supplementary material The online version of this article (10.1186/s12974-017-0958-7) contains supplementary material, which is available to authorized users.

Glioblastoma multiforme (GBM) is the most common and deadliest form of primary brain tumor. Despite treatment with surgery, radiotherapy, and chemotherapy with the drug temozolomide, the expected survival after diagnosis remains low. The median survival is only 14.6 months and the two-year survival is a mere 30%. One reason for this is the heterogeneity of GBM including the presence of glioblastoma cancer stem cells (GSCs). GSCs are a subset of cells with the unique ability to proliferate, differentiate, and create tumors. GSCs are resistant to chemotherapy and radiation and thought to play an important role in recurrence. In order to effectively treat GBM, a drug must be identified that can kill GSCs. The ionophore salinomycin has been shown to kill cancer stem cells and is therefore a promising future treatment for GBM. This study focuses on salinomycin's potential to treat GBM including its ability to reduce the CSC population, its toxicity to normal brain cells, its mechanism of action, and its potential for combination treatment.

Current methods for attachment of proteins to inorganic nanoparticle surfaces utilize strategies that can destroy the tertiary structure of large proteins and/or do not allow for directional attachment, thereby diminishing functionality. We report a novel approach for the conjugation of proteins to iron oxide nanoparticles (IONPs) through coiled-coil interactions. With this strategy, coiled-coiled interactions drive attachment of the desired protein, fused to a K-coil peptide, and E-coil peptides linked to the IONP surface via a nanoparticle attachment peptide (NAP). Importantly, this method allows for directional attachment of proteins without the use of either chemical conjugation or extreme pH solutions and, thus, allows maintenance of correct protein structure with functional orientation relative to the IONP surface. In our studies, monomeric red fluorescent protein (mRFP) and enhanced green fluorescent protein (EGFP) were used as model proteins to demonstrate the coiled-coil approach can be utilized to attach large proteins to IONP surfaces while maintaining complex tertiary structures. These proof-of-concept studies showed EGFP- and mRFP-K-coil fusion proteins interacted specifically with E-coil-coated nanoparticles and multiple different proteins could be displayed on individual IONPs. We did not observe fluorescence after conjugation of fluorescent proteins to either E-coil-coated IONPs or dopamine-coated IONPs, which was likely due to quenching or FRET activity induced by the iron oxide core. However, substituting an E-coil-bound chromatography resin for E-coil-coated IONPs allowed the preservation of protein fluorescence following coiled-coil attachment of K-coil-fused mRFP, demonstrating the coiled-coil attachment strategy provided maintenance of the mRFP tertiary structure required for fluorescence. The coiled-coil nanoparticle attachment strategy developed herein should have broad applications in nanomedicine.

Glioblastoma multiforme (GBM) is the deadliest and most common form of primary brain tumor. Conventional treatments are ineffective at treating GBM due to the heterogeneous cellular makeup of the tumors as well as the existence of drug‑resistant cells known as cancer stem cells (CSCs). CSCs have the ability to initiate tumorigenesis and self‑renew, which can lead to recurrence. Salinomycin, an antibiotic commonly used in agricultural feed, has been revealed to target CSCs in other cancer types. A few studies have suggested salinomycin can be effective at treating glioblastoma stem cells (GSCs); however, no study has examined the effect of salinomycin treatment on GSC markers. In the present study, flow cytometry, RT‑qPCR, and limiting dilution assays were used to further analyze the effects of salinomycin on GSCs. It was revealed that salinomycin decreased the expression of the GSC marker SOX2 at both the transcriptional and translational level. However, the effect of salinomycin on the GSC markers Nestin and CD133 was inconsistent between GBM subtypes. Additionally, the present findings provide initial evidence of caspase‑3‑dependent and independent apoptosis as the method by which salinomycin induces cell death in GBM. The present results indicated that salinomycin is an effective candidate as a chemotherapeutic agent that can treat GBM by targeting both bulk tumor cells as well as CSCs.

Human face recognition is one of the most important research areas in biometrics. However, the robust face recognition under a drastic change of the facial pose, expression, and illumination is a big challenging problem for its practical application. Such variations make face recognition more difficult. In this paper, we propose a novel face recognition method, called Attentional Feature-pair Relation Network (AFRN), which represents the face by the relevant pairs of local appearance block features with their attention scores. The AFRN represents the face by all possible pairs of the 9x9 local appearance block features, the importance of each pair is considered by the attention map that is obtained from the low-rank bilinear pooling, and each pair is weighted by its corresponding attention score. To increase the accuracy, we select top-K pairs of local appearance block features as relevant facial information and drop the remaining irrelevant. The weighted top-K pairs are propagated to extract the joint feature-pair relation by using bilinear attention network. In experiments, we show the effectiveness of the proposed AFRN and achieve the outstanding performance in the 1:1 face verification and 1:N face identification tasks compared to existing state-of-the-art methods on the challenging LFW, YTF, CALFW, CPLFW, CFP, AgeDB, IJB-A, IJB-B, and IJB-C datasets., To appear in ICCV 2019