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Cholinergic signaling is critical for an individual to react appropriately and adaptably to salient stimuli while navigating a complex environment. The cholinergic neurotransmitter system drives attention to salient stimuli, such as stressors, and aids in orchestrating the proper neural and behavioral response. Fine-tuned regulation of the cholinergic system has been linked to appropriate stress responses and subsequent mood regulation while dysregulation has been implicated in mood disorders. Among the multiple layers of regulation are cholinergic protein modulators. Here, we use validated models of experiential-based affective disorders to investigate differences in responses to stress in a genetic mouse model of cholinergic dysregulation based on the loss of protein modulator. The lynx2 nicotinic receptor modulatory protein provides negative cholinergic regulation within the amygdala, medial prefrontal cortex, and other brain regions. We discovered here that lynx2 knockout (KO) mice demonstrate an inability to update behavior with an inability to extinguish learned fear during a fear extinction test. We also observed, under an increased stress load following exposure to chronic social defeat stress (CSDS) paradigm, there was a unified resilience phenotype in lynx2KO mice, as opposed to the wild-type cohort which was split between resilience and susceptible phenotypes. Furthermore, we provide evidence for the functional role of α7 nicotinic receptor subtypes by phenotypic rescue with MLA or crossing with an α7 null mutant mouse (e.g. lynx2/α7 double KO mice). We demonstrate a direct physical interaction between lynx2 and α7 nAChR by co-immunoprecipitation of complexes from mouse BLA extracts. The genetic predisposition to heightened basal anxiety-like behavior and altered cholinergic signaling impairs individual behavior responses stressors. Together, these data indicate that the effects of social stress can be influenced by baseline genetic factors involved in anxiety regulation.

Nicotinic acetylcholine receptors (nAChRs) are broadly expressed in the central and peripheral nervous systems, playing essential roles in cholinergic neurotransmission. The lynx family proteins, a subset of the Ly6/uPAR superfamily expressed in multiple brain regions, have been shown to bind to nAChRs and modulate their function via allosteric regulation. The binding interactions between lynx and nAChRs, however, have not been systematically quantified and compared. In this work, we characterized the interactions between lynx1 or lynx2 and α3β4‐ or α7‐nAChRs using single‐molecule atomic force microscopy (AFM). The AFM technique allows the quantification of the off‐rate of lynx‐nAChR binding and of the energetic barrier width between the bound state and transition state, providing a biophysical means to compare the selectivity of lynx proteins for nAChR subtypes. Results indicate that lynx1 has a marginal preference for α7‐ over α3β4‐nAChRs. Strikingly, lynx2 exhibits a two order of magnitude stronger affinity for α3β4‐ compared to α7‐nAChRs. Together, the AFM assay serves as a valuable tool for the biophysical characterization of lynx‐nAChR binding affinities. Revealing the differential affinities of lynx proteins for nAChR subtypes will help elucidate how lynx regulates nAChR‐dependent functions in the brain, including nicotine addiction and other critical pathways.

The present paper investigates a portable eustachian-tube-function testing device by sonotubometry based on pure-tone sound transmission via the eustachian tube (ET). The measured results obtained by the proposed method were validated through comparison with the existing testing technique based on broadband sound inspection. The measurement results for the ET opening time (Topen) and the sound pressure difference in the ear canal between open and closed ETs (ΔL) obtained using pure-tone sounds with tonal frequency components of 7.0 and 9.5 kHz generally agreed with the results obtained by the existing technique with broadband testing sound.

The status of a project to measure spin-isospin responses of neutron drip-line nuclei using a new low-energy neutron detector, PANDORA (Particle Analyzer Neutron Detector Of Real-time Acquisition), is reported. The performance of PANDORA was characterized by the 6He(p,n)6Li reaction in inverse kinematics at the HIMAC facility in Chiba. Observation of the strong transition to the ground state in 6Li is discussed. Preliminary results of 11Li(p,n)11Be and 14Be(p,n)14B experiments in inverse kinematics at RI Beam Factory (RIBF) of RIKEN Nishina Center are also presented including the exotic decay channel of 11Be→9Li + d. Details of the experimental setup based on PANDORA and the SAMURAI large-acceptance magnetic spectrometer, as well as the combined data-acquisition system are described. The neutron-gamma discrimination capability of PANDORA was evaluated, Figure-of-Merit (FoM) values higher than those found in the literature for similar materials were derived from experimental data.

