Synaptotagmin-11 regulates immune functions of microglia in vivo.

Membrane trafficking pathways mediate key microglial activities such as cell migration, cytokine secretion, and phagocytosis. However, the underlying molecular mechanism remains poorly understood. Previously, we found that synaptotagmin-11 (Syt11), a non-Ca2+ -binding Syt associated with Parkinson's disease (PD) and schizophrenia, inhibits cytokine release and phagocytosis in primary microglia. Here we reported the in vivo function of Syt11 in microglial immune responses using an inducible microglia-specific Syt11-conditional-knockout (cKO) mouse strain. Syt11-cKO resulted in activation of microglia and elevated mRNA levels of IL-6, TNF-α, IL-1ß, and iNOS in various brain regions under both resting state and LPS-induced acute inflammation state in adult mice. In a PD mouse model generated by microinjection of preformed α-synuclein fibrils into the striatum, a reduced number of microglia migrated toward the injection sites and an enhanced phagocytosis of α-synuclein fibrils by microglia were found in Syt11-cKO mice. To understand the molecular mechanism of Syt11 function, we identified its direct binding proteins vps10p-tail-interactor-1a (vti1a) and vti1b. The linker domain of Syt11 interacted with both proteins and a peptide derived from it competitively inhibited the interaction of Syt11 with vti1a/vti1b in vitro and in cells. Importantly, application of this peptide induced more cytokine secretion in wild-type microglia upon LPS treatment, phenocopying defects in Syt11 knockdown cells. Altogether, we propose that Syt11 inhibits microglial activation in vivo and regulates cytokine secretion through interactions with vti1a and vti1b.

Double-sided telecentric zoom optical system using adaptive liquid lenses.

In this paper, the adaptive liquid (AL) lenses are introduced into the double-sided telecentric zoom system, which could greatly decrease the mechanical motion group compared with the traditional zoom system, and only one movable aperture stop (STO) is retained. Firstly, this paper derived the Gaussian brackets used in this system, and we found the appropriate screening method to get the suitable initial structure parameters from the solution space. Then we used the lens module design method to create the initial system. Finally, we used CODEV to further optimize the system, and we got an excellent design result, which controlled the telecentricity of both sides within 0.1°, the distortion was controlled within 0.5%, and the MTF of each zoom configuration above 0.3. This optical system has high application potential and value in the field of precision machine vision. The design method proposed in this article can effectively solve the problem that the zoom system using adaptive liquid lenses lacks the initial structure.

Optical system design of double-sided telecentric microscope with high numerical aperture and long working distance.

The working distance of the high numerical aperture visible video microscope is extremely short, which greatly limits its application scenarios. To solve this problem, this paper proposes an unobstructed design method of double-sided telecentric microscope with high numerical aperture and long working distance. First, aiming at the obstruction problem of the image-side telecentric catadioptric microscope objective, the structure of the catadioptric optical system is improved. Then, the aspheric design method based on the best aberration compensation is analyzed theoretically to better correct the primary aberration of the high-numerical aperture microscope objective. Finally, a double-sided telecentric microscope optical system with a numerical aperture (NA) of 0.8 and a working distance of 10.0 mm was designed, which is composed of a spherical reflector, a beam splitter plate, a collimating lens group, and an image-side telecentric eyepiece optical system. The design results show that the imaging resolution of this high numerical aperture video microscope is as high as 0.42 µm, and the microscope has a magnification of about 220× for the image with 1080P (1920 × 1080 pixels) resolution. This double-sided telecentric microscope has the advantages of a large field of view, compact structure, good stray light suppression ability, and manufacturability, and has high practical value in the field of high-precision measurement and detection.

Design method of dual-band synchronous zoom microscope optical system based on coaxial Kohler illumination.

