Seizure-induced LIN28A disrupts pattern separation via aberrant hippocampal neurogenesis.

Prolonged seizures can disrupt stem cell behavior in the adult hippocampus, an important brain structure for spatial memory. Here, using a mouse model of pilocarpine-induced status epilepticus (SE), we characterized spatiotemporal expression of Lin28a mRNA and proteins after SE. Unlike Lin28a transcripts, induction of LIN28A protein after SE was detected mainly in the subgranular zone, where immunoreactivity was found in progenitors, neuroblasts, and immature and mature granule neurons. To investigate roles of LIN28A in epilepsy, we generated Nestin-Cre:Lin28aloxP/loxP (conditional KO [cKO]) and Nestin-Cre:Lin28a+/+ (WT) mice to block LIN28A upregulation in all neuronal lineages after acute seizure. Adult-generated neuron- and hippocampus-associated cognitive impairments were absent in epileptic LIN28A-cKO mice, as evaluated by pattern separation and contextual fear conditioning tests, respectively, while sham-manipulated WT and cKO animals showed comparable memory function. Moreover, numbers of hilar PROX1-expressing ectopic granule cells (EGCs), together with PROX1+/NEUN+ mature EGCs, were significantly reduced in epileptic cKO mice. Transcriptomics analysis and IHC validation at 3 days after pilocarpine administration provided potential LIN28A downstream targets such as serotonin receptor 4. Collectively, our findings indicate that LIN28A is a potentially novel target for regulation of newborn neuron-associated memory dysfunction in epilepsy by modulating seizure-induced aberrant neurogenesis.

Enhanced Electrical and Thermal Conductivities of Polymer Composites with a Segregated Network of Graphene Nanoplatelets.

Introducing a segregated network constructed through the selective localization of small amounts of fillers can be a solution to overcome the limitations of the practical use of graphene-based conductive composites due to the high cost of fillers. In this study, polypropylene composites filled with randomly dispersed GNPs and a segregated GNP network were prepared, and their conductive properties were investigated according to the formation of the segregated structure. Due to the GNP clusters induced by the segregated structure, the electrical percolation threshold was 2.9 wt% lower than that of the composite incorporating randomly dispersed GNPs. The fully interconnected GNP cluster network inside the composite contributed to achieving the thermal conductivity of 4.05 W/m∙K at 10 wt% filler content. Therefore, the introduction of a segregated filler network was suitable to simultaneously achieve excellent electrical and thermal conductivities at a low content of GNPs.

Easy, Fast Self-Heating Polyurethane Nanocomposite with the Introduction of Thermally Annealed Carbon Nanotubes Using Near-Infrared Lased Irradiation.

In this study, high-crystallinity single walled carbon nanotubes (H-SWNTs) were prepared by high-temperature thermal annealing at 1800 °C and a self-heating shape memory polyurethane nanocomposite with excellent self-heating characteristics was developed within a few seconds by irradiation with near-infrared rays. With a simple method (heat treatment), impurities at the surface of H-SWNTs were removed and at the same time the amorphous structure converted into a crystalline structure, improving crystallinity. Therefore, high conductivity (electric, thermal) and interfacial affinity with PU were increased, resulting in improved mechanical, thermal and electric properties. The electrical conductivity of neat polyurethane was enhanced from ~10-11 S/cm to 4.72 × 10-8 S/cm, 1.07 × 10-6 and 4.66 × 10-6 S/cm, while the thermal conductivity was enhanced up to 60% from 0.21 W/mK, 0.265 W/mK and 0.338 W/mK for the composites of 1, 3 and 5 wt%, respectively. Further, to achieve an effective photothermal effect, H-SWNTs were selected as nanofillers to reduce energy loss while increasing light-absorption efficiency. Thereafter, near-infrared rays of 818 nm were directly irradiated onto the nanocomposite film to induce photothermal properties arising from the local surface plasmon resonance effect on the CNT surface. A self-heating shape memory composite material that rapidly heated to 270 °C within 1 min was developed, even when only 3 wt.% of H-SWNTs were added. The results of this study can be used to guide the development of heat-generating coating materials and de-icing materials for the wing and body structures of automobiles or airplanes, depending on the molding method.

