Roles and therapeutic potential of different extracellular vesicle subtypes on traumatic brain injury.

Traumatic brain injury (TBI) is a leading cause of injury-related disability and death around the world, but the clinical stratification, diagnosis, and treatment of complex TBI are limited. Due to their unique properties, extracellular vesicles (EVs) are emerging candidates for being biomarkers of traumatic brain injury as well as serving as potential therapeutic targets. However, the effects of different extracellular vesicle subtypes on the pathophysiology of traumatic brain injury are very different, or potentially even opposite. Before extracellular vesicles can be used as targets for TBI therapy, it is necessary to classify different extracellular vesicle subtypes according to their functions to clarify different strategies for EV-based TBI therapy. The purpose of this review is to discuss contradictory effects of different EV subtypes on TBI, and to propose treatment ideas based on different EV subtypes to maximize their benefits for the recovery of TBI patients. Video Abstract.

Hyperadhesive von Willebrand Factor Promotes Extracellular Vesicle-Induced Angiogenesis: Implication for LVAD-Induced Bleeding.

Bleeding associated with left ventricular assist device (LVAD) implantation has been attributed to the loss of large von Willebrand factor (VWF) multimers to excessive cleavage by ADAMTS-13, but this mechanism is not fully supported by the current evidence. We analyzed VWF reactivity in longitudinal samples from LVAD patients and studied normal VWF and platelets exposed to high shear stress to show that VWF became hyperadhesive in LVAD patients to induce platelet microvesiculation. Platelet microvesicles activated endothelial cells, induced vascular permeability, and promoted angiogenesis in a VWF-dependent manner. Our findings suggest that LVAD-driven high shear stress primarily activates VWF, rather than inducing cleavage in the majority of patients.

Anticoagulation targeting membrane-bound anionic phospholipids improves outcomes of traumatic brain injury in mice.

Severe traumatic brain injury (TBI) often causes an acute systemic hypercoagulable state that rapidly develops into consumptive coagulopathy. We have recently demonstrated that TBI-induced coagulopathy (TBI-IC) is initiated and disseminated by brain-derived extracellular vesicles (BDEVs) and propagated by extracellular vesicles (EVs) from endothelial cells and platelets. Here, we present results from a study designed to test the hypothesis that anticoagulation targeting anionic phospholipid-expressing EVs prevents TBI-IC and improves the outcomes of mice subjected to severe TBI. We evaluated the effects of a fusion protein (ANV-6L15) for improving the outcomes of TBI in mouse models combined with in vitro experiments. ANV-6L15 combines the phosphatidylserine (PS)-binding annexin V (ANV) with a peptide anticoagulant modified to preferentially target extrinsic coagulation. We found that ANV-6L15 reduced intracranial hematoma by 70.2%, improved neurological function, and reduced death by 56.8% in mice subjected to fluid percussion injury at 1.9 atm. It protected the TBI mice by preventing vascular leakage, tissue edema, and the TBI-induced hypercoagulable state. We further showed that the extrinsic tenase complex was formed on the surfaces of circulating EVs, with the highest level found on BDEVs. The phospholipidomic analysis detected the highest levels of PS on BDEVs, as compared with EVs from endothelial cells and platelets (79.1, 15.2, and 3.5 nM/mg of protein, respectively). These findings demonstrate that TBI-IC results from a trauma-induced hypercoagulable state and may be treated by anticoagulation targeting on the anionic phospholipid-expressing membrane of EVs from the brain and other cells.

Expression, structure and function analysis of the sperm-oocyte fusion genes Juno and Izumo1 in sheep (Ovis aries).