The intrinsic properties of the observed object are closely related to its spectral information, to extend the imaging spectral range of a continuous zoom microscope to obtain more detailed intrinsic properties of the object, this paper proposes a design method of dual-band simultaneous zoom microscope optical system based on the coaxial Koehler uniform illumination. First, the imaging principle of the dual-band simultaneous zoom microscope optical system is theoretically analyzed, and we propose to split the front fixed group of the zoom system into a collimation lens group and a converging lens group to realize the compact design of the system. Then, two different rear fixed groups are used to correct the residual aberration, and a method for solving the initial structure of the dual-band simultaneous zoom microscope optical system is proposed. Finally, a dual-band synchronous zoom microscope optical system is designed using the method proposed in this paper. The design results show that the imaging magnification of the visible (VIS) band is -0.4 to -4.0, the simultaneous imaging magnification ranges are -0.4 to -0.8 in the VIS and short-wave infrared (SWIR) bands, and the magnification difference of its simultaneous zoom imaging is less than 1.25%. In addition, the system has the advantages of good imaging quality, clever design of coaxial illumination, and compact structure, thus verifying the feasibility of the design method.

Design of the dual-band shared-aperture asynchronous zoom optical system using focus tunable lenses.

To meet the increasing demand of the current market and the diversity of application scenarios, combine the zoom system and the multi-band shared-aperture system, and fully harness their respective advantages, this paper proposes a dual-band shared-aperture asynchronous zoom optical system using focus tunable lenses (FTLs). To address the lack of available patents for such systems, we designed a sub-system simultaneous iterative optimization algorithm to calculate the initial structure parameters. This synchronous iterative optimization approach can strengthen the connection between sub-systems and compensate for the shortcomings of current mainstream design methods. The initial structure constructed in this way has a good performance in terms of structural stability and optimization potential. Based on these methods, we successfully designed an optical system that can work in both VIS and NIR bands, and the two sub-systems can zoom independently. The design results possess good performance in terms of distortion control, aberration correction, and volume control.

Cooper Pair Excitation Mediated by a Molecular Quantum Spin on a Superconducting Proximitized Gold Film.

Breaking a correlated pair in a superconductor requires an even number of fermions providing at least twice the pairing energy Δ. Here, we show that a single tunneling electron can also excite a pair breaking excitation in a proximitized gold film in the presence of magnetic impurities. Combining scanning tunneling spectroscopy with theoretical modeling, we map the excitation spectrum of an Fe-porphyrin molecule on the Au/V(100) proximitized surface into a manifold of entangled Yu-Shiba-Rusinov and spin excitations. Pair excitations emerge in the tunneling spectra as peaks outside the spectral gap only in the strong coupling regime, where the presence of a bound quasiparticle in the ground state ensures the even fermion parity of the excitation. Our results unravel the quantum nature of magnetic impurities on superconductors and demonstrate that pair excitations unequivocally reveal the parity of the ground state.

High-precision calibration to zoom lens of optical measurement machine based on FNN.

We proposed a calibration method for high-precision zoom lenses of optical measurement machines based on Fully Connected Neural Network (FNN), using a 5-layer neural network instead of a camera calibration model, to achieve continuous calibration of zoom lenses at any zoom setting by calibrating typical zooms. From the experimental verification, the average calibration error of this method is 9.83×10-4mm and the average measurement error at any zoom setting is 0.01317mm. The overall calibration precision is better than that of Zhang's calibration method and can meet the application requirements of a high-precision optical measurement machine. The method proposed in this paper provided a new solution and a new idea for the calibration of zoom lenses, which can be widely used in the fields of precision parts inspection and machine-vision measurement.

Activation of Topological Defects Induces a Brittle-to-Ductile Transition in Epithelial Monolayers.

Epithelial monolayers are subjected to various mechanical forces, such as stretching, shearing, and compression. Thus, its mechanical response to external loadings is essential for its biological functions. However, the mechanism of the fracture failure of the epithelial monolayer remains poorly understood. Here, by introducing a new type of topological transition, i.e., detach transition or T4 transition, we develop a modified cellular vertex model to investigate the rupture of the cell monolayer. Interestingly, we find a brittle-to-ductile transition in epithelial monolayers, which is controlled by the mechanical properties of single cells and cell-cell contacts. We reveal that the external loadings can activate cell rearrangement in ductile cell monolayers. The plastic deformation results from the nucleation and propagation of "pentagon-heptagon defects" in analogy with the topological defects commonly seen in 2D materials. By using a simplified four-cell model, we further demonstrate that the brittle-to-ductile transition is induced by the competition between cell rearrangement and cell detachment. Our work provides a new theoretical framework to study the rupture of living tissues and may have important implications for many other biological processes, such as wound healing and tissue morphogenesis.