Many tools that explore models of protein complexes are also able to analyze interactions between specific residues and atoms. A comprehensive exploration of these interactions can often uncover aspects of protein-protein recognition that are not obvious using other protein analysis techniques. This paper describes DiffBond, a novel method for searching for intermolecular interactions between protein complexes while differentiating between three different types of interaction: hydrogen bonds, ionic bonds, and salt bridges. DiffBond incorporates textbook definitions of these three interactions while contending with uncertainties that are inherent in computational models of interacting proteins. We used it to examine the barnase-barstar, Rap1a-raf, and Smad2-Smad4 complexes, as well as a subset of protein complexes formed between three-finger toxins and nAChRs. Based on electrostatic interactions established by previous experimental studies, DiffBond was able to identify ionic and hydrogen bonds with high precision and recall, and identify salt bridges with high precision. In combination with other electrostatic analysis methods, DiffBond can be a useful tool in helping predict influential amino acids in protein-protein interactions and characterizing the type of interaction.

Negative allosteric modulators, such as lynx1 and lynx2, directly interact with nicotinic acetylcholine receptors (nAChRs). The nAChRs are integral to cholinergic signaling in the brain and have been shown to mediate different aspects of cognitive function. Given the interaction between lynx proteins and these receptors, we examined whether these endogenous negative allosteric modulators are involved in cognitive behaviors associated with cholinergic function. We found both cell-specific and overlapping expression patterns of lynx1 and lynx2 mRNA in brain regions associated with cognition, learning, memory, and sensorimotor processing, including the prefrontal cortex (PFC), cingulate cortex, septum, hippocampus, amygdala, striatum, and pontine nuclei. Since lynx proteins are thought to play a role in conditioned associations and given the expression patterns across brain regions, we first assessed whether lynx knockout mice would differ in a cognitive flexibility task. We found no deficits in reversal learning in either the lynx1–/– or lynx2–/– knockout mice. Thereafter, sensorimotor gating was examined with the prepulse inhibition (PPI) assessment. Interestingly, we found that both male and female lynx1–/– mice exhibited a deficit in the PPI behavioral response. Given the comparable expression of lynx2 in regions involved in sensorimotor gating, we then examined whether removal of the lynx2 protein would lead to similar behavioral effects. Unexpectedly, we found that while male lynx2–/– mice exhibited a decrease in the baseline startle response, no differences were found in sensorimotor gating for either male or female lynx2–/– mice. Taken together, these studies provide insight into the expression patterns of lynx1 and lynx2 across multiple brain regions and illustrate the modulatory effects of the lynx1 protein in sensorimotor gating.

To extract information on elemental components and PM characteristics from smoke number (SN) filter paper, we were performed elemental analysis by micro-PIXE. As a result of elemental analysis of the SN filter, information on the elemental composition of jet engine exhaust PM was obtained. Regarding the SN filter, jet engine exhaust PM have been collected for each difference in engine thrust of various aircraft engines at aircraft turbine engines test cells around the world. By periodically analyzing the elemental composition of such the SN filter, it is possible to grasp the characteristics of the emission PM of each aircraft engine and the transformation of the elemental composition of the emission PM with the passage of time of the jet engine. This was thought to lead to the evaluation of jet engine.

The availability of pollen in urban-rural landscapes is an essential factor that influences the population dynamics of insect pollinators. The amount and diversity of pollen play a pivotal role in the foraging ecology of pollinators for their growth and health, but investigations on the spatio-temporal patterns of foraged plants remain rare, especially in cities as neo-ecosystems. Here, we explored the temporal foraging habits of a highly polylectic pollinator (Apis melliferaL.) in a study area, including different landscape classes from rural to urban areas. Mixed-pollen in each month and each location (N = 17) were analysed using DNA meta-barcoding to identify plants visited by honeybees. The results showed that the landscape class (rural, suburban and urban areas) explains spatial variations in the plant composition foraged by honeybees, but not in taxa richness. Furthermore, pollen diversity and plant composition showed a strong seasonal dependence. Furthermore, a higher plant richness and foraged woody taxa was found to occur in spring, which was mainly dominated by the generaPrunusandAcer. In summer and autumn, the generaTrifoliumandPlantagoof the herbaceous stratum were the most visited plants. The Fabaceae, Rosaceae, Brassicaceae, Plantaginaceae, and Onagraceae plant families were the most frequently observed in all combined samples. The present study contributes to a broader understanding of the ecology and floral preferences of honeybees, on which urban planning can rely to promote biodiversity in cities.

The tetraneutron has attracted the attention of nuclear physicists during the past decades, but there is still no unambiguous confirmation of its existence or non-existence. A new experiment based on $$^{8}$$ He(p, 2p) $$^{7}$$ H{t+ $$^{4}n$$ } reaction, with direct detection of the four neutrons, has been carried out at RIBF, which can hopefully help to draw a definite conclusion on the tetraneutron system.

