Desmosomal connectomics of all somatic muscles in an annelid larva.

Cells form networks in animal tissues through synaptic, chemical, and adhesive links. Invertebrate muscle cells often connect to other cells through desmosomes, adhesive junctions anchored by intermediate filaments. To study desmosomal networks, we skeletonised 853 muscle cells and their desmosomal partners in volume electron microscopy data covering an entire larva of the annelid Platynereis. Muscle cells adhere to each other, to epithelial, glial, ciliated, and bristle-producing cells and to the basal lamina, forming a desmosomal connectome of over 2000 cells. The aciculae - chitin rods that form an endoskeleton in the segmental appendages - are highly connected hubs in this network. This agrees with the many degrees of freedom of their movement, as revealed by video microscopy. Mapping motoneuron synapses to the desmosomal connectome allowed us to infer the extent of tissue influenced by motoneurons. Our work shows how cellular-level maps of synaptic and adherent force networks can elucidate body mechanics.

Effect of warm/cool white lights on visual perception and mood in warm/cool color environments.

Color and light are two ambient attributes for interior spaces that can be used in the design and modification of workspaces. The visual and psychological effects of color and light of each have been studied separately and widely. The aim of this study was to investigate the simultaneous effects of warm/cool white light on visual perception and mood in a simulated colored workspace. Thirty-three healthy male participants were recruited. They were asked to judge the visual perception and mood of three types of workspace that were designed by colors of white, red, blue, and lights of a cool and warm white in the random six sessions. The participants have experienced higher levels of tension, anger, depression, anxiety and lower levels of visual comfort, attractiveness, brightness and calmness of environment in the red condition than to white in both state of light. The blue wall reduced brightness and increased attractiveness of environment compared to white wall. Cool white light reduced the warmth of color and increased brightness in all three color environments compared to warm light. The preference for cool or warm light depends on the color of the environment's indoor surface. It seems that the combination of the white color and warm light or the blue color with cool light has a more favorable effect on visual perception and people's mood in workplaces.

Explanation and prediction of accidents using the path analysis approach in industrial units: The effect of safety performance and climate.

BACKGROUND: The safety climate in an organization depends on people's understanding of the safety policies and procedures, as well as the value, importance, and priority of safety in the workplace. OBJECTIVE: This study aimed to describe and predict accidents using the path analysis model (PAM) in industrial units though the analysis of the effect of safety performance and climate. METHODS: This cross-sectional study was conducted on 294 workers in industrial units in Hamadan, a province in the western part of Iran. The data on safety performance and climate was collected using a questionnaire. The first part of the questionnaire was a short version inventory (with 25 items on safety climate) that was used to assess five variables of management commitment, supportive environment, training, personal safety prioritization, and perceived work pressure. Moreover, the safety performance was measured using 10 items on safety rules and participation. The PAM was used to describe the effects of safety climate and performance on accidents. RESULTS: The results showed that the safety climate had the strongest negative impact on work pressure and safety compliance toward accident, followed by safety participation, and quality of training. Moreover, the negative influence of safety climate on accident was mainly mediated by two variables: work pressure and safety participation toward accident. The work pressure had the strongest indirect and total influence on accidents. However, none of the variables had a direct effect on accidents. Training was the most important direct cause of promoting personal safety priority. The safety compliance was more effective than safety participation in reducing accidents rates. CONCLUSIONS: Therefore, it seems that perceived work pressure has an indirect effect on accidents which is mediated by other variables, mainly personal safety priority and safety performance.

Content-aware image restoration for electron microscopy.

Multiple approaches to use deep neural networks for image restoration have recently been proposed. Training such networks requires well registered pairs of high and low-quality images. While this is easily achievable for many imaging modalities, e.g., fluorescence light microscopy, for others it is not. Here we summarize on a number of recent developments in the fast-paced field of Content-Aware Image Restoration (CARE), in particular, and the associated area of neural network training, more in general. We then give specific examples how electron microscopy data can benefit from these new technologies.

