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SARS-CoV-2 caused a large-scale global pandemic between 2020 and 2022. Despite efforts to understand its biology and mechanisms of pathogenicity, the viral impact on the neurological systems remains unclear. The main goal of this study was to quantify the neurological phenotypes induced by SARS-CoV-2 spike protein in neurons, measured byin-vitromulti-well micro-electrode arrays (MEAs). We extracted the whole-brain neurons from the newborn P1 mice and plated them on multi-well micro-electrode arrays and administered purified recombinant spike proteins (S1 and S2 subunits respectively) from the SARS-CoV-2 virus. The signals from the MEAs were transmitted from an amplifier to a high-performance computer for recording and analysis. We used an in-house developed algorithm to quantify neuronal phenotypes. Among all the phenotypic features analyzed, we discovered that the S1 protein of SARS-CoV-2 decreased the mean burst numbers observed on each electrode; This effect was not observed for the spike 2 protein (S2) and could be rescued by an anti-S1 antibody. Finally, our data strongly suggest that the receptor binding domain (RBD) of S1 is responsible for the reduction of burst activities in neurons. Overall, our results strongly indicate that spike proteins may play an important role in altering neuronal phenotypes, specifically the burst patterns, when neurons are exposed during early development.

Current situation and prospect of artificial intelligence application in endoscopic diagnosis of Helicobacter pylori infection

We propose an AgGeSe/Al2O3/Pt selector with great potential in a dense memory array. In terms of on-current drive, selectivity, and uniformity, devices with the mixed AgGeSe layer outperform the Ag/Al2O3/Pt device. The introduction of GeSe not only blocks the diffusion of Ag, but also facilitates the backflow of Ag to the active electrode, therefore increasing the on-current. The ultra-high on-current drive (3 mA and 12.7 MA/cm2), high selectivity (1010), small variation, high thermal stability, and steep switching slope (0.27 mV/dec) realized in our AgGeSe/Al2O3/Pt device render it a promising candidate for selector application.

Bjerkandera adusta degrades polycyclic aromatic compound. However, its effect on preventing plant disease is unknown. We carried out both the confrontation culture and greenhouse pot experiments to address effects of B. adusta M1 strain on the growth of pathogenic Fusarium oxysporum f. sp. conglutinans (FOC) and incidence of Fusarium wilt in Brassica napus L. The results showed the hyphal growth rate of B. adusta M1 was significantly greater than that of FOC. The B. adusta M1 fermentation broth also significantly inhibited the growth of FOC hyphae by 62.79 ± 1.80%, which was greater than an inhibition rate of 40.63 ± 1.68% by the fungicide, carbendazim. The electron microscopic image showed the FOC hyphae was directly penetrated by the B. adusta M1 hyphae, indicating a strong mycoparasitism by B. adusta M1. Both the B. adusta M1 and its fermentation broth reduced the incidence and disease index of the Fusarium wilt in Br. napus, and the control effects of B. adusta M1 and its fermentation broth on Fusarium wilt were comparable or better than carbendazim. Furthermore, both the B. adusta M1 and its fermentation broth increased the activity of the defense enzymes including superoxide dismutase (25.92% and 22.38%), catalase (78.80% and 82.28%), peroxidase (46.64% and 6.83%) and phenylalanine ammonia-lyase (44.23% and 5.77%). Similarly, both the B. adusta M1 and its fermentation broth decreased the cell membrane permeability (48.68% and 33.71%) and malondialdehyde concentration (71.04% and 17.51%) of plant leaf. Therefore, B. adusta M1 can be applied as a potential biocontrol fungus to against Fusarium wilt.

Threshold switching (TS) devices are promising candidates to build highly compact and energy efficient artificial neurons. Here, we present a Pt/Ag/TiN/HfAlOx/Pt (PATHP) device with excellent TS characteristics, including a large selectivity(1010), a wide range of operation current from 10 nA to 1 mA, an extremely steep slope (0.63 mV/dec) and fast turn-on speed (50 ns). The stable TS performance can be ascribed to the introduction of TiN buffer layer and the alternate atomic layer deposited HfAlOx layer. Further, we experimentally demonstrate the functions of leaky-integrate-and-fire neurons with low power feature based on a RC circuit and a single device, respectively, which are essential for constructing spiking neuromorphic systems.