BACKGROUND: JUNO and IZUMO1 are the first receptor-ligand protein pairs discovered to be essential for sperm-oocyte fusion; their interaction is indispensable for fertilization. METHODS: PCR was used to clone the full-length DNA sequence of the Juno gene in sheep. The single nucleotide polymorphism (SNP) loci of Juno were genotyped by Sequenom MassARRAY®. PCR combined with rapid amplification of cDNA Ends were used to clone the full-length cDNA sequence of Juno and Izumo1. Reverse transcriptase-PCR (RT-PCR) and real time-quantitative-PCR (RT-qPCR) were used to analyze the genes' expression in tissues of sheep, and single cell RNA-seq was used to analyze the genes' expression in oocytes, granulosa cells and follicular theca of polytocous and monotocous Small Tail Han ewes. Bioinformatics was used to analyze advanced structure and phylogeny of JUNO and IZUMO1 proteins. RESULTS: The full-length DNA sequence of the Juno gene in sheep was cloned and nine SNPs were screened. We found a significant association between the g.848253 C > A locus of Juno and litter size of Small Tail Han sheep (P < 0.05). The full-length cDNA sequence of Juno and Izumo1 genes from Small Tail Han sheep were obtained. We found a new segment of the Izumo1 CDS consisting of 35 bp, and we confirmed the Izumo1 gene has 9 exons, not 8. RT-qPCR showed that Juno and Izumo1 genes were highly expressed in ovarian and testicular tissues, respectively (P < 0.01). Single cell RNA-seq showed Juno was specifically expressed in oocytes, but not in granulosa cells or follicular theca, while Izumo1 displayed little to no expression in all three cell types. There was no difference in expression of the Juno gene in oocyte and ovarian tissue in sheep with different litter sizes, indicating expression of Juno is not related to litter size traits. Bioinformatic analysis revealed the g.848253 C > A locus of Juno results in a nonconservative missense point mutation leading to a change from Phe to Leu at position 219 in the amino acid sequence. CONCLUSIONS: For the first time, this study systematically analyzed the expression, structure and function of Juno and Izumo1 genes and their encoded proteins in Small Tail Han sheep, providing the basis for future studies of the regulatory mechanisms of Juno and Izumo1 genes.

Significant tunneling magnetoresistance and excellent spin filtering effect in CrI3-based van der Waals magnetic tunnel junctions.

van der Waals (vdW) heterojunctions stacked by different two-dimensional (2D) layered materials not only exhibit the complementary effect of short plates, but also harbor novel physical phenomena. In particular, the emergence of 2D magnetic vdW materials has provided novel opportunities for the application of these materials in spintronics. However, to the best of our knowledge, to date, the spin-related transport mechanism in magnetic tunnel junctions (MTJs) based on these 2D vdW magnetic materials and the effect of pinning layers on their transport properties have not been elucidated by the non-equilibrium state theory. Herein, based on first-principles calculations, we report the spin-polarized quantum transport properties of sandwich-type vdW magnetic tunnel junctions (CrI3/h-BN/n·CrI3) comprising monolayer CrI3, a hexagonal boron nitride (h-BN) spacer layer, and n-layer CrI3 (n = 1, 2, 3, and 4). Considering the inter-layer antiferromagnetic coupling in n-layer CrI3, a few layers of CrI3 can be regarded as its own natural pinning layers. Especially, when n is equal to 3, an almost fully spin-polarized current and large tunnel magnetoresistance ratio (3600%) are obtained in the equilibrium state. Excitingly, due to different numbers of pinning layers in MTJs, the transport properties of these MTJs at positive bias voltages exhibit an interesting odd-even effect within a limited thickness of these pinning layers. Moreover, an almost perfect spin filtering effect and remarkable negative differential resistance (NDR) were observed in the MTJs where n was odd (n = 1 and 3). The observed non-equilibrium quantum transport phenomenon is explained by spin-dependent transmission coefficient at different bias voltages. Our results provide effective guidance for the experimental studies of the MTJs based on 2D magnetic vdW materials.

An Improved Lagrangian-Inverse Method for Evaluating the Dynamic Constitutive Parameters of Concrete.

The dynamic constitutive behaviors of concrete-like materials are of vital importance for structure designing under impact loading conditions. This study proposes a new method to evaluate the constitutive behaviors of ordinary concrete at high strain rates. The proposed method combines the Lagrangian-inverse analysis method with optical techniques (ultra-high-speed camera and digital image correlation techniques). The proposed method is validated against finite-element simulation. Spalling tests were conducted on concretes where optical techniques were employed to obtain the high-frequency spatial and temporal displacement data. We then obtained stress-strain curves of concrete by applying the proposed method on the results of spalling tests. The results show non-linear constitutive behaviors in these stress-strain curves. These non-linear constitutive behaviors can be possibly explained by local heterogeneity of concrete. The proposed method provides an alternative mean to access the dynamic constitutive behaviors which can help future structure designing of concrete-like materials.