Design of a Concentric Multi-Scale Zoom Optical System Based on Wide Object Distance and High-Precision Imaging.

To effectively balance the trade-off between a large field of view (FOV) and high resolution of an optical system, as well as to solve the problem of image stitching misalignment after focusing, firstly, this paper conducts a theoretical analysis of the design principle of the concentric multi-scale optical system and the causes of image stitching misalignment after focusing. Secondly, the design idea of using a combination structure of a two-layer front concentric imaging group and an image-space telecentric relay imaging array and then a joint full-motion zoom relay imaging system is proposed. Finally, an image-space telecentric two-step zoom concentric multi-scale optical system with a 7 × 7 relay imaging array is designed. The FOV of this optical system is 60° × 45°; the focal lengths are 50 mm and 100 mm for the center channel and 50 mm for the other channels. This concentric multi-scale zoom system has the advantages of both high-precision imaging stitching with a wide object distance and high-resolution imaging, which makes up for the defects of the conventional concentric multi-scale optical system, making it a promising application in the fields of aviation and security.

Synaptotagmin-11 inhibits spontaneous neurotransmission through vti1a.

Recent work has revealed that spontaneous release plays critical roles in the central nervous system, but how it is regulated remains elusive. Here, we report that synaptotagmin-11 (Syt11), a Ca2+ -independent Syt isoform associated with schizophrenia and Parkinson's disease, suppressed spontaneous release. Syt11-knockout hippocampal neurons showed an increased frequency of miniature excitatory post-synaptic currents while over-expression of Syt11 inversely decreased the frequency. Neither knockout nor over-expression of Syt11 affected the average amplitude, suggesting the pre-synaptic regulation of spontaneous neurotransmission by Syt11. Glutathione S-transferase pull-down, co-immunoprecipitation, and affinity-purification experiments demonstrated a direct interaction of Syt11 with vps10p-tail-interactor-1a (vti1a), a non-canonical SNARE protein that maintains spontaneous release. Importantly, knockdown of vti1a reversed the phenotype of Syt11 knockout, identifying vti1a as the main target of Syt11 inhibition. Domain analysis revealed that the C2A domain of Syt11 bound vti1a with high affinity. Consistently, expression of the C2A domain alone rescued the phenotype of elevated spontaneous release in Syt11-knockout neurons similar to the full-length protein. Altogether, our results suggest that Syt11 inhibits vti1a-containing vesicles during spontaneous release.

Facile synthesis of rGO-MoS2-Ag nanocomposites with long-term antimicrobial activities.

The abuse of antibiotics has led to the emergence of numerous super resistant bacteria, which pose a serious threat to public health. Developing nanomaterials with novel modes of bactericidal activity offers the promise of fighting pathogens without the risk of causing drug resistances. Here, we used reduced graphene oxide (rGO), bulk molybdenum disulfide (MoS2) and silver nitrate (AgNO3) to synthesize a ternary nanocomposite, rGO-MoS2-Ag, via a simple one-pot method. The resulting rGO-MoS2-Ag presented as crumpled and sheet-like structures decorated with Ag nanoparticles. The minimum inhibitory concentration and minimum bactericidal concentration of rGO-MoS2-Ag against Escherichia coli were 50 and 100 µg ml-1, while Staphylococcus aureus reacted only to twice higher concentrations of 100 and 200 µg ml-1, respectively. Notably, rGO-MoS2-Ag exhibited better antibacterial activity towards E. coli and S. aureus than rGO, MoS2, or rGO-MoS2. This result can be attributed to the excellent performance of rGO-MoS2-Ag in destroying the bacterial cell membrane and inducing the generation of reactive oxygen species. The Ag+ ion release of rGO-MoS2-Ag was delayed, endowing the nanocomposite with long-term antibacterial capabilities and better biosafety. Our results indicate that the as-prepared rGO-MoS2-Ag has promising potential for application in biomedicine and public health.

Significant Effect of Evaporation Process on the Reaction of Sulfamethoxazole with Manganese Oxide.