Development of N,N,N',N'-tetra-2-ethylhexyl-thiodiglycolamide silica-based adsorbent to separate useful metals from simulated high-level liquid waste.

A novel silica-based adsorbent was synthesized by impregnating macroporous silica polymer composite (SiO2-P) particles with a mixture of N,N,N',N'-tetra-2-ethylhexyl-thiodiglycolamide (TEHTDGA) and tri-n-octylamine (TOA). Then, the possibility of separating Pd(II) and other metal ions from simulated high-level liquid waste (HLLW) using the newly synthesized adsorbent (TEHTDGA + TOA)/SiO2-P was evaluated based on various adsorption characteristics obtained via batch-adsorption experiments, such as the HNO3 concentration, contact time, reaction temperature, adsorption isotherm, and chemical stability of the adsorbent. Furthermore, column separation experiments were performed based on the characteristics obtained from the batch-adsorption experiment, and the possibility of simultaneous separation of multiple metal ions was examined. The experimental results revealed that (TEHTDGA + TOA)/SiO2-P performs well in the separation of multiple metal ions from simulated HLLW.

Extraction behavior of a novel functionalized ionic liquid for separation of platinum group metals from aqueous nitric acid solution.

A novel ionic liquid (IL) functionalized with thiodiglycol amic acid containing a soft S donor was synthesized for the effective and efficient extraction of platinum group metals (Ru, Rh, and Pd) from aqueous nitric acid solutions, such as high-level radioactive liquid waste (HLLW). The IL enabled rapid extraction of Pd(II) with an extraction ratio of approximately 100%. The extractions of Ru(III) and Rh(III) by the IL were slower than that of Pd(II), but the rates were accelerated by temperature elevation. The extractions of Ru(III) and Rh(III) at 50 °C reached equilibrium within 4 and 8 h, respectively, with the extraction ratios of over 90% without assisting agents or other methods for the extraction system. Furthermore, the IL could extract more than 90% of Ru(III), Rh(III), and Pd(II) from the simulated HLLW within 2 h at 50 °C.

Thermal Percolation Behavior in Thermal Conductivity of Polymer Nanocomposite with Lateral Size of Graphene Nanoplatelet.

In this study, the thermal percolation behavior for the thermal conductivity of nanocomposites according to the lateral size of graphene nanoplatelets (GNPs) was studied. When the amount of GNPs reached the critical concentration, a rapid increase in thermal conductivity and thermal percolation behavior of the nanocomposites were induced by the GNP network. Interestingly, as the size of GNPs increased, higher thermal conductivity and a lower percolation threshold were observed. The in-plane thermal conductivity of the nanocomposite containing 30 wt.% M25 GNP (the largest size) was 8.094 W/m·K, and it was improved by 1518.8% compared to the polymer matrix. These experimentally obtained thermal conductivity results for below and above the critical content were theoretically explained by applying Nan's model and the percolation model, respectively, in relation to the GNP size. The thermal percolation behavior according to the GNP size identified in this study can provide insight into the design of nanocomposite materials with excellent heat dissipation properties.

Adsorption Performances of an Acid-stable 2D Covalent Organic Framework towards Palladium(II) in Simulated High-level Liquid Waste.

An acid-stable 2D covalent organic framework (COF TpPa-1) was synthesized by a reversible Schiff-base reaction and the following irreversible enol-keto tautomerism. The adsorption behaviors of COF TpPa-1 towards Pd(II) in simulated high-level liquid waste (HLLW) were investigated under the effect of contact time, the concentration of nitric acid etc. The obtained experimental results supported that the utilization of this type of acid-stable COF in HLLW to recover metal ion was feasible.

Synergistic adsorption behavior of a silica-based adsorbent toward palladium, molybdenum, and zirconium from simulated high-level liquid waste.