Effects of Brain-Derived Mitochondria on the Function of Neuron and Vascular Endothelial Cell After Traumatic Brain Injury.

OBJECTIVE: Mitochondrial dysfunction plays an essential role in secondary brain injury following traumatic brain injury (TBI). Interestingly, accumulating evidence has shown that therapeutic benefits of mitochondrial transplantation exist. Therefore, we hypothesized that the injection of exogenous mitochondria would contribute to the mitigation of cellular energy metabolism disorders and neurologic functions after TBI. METHODS: We first extracted isolated mitochondria from fresh brain tissue using a kit and then identified their activity and purity. The role of exogenous mitochondria was assessed using the glucose oxygen deprivation-induced cellular damage model and controlled cortical impact-induced mice with TBI. RESULTS: The results showed that treatment with exogenous mitochondria improved the cellular respiratory control rate, the expression of tight junction-associated proteins, and synaptic plasticity-related proteins in vitro. Moreover, the application of exogenous mitochondria significantly reduced cellular apoptosis, promoted angiogenesis and alleviated brain edema and blood-brain barrier leakage in mice subjected to TBI. Additionally, exogenous mitochondria significantly reduced excessive inhibition of long-term depression in the hippocampus 7 days after TBI. CONCLUSIONS: Taken together, the data suggested that exogenous mitochondrial intervention ameliorated glucose oxygen deprivation-induced cell damage and controlled cortical impact-induced TBI in a mouse model. The new discovery in the current study inspires us to suggest that mitochondrial transplantation might serve as a new therapeutic strategy for TBI.

Effects of Cryopreservation on Microparticles Concentration, Procoagulant Function, Size Distribution, and Morphology.

BACKGROUND Research on microparticles is rapidly evolving and has extended to the field of many diseases. It is unclear whether microparticles can be stored frozen. In this study, our goal was to verify whether cryopreservation had an effect on the properties of the microparticles. MATERIAL AND METHODS We obtained C57BL/6J mouse-derived microparticles by grinding and gradient centrifugation. The specimens were divided into 2 groups: without dimethyl sulfoxide and with dimethyl sulfoxide. The microparticles were then stored at 25°C, 4°C, -20°C, -80°C, and -196°C for 0.5 days, 1 day, 3 days, 5 days, and 7 days. We tested whether the concentration, coagulation function, diameter distribution, and morphology of the microparticles in the 2 groups changed compared to those of a fresh sample. RESULTS We discovered that the concentrations of total microparticles, annexin V-positive microparticles, and brain-derived microparticles changed with freezing. The coagulation function, morphology, and size distribution of the microparticles were also affected by cryopreservation. Finally, there was no difference in the effects of cryopreservation on microparticles between the dimethyl sulfoxide group and the dimethyl sulfoxide-free group. CONCLUSIONS This study suggests that cryopreservation has diverse effects on microparticles within 1 week and that dimethyl sulfoxide has no protective effect on cryopreserved microparticles. Therefore, microparticles should be used fresh for future studies, and they should not be cryopreserved with or without dimethyl sulfoxide.

Behavior of Cross Arms Inserted in Concrete-Filled Circular GFRP Tubular Columns.

Fiber-reinforced polymer (FRP) materials nowadays have attracted much attention in both retrofitting of aged infrastructure and developing of new structural systems attributed to the outstanding mechanical properties. Extensive studies have been performed on concrete-filled glass FRP (GFRP) tubes for the potential application in piling, poles, highways overhead sign structures and bridge components. The new hybrid member also provides an alternative solution for traditional transmission structures. However, the connection between concrete-filled GFRP tubes and cross arms has not been fully understood. In this paper, an experimental study and theoretical analysis were conducted on the behavior of cross arms inserted in concrete-filled circular GFRP tubular columns. Steel bars with a larger stiffness in comparison with GFRP tubes were selected here for the cross arm to simulate a more severe scenario. The structural responses of the system when the cross arms were subjected to concentrated loads were carefully recorded. Experimental results showed that the concrete-filled GFRP tubes could offer a sufficient restraint to the deformation of the cross arm. No visible cracks were found on the GFRP tube at the corner of the cross arm where the stress and strain concentrated. Theoretical solutions based on available theories and equations were adopted to predict the displacement of the cross arms and a good agreement was achieved between the prediction results and experimental findings.