Soil in the vadose zone is an important sink for antibiotics. However, previous studies have examined only the degradation of antibiotics in soil slurry systems, which were largely different from real-world unsaturated soil environments. Whether the same transformation mechanisms apply to unsaturated soil systems has been a question. Here, the degradation of sulfamethoxazole (SMX) by manganese dioxide (γ-MnO2) in both suspension systems and evaporation processes were examined. Results show that the slow degradation of SMX in the suspension system can be significantly promoted as the water gradually evaporates. SMX degraded differently in evaporation as compared to suspension systems because of the quenching effect of generated Mn2+. Transformation products of SMX in both systems also showed different toxicity toward Escherichia coli because of different evolutions of intermediates. This study has strong implications for the assessment and prediction of the transformation and fate of antibiotics in natural soil environments.

Geometric Asymmetry Induces Upper Limit of Mitotic Spindle Size.

Proper organelle size is critical for many cell functions. However, how cells sense and control their organelle size remains elusive. Here, we develop a general model to study the size control of mitotic spindles by considering both extrinsic and intrinsic factors, such as the limited number of building blocks of the spindle, the interaction between the spindle and cell boundary, the DNA content, the forces generated by various molecular motors, and the dynamics of microtubules. We show that multiple pairs of chromatids, two centrosomes, and microtubules can self-assemble to form a mitotic spindle robustly. We also show that the boundary-sensing and volume-sensing mechanisms coexist in small cells, but both break down in large cells. Strikingly, we find that the upper limit of spindle length naturally arises from the geometric asymmetry of the spindle structure. Thus, our findings reveal, to our knowledge, a novel intrinsic mechanism that limits the organelle size.

Sinomenine activates astrocytic dopamine D2 receptors and alleviates neuroinflammatory injury via the CRYAB/STAT3 pathway after ischemic stroke in mice.

BACKGROUND: Astrocyte-mediated neuroinflammation plays a critical role in ischemic stroke-induced secondary cerebral injury. Previous studies have suggested that the dopamine D2 receptor (DRD2) acts as a key target in regulating the neuroinflammatory response. However, the underlying molecular mechanisms are still unknown, and effective DRD2 agonists are lacking. In the present study, we examined the anti-inflammatory and neuroprotective effects of sinomenine (Sino), a monomeric compound with potential immunoregulatory properties in nervous system. METHODS: TTC staining, apoptosis assay, evaluation of brain edema, and neurological assessment were performed in the middle cerebral artery occlusion (MCAO) mouse model. Primary astrocytes exposed to oxygen glucose deprivation (OGD) were used in the in vitro experiments. Quantitative PCR was applied to assess the levels of inflammatory cytokines. Multi-labeling immunofluorescence, Western blot, co-immunoprecipitation, and electrophoretic mobility shift assay (EMSA) were also used to investigate the molecular mechanisms underlying the Sino-mediated anti-inflammatory effects in vivo and in vitro. RESULTS: Sino remarkably attenuated the cerebral infarction and neuronal apoptosis, reduced the levels of inflammatory cytokines, and alleviated neurological deficiency in MCAO mice. Sino significantly inhibited astrocytic activation and STAT3 phosphorylation as well as increased DRD2 and αB-crystallin (CRYAB) expression after MCAO. In vitro, Sino blocked OGD-induced activation of STAT3 and generation of pro-inflammatory cytokines in primary astrocytes, and these effects were significantly abolished by either DRD2 or CRYAB knockdown. Additionally, Sino induced up-regulation and nuclear translocation of CRYAB in astrocytes and enhanced the interaction between CRYAB and STAT3, which further inhibited the activation and DNA-binding activity of STAT3. CONCLUSIONS: Our study demonstrates that Sino activates astrocytic DRD2 and thereby suppresses neuroinflammation via the CRYAB/STAT3 pathway, which sheds some light on a promising therapeutic strategy for ischemic stroke.

Epigenetic Suppression of GADs Expression is Involved in Temporal Lobe Epilepsy and Pilocarpine-Induced Mice Epilepsy.

