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The need for more in-depth exploration of the human immune system has moved the flow cytometry field forward with advances in instrumentation, reagent development and availability, and user-friendly implementation of data analysis methods. We developed a high-quality human 45-color panel, for comprehensive characterization of major cell lineages present in circulation including T cells, γδ T cells, NKT-like cells, B cells, NK cells, monocytes, basophils, dendritic cells, and ILCs. Assay optimization steps are described in detail to ensure that each marker in the panel was optimally resolved. In addition, we highlight the outstanding discernment of cell activation, exhaustion, memory, and differentiation states of CD4+ and CD8+ T cells using this 45-color panel. The panel enabled an in-depth description of very distinct phenotypes associated with the complexity of the T cell memory response. Furthermore, we present how this panel can be effectively used for cell sorting on instruments with a similar optical layout to achieve the same level of resolution. Functional evaluation of sorted specific rare cell subsets demonstrated significantly different patterns of immunological responses to stimulation, supporting functional and phenotypic differences within the T cell memory subsets. In summary, the combination of full spectrum profiling technology and careful assay design and optimization results in a high resolution multiparametric 45-color assay. This panel offers the opportunity to fully characterize immunological profiles present in peripheral blood in the context of infectious diseases, autoimmunity, neurodegeneration, immunotherapy, and biomarker discovery., (© 2024 The Author(s). Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.)

The centrosome of the amoebozoan model Dictyostelium discoideum provides the best-established model for an acentriolar centrosome outside the Opisthokonta . Dictyostelium exhibits an unusual centrosome cycle, in which duplication is initiated only at the G2/M transition and occurs entirely during the M phase. Little is known about the role of conserved centrosomal kinases in this process. Therefore, we have generated knock-in strains for Aurora (AurK), CDK1, cyclin B, Nek2, and Plk, replacing the endogenous genes with constructs expressing the respective green fluorescent Neon fusion proteins, driven by the endogenous promoters, and studied their behavior in living cells. Our results show that CDK1 and cyclin B arrive at the centrosome first, already during G2, followed by Plk, Nek2, and AurK. Furthermore, CDK1/cyclin B and AurK were dynamically localized at kinetochores, and AurK in addition at nucleoli. The putative roles of all four kinases in centrosome duplication, mitosis, cytokinesis, and nucleolar dynamics are discussed.

Background: SGLT2i reduce cardiac hypertrophy, but underlying mechanisms remain unknown. Here we explore a role for serine/threonine kinases (STK) and sodium hydrogen exchanger 1(NHE1) activities in SGLT2i effects on cardiac hypertrophy., Methods: Isolated hearts from db/db mice were perfused with 1 µM EMPA, and STK phosphorylation sites were examined using unbiased multiplex analysis to detect the most affected STKs by EMPA. Subsequently, hypertrophy was induced in H9c2 cells with 50 µM phenylephrine (PE), and the role of the most affected STK (p90 ribosomal S6 kinase (RSK)) and NHE1 activity in hypertrophy and the protection by EMPA was evaluated., Results: In db/db mice hearts, EMPA most markedly reduced STK phosphorylation sites regulated by RSKL1, a member of the RSK family, and by Aurora A and B kinases. GO and KEGG analysis suggested that EMPA inhibits hypertrophy, cell cycle, cell senescence and FOXO pathways, illustrating inhibition of growth pathways. EMPA prevented PE-induced hypertrophy as evaluated by BNP and cell surface area in H9c2 cells. EMPA blocked PE-induced activation of NHE1. The specific NHE1 inhibitor Cariporide also prevented PE-induced hypertrophy without added effect of EMPA. EMPA blocked PE-induced RSK phosphorylation. The RSK inhibitor BIX02565 also suppressed PE-induced hypertrophy without added effect of EMPA. Cariporide mimicked EMPA's effects on PE-treated RSK phosphorylation. BIX02565 decreased PE-induced NHE1 activity, with no further decrease by EMPA., Conclusions: RSK inhibition by EMPA appears as a novel direct cardiac target of SGLT2i. Direct cardiac effects of EMPA exert their anti-hypertrophic effect through NHE-inhibition and subsequent RSK pathway inhibition., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. On behalf of all authors, the corresponding author CJ Zuurbier declares that none of the authors have known competing interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Authors. Published by Elsevier Masson SAS.. All rights reserved.)

Purpose: Robot-assisted vitreoretinal surgery provides precise and consistent operations on the back of the eye. To perform this safely, knowledge of the surgical instrument's remote centre of motion (RCM) and the location of the insertion point into the eye (trocar) is required. This enables the robot to align both positions to pivot the instrument about the trocar, thus preventing any damaging lateral forces from being exerted., Methods: Building on a system developed in previous work, this study presents a trocar localisation method that uses a micro-camera mounted on a vitreoretinal surgical forceps, to track two ArUco markers attached on either side of a trocar. The trocar position is the estimated midpoint between the markers., Results: Experimental evaluation of the trocar localisation was conducted. Results showed an RMSE of 1.82 mm for the localisation of the markers and an RMSE of 1.24 mm for the trocar localisation., Conclusions: The proposed camera-based trocar localisation presents reasonable consistency and accuracy and shows improved results compared to other current methods. Optimum accuracy for this application would necessitate a 1.4 mm absolute error margin, which corresponds to the trocar's radius. The trocar localisation results are successfully found within this margin, yet the marker localisation would require further refinement to ensure consistency of localisation within the error margin. Further work will refine these position estimates and ensure the error stays consistently within this boundary., (© 2023. The Author(s).)

DNA-encoded libraries (DELs) provide unmatched chemical diversity and starting points for novel drug modalities. Here, we describe a workflow that exploits the bifunctional attributes of DEL ligands as a platform to generate BRET probes for live cell target engagement studies. To establish proof of concept, we performed a DEL screen using aurora kinase A and successfully converted aurora DEL ligands as cell-active BRET probes. Aurora BRET probes enabled the validation and stratification of the chemical series identified from primary selection data. Furthermore, we have evaluated the effective repurposing of pre-existing DEL screen data to find suitable leads for BRET probe development. Our findings support the use of DEL workflows as an engine to create cell-active BRET probes independent of structure or compound SAR. The combination of DEL and BRET technology accelerates hit-to-lead studies in a live cell setting., Competing Interests: Declaration of interests Authors in this manuscript are employed by Promega or WuXi Apptec. Promega owns patents related to Target Engagement using BRET., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Aim: While most patients with RET-altered cancer responded to the RET protein tyrosine kinase inhibitors (TKIs) pralsetinib (BLU667) and selpercatinib (LOXO292), few achieved a complete response. Heterogeneity in residual tumors makes it difficult to target their diverse genetic alterations individually. The aim of this study is to characterize the cancer cells that persist under continuous RET TKI treatment and identify the shared vulnerability of these cells., Methods: We analyzed residual RET-altered cancer cells under prolonged RET TKI treatment by whole exome sequencing (WES), RNA-seq analysis, and drug-sensitivity screening. These were followed by tumor xenograft experiments of mono- and combinational drug treatments., Results: BLU667- and LOXO292-tolerated persisters were cellularly heterogeneous, contained slowly proliferating cells, regained low levels of active ERK1/2, and displayed plasticity in growth rate, which we designated as in the transition state of resistance (TSR). TSR cells were genetically heterogeneous. Aurora A/B kinases were among the most significantly upregulated genes and that the MAPK pathway activity had significantly higher transcript footprints. MEK1/2 and Aurora kinase inhibitors were the most effective drugs when combined with a RET kinase inhibitor. In a TSR tumor model, combination of BLU667 with an Aurora kinase or a MEK1/2 kinase inhibitor caused TSR tumor regression., Conclusion: Our experiments reveal that the heterogeneous TSR cancer cells under continuous RET TKI treatment converge on the targetable ERK1/2-driven Aurora A/B kinases. The discovery of the targetable convergent point in the genetically heterogeneous TSR points to an effective combination therapy approach to eliminate the residual tumors., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper. Competing Interests The authors declare no competing interests., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