The design and application of a bacterial ghost vaccine to evaluate immune response and defense against avian pathogenic Escherichia coli O2:K1 serotype.

An Escherichia coli (E. coli) O2:K1 bacterial ghost was produced by controlled expression of bacteriophage PhiX 174 lysis gene E. Temperature controlled expression of this gene caused tunnels and holes in the cell wall of E. coli O2:K1 bacterium, leading to loss of cytoplasmic contents. Formation of E. coli O2:K1 ghost was confirmed by scanning electron microscopy and determination of colony forming units. To evaluate the efficiency of this bacterial ghost vaccine to elicit cellular and humoral immune responses, 85 one day old chickens from Ross 308 breed were divided into the following 5 groups; group 1 (non-immunized control), group 2 (vaccine administered by injection of E. coli O2:K1 killed vaccine), group 3 (vaccine administered by injection of E. coli O2:K1 ghost), group 4 (vaccine administered by inhalation of E. coli O2:K1 ghost), and group 5 (neither immunized, nor challenged as negative control). The groups of 2, 3, and 4 were received vaccines at days 7, 14, and 22. Groups 1 to 4 were challenged with the wild type at day 33. Evaluation of post-mortem lesions and immune responses in all groups showed that chicken injected with the killed vaccine and the bacterial ghost had the best protection. These findings suggest that this bacterial ghost has the potential to be used as a poultry colibacillosis vaccine.

Neural circuitry of a polycystin-mediated hydrodynamic startle response for predator avoidance.

Startle responses triggered by aversive stimuli including predators are widespread across animals. These coordinated whole-body actions require the rapid and simultaneous activation of a large number of muscles. Here we study a startle response in a planktonic larva to understand the whole-body circuit implementation of the behaviour. Upon encountering water vibrations, larvae of the annelid Platynereis close their locomotor cilia and simultaneously raise the parapodia. The response is mediated by collar receptor neurons expressing the polycystins PKD1-1 and PKD2-1. CRISPR-generated PKD1-1 and PKD2-1 mutant larvae do not startle and fall prey to a copepod predator at a higher rate. Reconstruction of the whole-body connectome of the collar-receptor-cell circuitry revealed converging feedforward circuits to the ciliary bands and muscles. The wiring diagram suggests circuit mechanisms for the intersegmental and left-right coordination of the response. Our results reveal how polycystin-mediated mechanosensation can trigger a coordinated whole-body effector response involved in predator avoidance.

Ciliary and rhabdomeric photoreceptor-cell circuits form a spectral depth gauge in marine zooplankton.

Ciliary and rhabdomeric photoreceptor cells represent two main lines of photoreceptor-cell evolution in animals. The two cell types coexist in some animals, however how these cells functionally integrate is unknown. We used connectomics to map synaptic paths between ciliary and rhabdomeric photoreceptors in the planktonic larva of the annelid Platynereis and found that ciliary photoreceptors are presynaptic to the rhabdomeric circuit. The behaviors mediated by the ciliary and rhabdomeric cells also interact hierarchically. The ciliary photoreceptors are UV-sensitive and mediate downward swimming in non-directional UV light, a behavior absent in ciliary-opsin knockout larvae. UV avoidance overrides positive phototaxis mediated by the rhabdomeric eyes such that vertical swimming direction is determined by the ratio of blue/UV light. Since this ratio increases with depth, Platynereis larvae may use it as a depth gauge during vertical migration. Our results revealed a functional integration of ciliary and rhabdomeric photoreceptor cells in a zooplankton larva.

Synaptic and peptidergic connectome of a neurosecretory center in the annelid brain.