Nicotinic receptors of the cholinergic system are ligand-gated ion channels, responding to the excitatory neurotransmitter, acetylcholine, and the addictive component of tobacco, nicotine. They help to transduce salient information in the environment by activating specific neural circuitry in normal and disease states. While nicotinic receptors are promising neurological and neuropsychiatric disorder targets, they have fallen out of favor after several late-stage clinical failures. Targeting the complex of the nicotinic receptor, including lynx1 accessory proteins, could be the key to unlocking the intractable nAChR for therapeutic development. Lynx1 binds to the extracellular face of the nAChR and acts as a critical modulator, suppressing memory, learning, and plasticity. Lynx1 removal in animal models leads to memory and plasticity enhancements, some of which have therapeutic relevance for neuropsychiatric and neurological disease. A review of the lynx1 accessory modulator and its role in modulating neuronal nAChRs will be discussed.

Nicotinic acetylcholine receptor (nAChR) participates in diverse biological processes, e.g., mood, learning, and addiction. Glycosylphosphatidylinositol (GPI)-linked lynx1 is an allosteric modulator of nAChR function, including shifts in agonist sensitivity, reduced desensitization, and slower recovery from desensitization. This modulation is thought to be achieved by lynx1’s interaction with nAChR subunits, particularly at the α4:α4 interface. In this study, we used molecular modeling and simulation to study the structure, dynamics, and interactions of lynx1 when bound to nAChRs, as well as unbound, monomeric lynx1, when embedded in membranes. Though lynx1 structures are similar in both states, lynx1 dynamics are more restricted in the bound state than in the unbound one. When bound, interactions between lynx1 and nAChR are observed to be maintained throughout the simulations. Of particular note, lynx1 demonstrates prolonged interactions with the receptor C-loop in one of the nAChR α4 subunits, a region important for agonist binding and possibly the transition between open/close states. During interactions with lynx1, an α4 C-loop tends to be restricted in either close or open state, whereas the C-loop state transitions are more evident in when the nAChR is unbound by lynx1. Interestingly, the conformational change of the C-loop is stochastic, suggesting that lynx1 can influence nAChR (critical for its multimodal action), for instance by shifting its agonist sensitivity and recovery from desensitization.

7 pags., 3 figs. -- 27th International Nuclear Physics Conference (INPC2019), The tetraneutron has attracted the attention of nuclear physicists during the past decades, but there is still no unambiguous confirmation of its existence or nonexistence. A new experiment based on 8 He(p,2p)7H{t+4n} reaction, with direct detection of the four neutrons, has been carried out at RIBF, which can hopefully help to draw a definite conclusion on the tetraneutron system., We acknowledge the support of the RIBF accelerator staff and the BigRIPS team for providing the high-quality beam. Z. H. Yang acknowledges the financial support from the Foreign Postdoctoral Researcher program of RIKEN. T. Aumann acknowledges the support by DFG via SFB 1245. P. Koseoglou acknowledges the support from BMBF (NUSTAR.DA grant No.05P 15RDFN1).

This study investigates—for the first time to our knowledge—the existence and mechanisms of functional interactions between the endogenous mammalian prototoxin, lynx1, and α3- and β4-subunit–containing human nicotinic acetylcholine receptors (α3β4*-nAChRs). Concatenated gene constructs were used to express precisely defined α3β4*-nAChR isoforms (α3β4)2β4-, (α3β4)2α3-, (α3β4)2α5(398D)-, and (α3β4)2α5(398N)-nAChR in Xenopus oocytes. In the presence or absence of lynx1, α3β4*-nAChR agonist responses were recorded by using 2-electrode voltage clamp and single-channel electrophysiology, whereas radioimmunolabeling measured cell-surface expression. Lynx1 reduced (α3β4)2β4-nAChR function principally by lowering cell-surface expression, whereas single-channel effects were primarily responsible for reducing (α3β4)2α3-nAChR function [decreased unitary conductance (≥50%), altered burst proportions (3-fold reduction in the proportion of long bursts), and enhanced closed dwell times (3- to 6-fold increase)]. Alterations in both cell-surface expression and single-channel properties accounted for the reduction in (α3β4)2α5-nAChR function that was mediated by lynx1. No effects were observed when α3β4*-nAChRs were coexpressed with mutated lynx1 (control). Lynx1 is expressed in the habenulopeduncular tract, where α3β4*-α5*-nAChR subtypes are critical contributors to the balance between nicotine aversion and reward. This gives our findings a high likelihood of physiologic significance. The exquisite isoform selectivity of lynx1 interactions provides new insights into the mechanisms and allosteric sites [α(−)-interface containing] by which prototoxins can modulate nAChR function.—George, A. A., Bloy, A., Miwa, J. M., Lindstrom, J. M., Lukas, R. J., Whiteaker, P. Isoform-specific mechanisms of α3β4*-nicotinic acetylcholine receptor modulation by the prototoxin lynx1.