Autonomous Underwater Helicopter (AUH) is a novel dish-shaped multi-functional submersible vehicle in the autonomous underwater vehicle (AUV) family. Numerical study of launch and recovery system for AUVs from a research vessel is significant in the overall design. Single-arm crane and air-launch way are the important ways of launching AUVs with the high expertise and security. However, AUVs are still easily subject to a huge impact load which can cause the body damage, malfunction of electronic components, and other serious accidents when AUVs are launching into water. In this paper, unsteady, incompressible, two-phase flow with identified boundary conditions was modeled in the computational domain using Volume of Fluid (VOF) technology, and different overset-grid sizes were set and analyzed to ascertain the accuracy of computational fluid dynamics (CFD) simulation first due to the setting of the grid which will directly affect the accuracy of the calculation results, especially the overset-grid convergence validation. Then, the change of water-entry impact force and velocity of AUH with different water-entry velocities and angles were calculated by CFD analysis software STAR-CCM + solver. The proper immersion angle of the AUH should be 75° under comprehensive analysis and comparison. Besides, the variations of pressure distribution of the AUH in the whole water-entry process were also obtained, which provides reference for the shape optimization in the future work.

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Amino acids (AAs) are important metabolites that are related with diabetes. However, their roles in the initiation and development of diabetes mellitus (DM), especially in the treatment of Ginkgo biloba leaves extract (GBE) have not been fully explored. Thus, we investigated the roles that AAs played in the progression and GBE supplementation of DM rat induced by streptozotocin. The rats were randomly divided into a normal control group treated with drug-free solution, a normal control group treated with GBE, a DM group treated with drug-free solution, and DM group treated with GBE; and maintained on this protocol for 9 weeks. Rat plasma was collected from the sixth week to the ninth week and then analyzed with the optimized hydrophilic interaction liquid chromatography coupled with tandem mass spectrometry method. A total of 17 AAs with differential levels were monitored to indicate dysfunction of AAs metabolism to confirm the occurrence and development of DM. Treatment with GBE partially reversed the changes seen in seven AAs including leucine, isoleucine, tyrosine, glutamic acid, asparagines, lysine and alanine in DM rats, indicating that GBE could prevent the occurrence and development of DM by acting on AAs metabolism. The improvement of those AAs metabolism disorders may play a considerable role in the treatment of GBE on the occurrence and development of DM. Those findings potentially promote the understanding of the pathogenic progression of DM and reveal the therapeutic mechanism of GBE against DM.

A sensor array mimicking a chemical nose/tongue based on bovine serum albumin nanoparticles (BSANsn) has been developed for the fluorescence pattern recognition of metal ions in biofluids. Three types of BSANsn (BSANs10, BSANs20, and BSANs40) show the same excitation/emission peak at 478/526 nm. According to the differential fluorescence variation, the sensor array shows particular fluorescence response patterns depending upon metal ions. Upon principal component analysis (PCA), it was found that the sensor array can distinguish 18 metal ions clearly at a concentration of as low as 10 μM. Moreover, different concentrations of metal ions and mixed metal ions of diverse kinds or valence states can be differentiated by the sensor in biofluids. In addition, the results were well consistent with those obtained with the traditional ICP-AES method.

This letter reports on a novel cost-efficient and multifunctional barcode-like sensors array (BLSA) printed with a conductive bioinspired smart ink. The conductive ink (P@G ink), which can be further chemically engineered with different organic ligands, was generated via facile one-pot hydrothermal reduction of graphene oxide (GO) in dopamine (DA) as coreductan Usingvarious chemical derivatives of the P@G inks on a flexible substrate (e.g., Kapton), a highly integrated BLSA as well as smart nose/tongue mimic array were generated for simultaneous sensing and distinguishing of complex physical and chemical stimuli, including temperature, light, air pressure, relative humidity, and volatile organic compounds (VOCs). Due to these very attractive features, the reported P@G ink-based BLSA would have the potential for unique opportunities regarding “all-in-one”—yet cost-effective—disposable electronics and sensors.