High-dimensional single-cell data has become an important tool in unraveling the complexity of the immune system and its involvement in homeostasis and a large array of pathologies. As technological tools are developed, researchers are adopting them to answer increasingly complex biological questions. Up until recently, mass cytometry (MC) has been the main technology employed in cytometric assays requiring more than 29 markers. Recently, however, with the introduction of full spectrum flow cytometry (FSFC), it has become possible to break the fluorescence barrier and go beyond 29 fluorescent parameters. In this study, in collaboration with the Stanford Human Immune Monitoring Center (HIMC), we compared five patient samples using an established immune panel developed by the HIMC using their MC platform. Using split samples and the same antibody panel, we were able to demonstrate highly comparable results between the two technologies using multiple data analysis approaches. We report here a direct comparison of two technology platforms (MC and FSFC) using a 32-marker flow cytometric immune monitoring panel that can identify all the previously described and anticipated immune subpopulations defined by this panel., (© 2022 The Authors. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.)

Autofluorescence (AF) is an intrinsic characteristic of cells caused by the presence of fluorescent biological compounds within the cell; these can include structural proteins (e.g., collagen and elastin), cellular organelles, and metabolites (e.g., aromatic amino acids). In flow cytometric studies, the presence of AF can lead to reduced antigen and population resolution, as well as the presence of artifacts due to false positive events. Here, we describe a methodology that uses the inherent ability of full spectrum cytometry to treat AF as a fluorochrome and to thereby separate it from the other fluorochromes of the assay. This method can be applied to complex inflamed tissues; for instance, in regenerating skeletal muscle we have developed a 16-color panel targeting highly autofluorescent myeloid cells. This represents a first step toward overcoming technological limitations in flow cytometry due to AF., (© 2022 International Society for Advancement of Cytometry.)

In recent years, neural networks have been increasingly used for classifying aurora images. In particular, convolutional neural networks have been actively studied. However, there are not many studies on the application of deep learning techniques that take into account the characteristics of aurora images. Therefore, in this study, we propose the use of deep metric learning as a suitable method for aurora image classification. Deep metric learning is one of the deep learning techniques. It was developed to distinguish human faces. Identifying human faces is a more difficult task than standard classification tasks because this task is characterized by a small number of sample images for each class and poor feature variation between classes. We thought that the face identification task is similar to aurora image classification in that the number of labeled images is relatively small and the feature differences between classes are small. Therefore, we studied the application of deep metric learning to aurora image classification. As a result, our experiments showed that deep metric learning improves the accuracy of aurora image classification by nearly 10% compared to previous studies.

The Juno spacecraft's polar orbits have enabled direct sampling of Jupiter's low-altitude auroral field lines. While various data sets have identified unique features over Jupiter's main aurora, they are yet to be analyzed altogether to determine how they can be reconciled and fit into the bigger picture of Jupiter's auroral generation mechanisms. Jupiter's main aurora has been classified into distinct "zones", based on repeatable signatures found in energetic electron and proton spectra. We combine fields, particles, and plasma wave data sets to analyze Zone-I and Zone-II, which are suggested to carry upward and downward field-aligned currents, respectively. We find Zone-I to have well-defined boundaries across all data sets. H + and/or H 3 + beams and downward energetic electron beams. Zone-II, on the other hand, does not have a clear poleward boundary with the polar cap, and its signatures are more sporadic. Large-amplitude solitary waves, which are reminiscent of those ubiquitous in Earth's downward current region, are a key feature of Zone-II. Alfvénic fluctuations are most prominent in the diffuse aurora and are repeatedly found to diminish in Zone-I and Zone-II, likely due to dissipation, at higher altitudes, to energize auroral electrons. Finally, we identify significant electron density depletions, by up to 2 orders of magnitude, in Zone-I, and discuss their important implications for the development of parallel potentials, Alfvénic dissipation, and radio wave generation.+ beams and downward energetic electron beams. Zone-II, on the other hand, does not have a clear poleward boundary with the polar cap, and its signatures are more sporadic. Large-amplitude solitary waves, which are reminiscent of those ubiquitous in Earth's downward current region, are a key feature of Zone-II. Alfvénic fluctuations are most prominent in the diffuse aurora and are repeatedly found to diminish in Zone-I and Zone-II, likely due to dissipation, at higher altitudes, to energize auroral electrons. Finally, we identify significant electron density depletions, by up to 2 orders of magnitude, in Zone-I, and discuss their important implications for the development of parallel potentials, Alfvénic dissipation, and radio wave generation., (© 2022. The Authors.)

The accurate determination of auroral precipitation in global models has remained a daunting and rather inexplicable obstacle. Understanding the calculation and balance of multiple sources that constitute the aurora, and their eventual conversion into ionospheric electrical conductance, is critical for improved prediction of space weather events. In this study, we present a semi-physical global modeling approach that characterizes contributions by four types of precipitation-monoenergetic, broadband, electron, and ion diffuse-to ionospheric electrodynamics. The model uses a combination of adiabatic kinetic theory and loss parameters derived from historical energy flux patterns to estimate auroral precipitation from magnetohydrodynamic (MHD) quantities. It then converts them into ionospheric conductance that is used to compute the ionospheric feedback to the magnetosphere. The model has been employed to simulate the 5-7 April 2010 Galaxy15 space weather event. Comparison of auroral fluxes show good agreement with observational data sets like NOAA-DMSP and OVATION Prime. The study shows a dominant contribution by electron diffuse precipitation, accounting for ∼74% of the auroral energy flux. However, contributions by monoenergetic and broadband sources dominate during times of active upstream solar conditions, providing for up to 61% of the total hemispheric power. The study also finds a greater role played by broadband precipitation in ionospheric electrodynamics which accounts for ∼31% of the Pedersen conductance., (©2022. The Authors.)