The cholinergic system modulates many biological functions, due to the widespread distribution of cholinergic neuronal terminals, and the diffuse release of its neurotransmitter, acetylcholine. Several layers of regulation help to refine and control the scope of this excitatory neurotransmitter system. One such regulatory mechanism is imparted through endogenous toxin-like proteins, prototoxins, which largely control the function of nicotinic receptors of the cholinergic system. Prototoxins and neurotoxins share the distinct three finger toxin fold, highly effective as a receptor binding protein, and the former are expressed in the mammalian brain, immune system, epithelium, etc. Prototoxins and elapid snake neurotoxins appear to be related through gene duplication and divergence from a common ancestral gene. Protein modulators can provide a graded response of the cholinergic system, and within the brain, stabilize neural circuitry through direct interaction with nicotinic receptors. Understanding the roles of each prototoxin (e.g., lynx1, lynx2/lypd1, PSCA, SLURP1, SLURP2, Lypd6, lypd6b, lypdg6e, PATE-M, PATE-B, etc.), their binding specificity and unique expression profile, has the potential to uncover many fascinating cholinergic-dependent mechanisms in the brain. Each family member can provide a spatially restricted level of control over nAChR function based on its expression in the brain. Due to the difficulty in the pharmacological targeting of nicotinic receptors in the brain as a result of widespread expression patterns and similarities in receptor sequences, unique interfaces between prototoxin and nicotinic receptor could provide more specific targeting than nicotinic receptors alone. As such, this family is intriguing from a long-term therapeutic perspective.

The spin-isospin responses of the 11Li drip-line nucleus has been measured. Preliminary results of the 11Li(p, n)11Be experiment in inverse kinematics at RI Beam Factory (RIBF) of RIKEN Nishina Center are presented including the observation of 1n, 2n, t, d, 2α and 6He+α decay channels of 11Be reaction product. Details of the experimental setup based on PANDORA (Particle Analyzer Neutron Detector Of Real-time Acquisition) low-energy neutron detector and the SAMURAI large-acceptance magnetic spectrometer are described.

The radioactive noble gas radon can be a serious background source in the underground particle physics experiments studying processes that deposit energy comparable to its decay products. Low energy solar neutrino measurements at Super-Kamiokande suffer from these backgrounds and therefore require precise characterization of the radon concentration in the detector's ultra-pure water. For this purpose, we have developed a measurement system consisting of a radon extraction column, a charcoal trap, and a radon detector. In this article we discuss the design, calibration, and performance of the radon extraction column. We also describe the design of the measurement system and evaluate its performance, including its background. Using this system we measured the radon concentration in Super-Kamiokande's water between May 2014 and October 2015. The measured radon concentration in the supply lines of the water circulation system was $1.74\pm0.14~\mathrm{mBq/m^{3}}$ and in the return line was $9.06\pm0.58~\mathrm{mBq/m^{3}}$. Water sampled from the center region of the detector itself had a concentration of $, 20 pages, 10 figures

Generally, uniformly-distributed irradiation defects such as artificial pinning centers improve the critical current, I c, of REBCO tapes in strong external field. On the other hand, at self-field or low field, irradiation is not effective for improving I c. However, if non-uniform pinning center distribution similar to vortex profile is assumed, this would increase I c at selffield. Therefore, in this study, we experimentally investigated the effect of partial irradiation on I c based on the assumption. We prepared REBCO tapes irradiated with trihydrogen cations in the c-axis direction. Three different patterns of defects were examined; overall, edge-only and center-only irradiations. We measured I c at 77 K, 25 K and 15 K using the four terminal method. Results showed that, while I c was not improved after the irradiations for each pattern, I c behaviour depends on the patterns; edge-only-irradiated tapes showed the highest I c and overall irradiation caused the largest I c degradation at 77 K. However, center-only-irradiated tapes showed the lowest critical current at 25 K and 15 K which may be due to enhancement of flux pinning effect at lower temperature. Moreover, pinning centers in the edge region might be more effective to increase I c, though further investigation on the effect of non-uniform pinning centers is required.

The tetraneutron has been drawing the attention of the nuclear physics community for decades, but a firm conclusion on its existence and properties is still far from being reached despite many experimental and theoretical efforts. New measurements have recently been performed at RIBF with the SAMURAI spectrometer by applying complementary reaction probes, which will help to pin down the properties of this four-neutron system., Comment: 5 pages, 2 figures, conference proceedings of XXII International Conference on Few-Body Problems in Physics (FB22), Caen, France, July 2018