Congenital disorders of glycosylation (CDG) and deglycosylation (CDDG) are a collection of rare pediatric disorders with symptoms that range from mild to life threatening. They typically affect multiple organ systems and usually present with neurological abnormalities including hypotonia, cognitive impairment, and intractable seizures. Several genes have been implicated in the thirty-six types of CDG, but currently NGLY1 is the only known CDDG gene. A common biological mechanism among CDG types and in CDDG is endoplasmic reticulum (ER) stress. Here, we develop two isogenic human cellular models of CDG (PMM2, the most prevalent type of CDG, and DPAGT1) and of the only CDDG (NGLY1) in an effort to identify drugs that can alleviate ER stress. Systematic phenotyping identified elevated ER stress and autophagy levels among other cellular and morphological phenotypes in each of the cellular models. We screened a complex drug library for compounds able to correct aberrant morphological phenotypes in each of the models using an agnostic phenotypic cell painting assay based on >300 cellular features. The image-based screen identified multiple candidate compounds able to correct aberrant morphology, and we show a subset of these are able to correct cellular and molecular defects in each of the models. These results provide new directions for the treatment of rare diseases of glycosylation and deglycosylation and a framework for new drug screening paradigms for more common neurodegenerative diseases characterized by ER stress.Summary sentenceNovel drug screening modality identifies compounds that correct aberrant molecular phenotypes in precision cellular models of glycosylation defects.

Very limited experimental data is available regarding the safe dosages related to synchrotron radiation (SR) procedures. We used young rats and macaques to address bone marrow and skin tolerance to various doses of synchrotron radiation.Rats were subjected to 0, 0.5, 2.5, 5, 25 or 100 Gy local SR X-ray irradiation at left hind limb. Rat blood samples were analyzed at 2-90 days after irradiation. The SR X-ray irradiated skin and tibia were sectioned for morphological examination. For non-human primate study, three male macaques were subjected to 0.5 or 2.5 Gy SR X-ray on crus. Skin responses of macaques were observed.All rats that received SR X-ray irradiation doses greater than 2.5 Gy experienced hair loss and bone-growth inhibition, which were accompanied by decreased number of follicles, thickened epidermal layer, and decreased density of bone marrow cells (p 0.05). Macaque skin could tolerate 0.5 Gy SR X-ray but showed significant hair loss when the dose was raised above 2.5 Gy.The safety threshold doses of SR X-ray for rat skin, bone marrow and macaque skin are between 0.5 and 2.5 Gy. Our study provided essential information regarding the biosafety of SR X-ray irradiation.

Owing to the superior hydrodynamic characteristics of an underwater glider designed with diamond-wings in comparison with the installation of conventional swept-wings. This article successively engages in the design and analysis of the hydrodynamic performance of the tail rudder adaptive to such kind new type underwater glider. Four kinds arrangements of X-shaped rudders have been analyzed and compared for the lifts, drags and lift-to-drag ratios by using the software package STAR-CCM+. The hydrodynamic performance of the simulation results showed that the 45° X-rudder is most conducive to the UG with diamond-wing.

Neural stem cell (NSC) transplantation is a promising treatment to improve the recovery after brain ischemia. However, how the survival, proliferation, migration, and differentiation of implanted NSC are influenced by endogenous neuronal activity remains unclear. In this work, we used optogenetic techniques to control the activity of striatal neurons and investigated how their activity affected the survival and migration of transplanted NSCs and overall neurological outcome after ischemic stroke. NSCs cultured from transgenic mice expressing fluorescent protein were transplanted into the peri-infarct region of the striatum after transient middle cerebral artery occlusion (tMCAO) surgery. The striatal neurons were excited or inhibited for 15 minutes daily via implanted optical fiber after tMCAO. The results revealed that mice which received NSC transplantation and optogenetic inhibition had smaller brain infarct volume and increased NSC migration compared to the NSC alone or PBS group (p<0.05). In contrast, mice which received NSC transplantation and optogenetic excitation showed no difference in infarct volume and neurological behavior improvement compared to the PBS control group. In vitro experiments further revealed that the conditioned media from excited GABAergic neurons reduced NSC viability through paracrine mechanisms.Conclusion. Optogenetic inhibition of striatal neuronal activity further improved neurological recovery after NSC transplantation at the subacute phase after brain ischemia.

