Probabilistic Fluorescence-Based Synapse Detection

Brain function results from communication between neurons connected by complex synaptic networks. Synapses are themselves highly complex and diverse signaling machines, containing protein products of hundreds of different genes, some in hundreds of copies, arranged in precise lattice at each individual synapse. Synapses are fundamental not only to synaptic network function but also to network development, adaptation, and memory. In addition, abnormalities of synapse numbers or molecular components are implicated in most mental and neurological disorders. Despite their obvious importance, mammalian synapse populations have so far resisted detailed quantitative study. In human brains and most animal nervous systems, synapses are very small and very densely packed: there are approximately 1 billion synapses per cubic millimeter of human cortex. This volumetric density poses very substantial challenges to proteometric analysis at the critical level of the individual synapse. The present work describes new probabilistic image analysis methods for single-synapse analysis of synapse populations in both animal and human brains.Comment: Current awaiting peer revie

Preparing UK students for the workplace: The Acceptability of a Gamified Cybersecurity Training

This pilot study aims to assess the acceptability of Open University’s training platform called Gamified Intelligent Cyber Aptitude and Skills Training course (GICAST), as a means of improving cybersecurity knowledge, attitudes, and behaviours in undergraduate students using both quantitative and qualitative methods. A mixed-methods, pre-post experimental design was employed. 43 self-selected participants were recruited via an online register and posters at the university (excluding IT related courses). Participants completed the Human Aspects of Information Security Questionnaire (HAIS-Q) and Fear of Missing Out (FoMO) Scale. They then completed all games and quizzes in the GICAST course before repeating the HAIS-Q and FoMO scales as well as several open-ended questions. Pre-training HAIS-Q Knowledge, Attitude and Behaviour all improved from ‘reasonable’ pre-training levels to become ‘very high’ following training with large effect sizes estimated. FoMO improved to a lesser degree but also predicted the degree of HAIS-Q improvement suggesting it is relevant to the impact of this training course. Qualitatively, five key themes were generated: enjoyment, engagement, usability of GICAST, content relevance, and perceived educational efficacy. Overall, sentiment towards training was very positive as an enjoyable engaging and usable course. GICAST was found to be a feasible course for a wide range of students at a UK university: overall the training improved cyber-security awareness on a well validated measure with outcomes comparable to information-security-trained employees of a secure workplace. Despite a diversity of views about content, the course appears to be well suited to the non-IT undergraduate sector and may suit wide uptake to enhance students’ employability in a wide range of cybersecurity relevant contexts

Sawflies (Hymenoptera, Symphyta) Newly Recorded from Washington State

Examination of museum specimens, unpublished collection data, and field surveys conducted between 2010 and 2014 resulted in records for 22 species of sawflies new to Washington State, seven of which are likely to be pest problems in ornamental landscapes. These data highlight the continued range expansion of exotic species across North America. These new records also indicate that our collective knowledge of Pacific Northwest arthropod biodiversity and biogeography is underdeveloped, even for a relatively well known and species-poor group of insects. Notable gaps in the knowledge of Washington State’s Symphyta remain for the Olympic Peninsula, the Cascade Mountain Range, and the arid interior of the state. Washington’s shrub-steppe appears to be particularly poorly surveyed for sawflies

Sawflies (Hymenoptera, Symphyta) Newly Recorded from Washington State

Examination of museum specimens, unpublished collection data, and field surveys conducted between 2010 and 2014 resulted in records for 22 species of sawflies new to Washington State, seven of which are likely to be pest problems in ornamental landscapes. These data highlight the continued range expansion of exotic species across North America. These new records also indicate that our collective knowledge of Pacific Northwest arthropod biodiversity and biogeography is underdeveloped, even for a relatively well known and species-poor group of insects. Notable gaps in the knowledge of Washington State’s Symphyta remain for the Olympic Peninsula, the Cascade Mountain Range, and the arid interior of the state. Washington’s shrub-steppe appears to be particularly poorly surveyed for sawflies