In this study, the synergistic adsorption behavior of palladium [Pd(II)], molybdenum [Mo(VI)], and zirconium [Zr(IV)] in simulated high-level liquid waste was systematically investigated based on various factors, such as the contact time, concentration of nitric acid, adsorption amount, and temperature using a silica-based adsorbent impregnated with N,N'-dimethyl-N,N'-di-n-hexyl-thiodiglycolamide (Crea) and 2, 2', 2' -nitrilotris[N,N-bis(2-ethylhexyl)acetamide] (TAMIA-EH). The adsorption rates of Pd(II), Mo(VI), and Zr(IV) in this synergistic adsorption system were high; thus, equilibrium states could be obtained in only 1 h with high uptake percentages of more than 90%. The adsorption abilities of Pd(II), Mo(VI), and Zr(IV) were only slightly affected by variation in the concentration of nitric acid in the range of 0.1-5 M and solution temperature in the range of 288-313 K. Selective stripping of the adsorbed Re(VII), Pd(II), Zr(IV), and Mo(VI) was successfully achieved under elution with 5 M HNO3, 0.2 M Tu (pH 1), 50 mM DTPA (pH 2), and 50 mM DTPA dissolved in 0.5 M Na2CO3 (pH 11) solutions using the chromatography method. In addition, the adsorption performance in solid-state was studied using the particle-induced X-ray emission (PIXE) method; the obtained results were in good agreement with the results obtained via column separation.

Vascular endothelial growth factor receptor-3 regulates astroglial glutamate transporter-1 expression via mTOR activation in reactive astrocytes following pilocarpine-induced status epilepticus.

Recent evidence has shown that the vascular endothelial growth factor (VEGF) system plays a crucial role in several neuropathological processes. We previously reported an upregulation of VEGF-C and its receptor, VEGFR-3, in reactive astrocytes after the onset of status epilepticus (SE). However, it remains unknown, which molecules act as downstream signals following VEGFR-3 upregulation, and are involved in reactive astrogliosis after SE. Therefore, we investigated whether VEGFR-3 upregulation within reactive astrocytes is associated with the activation of mammalian target of rapamycin (mTOR) signaling, which we confirmed by assaying for the phosphorylated form of S6 protein (pS6), and whether VEGFR-3-mediated mTOR activation induces astroglial glutamate transporter-1 (GLT-1) expression in the hippocampus after pilocarpine-induced SE. We found that spatiotemporal expression of pS6 was consistent with VEGFR-3 expression in the hippocampus after SE, and that both pS6 and VEGFR-3 were highly expressed in SE-induced reactive astrocytes. Treatment with the mTOR inhibitor rapamycin decreased astroglial VEGFR-3 expression and GLT-1 expression after SE. Treatment with a selective inhibitor for VEGFR-3 attenuated astroglial pS6 expression as well as suppressed GLT-1 expression and astroglial reactivity in the hippocampus after SE. These findings demonstrate that VEGFR-3-mediated mTOR activation could contribute to the regulation of GLT-1 expression in reactive astrocytes during the subacute phase of epilepsy. In conclusion, the present study suggests that VEGFR-3 upregulation in reactive astrocytes may play a role in preventing hyperexcitability induced by continued seizure activity.

Adsorption Behaviors of Palladium Ion from Nitric Acid Solution by a Silica-based Hybrid Donor Adsorbent.

The adsorption behaviors of a silica-based hybrid donor adsorbent (TAMIA-EH+1-dodecanol)/SiO2-P towards Pd(II) were investigated under the effect of the contact time, temperature etc. in simulated high-level liquid waste. The adsorption rates of Pd(II) and Re(VII) were fairly fast and could reach the equilibrium state in only 1 h compared with other co-existing metal ions. The adsorption kinetics of Pd(II) was found to fit well with the pseudo-first order model. Even though with increasing the concentration of HNO3 above 1 M, the adsorption performance of (TAMIA-EH+1-dodecanol)/SiO2-P decreased gradually; it still exhibited a better selectivity towards Pd(II) when [HNO3] > 0.5 M. The adsorption isotherms of Pd(II) and Re(VII) were well-described by the Langmuir isotherm model, while the Freundlich isotherm model was considered to be more suitable for the adsorption of Ru(III), Zr(IV) and Mo(VI). A high temperature of an aqueous solution was not good for the effective recovery of Pd(II). The calculated thermodynamic parameters revealed that the adsorption of Pd(II) was exothermic in nature.