We present 14 simultaneous Chandra X-ray Observatory (CXO)-Hubble Space Telescope (HST) observations of Jupiter's Northern X-ray and ultraviolet (UV) aurorae from 2016 to 2019. Despite the variety of dynamic UV and X-ray auroral structures, one region is conspicuous by its persistent absence of emission: the dark polar region (DPR). Previous HST observations have shown that very little UV emission is produced by the DPR. We find that the DPR also produces very few X-ray photons. For all 14 observations, the low level of X-ray emission from the DPR is consistent (within 2-standard deviations) with scattered solar emission and/or photons spread by Chandra's Point Spread Function from known X-ray-bright regions. We therefore conclude that for these 14 observations the DPR produced no statistically significant detectable X-ray signature., (© 2022. The Authors.)

Discrete aurora at Mars, characterized by their small spatial scale and tendency to form near strong crustal magnetic fields, are emissions produced by particle precipitation into the Martian upper atmosphere. Since 2014, Mars Atmosphere and Volatile EvolutioN's (MAVEN's) Imaging Ultraviolet Spectrograph (IUVS) has obtained a large collection of UV discrete aurora observations during its routine periapsis nightside limb scans. Initial analysis of these observations has shown that, near the strongest crustal magnetic fields in the southern hemisphere, the IUVS discrete aurora detection frequency is highly sensitive to the interplanetary magnetic field (IMF) clock angle. However, the role of other solar wind properties in controlling the discrete aurora detection frequency has not yet been determined. In this work, we use the IUVS discrete aurora observations, along with MAVEN observations of the upstream solar wind, to determine how the discrete aurora detection frequency varies with solar wind dynamic pressure, IMF strength, and IMF cone angle. We find that, outside of the strong crustal field region (SCFR) in the southern hemisphere, the aurora detection frequency is relatively insensitive to the IMF orientation, but significantly increases with solar wind dynamic pressure, and moderately increases with IMF strength. Interestingly however, although high solar wind dynamic pressures cause more aurora to form, they have little impact on the brightness of the auroral emissions. Alternatively, inside the SCFR, the detection frequency is only moderately dependent on the solar wind dynamic pressure, and is much more sensitive to the IMF clock and cone angles. In the SCFR, aurora are unlikely to occur when the IMF points near the radial or anti-radial directions when the cone angle (arccos( B x |)) is less than 30° or between 120° and 150°. Together, these results provide the first comprehensive characterization of how upstream solar wind conditions affect the formation of discrete aurora at Mars.B |)) is less than 30° or between 120° and 150°. Together, these results provide the first comprehensive characterization of how upstream solar wind conditions affect the formation of discrete aurora at Mars., (© 2022 The Authors.)

We develop an open source algorithm to apply T ransfer learning to A urora image classification and M agnetic disturbance E valuation (TAME). For this purpose, we evaluate the performance of 80 pretrained neural networks using the Oslo Auroral THEMIS (OATH) data set of all-sky images, both in terms of runtime and their features' predictive capability. From the features extracted by the best network, we retrain the last neural network layer using the Support Vector Machine (SVM) algorithm to distinguish between the labels "arc," "diffuse," "discrete," "cloud," "moon" and "clear sky/ no aurora". This transfer learning approach yields 73% accuracy in the six classes; if we aggregate the 3 auroral and 3 non-aurora classes, we achieve up to 91% accuracy. We apply our classifier to a new dataset of 550,000 images and evaluate the classifier based on these previously unseen images. To show the potential usefulness of our feature extractor and classifier, we investigate two test cases: First, we compare our predictions for the "cloudy" images to meteorological data and second we train a linear ridge model to predict perturbations in Earth's locally measured magnetic field. We demonstrate that the classifier can be used as a filter to remove cloudy images from datasets and that the extracted features allow to predict magnetometer measurements. All procedures and algorithms used in this study are publicly available, and the code and classifier are provided, which opens possibility for large scale studies of all-sky images., (© 2021. The Authors.)

Series of novel sulfonamide-based 3-indolinones 3a-m and 4a-f were designed, synthesized and then their cytotoxic activity was evaluated against a panel of sixty cancer cell lines. This screening indicated that 4-(2-(5-fluoro-2-oxoindolin-3-ylidene)acetyl)phenyl benzenesulfonate (4f) possessed promising cytotoxicity against CCRF-CEM and SR leukemia cell lines with IC 50 values 6.84 and 2.97 µM, respectively. Further investigation of the leukemic cytotoxicity of compound 4f was carried out by performing PDGFRα, VEGFR2, Aurora A/B and FLT3 enzyme assays and CCRF-CEM and SR cell cycle analysis. These investigations showed that compound 4f exhibited pronounced dual inhibition of both kinases PDGFRα and Aurora A with potency of 24.15 and 11.83 nM, respectively. The in vitro results were supported by molecular docking studies in order to explore its binding affinity and its key amino acids interactions. This work represents compound 4f as a promising anticancer agent against leukemia., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Flow cytometry is a powerful technology used in research, drug development and clinical sample analysis for cell identification and characterization, allowing for the simultaneous interrogation of multiple targets on various cell subsets from limited samples. Recent advancements in instrumentation and fluorochrome availability have resulted in significant increases in the complexity and dimensionality of flow cytometry panels. Though this increase in panel size allows for detection of a broader range of markers and sub-populations, even in restricted biological samples, it also comes with many challenges in panel design, optimization, and downstream data analysis and interpretation. In the current paper we describe the practices we established for development of high-dimensional panels on the Aurora spectral flow cytometer to aid clinical sample analysis.

We report the first simultaneous, common-volume lidar observations of thermosphere-ionosphere Fe (TIFe) and Na (TINa) layers in Antarctica. We also report the observational discovery of nearly one-to-one correspondence between TIFe and aurora activity, enhanced ionization layers, and converging electric fields. Distinctive TIFe layers have a peak density of ~384 cm -3 and the TIFe mixing ratio peaks around 123 km, ~5 times the mesospheric layer maximum. All evidence shows that Fe + ion-neutralization is the major formation mechanism of TIFe layers. The TINa mixing ratio often exhibits a broad peak at TIFe altitudes, providing evidence for in situ production via Na + neutralization. However, the tenuous TINa layers persist long beyond TIFe disappearance and reveal gravity wave perturbations, suggesting a dynamic background of neutral Na, but not Fe, above 110 km. The striking differences between distinct TIFe and diffuse TINa suggest differential transport between Fe and Na, possibly due to mass separation., (©2020. The Authors.)