Recent studies have shown that histone acetylation is involved with the regulation of enzyme glutamate decarboxylases (GADs), including GAD67 and GAD65. Here, we investigated the histone acetylation modifications of GADs in the pathogenesis of epilepsy and explored the therapeutic effect of a novel second-generation histone deacetylase inhibitor (HDACi) JNJ-26481585 in epilepsy animals. We revealed the suppression of GADs protein and mRNA level, and histone hypoacetylation in patients with temporal lobe epilepsy and pilocarpine-induced epilepsy mice model. Double-immunofluorescence also indicated that the hypoacetyl-H3 was located in hippocampal GAD67/GAD65 positive neurons in epilepsy mice. JNJ-26481585 significantly reversed the decrease of the GAD67/GAD65 both protein and mRNA levels, and the histone hypoacetylation of GABAergic neurons in epilepsy mice. Meanwhile, single-cell real-time PCR performed in GFP-GAD67/GAD65 transgenic mice demonstrated that JNJ-26481585 induced increase of GAD67/GAD65 mRNA level in GABAergic neurons. Furthermore, JNJ-26481585 significantly alleviated the epileptic seizures in mice model. Together, our findings demonstrate inhibition of GADs gene via histone acetylation plays an important role in the pathgenesis of epilepsy, and suggest JNJ-26481585 as a promising therapeutic strategy for epilepsy.

Blood-brain barrier permeability change and regulation mechanism after subarachnoid hemorrhage.

We aimed to investigate the blood brain barrier (BBB) change caused by subarachnoid hemorrhage (SAH) and to explore the molecular mechanisms of acute brain injury after SAH. The SD rat model of SAH was firstly established by endovascular filament perforation technique. The changes of regional cerebral blood flow (rCBF), BBB permeability and ultrastructure of brain tissue at different time points after SAH were respectively observed by Doppler flowmetry, evans blue extravasation and transmission electron microscopy. Meanwhile, the expression changes of Claudin-5, Occludin, Zo-1 and Caveolin-1 were detected by immunohistochemistry and Western blot. Furthermore, the expressions of Akt, P-Akt and Foxo1A were also measured by Western blot. The change of BBB permeability showed two peaks at 3 and 72 h after SAH, corresponding to the change of rCBF. The BBB tight junction opening can be observed after SAH, and the largest opening was occurred at 3 h and 72 h. There was no significant change in Caveolin-1, Claudin-5 and Akt expressions after SAH (P > 0.05), while Zo-1 and Occludin were significantly down-regulated (P < 0.05). The expression of P-Akt was obviously reduced at 30 min and then increased at 1 and 24 h, while Foxo1A was up-regulated at 1 and 24 h after SAH (P < 0.05). Down-regulated Zo-1 and Occludin, as well as Akt/FOXO signaling pathway may be involved in the regulation of tight junction opening and the BBB permeability in the early stage after SAH.

Unbiased organism-agnostic and highly sensitive signal peptide predictor with deep protein language model.

Signal peptides (SPs) are essential to target and transfer transmembrane and secreted proteins to the correct positions. Many existing computational tools for predicting SPs disregard the extreme data imbalance problem and rely on additional group information of proteins. Here we introduce Unbiased Organism-agnostic Signal Peptide Network (USPNet), an SP classification and cleavage-site prediction deep learning method. Extensive experimental results show that USPNet substantially outperforms previous methods on classification performance by 10%. An SP-discovering pipeline with USPNet is designed to explore unprecedented SPs from metagenomic data. It reveals 347 SP candidates, with the lowest sequence identity between our candidates and the closest SP in the training dataset at only 13%. In addition, the template modeling scores between candidates and SPs in the training set are mostly above 0.8. The results showcase that USPNet has learnt the SP structure with raw amino acid sequences and the large protein language model, thereby enabling the discovery of unknown SPs.

Eliminating Primacy Bias in Online Reinforcement Learning by Self-Distillation.

Excessive invalid explorations at the beginning of training lead deep reinforcement learning process to fall into the risk of overfitting, further resulting in spurious decisions, which obstruct agents in the following states and explorations. This phenomenon is termed primacy bias in online reinforcement learning. This work systematically investigates the primacy bias in online reinforcement learning, discussing the reason for primacy bias, while the characteristic of primacy bias is also analyzed. Besides, to learn a policy generalized to the following states and explorations, we develop an online reinforcement learning framework, termed self-distillation reinforcement learning (SDRL), based on knowledge distillation, allowing the agent to transfer the learned knowledge into a randomly initialized policy at regular intervals, and the new policy network is used to replace the original one in the following training. The core idea for this work is distilling knowledge from the trained policy to another policy can filter biases out, generating a more generalized policy in the learning process. Moreover, to avoid the overfitting of the new policy due to excessive distillations, we add an additional loss in the knowledge distillation process, using L2 regularization to improve the generalization, and the self-imitation mechanism is introduced to accelerate the learning on the current experiences. The results of several experiments in DMC and Atari 100k suggest the proposal has the ability to eliminate primacy bias for reinforcement learning methods, and the policy after knowledge distillation can urge agents to get higher scores more quickly.