Enhanced Tensile Properties of Multi-Walled Carbon Nanotubes Filled Polyamide 6 Composites Based on Interface Modification and Reactive Extrusion.

To induce uniform dispersion and excellent interfacial properties, we adopted a strategy of combining both polyamide 6 (PA6) grafting for multi-walled carbon nanotubes (MWCNTs) and reactive extrusion of PA6 matrix, based on anionic ring-opening polymerization of ε-caprolactam (CL). Compared to -COOH and -NCO treatments of MWCNTs, enhanced MWCNT dispersion and tensile properties of the composites were achieved using the applied strategy, and the tensile strength and modulus of the PA6-grafted MWCNT-filled PA6 composites were 5.3% and 20.5% higher than those of the purified MWCNT-filled PA6 composites, respectively. In addition, they were almost similar to the theoretical ones calculated by the modified Mori-Tanaka method (MTM) assuming a perfect interface, indicating that the tensile properties of MWCNT-filled PA6 composites can be optimized by PA6 grafting and reactive extrusion based on the anionic ring-opening polymerization of CL due to uniform MWCNT dispersion and excellent interfacial property.

Spatiotemporal expression of RCAN1 and its isoform RCAN1-4 in the mouse hippocampus after pilocarpine-induced status epilepticus.

Regulator of calcineurin 1 (RCAN1) can be induced by an intracellular calcium increase and oxidative stress, which are characteristic features of temporal lobe epilepsy. Thus, we investigated the spatiotemporal expression and cellular localization of RCAN1 protein and mRNA in the mouse hippocampus after pilocarpine-induced status epilepticus (SE). Male C57BL/6 mice were given pilocarpine hydrochloride (280 mg/kg, i.p.) and allowed to develop 2 h of SE. Then the animals were given diazepam (10 mg/kg, i.p.) to stop the seizures and sacrificed at 1, 3, 7, 14, or 28 day after SE. Cresyl violet staining showed that pilocarpine-induced SE resulted in cell death in the CA1 and CA3 subfields of the hippocampus from 3 day after SE. RCAN1 immunoreactivity showed that RCAN1 was mainly expressed in neurons in the shammanipulated hippocampi. At 1 day after SE, RCAN1 expression became detected in hippocampal neuropils. However, RCAN1 signals were markedly enhanced in cells with stellate morphology at 3 and 7 day after SE, which were confirmed to be reactive astrocytes, but not microglia by double immunofluorescence. In addition, real-time reverse transcriptase-polymerase chain reaction showed a significant upregulation of RCAN1 isoform 4 (RCAN1-4) mRNA in the SE-induced hippocampi. Finally, in situ hybridization with immunohistochemistry revealed astrocytic expression of RCAN1-4 after SE. These results demonstrate astrocytic upregulation of RCAN1 and RCAN1-4 in the mouse hippocampus in the acute and subacute phases of epileptogenesis, providing foundational information for the potential role of RCAN1 in reactive astrocytes during epileptogenesis.

Alteration of Gene Associated with Retinoid-interferon-induced Mortality-19-expressing Cell Types in the Mouse Hippocampus Following Pilocarpine-induced Status Epilepticus.