This 40-color flow cytometry-based panel was developed for in-depth immunophenotyping of the major cell subsets present in human peripheral blood. Sample availability can often be limited, especially in cases of clinical trial material, when multiple types of testing are required from a single sample or timepoint. Maximizing the amount of information that can be obtained from a single sample not only provides more in-depth characterization of the immune system but also serves to address the issue of limited sample availability. The panel presented here identifies CD4 T cells, CD8 T cells, regulatory T cells, γδ T cells, NKT-like cells, B cells, NK cells, monocytes and dendritic cells. For each specific cell type, the panel includes markers for further characterization by including a selection of activation and differentiation markers, as well as chemokine receptors. Moreover, the combination of multiple markers in one tube might lead to the discovery of new immune phenotypes and their relevance in certain diseases. Of note, this panel was designed to include only surface markers to avoid the need for fixation and permeabilization steps. The panel can be used for studies aimed at characterizing the immune response in the context of infectious or autoimmune diseases, monitoring cancer patients on immuno- or chemotherapy, and discovery of unique and targetable biomarkers. Different from all previously published OMIPs, this panel was developed using a full spectrum flow cytometer, a technology that has allowed the effective use of 40 fluorescent markers in a single panel. The panel was developed using cryopreserved human peripheral blood mononuclear cells (PBMC) from healthy adults (Table 1). Although we have not tested the panel on fresh PBMCs or whole blood, it is anticipated that the panel could be used in those sample preparations without further optimization. @ 2020 Cytek Biosciences, Inc. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry., (@ 2020 Cytek Biosciences, Inc. Cytometry Part A published by Wiley Periodicals LLC on behalf of International Society for Advancement of Cytometry.)

A distinct class of aurora, called transpolar auroral arc (TPA) (in some cases called "theta" aurora), appears in the extremely high-latitude ionosphere of the Earth when interplanetary magnetic field (IMF) is northward. The formation and evolution of TPA offers clues about processes transferring energy and momentum from the solar wind to the magnetosphere and ionosphere during a northward IMF. However, their formation mechanisms remain poorly understood and controversial. We report a mechanism identified from multiple-instrument observations of unusually bright, multiple TPAs and simulations from a high-resolution three-dimensional (3D) global MagnetoHydroDynamics (MHD) model. The observations and simulations show an excellent agreement and reveal that these multiple TPAs are generated by precipitating energetic magnetospheric electrons within field-aligned current (FAC) sheets. These FAC sheets are generated by multiple-flow shear sheets in both the magnetospheric boundary produced by Kelvin-Helmholtz instability between supersonic solar wind flow and magnetosphere plasma, and the plasma sheet generated by the interactions between the enhanced earthward plasma flows from the distant tail (less than -100 R E ) and the enhanced tailward flows from the near tail (about -20 R E ). The study offers insight into the complex solar wind-magnetosphere-ionosphere coupling processes under a northward IMF condition, and it challenges existing paradigms of the dynamics of the Earth's magnetosphere., Competing Interests: The authors declare no competing interest.

The most fantastic optical phenomena in the Earth's upper atmosphere are the auroras. They are highly informative indicators of solar activity, geomagnetic activity, upper atmospheric structures and dynamics, and magnetospheric energetic particles. An area where the geomagnetic field differs significantly from the expected symmetric dipole, such as at the South Atlantic Anomaly, where the magnetic field intensity is low, gives rise to stronger precipitation of energetic particles into the upper atmosphere. Impact excitation and the subsequent airglow emissions exhibit aurora-like dynamic signatures. Nomenclatures of nonpolar aurora or equatorial auroras are similar to those used with the polar auroras owing to their similar excitation mechanisms. This paper provides an overview of the knowledge and the challenges concerning auroral activity at the South Atlantic Anomaly, or more generally, at the negative magnetic anomaly. We emphasize systematic investigation of the equatorial auroras to reveal the temporal and spatial evolution of the magnetic anomaly and the behaviour of energetic particles in near-Earth space., (© The Author(s) 2020. Published by Oxford University Press on behalf of China Science Publishing & Media Ltd.)

The coordinated activities of many protein kinases, acting on multiple protein substrates, ensures the error-free progression through mitosis of eukaryotic cells. Enormous research effort has thus been devoted to studying the roles and regulation of these mitotic kinases, and to the identification of their physiological substrates. Central for the timely deployment of specific protein kinases to their appropriate substrates during the cell division cycle are the many anchoring proteins, which serve critical regulatory roles. Through direct association, anchoring proteins are capable of modulating the catalytic activity and/or sub-cellular distribution of the mitotic kinases they associate with. The key roles of some anchoring proteins in cell division are well-established, whilst others are still being unearthed. Here, we review the current knowledge on anchoring proteins for some mitotic kinases, and highlight how targeting anchoring proteins for inhibition, instead of the mitotic kinases themselves, could be advantageous for disrupting the cell division cycle.

The Jovian polar regions produce X-rays that are characteristic of very energetic oxygen and sulfur that become highly charged on precipitating into Jupiter's upper atmosphere. Juno has traversed the polar regions above where these energetic ions are expected to be precipitating revealing a complex composition and energy structure. Energetic ions are likely to drive the characteristic X-rays observed at Jupiter (Haggerty et al., 2017, https://doi.org/10.1002/2017GL072866; Houston et al., 2018, https://doi.org/10.1002/2017JA024872; Kharchenko et al., 2006, https://doi.org/10.1029/2006GL026039). Motivated by the science of X-ray generation, we describe here Juno Jupiter Energetic Particle Detector Instrument (JEDI) measurements of ions above 1 MeV and demonstrate the capability of measuring oxygen and sulfur ions with energies up to 100 MeV. We detail the process of retrieving ion fluxes from pulse width data on instruments like JEDI (called "puck's"; Clark, Cohen, et al., 2016, https://doi.org/10.1002/2017GL074366; Clark, Mauk, et al., 2016, https://doi.org/10.1002/2015JA022257; Mauk et al., 2013, https://doi.org/10.1007/s11214-013-0025-3) as well as details on retrieving very energetic particles (>20 MeV) above which the pulse width also saturates., (©2019. The Authors.)

The study of the composition of brown dwarf atmospheres helped to understand their formation and evolution. Similarly, the study of exoplanet atmospheres is expected to constrain their formation and evolutionary states. We use results from three-dimensional simulations, kinetic cloud formation and kinetic ion-neutral chemistry to investigate ionization processes that will affect their atmosphere chemistry: the dayside of super-hot Jupiters is dominated by atomic hydrogen, and not H 2 O. Such planetary atmospheres exhibit a substantial degree of thermal ionization and clouds only form on the nightside where lightning leaves chemical tracers (e.g. HCN) for possibly long enough to be detectable. External radiation may cause exoplanets to be enshrouded in a shell of highly ionized, H 3 -forming gas and a weather-driven aurora may emerge. Brown dwarfs enable us to study the role of electron beams for the emergence of an extrasolar, weather system-driven aurora-like chemistry, and the effect of strong magnetic fields on cold atmospheric gases. Electron beams trigger the formation of H + -forming gas and a weather-driven aurora may emerge. Brown dwarfs enable us to study the role of electron beams for the emergence of an extrasolar, weather system-driven aurora-like chemistry, and the effect of strong magnetic fields on cold atmospheric gases. Electron beams trigger the formation of H 3 + in the upper atmosphere of a brown dwarf (e.g. LSR-J1835), which may react with it to form hydronium, H 3 O + , as a longer lived chemical tracer. Brown dwarfs and super-hot gas giants may be excellent candidates to search for H 3 O + as an H 3 + product. This article is part of a discussion meeting issue 'Advances in hydrogen molecular ions: H 3 + , H 5 + and beyond'.