Neuronal nicotinic acetylcholine receptors (nAChRs) of the cholinergic system have been linked to antinociception, and therefore could be an alternative target for pain alleviation. nAChR activity has been shown to be regulated by the nicotinic modulator, lynx1, which forms stable complexes with nAChRs and has a negative allosteric action on their function. The objective in this study was to investigate the contribution of lynx1 to nicotine-mediated antinociception. Lynx1 contribution was investigated by mRNA expression analysis and electrophysiological responses to nicotine in the dorsal raphe nucleus (DRN), a part of the pain signaling pathway. In vivo antinociception was investigated in a test of nociception, the hot-plate analgesia assay with behavioral pharmacology. Lynx1/α4β2 nAChR interactions were investigated using molecular dynamics computational modeling. Nicotine evoked responses in serotonergic and GABAergic neurons in the DRN are augmented in slices lacking lynx1 (lynx1KO). The antinociceptive effect of nicotine and epibatidine is enhanced in lynx1KO mice and blocked by mecamylamine and DHβE. Computer simulations predict preferential binding affinity of lynx1 to the α:α interface that exists in the stoichiometry of the low sensitivity (α4)3(β2)2 nAChRs. Taken together, these data point to a role of lynx1 in mediating pain signaling in the DRN through preferential affinity to the low sensitivity α4β2 nAChRs. This study suggests that lynx1 is a possible alternative avenue for nociceptive modulation outside of opioid-based strategies.

The α6 nicotinic acetylcholine receptor (nAChR) subunit is an attractive drug target for treating nicotine addiction because it is present at limited sites in the brain including the reward pathway. Lynx1 modulates several nAChR subtypes; lynx1-nAChR interaction sites could possibly provide drug targets. We found that dopaminergic cells from the substantia nigra pars compacta (SNc) express lynx1 mRNA transcripts and, as assessed by co-immunoprecipitation, α6 receptors form stable complexes with lynx1 protein, although co-transfection with lynx1 did not affect nicotine-induced currents from cell lines transfected with α6 and β2. To test whether lynx1 is important for the function of α6 nAChRs in vivo, we bred transgenic mice carrying a hypersensitive mutation in the α6 nAChR subunit (α6L9'S) with lynx1 knockout mice, providing a selective probe of the effects of lynx1 on α6* nAChRs. Lynx1 removal reduced the α6 component of nicotine-mediated rubidium efflux and dopamine (DA) release from synaptosomal preparations with no effect on numbers of α6β2 binding sites, indicating that lynx1 is functionally important for α6* nAChR activity. No effects of lynx1 removal were detected on nicotine-induced currents in slices from SNc, suggesting that lynx1 affects presynaptic α6* nAChR function more than somatic function. In the absence of agonist, lynx1 removal did not alter DA release in dorsal striatum as measured by fast scan cyclic voltammetry. Lynx1 removal affected some behaviors, including a novel-environment assay and nicotine-stimulated locomotion. Trends in 24-hour home-cage behavior were also suggestive of an effect of lynx1 removal. Conditioned place preference for nicotine was not affected by lynx1 removal. The results show that some functional and behavioral aspects of α6-nAChRs are modulated by lynx1.

The glutamatergic subthalamic nucleus (STN) exerts control over motor output through nuclei of the basal ganglia. High-frequency electrical stimuli in the STN effectively alleviate motor symptoms in movement disorders, and cholinergic stimulation boosts this effect. To gain knowledge about the mechanisms of cholinergic modulation in the STN, we studied cellular and circuit aspects of nicotinic acetylcholine receptors (nAChRs) in mouse STN. We discovered two largely divergent microcircuits in the STN; these are regulated in part by either α4β2 or α7 nAChRs. STN neurons containing α4β2 nAChRs (α4β2 neurons) received more glutamatergic inputs, and preferentially innervated GABAergic neurons in the substantia nigra pars reticulata. In contrast, STN neurons containing α7 nAChRs (α7 neurons) received more GABAergic inputs, and preferentially innervated dopaminergic neurons in the substantia nigra pars compacta. Interestingly, local electrical stimuli excited a majority (79%) of α4β2 neurons but exerted strong inhibition in 58% of α7 neurons, indicating an additional diversity of STN neurons: responses to electrical stimulation. Chronic exposure to nicotine selectively affects α4β2 nAChRs in STN: this treatment increased the number of α4β2 neurons, upregulated α4-containing nAChR number and sensitivity, and enhanced the basal firing rate of α4β2 neurons bothex vivoandin vivo. Thus, chronic nicotine enhances the function of the microcircuit involving α4β2 nAChRs. This indicates chronic exposure to nicotinic agonist as a potential pharmacological intervention to alter selectively the balance between these two microcircuits, and may provide a means to inhibit substantia nigra dopaminergic neurons.

TRPV1 is an ion channel activated by heat and pungent agents including capsaicin, and has been extensively studied in nociception of sensory neurons. However, the location and function of TRPV1 in the hippocampus is debated. We found that TRPV1 is expressed in oriens-lacunosum-moleculare (OLM) interneurons in the hippocampus, and promotes excitatory innervation. TRPV1 knockout mice have reduced glutamatergic innervation of OLM neurons. When activated by capsaicin, TRPV1 recruits more glutamatergic, but not GABAergic, terminals to OLM neurons in vitro. When TRPV1 is blocked, glutamatergic input to OLM neurons is dramatically reduced. Heterologous expression of TRPV1 also increases excitatory innervation. Moreover, TRPV1 knockouts have reduced Schaffer collateral LTP, which is rescued by activating OLM neurons with nicotine—via α2β2-containing nicotinic receptors—to bypass innervation defects. Our results reveal a synaptogenic function of TRPV1 in a specific interneuron population in the hippocampus, where it is important for gating hippocampal plasticity., The role of TRPV1 in the CNS is not fully understood. Here the authors show that TRPV1 is expressed specifically in somatostatin-positive OLM interneurons of the hippocampus, where it promotes excitatory innervation of these cells.