Pink1 deficiency enhances neurological deficits and inflammatory responses after intracerebral hemorrhage in mice.

Pink1 (PTEN-induced putative kinase 1) is a protein associated with maintaining mitochondrial function and integrity and has been reported to mediate neurodegeneration and neuroinflammation. While the role of Pink1 in intracerebral hemorrhage (ICH)-related neurological deficits and inflammatory responses is not deciphered. Congenic blood was transfused into the left corpus striatum to construct the ICH model in C57/BL6 wild-type (WT) and Pink1-/- mice. The relative expression of Pink1, monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein (MIP)-2, tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, Cd86, nitric oxide synthase 2 (Nos2), Cd206, arginase 1 (Arg-1), and IL-10 was detected with qRT-PCR, Western blotting, or ELISA. Mouse neurological deficit scores (mNSS) and water content were detected, and an open-field test was performed to assay anxiety-like behavior. Remarkably decreased Pink1 expression and increased MIP-2, IL-1ß, MCP-1, and TNF-α expression were observed after 12 â€‹h, 24 â€‹h, 48 â€‹h, 72 â€‹h, and 7 â€‹d post-ICH induction in the ipsilateral injury hemispheres. Pink1 deficiency could further up-regulate mNSS scores, brain water content, MIP-2, MCP-1, IL-1ß, and TNF-α in the ipsilateral injury hemispheres. On the other hand, Pink1 deficiency could decrease the number of center cross, the velocity, and the total distance traveled in open field test. Pink1 deficiency could further up-regulate the mRNA levels of pro-inflammatory (M1) molecules (Cd86, Nos2), and down-regulate the relative expression of anti-inflammatory (M2) molecules (Cd206, Arg-1, and IL-10). Pink1 deficiency deteriorates neurological deficits and inflammatory responses after ICH, which can be considered as a treatment target.

Serotonergic neurotransmission mediated cognitive dysfunction in two mouse models of sepsis-associated encephalopathy.

BACKGROUND: Patients with sepsis-associated encephalopathy (SAE) often exhibit cognitive impairments. Despite this, the underlying mechanisms of SAE remain largely unexplored. Here, we explored the role of serotonergic neurotransmission in cognitive dysfunction of two mouse models of SAE. METHODS: The mouse models of SAE were established by injection of lipopolysaccharide (LPS, 10 mg/kg, intraperitoneal) and cecal ligation puncture (CLP) respectively. Barnes maze, new object recognition test and open field test were used to evaluate the effects of fluoxetine (selective serotonin reuptake inhibitor) and cyproheptadine (nonselective 5-HT2 receptor antagonist) on cognition and motor activity of mice. Additionally, WAY100635 (5-HT1A receptor antagonist) was co-administered with fluoxetine to explore the mechanism underlying effect of fluoxetine on cognitive impairments of SAE. Enzyme-linked immunosorbent assay (ELISA) was performed to determine 5-HT levels in hippocampus, brainstem and frontal lobe of experimental groups. RESULTS: Both LPS-induced sepsis and CLP induced sepsis resulted in a notable learning deficit. Fluoxetine ameliorated, while cyproheptadine aggravated, cognitive impairment in two classic mouse models of SAE. The cognition-enhancing effect of fluoxetine is reversed by WAY100635. Decreased 5-HT levels in hippocampus, brainstem and frontal lobe were observed in LPS septic model and CLP septic model. Notably, both fluoxetine and cyproheptadine significantly increased 5-HT levels in those brain regions in LPS septic model. Additionally, fluoxetine significantly increased 5-HT levels in frontal lobe of CLP septic model. CONCLUSIONS: Our study demonstrated that serotonergic neurotransmission plays a significant role in mechanisms underlying cognitive impairment in SAE. These findings contribute to identification of novel targets to prevent and arrest cognitive impairment in SAE.