The 4-aminoquinazoline core is an interesting pharmacophore and its applications in medicinal chemistry are wide spread. The core has been used for making many kinase inhibitors in past few years. Many researcher demonstrated 4-aminoquinazoline derivatives as specific kinase inhibitors, including tyrosine kinase and serine/theronine kinases. A number of anticancer drugs with 4-aminoquinazoline core are in the market, e.g. gefitinib, erlotinib, afatinib, lapatinib, decomitinib etc. 4-aminoquinazoline derivatives are applied for target specific treatment of lung, breast, colon, prostate cancers. In this review, we discussed the current development of 4-aminoquinazoline derivatives as kinase inhibitors and their uses as anticancer agents in recent years., (Copyright © 2019 Elsevier Masson SAS. All rights reserved.)

Substorm onset has originally been defined as a longitudinally extended sudden auroral brightening (Akasofu initial brightening: AIB) followed a few minutes later by an auroral poleward expansion in ground-based all-sky images (ASIs). In contrast, such clearly marked two-stage development has not been evident in satellite-based global images (GIs). Instead, substorm onsets have been identified as localized sudden brightenings that expand immediately poleward. To resolve these differences, optical substorm onset signatures in GIs and ASIs are compared in this study for a substorm that occurred on December 7, 1999. For this substorm, the Polar satellite ultraviolet global imager was operated with a fixed-filter (170 nm) mode, enabling a higher time resolution (37 s) than usual to resolve the possible two-stage development. These data were compared with 20-s resolution green-line (557.7 nm) ASIs at Muonio in Finland. The ASIs revealed the AIB at 2124:50 UT and the subsequent poleward expansion at 2127:50 UT, whereas the GIs revealed only an onset brightening that started at 2127:49 UT. Thus, the onset in the GIs was delayed relative to the AIB and in fact agreed with the poleward expansion in the ASIs. The fact that the AIB was not evident in the GIs may be attributed to the limited spatial resolution of GIs for thin auroral arc brightenings. The implications of these results for the definition of substorm onset are discussed herein., Competing Interests: The authors declare that they have no competing interests.

Both mitosis and autophagy are highly regulated dynamic cellular processes and involve various phosphorylation events catalysed by kinases, which play vital roles in almost all physiological and pathological conditions. Mitosis is a key event during the cell cycle, in which the cell divides into two daughter cells. Autophagy is a process in which the cell digests its own cellular contents. Although autophagy regulation has mainly been studied in asynchronous cells, increasing evidence indicates that autophagy is in fact tightly regulated in mitosis. Here in this review, we will discuss kinases that were originally identified to be involved in only one of either mitosis or autophagy, but were later found to participate in both processes, such as CDKs (cyclin-dependent kinases), Aurora kinases, PLK-1 (polo-like kinase 1), BUB1 (budding uninhibited by benzimidazoles 1), MAPKs (mitogen-activated protein kinases), mTORC1 (mechanistic target of rapamycin complex 1), AMPK (AMP-activated protein kinase), PI3K (phosphoinositide-3 kinase) and protein kinase B (AKT). By focusing on kinases involved in both autophagy and mitosis, we will get a more comprehensive understanding about the reciprocal regulation between the two key cellular events, which will also shed light on their related therapeutic investigations., Competing Interests: The authors declare no conflict of interest.

Juno ultraviolet spectrograph (UVS) observations of Jupiter's aurora obtained during approach are presented. Prior to the bow shock crossing on 24 June 2016, the Juno approach provided a rare opportunity to correlate local solar wind conditions with Jovian auroral emissions. Some of Jupiter's auroral emissions are expected to be controlled or modified by local solar wind conditions. Here we compare synoptic Juno-UVS observations of Jupiter's auroral emissions, acquired during 3-29 June 2016, with in situ solar wind observations, and related Jupiter observations from Earth. Four large auroral brightening events are evident in the synoptic data, in which the total emitted auroral power increases by a factor of 3-4 for a few hours. Only one of these brightening events correlates well with large transient increases in solar wind ram pressure. The brightening events which are not associated with the solar wind generally have a risetime of ~2 h and a decay time of ~5 h.

Aurora kinases control multiple events during cell cycle progression and are essential for mitotic and meiotic bipolar spindle assembly and function. There are three Aurora kinases in mammals, some of which have oncogenic properties and all of which are overexpressed in multiple cancers. Pharmaceutical companies quickly made these kinases priority targets for the development of inhibitors to be used as cancer treatments. In this review, we focus on Aurora A, against which several inhibiting compounds have been discovered and made available; however, even though some of these compounds underwent clinical trials, none have yet gone beyond Phase III trials. The varying efficiencies and particularities of these drugs raise several questions that are explored in this review: is Aurora A even a good target? What biomarkers can we use to measure its activity in vivo? How can we improve the Aurora A-inhibiting drugs?, (Copyright © 2017 Elsevier Ltd. All rights reserved.)

We have developed a method for reprocessing the multi-decadal, multi-spacecraft Defense Meteorological Satellite Program Magnetometer (DMSP SSM) dataset and have applied it to fifteen spacecraft-years of data (DMSP Flight 16-18, 2010-2014). This Level-2 dataset improves on other available SSM datasets with recalculated spacecraft locations and magnetic perturbations, artifact signal removal, representations of the observations in geomagnetic coordinates, and in-situ auroral boundaries. Spacecraft locations have been recalculated using ground-tracking information. Magnetic perturbations (measured field minus modeled main-field) are recomputed. The updated locations ensure the appropriate model field is used. We characterize and remove a slow-varying signal in the magnetic field measurements. This signal is a combination of ring current and measurement artifacts. A final artifact remains after processing: step-discontinuities in the baseline caused by activation/deactivation of spacecraft electronics. Using coincident data from the DMSP precipitating electrons and ions instrument (SSJ4/5), we detect the in-situ auroral boundaries with an improvement to the Redmon et al. [2010] algorithm. We embed the location of the aurora and an accompanying figure of merit in the Level-2 SSM data product. Finally, we demonstrate the potential of this new dataset by estimating field-aligned current (FAC) density using the Minimum Variance Analysis (MVA) technique. The FAC estimates are then expressed in dynamic auroral boundary coordinates using the SSJ-derived boundaries, demonstrating a dawn-dusk asymmetry in average FAC location relative to the equatorward edge of the aurora. The new SSM dataset is now available in several public repositories.