The estimation of energy expenditure (EE) of grazing animals is of great importance for efficient animal management on pasture. In the present study, a method is proposed to estimate EE in grazing animals based on measurements of body acceleration of animals in combination with the conventional Agricultural and Food Research Council (AFRC) energy requirement system. Three-dimensional body acceleration and heart rate were recorded for tested animals under both grazing and housing management. An acceleration index, vectorial dynamic body acceleration (VeDBA), was used to calculate activity allowance (AC) during grazing and then incorporate it into the AFRC system to estimate the EE (EE derived from VeDBA [EE]) of the grazing animals. The method was applied to 3 farm ruminant species (7 cattle, 6 goats, and 4 sheep). Energy expenditure based on heart rate (EE) was also estimated as a reference. The result showed that larger VeDBA and heart rate values were obtained under grazing management, resulting in greater EE and EE under grazing management than under housing management. There were large differences between the EE estimated from the 2 methods, where EE values were greater than EE (averages of 163.4 and 142.5% for housing and grazing management, respectively); the EE was lower than the EE, whereas the increase in EE under grazing in comparison with housing conditions was larger than that in EE. These differences may have been due to the use of an equation for estimating EE derived under laboratory conditions and due to the presence of the effects of physiological, psychological, and environmental factors in addition to physical activity being included in measurements for the heart rate method. The present method allowed us to separate activity-specific EE (i.e., AC) from overall EE, and, in fact, AC under grazing management were about twice times as large as those under housing management for farm ruminant animals. There is evidence that the conventional energy system can predict fasting metabolism and the AC of housed animals based on accumulated research on energy metabolism and that VeDBA can quantify physical activity separately from other factors in animals on pasture. Therefore, the use of the VeDBA appears to be a precise way to predict activity-specific EE under grazing conditions, and the method incorporating acceleration index data with a conventional energy system can be a simple and useful method for estimation of EE in farm ruminants on pastures.

The incidence of non-alcoholic fatty liver disease (NAFLD) increases with age. Cellular senescence refers to a state of irreversible cell-cycle arrest combined with the secretion of proinflammatory cytokines and mitochondrial dysfunction. Senescent cells contribute to age-related tissue degeneration. Here we show that the accumulation of senescent cells promotes hepatic fat accumulation and steatosis. We report a close correlation between hepatic fat accumulation and markers of hepatocyte senescence. The elimination of senescent cells by suicide gene-meditated ablation of p16Ink4a-expressing senescent cells in INK-ATTAC mice or by treatment with a combination of the senolytic drugs dasatinib and quercetin (D+Q) reduces overall hepatic steatosis. Conversely, inducing hepatocyte senescence promotes fat accumulation in vitro and in vivo. Mechanistically, we show that mitochondria in senescent cells lose the ability to metabolize fatty acids efficiently. Our study demonstrates that cellular senescence drives hepatic steatosis and elimination of senescent cells may be a novel therapeutic strategy to reduce steatosis. © The Author(s) 2017.

Neurological disorders affect millions of Americans and this number is expected to rise with the aging population. Development of drugs to treat these disorders may be facilitated by improved in vitro models that faithfully reproduce salient features of the relevant brain regions. Current 3D culture methods face challenges with reliably reproducing microarchitectural features of brain morphology such as cortical or hippocampal layers. In this work, polydimethylsiloxane (PDMS) mini-wells were used to create low aspect ratio, adherent 3D constructs where neurons self-assemble into layers. Layer self-assembly was determined to depend on the size of the PDMS mini-well. Layer formation occurred in cultures composed of primary rat cortical neurons or human induced pluripotent stem cell - derived neurons and astrocytes and was robust and reproducible. Layered 3D constructs were found to have spontaneous neural activity characterized by long bursts similar to activity in the developing cortex. The responses of layered 3D cultures to drugs were more similar to in vivo data than those of 2D cultures. 3D constructs created with this method may be thus suitable as in vitro models for drug discovery for neurological disorders.