In this work, we enhance the DC/AC resistive switching performance in AlOx memristors by using a two-technique bilayer approach. Compared to the single-layer memristors (W/AlOx or Al2O3/Pt), the dual-layer memristor (W/AlOx/AlOy/Pt) shows high uniformity in DC cycling (σ/μ 100), fast switching speed (20 ns), high endurance (107 cycles), and high-temperature stability (104 s at 125 °C). These performance enhancements are attributed to the localization of the conductive region after using a dual layer with different defect concentrations. Moreover, the W/AlOx/AlOy/Pt memristor exhibits stable III-bit multilevel storage capability by varying the amplitude of negative pulses. Our results provide an effective strategy to develop high-performance memristors for future memory and computing applications.

Here we present an open-source R package ‘meaRtools’ that provides a platform for analyzing neuronal networks recorded on Microelectrode Arrays (MEAs). Cultured neuronal networks monitored with MEAs are now being widely used to characterize in vitro models of neurological disorders and to evaluate pharmaceutical compounds. meaRtools provides core algorithms for MEA spike train analysis, feature extraction, statistical analysis and plotting of multiple MEA recordings with multiple genotypes and treatments. meaRtools functionality covers novel solutions for spike train analysis, including algorithms to assess electrode cross-correlation using the spike train tiling coefficient (STTC), mutual information, synchronized bursts and entropy within cultured wells. Also integrated is a solution to account for bursts variability originating from mixed-cell neuronal cultures. The package provides a statistical platform built specifically for MEA data that can combine multiple MEA recordings and compare extracted features between different genetic models or treatments. We demonstrate the utilization of meaRtools to successfully identify epilepsy-like phenotypes in neuronal networks from Celf4 knockout mice. The package is freely available under the GPL license (GPL>=3) and is updated frequently on the CRAN web-server repository. The package, along with full documentation can be downloaded from: https://cran.r-project.org/web/packages/meaRtools/.Author summaryCultured neuronal networks are widely used to study and characterize neuronal network activity. Among the many uses of neuronal cultures are the capabilities to evaluate neurotoxicity and the effects of pharmacological compounds on cellular physiology. Multi-well microelectrode arrays (MEAs) can collect high-throughput data from multiple neuronal cultures simultaneously, and thereby make possible hypotheses-driven inquiries into neurobiology and neuropharmacology. The analysis of MEA-derived information presents many computational challenges. High frequency data recorded simultaneously from hundreds of electrodes can be difficult to handle. The need to compare network activity across various drug treatments or genotypes recorded on the same plate from experiments lasting several weeks presents another challenge. These challenges inspired us to develop meaRtools; an MEA data analysis package that contains new methods to characterize network activity patterns, which are illustrated here using examples from a genetic mouse model of epilepsy. Among the highlights of meaRtools are novel algorithms designed to characterize neuronal activity dynamics and network properties such as bursting and synchronization, options to combine multiple recordings and use a robust statistical framework to draw appropriate statistical inferences, and finally data visualizations and plots. In summary, meaRtools provides a platform for the analyses of singular and longitudinal MEA experiments.

© 2018 Elsevier Inc., To identify novel genes associated with ALS, we undertook two lines of investigation. We carried out a genome-wide association study comparing 20,806 ALS cases and 59,804 controls. Independently, we performed a rare variant burden analysis comparing 1,138 index familial ALS cases and 19,494 controls. Through both approaches, we identified kinesin family member 5A (KIF5A) as a novel gene associated with ALS. Interestingly, mutations predominantly in the N-terminal motor domain of KIF5A are causative for two neurodegenerative diseases: hereditary spastic paraplegia (SPG10) and Charcot-Marie-Tooth type 2 (CMT2). In contrast, ALS-associated mutations are primarily located at the C-terminal cargo-binding tail domain and patients harboring loss-of-function mutations displayed an extended survival relative to typical ALS cases. Taken together, these results broaden the phenotype spectrum resulting from mutations in KIF5A and strengthen the role of cytoskeletal defects in the pathogenesis of ALS.

The lenticulostriate artery plays a vital role in the onset and development of cerebral ischemia. However, current imaging techniques cannot assess the in vivo functioning of small arteries such as the lenticulostriate artery in the brain of rats. Here, we report a novel method to achieve a high resolution multi-functional imaging of the cerebrovascular system using synchrotron radiation angiography, which is based on spatio-temporal analysis of contrast density in the arterial cross section. This method provides a unique tool for studying the sub-cortical vascular elasticity after cerebral ischemia in rats. Using this technique, we demonstrated that the vascular elasticity of the lenticulostriate artery decreased from day 1 to day 7 after transient middle cerebral artery occlusion in rats and recovered from day 7 to day 28 compared to the controls (p < 0.001), which paralleled with brain edema formation and inversely correlated with blood flow velocity (p < 0.05). Our results demonstrated that the change of vascular elasticity was related to the levels of brain edema and the velocity of focal blood flow, suggesting that reducing brain edema is important for the improvement of the function of the lenticulostriate artery in the ischemic brain.