The gene associated with retinoid-interferon-induced mortality-19 (GRIM-19) plays several significant roles in cellular processes, including ATP synthesis, reactive oxygen species formation, and the regulation of glycolytic enzyme activity, which are closely related to the pathophysiological mechanisms of epilepsy. Therefore, we investigated the expression pattern of GRIM-19 in the CA1 area of the hippocampus in 8-week-old male C57BL/6 mice following pilocarpine-induced status epilepticus (SE). Neuronal death in the hippocampal CA1 area was prominently observed at 4 and 7 days after SE, and astrocytes and microglia became progressively activated beginning at 1 day after SE. GRIM-19 immunoreactivity was decreased in the damaged pyramidal cell layer but markedly increased in the stratum radiatum and stratum lacunosum-moleculare of the hippocampus at 4 and 7 days after SE. In addition, the cell types of GRIM-19-expressing cells in the epileptic hippocampus were identified. GRIM-19 was mainly co-localized in neurons but only slightly expressed in glia in the normal hippocampus. Most of the GRIM-19-positive cells induced by SE in the stratum radiatum and stratum lacunosum-moleculare were glial fibrillary acidic protein-expressing reactive astrocytes. Moreover, we observed that both GRIM-19 and pyruvate kinase isozyme M2, a glycolytic enzyme, were highly expressed in reactive astrocytes after SE. These results indicate that expression of GRIM-19 in the hippocampus is mainly observed in neurons under normal conditions but is altered in the SE mouse model as evidenced by its increased expression in reactive astrocytes. The possible role of GRIM-19 in the glycolytic activity of reactive astrocytes is also discussed.

Increased expression of vascular endothelial growth factor-C and vascular endothelial growth factor receptor-3 after pilocarpine-induced status epilepticus in mice.

Vascular endothelial growth factor (VEGF)-C and its receptor, vascular endothelial growth factor receptor (VEGFR)-3, are responsible for lymphangiogenesis in both embryos and adults. In epilepsy, the expression of VEGF-C and VEGFR-3 was significantly upregulated in the human brains affected with temporal lobe epilepsy. Moreover, pharmacologic inhibition of VEGF receptors after acute seizures could suppress the generation of spontaneous recurrent seizures, suggesting a critical role of VEGF-related signaling in epilepsy. Therefore, in the present study, the spatiotemporal expression of VEGF-C and VEGFR-3 against pilocarpine-induced status epilepticus (SE) was investigated in C57BL/6N mice using immunohistochemistry. At 1 day after SE, hippocampal astrocytes and microglia were activated. Pyramidal neuronal death was observed at 4 days after SE. In the subpyramidal zone, VEGF-C expression gradually increased and peaked at 7 days after SE, while VEGFR-3 was significantly upregulated at 4 days after SE and began to decrease at 7 days after SE. Most VEGF-C/VEGFR-3-expressing cells were pyramidal neurons, but VEGF-C was also observed in some astrocytes in sham-manipulated animals. However, at 4 days and 7 days after SE, both VEGFR-3 and VEGF-C immunoreactivities were observed mainly in astrocytes and in some microglia of the stratum radiatum and lacunosum-moleculare of the hippocampus, respectively. These data indicate that VEGF-C and VEGFR-3 can be upregulated in hippocampal astrocytes and microglia after pilocarpine-induced SE, providing basic information about VEGF-C and VEGFR-3 expression patterns following acute seizures.

Heterogeneity of GRIM-19 Expression in the Adult Mouse Brain.

Gene associated with retinoid-interferon-induced mortality-19 (GRIM-19) is a subunit of the mitochondrial respiratory chain complex I that has a significant effect on ATP production. The brain is particularly susceptible to ATP deficiency due to its limited energy storage capability and its high rate of oxygen consumption. Thus, GRIM-19 might be involved in regulating ATP level in the brain or cell death caused by several neurological disorders. To understand the physiological and pathophysiological roles of GRIM-19 in the brain, a thorough investigation of the neuroanatomic distribution of GRIM-19 in the normal brain is necessary. Therefore, the present study examined the distribution patterns of GRIM-19 in the adult C57BL/6 mouse brain using immunohistochemistry and identified cell types expressing GRIM-19 using double immunofluorescence staining. We found that GRIM-19 was ubiquitously but not homogenously expressed throughout the brain. GRIM-19 immunoreactivity was predominantly observed in neurons, but not in astrocytes, microglia, or oligodendrocytes under normal physiological conditions. Following transient global cerebral ischemia, GRIM-19-positive immunoreactivity was, however, observed in neurons as well as glial cells including astrocytes in the hippocampus. Furthermore, GRIM-19 was weakly expressed in the hippocampal subgranular zone, in which neural stem and progenitor cells are abundant, but highly expressed in the immature and mature neuronal cells in the granular cell layer of the normal brain, suggesting an inverse correlation between expression of GRIM-19 and stemness activity. Collectively, our study demonstrating widespread and differential distribution of GRIM-19 in the adult mouse brain contributes to investigating the functional and pathophysiological roles of this protein.