Accurate oestrus detection is important in order to perform AI and embryo transfer procedures at appropriate times. Pedometers and accelerometers have been utilised in the livestock industry to detect oestrus. However, the efficacy of these devices in beef cattle production systems remains unclear. In this preliminary study, we evaluated the efficacy of a commercially available accelerometer device for oestrus detection in nonlactating Holstein×Japanese Black crossbred cows (n=14). Accelerometer devices (Farmnote Color, Farmnote Inc., Hokkaido, Japan) were placed on the neck of each animal and remained in place for 3 months. Cows were kept in a pen during the day (8-9 cows/pen) and a pasture during the night. Cows were visually monitored for oestrus activity twice daily in the morning and evening for 30min. The activity of cows detected by the sensor was recorded every hour. Oestrus occurrence and duration were automatically detected and calculated by the device. Alerts generated by the device were categorized as either true positives (TP), false positives (FP) or false negatives (FN). True positives were defined as the observation of oestrus in conjunction with a device-generated oestrus alert or confirmation of the presence of a corpus luteum (CL) by ovarian ultrasonography 5 days following an oestrus alert. False positives were defined as events in which an oestrus alert was generated by the device, but no corresponding visual observation of oestrus occurred or a CL was not present 5 days following the alert. False negatives were cases where oestrus was observed visually but there was no corresponding device-generated oestrus alert. Reasons for FP were categorized into 1 of 3 categories defined as (1) other cows in oestrus in the same pen, (2) cows that were moved from other herds, (3) ovarian disorder such as follicular cyst, and (4) unknown. Statistical analyses were conducted using Student’s t-test. Results were considered significant at P

We report the first observation that endoplasmic reticulum (ER) stress and the unfolded protein response (UPR) can decrease when a central nervous system drug acts as an intracellular pharmacological chaperone for its classic receptor. Transient expression of α4β2 nicotinic receptors (nAChRs) in Neuro-2a cells induced the nuclear translocation of activating transcription factor 6 (ATF6), which is part of the UPR. Cells were exposed for 48 h to the full agonist nicotine, the partial agonist cytisine, or the competitive antagonist dihydro-β-erythroidine; we also tested mutant nAChRs that readily exit the ER. Each of these four manipulations increased Sec24D-enhanced green fluorescent protein fluorescence of condensed ER exit sites and attenuated translocation of ATF6-enhanced green fluorescent protein to the nucleus. However, we found no correlation among the manipulations regarding other tested parameters [i.e., changes in nAChR stoichiometry (α4(2)β2(3) versus α4(3)β2(2)), changes in ER and trans-Golgi structures, or the degree of nAChR up-regulation at the plasma membrane]. The four manipulations activated 0 to 0.4% of nAChRs, which shows that activation of the nAChR channel did not underlie the reduced ER stress. Nicotine also attenuated endogenously expressed ATF6 translocation and phosphorylation of eukaryotic initiation factor 2α in mouse cortical neurons transfected with α4β2 nAChRs. We conclude that, when nicotine accelerates ER export of α4β2 nAChRs, this suppresses ER stress and the UPR. Suppression of a sustained UPR may explain the apparent neuroprotective effect that causes the inverse correlation between a person's history of tobacco use and susceptibility to developing Parkinson's disease. This suggests a novel mechanism for neuroprotection by nicotine.

In this study, we investigate the effect of eddy current loss reduction by segmented rare-earth magnets that are used in synchronous motors driven by inverters. First, the difference in the loss-reduction effect due to the rotor shape is estimated by the 3-D finite-element analysis that considers the carrier harmonics of the inverter. The results are compared to the theoretical solution. Next, a basic experiment using magnet specimens is carried out in order to confirm the calculated results. It is clarified that most of the magnet eddy current losses in the analyzed motors are caused by the carrier harmonics of the inverter and that the loss-reduction effect of the segmented magnet in the case of the interior permanent magnet motor decreases when it is compared to that of the surface permanent magnet motor. This phenomenon is mainly caused by the increase in the reaction field by the high-frequency harmonic eddy currents.

Decoding performance of linear programming (LP) decoding is closely related to geometrical properties of a fundamental polytope: fractional distance, pseudo codeword, etc. In this paper, an idea of the cutting-plane method is employed to improve the fractional distance of a given binary parity-check matrix. The fractional distance is the minimum weight (with respect to lscr1-distance) of nonzero vertices of the fundamental polytope. The cutting polytope is defined based on redundant rows of the parity-check matrix. The redundant rows are codewords of the dual code not yet appearing as rows in the parity-check matrix. The cutting polytope plays a key role to eliminate unnecessary fractional vertices in the fundamental polytope. We propose a greedy algorithm and its efficient implementation based on the cutting-plane method. It has been confirmed that the fractional distance of some parity-check matrices are actually improved by using the algorithm.

In this paper, we investigate eddy current loss in divided Nd-Fe-B sintered magnets for synchronous motors, from both sides of measurement and finite element analysis. First, the basic experiments with an exciting coil and divided magnets are carried out. The results are compared with the 3-D finite element analysis to clarify the effects of insulation resistance between divided magnets, number of magnet divisions, and frequency. Next, a permanent magnet synchronous motor is analyzed to understand the eddy current generation in the magnets applied to the motors. It is clarified that the effect of the eddy current loss reduction by the magnet division is weakened in the case of high frequency harmonic magnetic fields. The critical insulation resistance between the divided magnets is also clarified.