Since the mid 1970's, the Defense Meteorological Satellite Program (DMSP) spacecraft have operated instruments for monitoring the space environment from low earth orbit. As the program evolved, so to have the measurement capabilities such that modern DMSP spacecraft include a comprehensive suite of instruments providing estimates of precipitating electron and ion fluxes, cold/bulk plasma composition and moments, the geomagnetic field, and optical emissions in the far and extreme ultraviolet. We describe the creation of a new public database of precipitating electrons and ions from the Special Sensor J (SSJ) instrument, complete with original counts, calibrated differential fluxes adjusted for penetrating radiation, estimates of the total kinetic energy flux and characteristic energy, uncertainty estimates, and accurate ephemerides. These are provided in a common and self-describing format that covers 30+ years of DMSP spacecraft from F06 (launched in 1982) through F18 (launched in 2009). This new database is accessible at the National Centers for Environmental Information (NCEI) and the Coordinated Data Analysis Web (CDAWeb). We describe how the new database is being applied to high latitude studies of: the co-location of kinetic and electromagnetic energy inputs, ionospheric conductivity variability, field aligned currents and auroral boundary identification. We anticipate that this new database will support a broad range of space science endeavors from single observatory studies to coordinated system science investigations.

Past observations and modeling of Jupiter's thermosphere have, due to their limited resolution, suggested that heat generated by the aurora near the poles results in a smooth thermal gradient away from these aurorae, indicating a quiescent and diffuse flow of energy within the subauroral thermosphere. Here we discuss Very Large Telescope-Cryogenic High-Resolution IR Echelle Spectrometer observations that reveal a small-scale localized cooling of ~200 K within the nonauroral thermosphere. Using Infrared Telescope Facility NSFCam images, this feature is revealed to be quasi-stable over at least a 15 year period, fixed in magnetic latitude and longitude. The size and shape of this "Great Cold Spot" vary significantly with time, strongly suggesting that it is produced by an aurorally generated weather system: the first direct evidence of a long-term thermospheric vortex in the solar system. We discuss the implications of this spot, comparing it with short-term temperature and density variations at Earth.

Allostery is a phenomenon observed in many proteins where binding of a macromolecular partner or a small-molecule ligand at one location leads to specific perturbations at a site not in direct contact with the region where the binding occurs. The list of proteins under allosteric regulation includes AGC protein kinases. AGC kinases have a conserved allosteric site, the phosphoinositide-dependent protein kinase 1 (PDK1)-interacting fragment (PIF) pocket, which regulates protein ATP-binding, activity, and interaction with substrates. In this study, we identify small molecules that bind to the ATP-binding site and affect the PIF pocket of AGC kinase family members, PDK1 and Aurora kinase. We describe the mechanistic details and show that although PDK1 and Aurora kinase inhibitors bind to the conserved ATP-binding site, they differentially modulate physiological interactions at the PIF-pocket site. Our work outlines a strategy for developing bidirectional small-molecule allosteric modulators of protein kinases and other signaling proteins., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

We report the first Jupiter X-ray observations planned to coincide with an interplanetary coronal mass ejection (ICME). At the predicted ICME arrival time, we observed a factor of ∼8 enhancement in Jupiter's X-ray aurora. Within 1.5 h of this enhancement, intense bursts of non-Io decametric radio emission occurred. Spatial, spectral, and temporal characteristics also varied between ICME arrival and another X-ray observation two days later. Gladstone et al. (2002) discovered the polar X-ray hot spot and found it pulsed with 45 min quasiperiodicity. During the ICME arrival, the hot spot expanded and exhibited two periods: 26 min periodicity from sulfur ions and 12 min periodicity from a mixture of carbon/sulfur and oxygen ions. After the ICME, the dominant period became 42 min. By comparing Vogt et al. (2011) Jovian mapping models with spectral analysis, we found that during ICME arrival at least two distinct ion populations, from Jupiter's dayside, produced the X-ray aurora. Auroras mapping to magnetospheric field lines between 50 and 70  R J were dominated by emission from precipitating sulfur ions (S 7+,…,14+ ). Emissions mapping to closed field lines between 70 and 120  R J and to open field lines were generated by a mixture of precipitating oxygen (O 7+,8+ ) and sulfur/carbon ions, possibly implying some solar wind precipitation. We suggest that the best explanation for the X-ray hot spot is pulsed dayside reconnection perturbing magnetospheric downward currents, as proposed by Bunce et al. (2004). The auroral enhancement has different spectral, spatial, and temporal characteristics to the hot spot. By analyzing these characteristics and coincident radio emissions, we propose that the enhancement is driven directly by the ICME through Jovian magnetosphere compression and/or a large-scale dayside reconnection event.

Bipolar Disorder is a severe disease characterized by pathological mood swings from major depressive episodes to manic ones and vice versa. The biological underpinnings of Bipolar Disorder have yet to be defined. As a consequence, pharmacological treatments are suboptimal. In the present paper we test the hypothesis that the molecular pathways involved with the direct targets of lithium, hold significantly more genetic variations associated with BD. A molecular pathway approach finds its rationale in the polygenic nature of the disease. The pathways were tested in a sample of ∼ 7,000 patients and controls. Data are available from the public NIMH database. The definition of the pathways was conducted according to the National Cancer Institute (http://pid.nci.nih.gov/). As a result, 3 out of the 18 tested pathways related to lithium action resisted the permutation analysis and were found to be associated with BD. These pathways were related to Reelin, Integrins and Aurora. A pool of genes selected from the ones linked with the above pathways was further investigated in order to identify the fine molecular mechanics shared by our significant pathways and also their link with lithium mechanism of action. The data obtained point out to a possible involvement of microtubule-related mechanics., (Copyright © 2015 Elsevier B.V. All rights reserved.)

We present the first multievent study of the spatial and temporal structuring of the aurora to provide statistical evidence of the near-Earth plasma instability which causes the substorm onset arc. Using data from ground-based auroral imagers, we study repeatable signatures of along-arc auroral beads, which are thought to represent the ionospheric projection of magnetospheric instability in the near-Earth plasma sheet. We show that the growth and spatial scales of these wave-like fluctuations are similar across multiple events, indicating that each sudden auroral brightening has a common explanation. We find statistically that growth rates for auroral beads peak at low wave number with the most unstable spatial scales mapping to an azimuthal wavelength λ ≈ 1700-2500 km in the equatorial magnetosphere at around 9-12  R E . We compare growth rates and spatial scales with a range of theoretical predictions of magnetotail instabilities, including the Cross-Field Current Instability and the Shear Flow Ballooning Instability. We conclude that, although the Cross-Field Current instability can generate similar magnitude of growth rates, the range of unstable wave numbers indicates that the Shear Flow Ballooning Instability is the most likely explanation for our observations.

