Wake-like skin patterning and neural activity during octopus sleep.

While sleeping, many vertebrate groups alternate between at least two sleep stages: rapid eye movement and slow wave sleep1-4, in part characterized by wake-like and synchronous brain activity, respectively. Here we delineate neural and behavioural correlates of two stages of sleep in octopuses, marine invertebrates that evolutionarily diverged from vertebrates roughly 550 million years ago (ref. 5) and have independently evolved large brains and behavioural sophistication. 'Quiet' sleep in octopuses is rhythmically interrupted by approximately 60-s bouts of pronounced body movements and rapid changes in skin patterning and texture6. We show that these bouts are homeostatically regulated, rapidly reversible and come with increased arousal threshold, representing a distinct 'active' sleep stage. Computational analysis of active sleep skin patterning reveals diverse dynamics through a set of patterns conserved across octopuses and strongly resembling those seen while awake. High-density electrophysiological recordings from the central brain reveal that the local field potential (LFP) activity during active sleep resembles that of waking. LFP activity differs across brain regions, with the strongest activity during active sleep seen in the superior frontal and vertical lobes, anatomically connected regions associated with learning and memory function7-10. During quiet sleep, these regions are relatively silent but generate LFP oscillations resembling mammalian sleep spindles11,12 in frequency and duration. The range of similarities with vertebrates indicates that aspects of two-stage sleep in octopuses may represent convergent features of complex cognition.

Drosophila CORL is required for Smad2-mediated activation of Ecdysone Receptor expression in the mushroom body.

CORL proteins (FUSSEL/SKOR proteins in humans) are related to Sno/Ski oncogenes but their developmental roles are unknown. We have cloned Drosophila CORL and show that its expression is restricted to distinct subsets of cells in the central nervous system. We generated a deletion of CORL and noted that homozygous individuals rarely survive to adulthood. Df(4)dCORL adult escapers display mushroom body (MB) defects and Df(4)dCORL larvae are lacking Ecdysone Receptor (EcR-B1) expression in MB neurons. This is phenocopied in CORL-RNAi and Smad2-RNAi clones in wild-type larvae. Furthermore, constitutively active Baboon (type I receptor upstream of Smad2) cannot stimulate EcR-B1 MB expression in Df(4)dCORL larvae, which demonstrates a formal requirement for CORL in Smad2 signaling. Studies of mouse Corl1 (Skor1) revealed that it binds specifically to Smad3. Overall, the data suggest that CORL facilitates Smad2 activity upstream of EcR-B1 in the MB. The conservation of neural expression and strong sequence homology of all CORL proteins suggests that this is a new family of Smad co-factors.

Role of Quenching Temperature Selection in the Improvement of the Abrasive (Al2O3) Wear Resistance of Hybrid Multi-Component Cast Irons.

In this paper, enhancing the tribological characteristics of novel cast metallic materials-hybrid multi-component cast irons-by applying a strengthening heat treatment is described. The experimental materials were the cast alloys of a nominal composition (5 wt.% W, 5 wt.% Mo, 5 wt.% V, 10 wt.% Cr, 2.5 wt.% Ti, Fe is a balance) supplemented with 0.3-1.1 wt.% C and 1.5-2.5 wt.% B (total of nine alloys). The heat treatment was oil-quenching followed by 200 °C tempering. The quench temperature (QT) varied in the range of 900-1200 °C, with a step of 50 °C (with a 2-h holding at QT). The correlation of the QT with microstructure and properties was estimated using microstructure/worn surface characterization, differential scanning calorimetry, hardness measurement, and three-body-abrasive wear testing (using Al2O3 particles). The as-cast alloys had a multi-phase structure consisting of primary and/or eutectic borocarbide M2(B,C)5, carboborides M(C,B), M7(C,B)3, M3(C,B), and the matrix (ferrite, martensite, pearlite/bainite) in different combinations and volume fractions. Generally, the increase in the quenching temperature resulted in a gradual increase in hardness (maximally to 66-67 HRC) and a decrease in the wear rate in most alloys. This was due to the change in the phase-structure state of the alloys under quenching, namely, the secondary carboboride precipitation, and replacing ferrite and pearlite/bainite with martensite. The wear rate was found to be inversely proportional to bulk hardness. The maximum wear resistance was attributed to QT = 1150-1200 °C, when the wear rate of the alloys was lowered by three to six times as compared to the as-cast state. With the QT increase, the difference in the wear rate of the alloys decreased by three times. The highest abrasive resistance was attributed to the alloys with 1.1 wt.% C, which had a 2.36-3.20 times lower wear rate as compared with that of the reference alloy (13 wt.% Cr cast iron, hardness of 66 HRC). The effects of carbon and boron on hardness and wear behavior are analyzed using the regression models developed according to the factorial design procedure. The wear mechanisms are discussed based on worn surface characterization.