Background and Purpose— Striatal GABAergic neuron is known as a key regulator in adult neurogenesis. However, the specific role of striatal GABAergic neuronal activity in the promotion of neurological recovery after ischemic stroke remains unknown. Here, we used optogenetic approach to investigate these effects and mechanism. Methods— Laser stimulation was delivered via an implanted optical fiber to inhibit or activate the striatal GABAergic neurons in Gad2-Arch-GFP or Gad2-ChR2-tdTomato mice (n=80) 1 week after 60-minute transient middle cerebral artery occlusion. Neurological severity score, brain atrophy volume, microvessel density, and cell morphological changes were examined using immunohistochemistry. Gene expression and protein levels of related growth factors were further examined using real-time polymerase chain reaction and Western blotting. Results— Inhibiting striatal GABAergic neuronal activity improved functional recovery, reduced brain atrophy volume, and prohibited cell death compared with the control ( P P P Conclusions— After ischemic stroke, optogenetic inhibition of GABAergic neurons upregulated bFGF expression by endothelial cells and promoted neurobehavioral recovery, possibly orchestrated by astrocytes. Optogenetically inhibiting neuronal activity provides a novel approach to promote neurological recovery.

Rationale: Cerebral ischemia upregulates aquaporin-4 expression, increases blood-brain barrier (BBB) permeability, and induces brain edema. Mesenchymal stem cells (MSCs) can repress inflammatory cytokines and show great potential for ischemic stroke therapy. However, the effect of MSCs regarding the protection of ischemia-induced BBB break down is unknown. Objective: We test whether MSCs therapy protects BBB integrity and explore the molecular mechanisms of aquaporin-4 on BBB integrity. Methods and Results: Two hundred and twenty-eight adult CD1 male mice underwent 90 minutes transient middle cerebral artery occlusion and received 2 × 105 MSCs intracranial transplantation. The neurological severity score was improved and both ischemia-induced brain edema and BBB leakage were reduced in MSC-treated mice. MSCs therapy reduced astrocyte apoptosis and inhibited ischemia-induced aquaporin-4 upregulation. In addition, small-interfering RNA knockdown of aquaporin-4 after cerebral ischemia effectively reduced aquaporin-4 expression, brain edema, BBB leakage, and astrocyte apoptosis. Conditional medium from lipopolysaccharide (LPS)-activated microglia enhanced aquaporin-4 expression, p38 and JNK phosphorylation, and apoptosis of cultured astrocytes. MSC treatment reduced the expression of inflammatory cytokines in LPS-activated microglia, and subsequently reduced aquaporin-4 expression and apoptosis of astrocytes. Knockdown of aquaporin-4 in cultured astrocytes also reduced apoptosis. Treatment with p38 and JNK inhibitors showed that p38, but not the JNK signaling pathway, was responsible for the aquaporin-4 upregulation. Conclusion: MSCs protected BBB integrity by reducing the apoptosis of astrocytes after ischemic attack, which was due to the attenuation of inflammatory response and downregulation of aquaporin-4 expression via p38 signaling pathway. Stem Cells 2014;32:3150–3162

ultrasound characteristics of the material microstructure nondestructive evaluation of its experimental sample TC4 titanium alloy microstructure of two-phase microstructure:. Α-phase and α + β phase. Phase material microstructure and grain area ratio directly affects the material uniformity, uniformity of material differences caused by different grain characteristics which ultrasound parameters. Flooding experiments using ultrasonic method for measuring the material longitudinal wave velocity; measuring the sound attenuation coefficient, combined with the area ratio of the sample α using multiple pulse method. Associated with the sample area ratio between the speed of sound and sound attenuation analysis of α-phase titanium alloy TC4 through the material properties of ultrasound to characterize the degree of uniformity of the material microstructure, nondestructive evaluation of materials in order to achieve the purpose of the microstructure.