In just over two decades, structure based protein kinase inhibitor discovery has grown from trial and error approaches, using individual target structures, to structure and data driven approaches that may aim to optimize inhibition properties across several targets. This is increasingly enabled by the growing availability of potent compounds and kinome-wide binding data. Assessing the prospects for adapting known compounds to new therapeutic uses is thus a key priority for current drug discovery efforts. Tools that can successfully link the diverse information regarding target sequence, structure, and ligand binding properties now accompany a transformation of protein kinase inhibitor research, away from single, block-buster drug models, and toward "personalized medicine" with niche applications and highly specialized research groups. Major hurdles for the transformation to data driven drug discovery include mismatches in data types, and disparities of methods and molecules used; at the core remains the problem that ligand binding energies cannot be predicted precisely from individual structures. However, there is a growing body of experimental data for increasingly successful focussing of efforts: focussed chemical libraries, drug repurposing, polypharmacological design, to name a few. Protein kinase target similarity is easily quantified by sequence, and its relevance to ligand design includes broad classification by key binding sites, evaluation of resistance mutations, and the use of surrogate proteins. Although structural evaluation offers more information, the flexibility of protein kinases, and differences between the crystal and physiological environments may make the use of crystal structures misleading when structures are considered individually. Cheminformatics may enable the "calibration" of sequence and crystal structure information, with statistical methods able to identify key correlates to activity but also here, "the devil is in the details." Examples from specific repurposing and polypharmacology applications illustrate these points. This article is part of a Special Issue entitled: Inhibitors of Protein Kinases., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Alisertib (MLN8237) is a selective small molecule inhibitor of Aurora A kinase that is being developed in multiple cancer indications as a single agent and in combination with other therapies. A significant amount of research has elucidated a role for Aurora A in orchestrating numerous activities of cells transiting through mitosis and has begun to shed light on potential non-mitotic roles for Aurora A as well. These biological insights laid the foundation for multiple clinical trials evaluating the antitumor activity of alisertib in both solid cancers and heme-lymphatic malignancies. Several key facets of Aurora A biology as well as empirical data collected in experimental systems and early clinical trials have directed the development of alisertib toward certain cancer types, including neuroblastoma, small cell lung cancer, neuroendocrine prostate cancer, atypical teratoid/rhabdoid tumors, and breast cancer among others. In addition, these scientific insights provided the rationale for combining alisertib with other therapies, including microtubule perturbing agents, such as taxanes, EGFR inhibitors, hormonal therapies, platinums, and HDAC inhibitors among others. Here, we link the key aspects of the current clinical development of alisertib to the originating scientific rationale and provide an overview of the alisertib clinical experience to date.

We previously identified Aurora B kinase as the only independent factor predictive of the aggressive recurrence of hepatocellular carcinoma (HCC). In this preclinical study, JNJ-28841072, a novel Aurora/vascular endothelial growth factor receptor dual kinase inhibitor, was evaluated for treatment of HCC. In vitro and in vivo effects of JNJ-28841072 were analyzed using human HCC cell cultures and xenograft models. An orthotopic liver xenograft model was used for the pharmacobiological effects on Aurora kinase and vascularization in hepatic tumors. JNJ-28841072 suppressed in vitro phosphorylation of histone H3 with induction of cell polyploidy and death in a dose-dependent manner (IC50  = 0.8-1.2 μM). In s.c. human HCC xenografts, remarkable inhibition of tumor growth was observed after JNJ-28841072 treatment (P = 0.0005). In orthotopic liver xenografts, the treatment with JNJ-28841072 significantly suppressed in vivo phosphorylation of histone H3 (P = 0.0008), vessel formation (P = 0.018), normoxic area (P = 0.0001), and hepatoma growth (P = 0.038). Our preclinical studies indicate that JNJ-28841072 is a promising novel therapeutic approach for the treatment of HCC. It might be worthy of evaluation in further studies., (© 2015 The Authors. Cancer Science published by Wiley Publishing Asia Pty Ltd on behalf of Japanese Cancer Association.)

We introduce and analyze the first data from an array of closely spaced Global Positioning System (GPS) scintillation receivers established in the auroral zone in late 2013 to measure spatial and temporal variations in L band signals at 100-1000 m and subsecond scales. The seven receivers of the Scintillation Auroral GPS Array (SAGA) are sited at Poker Flat Research Range, Alaska. The receivers produce 100 s scintillation indices and 100 Hz carrier phase and raw in-phase and quadrature-phase samples. SAGA is the largest existing array with baseline lengths of the ionospheric diffractive Fresnel scale at L band. With an initial array of five receivers, we identify a period of simultaneous amplitude and phase scintillation. We compare SAGA power and phase data with collocated 630.0 nm all-sky images of an auroral arc and incoherent scatter radar electron precipitation measurements, to illustrate how SAGA can be used in multi-instrument observations for subkilometer-scale studies., Key Points: A seven-receiver Scintillation Auroral GPS Array (SAGA) is now at Poker Flat, Alaska SAGA is the largest subkilometer array to enable phase/irregularities studies Simultaneous scintillation, auroral arc, and electron precipitation are observed.

We have produced the first series of spherical harmonic, numerical maps of the time-dependent surface perturbations in the Earth's magnetic field following the onset of substorms. Data from 124 ground magnetometer stations in the Northern Hemisphere at geomagnetic latitudes above 33° were used. Ground station data averaged over 5 min intervals covering 8 years (1998-2005) were used to construct pseudo auroral upper, auroral lower, and auroral electrojet ( AU *, AL *, and AE *) indices. These indices were used to generate a list of substorms that extended from 1998 to 2005, through a combination of automated processing and visual checks. Events were sorted by interplanetary magnetic field (IMF) orientation (at the Advanced Composition Explorer (ACE) satellite), dipole tilt angle, and substorm magnitude. Within each category, the events were aligned on substorm onset. A spherical cap harmonic analysis was used to obtain a least error fit of the substorm disturbance patterns at 5 min intervals up to 90 min after onset. The fits obtained at onset time were subtracted from all subsequent fits, for each group of substorm events. Maps of the three vector components of the averaged magnetic perturbations were constructed to show the effects of substorm currents. These maps are produced for several specific ranges of values for the peak | AL *| index, IMF orientation, and dipole tilt angle. We demonstrate an influence of the dipole tilt angle on the response to substorms. Our results indicate that there are downward currents poleward and upward currents just equatorward of the peak in the substorms' westward electrojet., Key Points: Show quantitative maps of ground geomagnetic perturbations due to substorms Three vector components mapped as function of time during onset and recovery Compare/contrast results for different tilt angle and sign of IMF Y-component.