A Study of the Three-Body Abrasive Wear Resistance of 5V/5Nb-5Cr-5Mo-5W-5Co-Fe Multicomponent Cast Alloys with Different Carbon Percentages.

Since it is well known in the literature that transition metals can form extremely hard carbides and effectively strengthen a material's matrix, recently, some of them, such as V, Nb, Cr, Mo, and W, have been simultaneously added to cast iron. In addition, it is common to add Co to cast iron to strengthen the material's matrix. However, the wear resistance of cast iron can also be considerably affected by the addition of C, which is rarely discussed in the literature by the experts. Therefore, the effect of C content (1.0; 1.5; 2.0 wt.%) on the abrasive wear behavior of 5 wt.% V/Nb, Cr, Mo, W, and Co alloys was investigated in this study. An evaluation was conducted using a rubber wheel abrasion testing machine in accordance with ASTM G65 with silica sand (1100 HV; 300 µm) as abrasive particles. The results show that plural carbides (MC, M2C, and M7C3) precipitated on the microstructure of the material, which is not unlike the behavior of other types of carbides as the quantity of C increases. The hardness and wear resistance properties of 5V-5Cr-5Mo-5W-5Co-Fe and 5Nb-5Cr-5Mo-5W-5Co-Fe multicomponent cast alloys increased as the quantity of C increased. However, we observed no significant difference in the hardness between the two materials with the same C additions, while 5Nb presented better wear resistance properties compared to the 5V sample due to the larger size of NbC compared to VC. Therefore, it can be determined that, in this study, the size of the carbide plays a more important role than its volume fraction and hardness.

Investigations of Abrasive Wear Behaviour of Hybrid High-Boron Multi-Component Alloys: Effect of Boron and Carbon Contents by the Factorial Design Method.

This paper is devoted to the evaluation of the "three-body-abrasion" wear behaviour of (wt.%) 5W-5Mo-5V-10Cr-2.5Ti-Fe (balance) multi-component (C + B)-added alloys in the as-cast condition. The carbon (0.3 wt.%, 0.7 wt.%, 1.1 wt.%) and boron (1.5 wt.%, 2.5 wt.%, 3.5 wt.%) contents were selected using a full factorial (32) design method. The alloys had a near-eutectic (at 1.5 wt.% B) or hyper-eutectic (at 2.5-3.5 wt.% B) structure. The structural micro-constituents were (in different combinations): (a) (W, Mo, and V)-rich borocarbide M2(B,C)5 as the coarse primary prismatoids or as the fibres of a "Chinese-script" eutectic, (b) Ti-rich carboboride M(C,B) with a dispersed equiaxed shape, (c) Cr-rich carboboride M7(C,B)3 as the plates of a "rosette"-like eutectic, and (d) Fe-rich boroncementite (M3(C,B)) as the plates of "coarse-net" and ledeburite eutectics. The metallic matrix was ferrite (at 0.3-1.1 wt.% C and 1.5 wt.% B) and "ferrite + pearlite" or martensite (at 0.7-1.1 wt.% C and 2.5-3.5 wt.% B). The bulk hardness varied from 29 HRC (0.3 wt.% C-1.5 wt.% B) to 53.5 HRC (1.1 wt.% C-3.5 wt.% B). The wear test results were mathematically processed and the regression equation of the wear rate as a function of the carbon and boron contents was derived and analysed. At any carbon content, the lowest wear rate was attributed to the alloy with 1.5 wt.% B. Adding 2.5 wt.% B led to an increase in the wear rate because of the appearance of coarse primary borocarbides (M2(B,C)5), which were prone to chipping and spalling-off under abrasion. At a higher boron content (3.5 wt.%), the wear rate decreased due to the increase in the volume fraction of the eutectic carboborides. The optimal chemical composition was found to be 1.1 wt.% C-1.5 wt.% B with a near-eutectic structure with about 35 vol.% of hard inclusions (M2(B,C)5, M(C,B), M3(C,B), and M7(C,B)3) in total. The effect of carbon and boron on the abrasive behaviour of the multi-component cast alloys with respect to the alloys' structure is discussed, and the mechanism of wear for these alloys is proposed.

The Wnt5/planar cell polarity pathway regulates axonal development of the Drosophila mushroom body neuron.