Neurosecretory centers in animal brains use peptidergic signaling to influence physiology and behavior. Understanding neurosecretory center function requires mapping cell types, synapses, and peptidergic networks. Here we use transmission electron microscopy and gene expression mapping to analyze the synaptic and peptidergic connectome of an entire neurosecretory center. We reconstructed 78 neurosecretory neurons and mapped their synaptic connectivity in the brain of larval Platynereis dumerilii, a marine annelid. These neurons form an anterior neurosecretory center expressing many neuropeptides, including hypothalamic peptide orthologs and their receptors. Analysis of peptide-receptor pairs in spatially mapped single-cell transcriptome data revealed sparsely connected networks linking specific neuronal subsets. We experimentally analyzed one peptide-receptor pair and found that a neuropeptide can couple neurosecretory and synaptic brain signaling. Our study uncovered extensive networks of peptidergic signaling within a neurosecretory center and its connection to the synaptic brain.

Ciliomotor circuitry underlying whole-body coordination of ciliary activity in the Platynereis larva.

Ciliated surfaces harbouring synchronously beating cilia can generate fluid flow or drive locomotion. In ciliary swimmers, ciliary beating, arrests, and changes in beat frequency are often coordinated across extended or discontinuous surfaces. To understand how such coordination is achieved, we studied the ciliated larvae of Platynereis dumerilii, a marine annelid. Platynereis larvae have segmental multiciliated cells that regularly display spontaneous coordinated ciliary arrests. We used whole-body connectomics, activity imaging, transgenesis, and neuron ablation to characterize the ciliomotor circuitry. We identified cholinergic, serotonergic, and catecholaminergic ciliomotor neurons. The synchronous rhythmic activation of cholinergic cells drives the coordinated arrests of all cilia. The serotonergic cells are active when cilia are beating. Serotonin inhibits the cholinergic rhythm, and increases ciliary beat frequency. Based on their connectivity and alternating activity, the catecholaminergic cells may generate the rhythm. The ciliomotor circuitry thus constitutes a stop-and-go pacemaker system for the whole-body coordination of ciliary locomotion.

Introducing oriented Laplacian diffusion into a variational decomposition model.

The decomposition model proposed by Osher, Solé and Vese in 2003 (the OSV model) is known for its good denoising performance. This performance has been found to be due to its higher weighting of lower image frequencies in the H -1-norm modeling the noise component in the model. However, the OSV model tends to also move high-frequency texture into this noise component. Diffusion with an oriented Laplacian for oriented texture is introduced in this paper, in lieu of the usual Laplacian operator used to solve the OSV model, thereby significantly reducing the presence of such texture in the noise component. Results obtained from the proposed oriented Laplacian model for test images with oriented texture are given, and compared to those from the OSV model as well as the Mean Curvature model (MCM). In general, the proposed oriented Laplacian model yields higher signal-to-noise ratios and visually superior denoising results than either the OSV or the MCM models. We also compare the proposed method to a non-local means model and find that although the proposed method generally yields slightly lower signal-to-noise ratios, it generally gives results of better perceptual visual quality.

A serial multiplex immunogold labeling method for identifying peptidergic neurons in connectomes.

Electron microscopy-based connectomics aims to comprehensively map synaptic connections in neural tissue. However, current approaches are limited in their capacity to directly assign molecular identities to neurons. Here, we use serial multiplex immunogold labeling (siGOLD) and serial-section transmission electron microscopy (ssTEM) to identify multiple peptidergic neurons in a connectome. The high immunogenicity of neuropeptides and their broad distribution along axons, allowed us to identify distinct neurons by immunolabeling small subsets of sections within larger series. We demonstrate the scalability of siGOLD by using 11 neuropeptide antibodies on a full-body larval ssTEM dataset of the annelid Platynereis. We also reconstruct a peptidergic circuitry comprising the sensory nuchal organs, found by siGOLD to express pigment-dispersing factor, a circadian neuropeptide. Our approach enables the direct overlaying of chemical neuromodulatory maps onto synaptic connectomic maps in the study of nervous systems.

Inter-individual stereotypy of the Platynereis larval visual connectome.