We report far-ultraviolet observations of Jupiter's moon Europa taken by Space Telescope Imaging Spectrograph (STIS) of the Hubble Space Telescope (HST) in January and February 2014 to test the hypothesis that the discovery of a water vapor aurora in December 2012 by local hydrogen (H) and oxygen (O) emissions with the STIS originated from plume activity possibly correlated with Europa's distance from Jupiter through tidal stress variations. The 2014 observations were scheduled with Europa near the apocenter similar to the orbital position of its previous detection. Tensile stresses on south polar fractures are expected to be highest in this orbital phase, potentially maximizing the probability for plume activity. No local H and O emissions were detected in the new STIS images. In the south polar region where the emission surpluses were observed in 2012, the brightnesses are sufficiently low in the 2014 images to be consistent with any H2O abundance from (0-5)×10(15) cm(-2). Large high-latitude plumes should have been detectable by the STIS, independent of the observing conditions and geometry. Because electron excitation of water vapor remains the only viable explanation for the 2012 detection, the new observations indicate that although the same orbital position of Europa for plume activity may be a necessary condition, it is not a sufficient condition. However, the December 2012 detection of coincident HI Lyman-α and OI 1304-Å emission surpluses in an ∼200-km high region well separated above Europa's limb is a firm result and not invalidated by our 2014 STIS observations.

The substorm current wedge (SCW) is a fundamental component of geomagnetic substorms. Models tend to describe the SCW as a simple line current flowing into the ionosphere toward dawn and out of the ionosphere toward dusk, linked by a westward electrojet. We use multispacecraft observations from perigee passes of the Cluster 1 and 4 spacecraft during a substorm on 15 January 2010, in conjunction with ground-based observations, to examine the spatial structuring and temporal variability of the SCW. At this time, the spacecraft traveled east-west azimuthally above the auroral region. We show that the SCW has significant azimuthal substructure on scales of 100 km at altitudes of 4000-7000 km. We identify 26 individual current sheets in the Cluster 4 data and 34 individual current sheets in the Cluster 1 data, with Cluster 1 passing through the SCW 120-240 s after Cluster 4 at 1300-2000 km higher altitude. Both spacecraft observed large-scale regions of net upward and downward field-aligned current, consistent with the large-scale characteristics of the SCW, although sheets of oppositely directed currents were observed within both regions. We show that the majority of these current sheets were closely aligned to a north-south direction, in contrast to the expected east-west orientation of the preonset aurora. Comparing our results with observations of the field-aligned current associated with bursty bulk flows (BBFs), we conclude that significant questions remain for the explanation of SCW structuring by BBF-driven "wedgelets." Our results therefore represent constraints on future modeling and theoretical frameworks on the generation of the SCW., Key Points: The substorm current wedge (SCW) has significant azimuthal structureCurrent sheets within the SCW are north-south alignedThe substructure of the SCW raises questions for the proposed wedgelet scenario.

Mitosis has been studied since the early 1880s as a key event of the cell division cycle where remarkable changes in cellular architecture take place and ultimately lead to an equal segregation of duplicated chromosomes into two daughter cells. A detailed description of the complex and highly ordered cellular events taking place is now available. Many regulators involved in key steps including entry into mitosis, nuclear envelope breakdown, microtubule (MT) spindle formation, and chromosome attachment, as well as mitotic exit and cytokinesis, have also been identified. However, understanding the precise spatio-temporal contribution of each regulator in the cell reorganization process has been technically challenging. This review will focus on a number of recent advances in our understanding of the spatial distribution of protein activities and the temporal regulation of their activation and inactivation during entry and progression through mitosis by the use of intramolecular Förster resonance energy transfer (FRET)-based biosensors., (Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

During mitotic entry, the centrosomes provide a scaffold for initial activation of the CyclinB/Cdk1 complex, the mitotic kinase Aurora A, and the Aurora A-activating kinase p21-activated kinase (PAK). The activation of PAK at the centrosomes is yet regarded to happen independently of the Rho-GTPases Rac/Cdc42. In this study, Rac1 (but not RhoA or Cdc42) is presented to associate with the centrosomes from early G2 phase until prometaphase in a cell cycle-dependent fashion, as evidenced by western blot analysis of prepared centrosomes and by immunolabeling. PAK associates with the G2/M-phase centrosomes in a Rac1-dependent fashion. Furthermore, specific inhibition of Rac1 by C. difficile toxinB-catalyzed glucosylation or by knockout results in inhibited activation of PAK1/2, Aurora A, and the CyclinB/Cdk1 complex in late G2 phase/prophase and delayed mitotic entry. Inhibition of PAK activation at late G2-phase centrosomes caused by Rac1 inactivation coincides with impeded activation of Aurora A and the CyclinB/Cdk1 complex and delayed mitotic entry.

Animal cell division ends with the cutting of the microtubule and membrane intercellular bridge connecting the 2 daughter cells. This process, known as cytokinetic abscission (abscission), is widely regarded as the last step of cytokinesis, i.e., the last step of the cell cycle. Major breakthroughs have been recently achieved, illuminating mechanistic aspects of abscission; however, the timing of abscission with respect to the mammalian cell cycle remains unclear. In this study, we carefully measured the onset and progression of abscission in dividing cells expressing a G1 reporter. We conclude that abscission commences long after cells enter the G1 phase. Affiliating abscission with G1 is beyond semantics since it essentially postulates that the last step of the cell cycle is regulated in, and probably by, the following cycle.

Aurora kinases are key regulators of cell mitosis and have been implicated in the process of tumorigenesis. In recent years, the Aurora kinases have attracted much interest as promising targets for cancer treatment. Here we report on the roles of Aurora A and Aurora B kinases in clear cell renal cell carcinoma (ccRCC). Using genomewide expression array analysis of 174 patient samples of ccRCC, we found that expression levels of Aurora A and B were significantly elevated in ccRCC compared to normal kidney samples. High expression levels of Aurora A and Aurora B were significantly associated with advanced tumor stage and poor patient survival. Inhibition of Aurora kinase activity with the drug VX680 (also referred to as MK-0457) inhibited ccRCC cell growth in vitro and led to ccRCC cell accumulation in the G2/M phase and apoptosis. Growth of ccRCC xenograft tumors was also inhibited by VX680 treatment, accompanied by a reduction of tumor microvessel density. Analysis of endothelial cell lines demonstrated that VX680 inhibits endothelial cell growth with effects similar to that seen in ccRCC cells. Our findings suggest that VX680 inhibits the growth of ccRCC tumors by targeting the proliferation of both ccRCC tumor cells and tumor-associated endothelial cells. Aurora kinases and their downstream cell cycle proteins have an important role in ccRCC and may be potent prognostic markers and therapy targets for this disease.