Probabilistic fluorescence-based synapse detection

Deeper exploration of the brain’s vast synaptic networks will require new tools for high-throughput structural and molecular profiling of the diverse populations of synapses that compose those networks. Fluorescence microscopy (FM) and electron microscopy (EM) offer complementary advantages and disadvantages for single-synapse analysis. FM combines exquisite molecular discrimination capacities with high speed and low cost, but rigorous discrimination between synaptic and non-synaptic fluorescence signals is challenging. In contrast, EM remains the gold standard for reliable identification of a synapse, but offers only limited molecular discrimination and is slow and costly. To develop and test single-synapse image analysis methods, we have used datasets from conjugate array tomography (cAT), which provides voxel-conjugate FM and EM (annotated) images of the same individual synapses. We report a novel unsupervised probabilistic method for detection of synapses from multiplex FM (muxFM) image data, and evaluate this method both by comparison to EM gold standard annotated data and by examining its capacity to reproduce known important features of cortical synapse distributions. The proposed probabilistic model-based synapse detector accepts molecular-morphological synapse models as user queries, and delivers a volumetric map of the probability that each voxel represents part of a synapse. Taking human annotation of cAT EM data as ground truth, we show that our algorithm detects synapses from muxFM data alone as successfully as human annotators seeing only the muxFM data, and accurately reproduces known architectural features of cortical synapse distributions. This approach opens the door to data-driven discovery of new synapse types and their density. We suggest that our probabilistic synapse detector will also be useful for analysis of standard confocal and super-resolution FM images, where EM cross-validation is not practical

Rhodium(I) and Iridium(I) complexes of the conformationally rigid IBioxMe4Ligand : computational and experimental studies of unusually tilted NHC coordination geometries

Computational methods have been used to analyze distorted coordination geometries in a coherent range of known and new rhodium(I) and iridium(I) complexes containing bioxazoline-based NHC ligands (IBiox). Such distortions are readily placed in context of the literature through measurement of the Cnt(NHC)–CNCN–M angle (ΘNHC; Cnt = ring centroid). On the basis of restricted potential energy calculations using cis-[M(IBioxMe4)(CO)2Cl] (M1; M = Rh, Ir), in-plane (yawing) tilting of the NHC was found to incur significantly steeper energetic penalties than orthogonal out-of-plane (pitching) movement, which is characterized by noticeably flat potential energy surfaces. Energy decomposition analysis (EDA) of the ground-state and pitched structures of M1 indicated only minor differences in bonding characteristics. In contrast, yawing of the NHC ligand is associated with a significant increase in Pauli repulsion (i.e., sterics) and reduction in M→NHC π back donation, but is counteracted by supplemental stabilizing bonding interactions only possible due to the closer proximity of the methyl substituents with the metal and ancillary ligands. Aided by this analysis, comparison with a range of carefully selected model systems and EDA, distorted coordination modes in trans-[M(IBioxMe4)2(COE)Cl] (M2; M = Rh, Ir) and [M(IBioxMe4)3]+ (M3; M = Rh, Ir) have been rationalized. Steric interactions were identified as the major contributing factor and are associated with a high degree of NHC pitching. In the case of Rh3, weak agostic interactions also contribute to the distortions, particularly with respect to NHC yawing, and are notable for increasing the bond dissociation energy of the distorted ligands. Supplementing the computational analysis, an analogue of the formally 14 VE Rh(I) species Rh3 bearing the cyclohexyl-functionalized IBiox6 ligand ([Rh(IBiox6)3]+, Rh3-Cy) was prepared and found to exhibit an exceptionally distorted NHC ligand (ΘNHC = 155.7(2)°) in the solid state

Mapping Synapses by Conjugate Light-Electron Array Tomography

Synapses of the mammalian CNS are diverse in size, structure, molecular composition, and function. Synapses in their myriad variations are fundamental to neural circuit development, homeostasis, plasticity, and memory storage. Unfortunately, quantitative analysis and mapping of the brain's heterogeneous synapse populations has been limited by the lack of adequate single-synapse measurement methods. Electron microscopy (EM) is the definitive means to recognize and measure individual synaptic contacts, but EM has only limited abilities to measure the molecular composition of synapses. This report describes conjugate array tomography (AT), a volumetric imaging method that integrates immunofluorescence and EM imaging modalities in voxel-conjugate fashion. We illustrate the use of conjugate AT to advance the proteometric measurement of EM-validated single-synapse analysis in a study of mouse cortex