Background and Purpose— Acute interventions of stroke are often challenged by a narrow treatment window. In this study, we explore treatments in the postacute phase of stroke with wider windows of opportunity. We investigated the effects of stromal cell–derived factor (SDF-1α) in neurovascular recovery during the postacute phase and downstream signaling pathways, underlying SDF-1α–mediated neurovascular recovery. Methods— Adult male Institute of Cancer Research (ICR) mice underwent middle cerebral artery occlusion. One week after middle cerebral artery occlusion, the animals received stereotactic injection of adenoassociated virus (AAV) carrying SDF-1α gene as treatment or AAV-green fluorescent protein as control and were monitored for 5 weeks. Neurobehavioral outcomes were evaluated, and brain atrophy was measured. Neurogenesis and angiogenesis were examined. The proliferation and migration of neural progenitor cells were evaluated. Downstream pathways of SDF-1α were investigated. Inflammatory response was monitored. Results— Neurobehavioral outcomes were improved, and brain atrophy was greatly reduced for ≤5 weeks in AAV-SDF-1α groups when compared with the control. SDF-1 receptor CXCR4 was upregulated and colocalized with neural and endothelial progenitor cells. The number of nestin + and doublecortin + /bromodeoxyuridine + cells in the subventricular zone, doublecortin + and neuron + /bromodeoxyuridine + cells in the perifocal region, and cluster of differentiation (CD)31 + and bromodeoxyuridine + /CD31 + microvessels are also significantly increased in AAV-SDF-1α groups. Administration of CXCR4 antagonist AMD3100 eliminated the beneficial effects of SDF-1α. SDF-1α/CXCR4 interaction activated AKT, extracellular signal-regulated kinases (ERK), and P38 mitogen-activated protein kinase (MAPK) signaling pathways but not the c-Jun N-terminal kinase (JNK) pathway. Conclusions— SDF-1α promoted neurogenesis and angiogenesis during the postacute phase of ischemia without eliciting an inflammatory response. AAV-SDF-1α expression represents a promising avenue for ischemic stroke therapy with a wider treatment window.

The Flavivirus genus contains several arthropod-borne viruses that pose global health threats, including dengue viruses (DENV), yellow fever virus (YFV), and Zika virus (ZIKV). In order to understand how these viruses replicate in human cells, we previously conducted genome-scale RNA interference screens to identify candidate host factors. In these screens, we identified ribosomal proteins RPLP1 and RPLP2 (RPLP1/2) to be among the most crucial putative host factors required for DENV and YFV infection. RPLP1/2 are phosphoproteins that bind the ribosome through interaction with another ribosomal protein, RPLP0, to form a structure termed the ribosomal stalk. RPLP1/2 were validated as essential host factors for DENV, YFV, and ZIKV infection in two human cell lines: A549 lung adenocarcinoma and HuH-7 hepatoma cells, and for productive DENV infection of Aedes aegypti mosquitoes. Depletion of RPLP1/2 caused moderate cell-line-specific effects on global protein synthesis, as determined by metabolic labeling. In A549 cells, global translation was increased, while in HuH-7 cells it was reduced, albeit both of these effects were modest. In contrast, RPLP1/2 knockdown strongly reduced early DENV protein accumulation, suggesting a requirement for RPLP1/2 in viral translation. Furthermore, knockdown of RPLP1/2 reduced levels of DENV structural proteins expressed from an exogenous transgene. We postulate that these ribosomal proteins are required for efficient translation elongation through the viral open reading frame. In summary, this work identifies RPLP1/2 as critical flaviviral host factors required for translation. IMPORTANCE Flaviviruses cause important diseases in humans. Examples of mosquito-transmitted flaviviruses include dengue, yellow fever and Zika viruses. Viruses require a plethora of cellular factors to infect cells, and the ribosome plays an essential role in all viral infections. The ribosome is a complex macromolecular machine composed of RNA and proteins and it is responsible for protein synthesis. We identified two specific ribosomal proteins that are strictly required for flavivirus infection of human cells and mosquitoes: RPLP1 and RPLP2 (RPLP1/2). These proteins are part of a structure known as the ribosomal stalk and help orchestrate the elongation phase of translation. We show that flaviviruses are particularly dependent on the function of RPLP1/2. Our findings suggest that ribosome composition is an important factor for virus translation and may represent a regulatory layer for translation of specific cellular mRNAs.