Developmental programs have the fidelity to form neural circuits with the same structure and function among individuals of the same species. It is less well understood, however, to what extent entire neural circuits of different individuals are similar. Previously, we reported the neuronal connectome of the visual eye circuit from the head of a Platynereis dumerilii larva (Randel et al., 2014). We now report a full-body serial section transmission electron microscopy (ssTEM) dataset of another larva of the same age, for which we describe the connectome of the visual eyes and the larval eyespots. Anatomical comparisons and quantitative analyses of the two circuits reveal a high inter-individual stereotypy of the cell complement, neuronal projections, and synaptic connectivity, including the left-right asymmetry in the connectivity of some neurons. Our work shows the extent to which the eye circuitry in Platynereis larvae is hard-wired.

Neuronal connectome of a sensory-motor circuit for visual navigation.

Animals use spatial differences in environmental light levels for visual navigation; however, how light inputs are translated into coordinated motor outputs remains poorly understood. Here we reconstruct the neuronal connectome of a four-eye visual circuit in the larva of the annelid Platynereis using serial-section transmission electron microscopy. In this 71-neuron circuit, photoreceptors connect via three layers of interneurons to motorneurons, which innervate trunk muscles. By combining eye ablations with behavioral experiments, we show that the circuit compares light on either side of the body and stimulates body bending upon left-right light imbalance during visual phototaxis. We also identified an interneuron motif that enhances sensitivity to different light intensity contrasts. The Platynereis eye circuit has the hallmarks of a visual system, including spatial light detection and contrast modulation, illustrating how image-forming eyes may have evolved via intermediate stages contrasting only a light and a dark field during a simple visual task.

Study on the performance of wet electroscrubber in purifying airborne particles.

BACKGROUND: In this study, an electroscrubber was designed and experimented for evaluation of integrating particle and droplet charging effects separately and jointly on collection efficiency of a spray tower and also to discover the optimal condition. METHODS: A homogenous concentration of relatively fine particles was introduced to influent air stream and electroscrubber efficiency in purifying them was determined through the measurement of input and output particles concentration. The effect of various conditions such as particles and droplets charging alone and together (bipolar) for several applied voltages has been studied. RESULTS: In all of experiments, the applied charging voltage has a key role in promotion of electroscrubber efficiency. Maximum collection efficiency has achieved for 15 Kilovolt (Kv). The effectiveness of bipolar charging of particles and droplets with 15 Kv was higher than that of no-charging and singly charging. In other words, efficiency can be increased from 84.43% to 93.22 for total particles and from 50.8% to 75.16% for submicron particles. The maximum improvement of collection efficiency (42.2%) relates to bipolar charging of the initial size group with diameter smaller than 0.3 micrometer (µm) and the minimum (0.5%) to sizing group of 11 with diameter 4-5 µm. CONCLUSIONS: This approach can be an appropriate option for the purpose of purifying submicron particles in spray tower scrubbers.

Expression dynamics and protein localization of rhabdomeric opsins in Platynereis larvae.

The larval stages of polychaete annelids are often responsive to light and can possess one to six eyes. The early trochophore larvae of the errant annelid Platynereis dumerilii have a single pair of ventral eyespots, whereas older nectochaete larvae have an additional two pairs of dorsal eyes that will develop into the adult eyes. Early Platynereis trochophores show robust positive phototaxis starting on the first day of development. Even though the mechanism of phototaxis in Platynereis early trochophore larvae is well understood, no photopigment (opsin) expression has yet been described in this stage. In late trochophore larvae, a rhabdomeric-type opsin, r-opsin1, expressed in both the eyespots and the adult eyes has already been reported. Here, we identify another Platynereis rhabdomeric opsin, r-opsin3, that is expressed in a single photoreceptor in the eyespots in early trochophores, suggesting that it mediates early larval phototaxis. We also show that r-opsin1 and r-opsin3 are expressed in adjacent photoreceptor cells in the eyespots in later stages, indicating that a second eyespot-photoreceptor differentiates in late trochophore larvae. Using serial transmission electron microscopy (TEM), we identified and reconstructed both photoreceptors and a pigment cell in the late larval eyespot. We also characterized opsin expression in the adult eyes and found that the two opsins co-express there in several photoreceptor cells. Using antibodies recognizing r-opsin1 and r-opsin3 proteins, we demonstrate that both opsins localize to the rhabdomere in all six eyes. In addition, we found that r-opsin1 mRNA is localized to, and translated in, the projections of the adult eyes. The specific changes we describe in opsin transcription and translation and in the cellular complement suggest that the six larval eyes undergo spectral and functional maturation during the early planktonic phase of the Platynereis life cycle.