Alpha6-containing (alpha6*) nicotinic ACh receptors (nAChRs) are selectively expressed in dopamine (DA) neurons and participate in cholinergic transmission. We generated and studied mice with gain-of-function alpha6* nAChRs, which isolate and amplify cholinergic control of DA transmission. In contrast to gene knockouts or pharmacological blockers, which show necessity, we show that activating alpha6* nAChRs and DA neurons is sufficient to cause locomotor hyperactivity. alpha6(L9'S) mice are hyperactive in their home cage and fail to habituate to a novel environment. Selective activation of alpha6* nAChRs with low doses of nicotine, by stimulating DA but not GABA neurons, exaggerates these phenotypes and produces a hyperdopaminergic state in vivo. Experiments with additional nicotinic drugs show that altering agonist efficacy at alpha6* provides fine tuning of DA release and locomotor responses. alpha6*-specific agonists or antagonists may, by targeting endogenous cholinergic mechanisms in midbrain or striatum, provide a method for manipulating DA transmission in neural disorders.

Nicotinic acetylcholine receptors (nAChRs) affect a wide array of biological processes, including learning and memory, attention, and addiction. lynx1, the founding member of a family of mammalian prototoxins, modulates nAChR function in vitro by altering agonist sensitivity and desensitization kinetics. Here we demonstrate, through the generation of lynx1 null mutant mice, that lynx1 modulates nAChR signaling in vivo. Its loss decreases the EC(50) for nicotine by approximately 10-fold, decreases receptor desensitization, elevates intracellular calcium levels in response to nicotine, and enhances synaptic efficacy. lynx1 null mutant mice exhibit enhanced performance in specific tests of learning and memory. Consistent with reports that mutations resulting in hyperactivation of nAChRs can lead to neurodegeneration, aging lynx1 null mutant mice exhibit a vacuolating degeneration that is exacerbated by nicotine and ameliorated by null mutations in nAChRs. We conclude that lynx1 functions as an allosteric modulator of nAChR function in vivo, balancing neuronal activity and survival in the CNS.

Glycosylphosphatidylinositol-anchored neurotoxin-like receptor binding proteins, such as lynx modulators, are topologically positioned to exert pharmacological effects by binding to the extracellular portion of nAChRs. These actions are generally thought to proceed when both lynx and the nAChRs are on the plasma membrane. Here, we demonstrate that lynx1 also exerts effects on α4β2 nAChRs within the endoplasmic reticulum. Lynx1 affects assembly of nascent α4 and β2 subunits and alters the stoichiometry of the receptor population that reaches the plasma membrane. Additionally, these data suggest that lynx1 shifts nAChR stoichiometry to low sensitivity (α4)3(β2)2 pentamers primarily through this interaction in the endoplasmic reticulum, rather than solely via direct modulation of activity on the plasma membrane. To our knowledge, these data represent the first test of the hypothesis that a lynx family member, or indeed any glycosylphosphatidylinositol-anchored protein, could act within the cell to alter assembly of a multisubunit protein.

Soft computing techniques, e.g., neural networks, fuzzy inference, evolutionary computation, and chaos theory, have been applied to a wide variety of control systems in industry because of their control capability and flexibility. They are also powerful to handle the complicated mechatronic systems with various nonlinearities which are difficult to model using mathematical formulas. In order to achieve the system identification of unknown structured mechatronic systems, This work presents a novel evolutionary algorithm using genetic algorithms (GAs), where the optimal mathematical structure of plant mechanisms and the combination of parameters can be autonomously determined by means of the optimization ability of the GA. The effectiveness of the proposed identification has been verified by experiments with comparative studies, using the typical mechanical systems with velocity controller.

The physiologies of cells depend on electrochemical signals carried by ion channels and receptors. Venomous animals produce an enormous variety of peptide toxins with high affinity for specific ion channels and receptors. The mammalian prototoxin lynx1 shares with α-bungarotoxin the ability to bind and modulate nicotinic receptors (nAChRs); however, lynx1 is tethered to the membrane via a GPI anchor. We show here that several classes of neurotoxins, including bungarotoxins and cobratoxins, retain their selective antagonistic properties when tethered to the membrane. Targeted elimination of nAChR function in zebrafish can be achieved with tethered α-bungarotoxin, silencing synaptic transmission without perturbing synapse formation. These studies harness the pharmacological properties of peptide toxins for use in genetic experiments. When combined with specific methods of cell and temporal expression, the extension of this approach to hundreds of naturally occurring peptide toxins opens a new landscape for cell-autonomous regulation of cellular physiology in vivo.