Striatal neurons regulate the activity of neural progenitor cells in the subventricular zone, but the effect of striatal neuronal activity on neurogenesis after ischemic stroke is unclear. In this study, we used optogenetic tools to investigate the impact of striatal neuronal activity on the neurogenesis and functional recovery after cerebral ischemia. We transfected striatal neurons with channelrhodopsin-2 or halorhodopsin from Natronomonas so that they can be excited by 473 nm laser or inhibited by 594 nm laser, respectively. Neural inhibition but not excitation at 4–7 days after middle cerebral artery occlusion resulted in reduced atrophy volume (6.8 ± 0.7 vs 8.5 ± 1.2 mm3, p +, BrdU+/doublecortin+ and BrdU+/NeuN+ cells ( p

Microelectrode array (MEA) technology enables advanced drug screening and “disease-in-a-dish” modeling by measuring the electrical activity of cultured networks of neural or cardiac cells. Recent developments in human stem cell technologies, advancements in genetic models, and regulatory initiatives for drug screening have increased the demand for MEA-based assays. In response, Axion Biosystems previously developed a multiwell MEA platform, providing up to 96 MEA culture wells arrayed into a standard microplate format. Multiwell MEA-based assays would be further enhanced by optogenetic stimulation, which enables selective excitation and inhibition of targeted cell types. This capability for selective control over cell culture states would allow finer pacing and probing of cell networks for more reliable and complete characterization of complex network dynamics. Here we describe a system for independent optogenetic stimulation of each well of a 48-well MEA plate. The system enables finely graded control of light delivery during simultaneous recording of network activity in each well. Using human induced pluripotent stem cell (hiPSC) derived cardiomyocytes and rodent primary neuronal cultures, we demonstrate high channel-count light-based excitation and suppression in several proof-of-concept experimental models. Our findings demonstrate advantages of combining multiwell optical stimulation and MEA recording for applications including cardiac safety screening, neural toxicity assessment, and advanced characterization of complex neuronal diseases.

Polymorphisms near the interferon lambda 3 (IFNL3) gene strongly predict clearance of hepatitis C virus (HCV) infection. We analyzed a variant (rs4803217 G/T) located within the IFNL3 mRNA 3′ untranslated region (UTR); the G allele (protective allele) is associated with elevated therapeutic HCV clearance. We show that the IFNL3 3′ UTR represses mRNA translation and the rs4803217 allele modulates the extent of translational regulation. We analyzed the structures of IFNL3 variant mRNAs at nucleotide resolution by SHAPE-MaP. The rs4803217 G allele mRNA forms well-defined 3′ UTR structure while the T allele mRNA is more dynamic. The observed differences between alleles are among the largest possible RNA structural alterations that can be induced by a single nucleotide change and transform the UTR from a single well-defined conformation to one with multiple dynamic interconverting structures. These data illustrate that non-coding genetic variants can have significant functional effects by impacting RNA structure.

Effective setting strategies using Monte Carlo simulation are presented to mitigate the irradiation damage in synchrotron radiation microangiography (SRA). A one-dimensional mouse head model and a segmented voxel phantom mouse head were simulated using theEGSnrc/DOSXYZnrccode to investigate the dose enhancement effect of an iodine contrast agent irradiated by a monochromatic synchrotron radiation source. The influence of the iodine concentration, vessel width and depth, protection with and without the skull layer, and various incident X-ray energies were all simulated. The dose enhancement effect and the absolute dose based on the segmented voxel mouse head phantom were evaluated. The dose enhancement ratio depended little on the irradiation depth, but strongly and linearly increasing on iodine concentration. The protection given by the skull layer cannot be ignored in SRA because a 700 µm-thick skull can decrease the dose by 10%. The incident X-ray energy can affect the dose significantly. Compared with a dose of 33.2 keV for 50 mgI ml−1, a dose of 32.7 keV decreased by 38%, whereas a dose of 33.7 keV increased by 69.2% and the variation strengthened more with enhanced iodine concentration. The segmented voxel mouse head phantom also showed that the average dose enhancement effect and the maximal voxel dose per photon depended little on the iodine voxel volume ratio but strongly on the iodine concentration. To decrease the damage caused by the dose in SRA, a high-Zcontrast agent should be used as little as possible and irradiation of the injection site of the contrast agent should be avoided immediately after the injection. The fragile vessel containing iodine should avoid being closely irradiated. Avoiding irradiating through a thin (or no) skull region, or attaching a thin equivalent material on the outside for protection are better methods. An incident X-ray energy as low as possible should be used as long as the SRA image quality is ensured. The use of the synergetic and synchronous shuttering technique in SRA is also very critical in order to effectively shorten the accumulative irradiation time inin vivoanimal irradiation experiments.