Axonal development is a fundamental process for circuit formation in the nervous system and is dependent on many cellular events, including axon initiation, elongation, guidance, and branching. The molecular mechanisms underlying these events have been well studied, especially for axon guidance. In the presence of a guidance cue, the polarization of a growth cone precedes the turning response, which is one example of the diverse forms of cell polarity. Planar cell polarity (PCP) is another example of cell polarity. Although some PCP genes are required for axonal tract formation in vertebrates, it remains elusive whether these genes participate in a common PCP pathway concertedly. Here, we show that essential PCP signaling components, encoded by frizzled (fz), strabismus (stbm), flamingo (fmi), and dishevelled (dsh), are cooperatively required for axonal targeting and branching of the Drosophila mushroom body (MB) neurons. A detailed analysis of these mutants revealed that these components were required for the correct targeting and bifurcation of axons. In addition, we suggest that Wnt5 functions as a ligand in the PCP pathway in this process. Wnt5 mutants showed similar phenotypes to PCP mutants at the single-cell level and genetically interacted with PCP genes. Wnt5 was broadly expressed in the developing brain. We propose that Wnt5 and the PCP pathway concertedly regulate axonal development of the MB.

Abrasive Wear of High-Carbon Low-Alloyed Austenite Steel: Microhardness, Microstructure and X-ray Characteristics of Worn Surface.

A high-carbon, high-silicon steel (1.21 wt% C, 2.56 wt% Mn, 1.59 wt% Si) was subjected to quenching from 900 and 1000 °C, resulting in microstructures containing 60 and 94% of retained austenite, respectively. Subsequent abrasive wear tests of quenched samples were performed using two-body abrasion and three-body abrasion testing machines. Investigations on worn surface and subsurface were carried out using SEM, XRD, and microhardness measurement. It was found that the highest microhardness of worn surface (about 1400 HV0.05) was achieved on samples quenched from 900 °C after three-body abrasion. Microhardness of samples after two-body abrasion was noticeably smaller. with a maximum of about 1200 HV0.05. This difference correlates with microstructure investigations along with XRD results. Three-body abrasion has produced a significantly deeper deformed layer; corresponding diffractograms show bigger values of the full width at half maximum parameter (FWHM) for both α and γ alone standing peaks. The obtained results are discussed in the light of possible differences in abrasive wear conditions and differing stability of retained austenite after quenching from different temperatures. It is shown that a structure of metastable austenite may be used as a detector for wear conditions, as the sensitivity of such austenite to phase transformation strongly depends on wear conditions, and even small changes in the latter lead to significant differences in the properties of the worn surface.

Evaluation of the Microstructure, Tribological Characteristics, and Crack Behavior of a Chromium Carbide Coating Fabricated on Gray Cast Iron by Pulsed-Plasma Deposition.

The structural and tribological properties of a protective high-chromium coating synthesized on gray cast iron by air pulse-plasma treatments were investigated. The coating was fabricated in an electrothermal axial plasma accelerator equipped with an expandable cathode made of white cast iron (2.3 wt.% C-27.4 wt.% Cr-3.1 wt.% Mn). Optical microscopy, scanning electron microscopy, energy-dispersive spectroscopy, X-ray diffraction analysis, microhardness measurements, and tribological tests were conducted for coating characterizations. It was found that after ten plasma pulses (under a discharge voltage of 4 kV) and post-plasma heat treatment (two hours of holding at 950 °C and oil-quenching), a coating (thickness = 210-250 µm) consisting of 48 vol.% Cr-rich carbides (M7C3, M3C), 48 vol.% martensite, and 4 vol.% retained austenite was formed. The microhardness of the coating ranged between 980 and 1180 HV. The above processes caused a gradient in alloying elements in the coating and the substrate due to the counter diffusion of C, Cr, and Mn atoms during post-plasma heat treatments and led to the formation of a transitional layer and different structural zones in near-surface layers of cast iron. As compared to gray cast iron (non-heat-treated and heat-treated), the coating had 3.0-3.2 times higher abrasive wear resistance and 1.2-1208.8 times higher dry-sliding wear resistance (depending on the counter-body material). The coating manifested a tendency of solidification cracking caused by tensile stress due to the formation of a mostly austenitic structure with a lower specific volume. Cracks facilitated abrasive wear and promoted surface spalling under dry-sliding against the diamond cone.

Corrigendum: A Population of Projection Neurons that Inhibits the Lateral Horn but Excites the Antennal Lobe through Chemical Synapses in Drosophila.

[This corrects the article on p. 30 in vol. 11, PMID: 28515683.].

Two Parallel Pathways Assign Opposing Odor Valences during Drosophila Memory Formation.