Genetic polymorphisms and transcription profiles associated with intracranial aneurysm: a key role for NOTCH3.

Intracranial aneurysm (IA) incidence is about 1~2%. However, the specific mechanisms of IA onset and development need further study. Our objective was to discover novel IA-related genes to determine possible etiologies further. We performed next-generation sequencing on nineteen Chinese patients with familial IA and one patient with sporadic IA. We obtained mRNA expression data of 129 samples from Gene Expression Omnibus (GEO) and made statistical computing to discover differentially expressed genes (DEGs). The screened IA-related gene NOTCH3 was determined by bioinformatic data mining. We verified the IA-related indicators of NOTCH3. Association was found between IA and the NOTCH3 SNPs rs779314594, rs200504060 and rs2285981. Levels of NOTCH3 mRNA were lower in IA tissue than in control tissue, but higher in peripheral blood neutrophils from IA patients than in neutrophils from controls. Levels of NOTCH3 protein were lower in IA tissue than in cerebral artery tissue. NOTCH3 also decreased the expression of angiogenesis factors in human umbilical vein endothelial cells. Variation in NOTCH3 and alteration of its expression in cerebral artery or neutrophils may contribute to IA. Our findings also describe a bioinformatic-experimental approach that may prove useful for probing the pathophysiology of other complex diseases.

Meta-Analysis of Prognosis of Different Treatments for Symptomatic Moyamoya Disease.

OBJECTIVE: The purpose of this study was to evaluate the efficacy of surgical revascularization versus conservative treatment and different surgical modalities, in order to provide evidence for the patient with moyamoya disease (MMD) to choose the appropriate treatment. METHODS: We comprehensively searched PubMed, Embase, Web of Science, and the Cochrane Library for articles published regarding MMD treatment. If the I2 value, which evaluated the heterogeneity, was <50%, a fixed-effect model was used; if not, a random effect model was applied. RESULTS: Twenty-seven articles were included in the meta-analysis. The surgery group is more advantageous in reducing the risk of future stroke events than conservative treatment in MMD patients (odds ratio [OR] 0.26, 95% confidence interval [CI] 0.20-0.33, P < 0.001). In addition, the surgical group also had an advantage in terms of increased cerebral perfusion (OR 7.16, 95% CI 3.28-15.64, P < 0.001) and death due to rebleeding (OR 0.27, 95% CI 0.10-0.72, P < 0.01). Direct surgery showed a significant efficacy over indirect surgery (OR 2.03, 95% CI 1.32-3.13, P < 0.01). No obvious difference was found between the direct and indirect bypass subset (OR 0.76, 95% CI 0.51-1.14, P = 0.185). Angiographic results in patients undergoing direct bypass surgery are more pronounced (OR 0.20, 95% CI 0.06-0.67, P < 0.01). CONCLUSIONS: In patients with symptomatic moyamoya disease, bypass surgery is more effective than conservative treatment to prevent future strokes. In surgical patients, direct bypass seems to reduce the risk of stroke more than an indirect bypass.

Spin dependence of crossed Andreev reflection and electron tunneling induced by Majorana fermions.

We investigate spin dependence of the nonlocal transport induced by Majorana fermions in a one-dimensional ferromagnet-ferromagnetic-insulator-superconductor-ferromagnetic-insulator-ferromagnet junction on the edge of a two-dimensional topological insulator. The results show that coupled Majorana fermions lead to the nonlocal transport processes including electron tunneling and crossed Andreev reflection, which can be tuned by adjusting the spin polarizations of the Majorana fermions. By manipulating the bands in the two ferromagnets, the nonlocal transport can be selected as either pure electron tunneling or pure crossed Andreev reflection, the transmission probability of which could be 100%. Furthermore, the pure electron tunneling and the pure crossed Andreev reflection are well controlled by the spin directions of the electron states in the two ferromagnets.