induced with an intravenous injection of 50 μg of fentanyl and a target-controlled infusion of propofol titrated to a bispectral index value between 40 and 60. A nasopharyngeal airway was inserted to maintain the patent airway and to monitor the respiratory rate. The patient breathed spontaneously through a facemask, with supplemental oxygen to keep his oxygen saturation values above 90% during the procedure. Subsequently, he was placed in the right lateral decubitus position, with slight flexion at the level of the midchest. A 3-port thoracoscopic approach was used. A 1.5-cm thoracoscopic port was first created at the seventh intercostal space in the midaxillary line after infiltrating with 2% lidocaine. The operated lung collapsed spontaneously after introducing an iatrogenic open pneumothorax. Then, regional anesthesia was completed by injecting 0.5% bupivacaine through the second to the eighth intercostal spaces (1.5 mL for each intercostal nerve) under direct thoracoscopic vision. A left-sided vagal block was induced by infiltrating the aortopulmonary window with 3 mL of 0.5% bupivacaine, which effectively inhibited the cough reflex during surgery. Although the operated lung was not fully collapsed due to residual trapped air, it did not preclude the ongoing procedure. Pulmonary vessels, bronchus, and incomplete fissure of the lingual segment then were divided with endoscopic staplers. The patient’s ventilation was assisted with a facemask with positive pressure to partially reinflate the operated lung and to confirm the lingual segmental plan immediately before the lingual bronchus and the incomplete fissure were divided. The procedure was completed after mediastinal nodal dissection, with a 28-French chest tube placed for pleural drainage. During the surgery, the patient remained hemodynamically stable, with an undisturbed breathing pattern. Although his respiratory rate transiently increased from 18 to 28 breaths/min after introducing the open pneumothorax, it was soon ameliorated by the administration of an additional 25 μg of fentanyl and vagal blockade without jeopardizing the planned procedure and his respiratory sufficiency. The peak partial pressure of arterial carbon dioxide during single-lung breathing was 46 mmHg, which returned to 35 mmHg at the end of surgery while he was fully awake. The partial pressures of arterial oxygen was 381 mmHg before single-lung breathing and 98 mmHg during single-lung breathing under the same supplemental oxygen fraction (FIO2:0.5). Pulse oxygen saturation was above 98% for the entire 100-minute procedure. The final pathologic report revealed a second primary lung adenocarcinoma without nodal involvement (T1aN0), consistent with stage IA. The patient recovered well and was discharged uneventfully on postoperative day 5.

The receptor for advanced glycation end products (RAGE) and its proinflammatory S100/calgranulin ligands are enriched in joints of subjects with rheumatoid arthritis (RA) and amplify the immune/inflammatory response. In a model of inflammatory arthritis, blockade of RAGE in mice immunized and challenged with bovine type II collagen suppressed clinical and histologic evidence of arthritis, in parallel with diminished levels of TNF-alpha, IL-6, and matrix metalloproteinases (MMP) 3, 9 and 13 in affected tissues. Allelic variation within key domains of RAGE may influence these proinflammatory mechanisms, thereby predisposing individuals to heightened inflammatory responses. A polymorphism of the RAGE gene within the ligand-binding domain of the receptor has been identified, consisting of a glycine to serine change at position 82. Cells bearing the RAGE 82S allele displayed enhanced binding and cytokine/MMP generation following ligation by a prototypic S100/calgranulin compared with cells expressing the RAGE 82G allele. In human subjects, a case-control study demonstrated an increased prevalence of the 82S allele in patients with RA compared with control subjects. These data suggest that RAGE 82S upregulates the inflammatory response upon engagement of S100/calgranulins, and, thereby, may contribute to enhanced proinflammatory mechanisms in immune/inflammatory diseases.