During olfactory associative learning in Drosophila, odors activate specific subsets of intrinsic mushroom body (MB) neurons. Coincident exposure to either rewards or punishments is thought to activate extrinsic dopaminergic neurons, which modulate synaptic connections between odor-encoding MB neurons and MB output neurons to alter behaviors. However, here we identify two classes of intrinsic MB γ neurons based on cAMP response element (CRE)-dependent expression, γCRE-p and γCRE-n, which encode aversive and appetitive valences. γCRE-p and γCRE-n neurons act antagonistically to maintain neutral valences for neutral odors. Activation or inhibition of either cell type upsets this balance, toggling odor preferences to either positive or negative values. The mushroom body output neurons, MBON-γ5ß'2a/ß'2mp and MBON-γ2α'1, mediate the actions of γCRE-p and γCRE-n neurons. Our data indicate that MB neurons encode valence information, as well as odor information, and this information is integrated through a process involving MBONs to regulate learning and memory.

A Population of Projection Neurons that Inhibits the Lateral Horn but Excites the Antennal Lobe through Chemical Synapses in Drosophila.

In the insect olfactory system, odor information is transferred from the antennal lobe (AL) to higher brain areas by projection neurons (PNs) in multiple AL tracts (ALTs). In several species, one of the ALTs, the mediolateral ALT (mlALT), contains some GABAergic PNs; in the Drosophila brain, the great majority of ventral PNs (vPNs) are GABAergic and project through this tract to the lateral horn (LH). Most excitatory PNs (ePNs), project through the medial ALT (mALT) to the mushroom body (MB) and the LH. Recent studies have shown that GABAergic vPNs play inhibitory roles at their axon terminals in the LH. However, little is known about the properties and functions of vPNs at their dendritic branches in the AL. Here, we used optogenetic and patch clamp techniques to investigate the functional roles of vPNs in the AL. Surprisingly, our results show that specific activation of vPNs reliably elicits strong excitatory postsynaptic potentials (EPSPs) in ePNs. Moreover, the connections between vPNs and ePNs are mediated by direct chemical synapses. Neither pulses of GABA, nor pharmagological, or genetic blockade of GABAergic transmission gave results consistent with the involvement of GABA in vPN-ePN excitatory transmission. These unexpected results suggest new roles for the vPN population in olfactory information processing.

Gap junctions.

In vertebrates and invertebrates, signaling among neurons is most commonly mediated by chemical synapses. At these synapses neurotransmitter released by presynaptic neurons is detected by receptors on the postsynaptic neurons, leading to an influx of ions through the receptors themselves or through channels activated by intracellular signaling downstream of the receptors. But neurons can communicate with each other in a more direct way, by passing signals composed of small molecules and ions through pores called gap junctions. Gap junctions that transmit electrical signals are called electrical synapses. Unlike most chemical synapses, electrical synapses interact through axon-to-axon or dendrite-to-dendrite contacts. Found throughout the nervous system, they are probably best known for linking the relatively few inhibitory, GABAergic, neurons into large, effective networks within vertebrate brains. They are particularly important early in development before the formation of most chemical synapses, but recent work shows gap junctions play important roles in the adult nervous system, too. Gap junctions are sometimes thought to be mere passageways between cells. But, as recent work shows, their properties can be complex and surprising. Gap junctions help generate, propagate, and regulate neural oscillations, can filter electrical signals, and can be modulated in a variety of ways. Here we discuss recent work highlighting the diversity and importance of gap junctions throughout the nervous system.

Antigen-receptor genes of the agnathan lamprey are assembled by a process involving copy choice.

Jawless vertebrates have acquired immunity but do not have immunoglobulin-type antigen receptors. Variable lymphocyte receptors (VLRs) have been identified in lamprey that consist of multiple leucine-rich repeat (LRR) modules. An active VLR gene is generated by the assembly of a series of variable gene segments, including many that encode LRRs. Stepwise assembly of the gene segments seems to occur by replacement of the intervening DNA between the 5' and 3' constant-region genes. Here we report that lamprey (Lethenteron japonicum) assemble their VLR genes by a process involving 'copy choice'. Regions of short homology seemed to prime copying of donor LRR-encoding sequences into the recipient gene. Those LRR-encoding germline sequences were abundant and shared extensive sequence homologies. Such genomic organization permits initiation of copying anywhere in an LRR-encoding module for the generation of various hybrid LRRs. Thus, a vast repertoire of recombinant VLR genes could be generated not only by copying of various LRR segments in diverse combinations but also by the use of multiple sites in an LRR gene segment for priming.