Whole-genome sequences of 89 Chinese sheep suggest role of RXFP2 in the development of unique horn phenotype as response to semi-feralization.

Background: Animal domestication has been extensively studied, but the process of feralization remains poorly understood. Results: Here, we performed whole-genome sequencing of 99 sheep and identified a primary genetic divergence between 2 heterogeneous populations in the Tibetan Plateau, including 1 semi-feral lineage. Selective sweep and candidate gene analysis revealed local adaptations of these sheep associated with sensory perception, muscle strength, eating habit, mating process, and aggressive behavior. In particular, a horn-related gene, RXFP2, showed signs of rapid evolution specifically in the semi-feral breeds. A unique haplotype and repressed horn-related tissue expression of RXFP2 were correlated with higher horn length, as well as spiral and horizontally extended horn shape. Conclusions: Semi-feralization has an extensive impact on diverse phenotypic traits of sheep. By acquiring features like those of their wild ancestors, semi-feral sheep were able to regain fitness while in frequent contact with wild surroundings and rare human interventions. This study provides a new insight into the evolution of domestic animals when human interventions are no longer dominant.

Circular Ribonucleic Acid Expression Alteration in Exosomes from the Brain Extracellular Space after Traumatic Brain Injury in Mice.

Traumatic brain injury (TBI) has high morbidity and mortality rates. The mechanisms underlying TBI are unclear and may include the change in biological material in exosomes. Circular ribonucleic acids (circRNAs) are enriched and stable in exosomes, which can function as microRNA (miRNA) sponges to regulate gene expression levels. Therefore, we speculated that circRNAs in exosomes might play an important role in regulating gene expression after TBI and then regulate specific signaling pathways, which may protect the brain. We first isolated exosomes from the brain extracellular space in mice with TBI by digestion. We then investigated the alterations in circRNA expression in exosomes by high-throughput whole transcriptome sequencing, analyzed the data by gene ontology (GO) and pathway analysis, and constructed the circRNA-miRNA network. In this study, we identified 231 significantly and differentially expressed circRNAs, including 155 that were upregulated and 76 that were downregulated. GO analysis showed that these differentially expressed circRNAs might be related to the growth and repair of neurons, the development of the nervous system, and the transmission of nerve signals. The most highly correlated pathways that we identified were involved primarily with glutamatergic synapse and the cyclic guanosine monophosphate-protein kinase G signaling pathway. The circRNA-miRNA network predicted the potential roles of these differentially expressed circRNAs and the interaction of circRNAs with miRNAs. Our study broadens the horizon of research on gene regulation in exosomes from the brain extracellular space after TBI and provides novel targets for further research on both the molecular mechanisms of TBI and the potential intervention therapy targets.

Flexural Behavior of GFRP Tubes Filled with Magnetically Driven Concrete.

Experimental investigation of GFRP (glass fiber reinforced polymer) tubes that were filled with magnetically driven concrete was carried out to study the flexural behavior of specimens under bending. Specimens having different cross section and lengths were tested. The test specimens were fabricated by filling magnetically driven concrete into the GFRP tubes and the concrete was vibrated using magnetic force. Specimens vibrated using vibrating tube were also tested for comparison. In addition, specimens having steel reinforcing bars and GFRP bars were both tested to study the effect of reinforcing bars on the magnetically driven concrete. The load-displacement curves, load-strain curves, failure mode, and ultimate strengths of test specimens were obtained. Design methods for the flexural stiffness of test specimens were also discussed in this study.

Electrical stimulation improved cognitive deficits associated with traumatic brain injury in rats.