The Neuroprotective Effect of Hericium erinaceus Extracts in Mouse Hippocampus after Pilocarpine-Induced Status Epilepticus.

Hericium erinaceus (HE), a culinary-medicinal mushroom, has shown therapeutic potential in many brain diseases. However, the role of HE in status epilepticus (SE)-mediated neuronal death and its underlying mechanisms remain unclear. We investigated the neuroprotective effects of HE using a pilocarpine-induced SE model. Male C57BL/6 mice received crude extracts of HE (60 mg/kg, 120 mg/kg, or 300 mg/kg, p.o.) for 21 d from 14 d before SE to 6 d after SE. At 7 d after SE, cresyl violet and immunohistochemistry of neuronal nuclei revealed improved hippocampal neuronal survival in animals treated with 60 mg/kg and 120 mg/kg of HE, whereas those treated with 300 mg/kg of HE showed similar neuronal death to that of vehicle-treated controls. While seizure-induced reactive gliosis, assessed by immunohistochemistry, was not altered by HE, the number of hippocampal cyclooxygenase 2 (COX2)-expressing cells was significantly reduced by 60 and 120 mg/kg of HE. Triple immunohistochemistry demonstrated no overlap of COX2 labeling with Ox42, in addition to a decrease in COX2/GFAP-co-immunoreactivity in the group treated with 60 mg/kg HE, suggesting that the reduction of COX2 by HE promotes neuroprotection after SE. Our findings highlight the potential application of HE for preventing neuronal death after seizures.

Persistent Lin28 Expression Impairs Neurite Outgrowth and Cognitive Function in the Developing Mouse Neocortex.

Many neurodevelopmental disorders feature learning and memory difficulties. Regulation of neurite outgrowth during development is critical for neural plasticity and memory function. Here, we show a novel regulator of neurite outgrowth during cortical neurogenesis, Lin28, which is an RNA-binding protein. Persistent Lin28 upregulation by in utero electroporation at E14.5 resulted in neurite underdevelopment during cortical neurogenesis. We also showed that Lin28-overexpressing cells had an attenuated response to excitatory inputs and altered membrane properties including higher input resistance, slower action potential repolarization, and smaller hyperpolarization-activated cation currents, supporting impaired neuronal functionality in Lin28-electroporated mice. When we ameliorated perturbed Lin28 expression by siRNA, Lin28-induced neurite underdevelopment was rescued with reduction of Lin28-downstream molecules, high mobility group AT-Hook 2, and insulin-like growth factor 1 receptor. Finally, Lin28-electroporated mice showed significant memory deficits as assessed by the Morris water maze test. Taken together, these findings demonstrate a new role and the essential requirement of Lin28 in developmental control of neurite outgrowth, which has an impact on synaptic plasticity and spatial memory. These findings suggest that targeting Lin28 may attenuate intellectual disabilities by correction of impaired dendritic complexity, providing a novel therapeutic candidate for treating neurodevelopmental disorders.

Prediction of Defect Formation during Resin Impregnation Process through a Multi-Layered Fiber Preform in Resin Transfer Molding by a Proposed Analytical Model.

It is very important to predict any defects occurring by undesired fiber deformations to improve production yields of resin transfer molding, which has been widely used for mass production of carbon fiber reinforced composite parts. In this study, a simple and efficient analytic scheme was proposed to predict deformations of a multi-layered fiber preform by comparing the forces applied to the preform in a mold of resin transfer molding. Friction coefficient of dry and wet states, permeability, and compressive behavior of unidirectional (UD) and plain woven (PW) carbon fabrics were measured, which were used to predict deformations of the multi-layered fiber preforms with changing their constitution ratios. The model predicted the occurrence, type, and position of fiber deformation, which agreed with the experimental results of the multi-layered preforms.