Conserved MIP receptor-ligand pair regulates Platynereis larval settlement.

Life-cycle transitions connecting larval and juvenile stages in metazoans are orchestrated by neuroendocrine signals including neuropeptides and hormones. In marine invertebrate life cycles, which often consist of planktonic larval and benthic adult stages, settlement of the free-swimming larva to the sea floor in response to environmental cues is a key life cycle transition. Settlement is regulated by a specialized sensory-neurosecretory system, the larval apical organ. The neuroendocrine mechanisms through which the apical organ transduces environmental cues into behavioral responses during settlement are not yet understood. Here we show that myoinhibitory peptide (MIP)/allatostatin-B, a pleiotropic neuropeptide widespread among protostomes, regulates larval settlement in the marine annelid Platynereis dumerilii. MIP is expressed in chemosensory-neurosecretory cells in the annelid larval apical organ and signals to its receptor, an orthologue of the Drosophila sex peptide receptor, expressed in neighboring apical organ cells. We demonstrate by morpholino-mediated knockdown that MIP signals via this receptor to trigger settlement. These results reveal a role for a conserved MIP receptor-ligand pair in regulating marine annelid settlement.

Decorrelating the structure and texture components of a variational decomposition model.

The observation has been made by Aujol and Gilboa that the cartoon and texture components of the decomposition of an image should not be correlated, as they are generated from independent processes. They use this observation in order to choose an optimal fidelity parameter lambda for the decomposition process. However, this determination can be quite inefficient since a wide range of parameters lambda must be searched through before an estimated optimal parameter can be found. In the present paper, we take a different approach, in which the cartoon and texture components are explicitly decorrelated by adding a decorrelation term to the energy functional of the decomposition model of Osher, SolE, and Vese (the OSV model). Decomposition results of improved quality over those from the OSV model are obtained, as quantified by a series of new decomposition quality measures, with cartoon and texture information better separated into their respective components. A new derivation of the OSV model is developed which maintains the texture subcomponents g(1) and g(2) so that discrimination results similar to those from other decomposition models (e.g., from the model of Vese and Osher and Improved Edge Segregation) may be obtained. This derivation is extended to the proposed model, for which discrimination results are obtained in a substantially smaller number of iterations.

Notch activation results in phenotypic and functional changes consistent with endothelial-to-mesenchymal transformation.

Various studies have identified a critical role for Notch signaling in cardiovascular development. In this and other systems, Notch receptors and ligands are expressed in regions that undergo epithelial-to-mesenchymal transformation. However, there is no direct evidence that Notch activation can induce mesenchymal transdifferentiation. In this study we show that Notch activation in endothelial cells results in morphological, phenotypic, and functional changes consistent with mesenchymal transformation. These changes include downregulation of endothelial markers (vascular endothelial [VE]-cadherin, Tie1, Tie2, platelet-endothelial cell adhesion molecule-1, and endothelial NO synthase), upregulation of mesenchymal markers (alpha-smooth muscle actin, fibronectin, and platelet-derived growth factor receptors), and migration toward platelet-derived growth factor-BB. Notch-induced endothelial-to-mesenchymal transformation does not seem to require external regulation and is restricted to cells expressing activated Notch. Jagged1 stimulation of endothelial cells induces a similar mesenchymal transformation, and Jagged1, Notch1, and Notch4 are expressed in the ventricular outflow tract during stages of endocardial cushion formation. This is the first evidence that Jagged1-Notch interactions induce endothelial-to-mesenchymal transformation, and our findings suggest that Notch signaling may be required for proper endocardial cushion differentiation and/or vascular smooth muscle cell development.