Introduction: Cognitive deficits associated with traumatic brain injury (TBI) reduce patient quality of life. However, to date, there have been no effective treatments for TBI-associated cognitive deficits. In this study, we aimed to determine whether electrical stimulation (ES) improves cognitive deficits in TBI rats. Methods: Rats were randomly divided into three groups: the Sham control group, electrical stimulation group (ES group), and No electrical stimulation control group (N-ES group). Following fluid percussion injury, the rats in the ES group received ES treatment for 3 weeks. Potent cognitive function-relevant factors, including the escape latency, time percentage in the goal quadrant, and numbers of CD34+ cells, von Willebrand Factor+ (vWF +) vessels, and circulating endothelial progenitor cells (EPCs), were subsequently assessed using the Morris water maze (MWM) test, immunohistochemical staining, and flow cytometry. Results: Compared with the rats in the N-ES group, the rats in the ES group exhibited a shorter escape latency on day 3 (p = .025), day 4 (p = .011), and day 5 (p = .003), as well as a higher time percentage in the goal quadrant (p = .025) in the MWM test. After 3 weeks of ES, there were increased numbers of CD34+ cells (p = .008) and vWF + vessels (p = .000) in the hippocampus of injured brain tissue in the ES group compared with those in the N-ES group. Moreover, ES also significantly increased the number of EPCs in the peripheral blood from days 3 to 21 after TBI in the ES group (p < .05). Conclusions: Taken together, these findings suggest that ES may improve cognitive deficits induced by TBI, and this protective effect may be a result, in part, of enhanced angiogenesis, which may be attributed to the increased mobilization of EPCs in peripheral blood.

Reduction of Surgical Site Infections After Cranioplasty With Perioperative Bundle.

BACKGROUND: Surgical site infections (SSI) are the most common complication after cranioplasty and it is associated with poor prognosis. The aim of this study was to identify the risk factors that triggered the development of SSI after cranioplasty and establish a new perioperative bundle and monitoring system to reduce SSI. METHODS: A retrospective review of a database that included all cranioplasty patients from 2001 to 2007 was carried out to determine the prevalence of infection. A surveillance team was set up, which assessed the clinical practice and led to the development of a new perioperative bundle and supervision system. A prospective study between 2008 and 2014 was carried out to observe whether infection rates had changed and whether an active surveillance program can change clinical practice. RESULTS: Retrospective period included 86 adult patients. The overall rate of infection was 10.5% (9 SSI in 86 patients). Age ≥60 years (OR 1.05; 95% CI: 1.00-1.10; P = 0.04), smoking (OR 9.13; 95% CI: 1.65-50.60; P = 0.01), and duration of operation ≥180 minutes (OR1.19; 95% CI: 1.06-1.34; P < 0.01) as significant predictors of postcranioplasty SSI development. Length of preoperative stay and length of hospitalization was significantly longer among infected patients compared with uninfected patients (P < 0.01, respectively). In prospective period, the general SSI rate was reduced to 1.8% (2 SSI in 113 patients). The difference was statistically significant (P = 0.01). CONCLUSIONS: A perioperative bundle and monitoring system may help to reduce SSI rates after cranioplasty. This work also indicates how an active surveillance program can successfully change clinical practice.

Removal of organic matter, nitrogen and faecal indicators from diluted anaerobically digested slurry using tidal flow constructed wetlands.

The rapid implementation of anaerobic digestion for renewable energy production has resulted in increased generation of anaerobically digested slurry, which contains a variety of pollutants and therefore has the potential to cause serious environmental problems. Tidal flow constructed wetlands, which could generate beneficial oxygen conditions, were investigated for their success in removing nitrogen, organic matter and pathogens in anaerobically digested slurry. The results indicated that tidal operation had a positive effect on promoting NH4+-N and organic matter (chemical oxygen demand (COD)) removal. With an average influent NH4+-N concentration of 288 mg/L and COD concentration of 839 mg/L, the average removal efficiency reached up to 93% (325 g/m2 day) for NH4+-N and 53% (603 g/m2 day) for COD, with total inorganic nitrogen (TIN) removal efficiency of 51% (226 g/m2 day). The nitrogen removal in the tidal-operated CWs is highly dependent on the flooded and drained (F/D) time ratio. Changing flooded time from 3 to 5 h enhanced denitrification (nitrite reductase-K (nirK) abundance) and further resulted in improved TIN removal efficiency of 62% (237 g/m2 day). The removal of faecal indicators was also examined, with reduction rate of approximately 0.9 log10 CFU/100 mL for both Escherichia coli and total coliforms, which was independent of the influent loadings and differing flooded/drained time ratio. Tidal flow CWs were demonstrated to have the high potential to treat diluted anaerobically digested slurry.