Heterozygous variants in USP25 cause genetic generalized epilepsy.

USP25 encodes ubiquitin-specific proteases 25, a key member of deubiquitinating enzyme family and is involved in neural fate determination. Although abnormal expression in Down's syndrome was reported previously, the specific role of USP25 in human diseases has not been defined. In this study, we performed trio-based whole exome sequencing in a cohort of 319 cases (families) with generalized epilepsy of unknown etiology. Five heterozygous USP25 variants including two de novo and three co-segregated variants were determined in eight individuals affected by generalized seizures and/or febrile seizures from five unrelated families. The frequency of USP25 variants showed a significantly high aggregation in this cohort compared to the East Asian population and all populations in the gnomAD database. The mean onset ages of febrile and afebrile seizures were 10 months (infancy) and 11.8 years (juvenile), respectively. The patients achieved seizure freedom except one had occasional nocturnal seizures at the last follow-up. Two patients exhibited intellectual disability. Usp25 was ubiquitously expressed in mouse brain with two peaks on embryonic days (E14‒E16) and postnatal day 21, respectively. Similarly, USP25 expressed in fetus/early childhood stage with a second peak at approximately 12‒20 years old in human brain, consistent with the seizure onset age at infancy and juvenile in the patients. To investigate the functional impact of USP25 deficiency in vivo, we established Usp25 knock-out mice, which showed increased seizure susceptibility compared to wild-type mice in pentylenetetrazol-induced seizure test. To explore the impact of USP25 variants, we employed multiple functional detections. In HEK293T cells, the severe phenotype associated variant (p.Gln889Ter) led to a significant reduction of mRNA and protein expressions but formed a stable truncated dimers with increment of deubiquitinating enzyme activities and abnormal cellular aggregations, indicating a gain-of-function effect. The p.Gln889Ter and p.Leu1045del increased neuronal excitability in mice brain, with a higher firing ability in p.Gln889Ter. These functional impairments align with the severity of the observed phenotypes, suggesting a genotype-phenotype correlation. Hence, a moderate association between USP25 and epilepsy was noted, indicating USP25 is potentially a predisposing gene for epilepsy. Our results from Usp25 null mice and the patient-derived variants indicated that USP25 would play epileptogenic role via loss-of-function or gain-of-function effects. The truncated variant p.Gln889Ter would have profoundly different effect on epilepsy. Together, our results underscore the significance of USP25 heterozygous variants in epilepsy, thereby highlighting the critical role of USP25 in the brain.

Dual Surface Architectonics for Directed Self-Assembly of Ultrahigh-Resolution Electronics.

The directed self-assembly of electronic circuits using functional metallic inks has attracted intensive attention because of its high compatibility with extensive applications ranging from soft printed circuits to wearable devices. However, the typical resolution of conventional self-assembly technologies is not sufficient for practical applications in the rapidly evolving additively manufactured electronics (AMEs) market. Herein, an ultrahigh-resolution self-assembly strategy is reported based on a dual-surface-architectonics (DSA) process. Inspired by the Tokay gecko, the approach is to endow submicrometer-scale surface regions with strong adhesion force toward metallic inks via a series of photoirradiation and chemical polarization treatments. The prepared DSA surface enables the directed self-assembly of electronic circuits with unprecedented 600 nm resolution, suppresses the coffee-ring effect, and results in a reliable conductivity of 14.1 ± 0.6 µΩ cm. Furthermore, the DSA process enables the layer-by-layer fabrication of fully printed organic thin-film transistors with a short channel length of 1 µm, which results in a large on-off ratio of 106 and a high field-effect mobility of 0.5 cm2  V-1  s-1 .

NUS1 Variants Cause Lennox-Gastaut Syndrome Related to Unfolded Protein Reaction Activation.

NUS1 encodes the Nogo-B receptor, a critical regulator for unfolded protein reaction (UPR) signaling. Although several loss-of-function variants of NUS1 have been identified in patients with developmental and epileptic encephalopathy (DEE), the role of the NUS1 variant in Lennox-Gastaut syndrome (LGS), a severe child-onset DEE, remains unknown. In this study, we identified two de novo variants of NUS1, a missense variant (c.868 C > T/p.R290C) and a splice site variant (c.792-2 A > G), in two unrelated LGS patients using trio-based whole-exome sequencing performed in a cohort of 165 LGS patients. Both variants were absent in the gnomAD population and showed a significantly higher observed number of variants than expected genome-wide. The R290C variant was predicted to damage NUS1 and decrease its protein stability. The c.792-2 A > G variant caused premature termination of the protein. Knockdown of NUS1 activated the UPR pathway, resulting in apoptosis of HEK293T cells. Supplementing cells with expression of wild-type NUS1, but not the mutant (R290C), rescued UPR activation and apoptosis in NUS1 knockdown cells. Compared to wild-type Drosophila, seizure-like behaviors and excitability in projection neurons were significantly increased in Tango14 (homolog of human NUS1) knockdown and Tango14R290C/+ knock-in Drosophila. Additionally, abnormal development and a small body size were observed in both mutants. Activated UPR signaling was also detected in both mutants. Thus, NUS1 is a causative gene for LGS with dominant inheritance. The pathogenicity of these variants is related to the UPR signaling activation, which may be a common pathogenic mechanism of DEE.

The role of muscle spindles in the development of the monosynaptic stretch reflex.

Muscle sensory axons induce the development of specialized intrafusal muscle fibers in muscle spindles during development, but the role that the intrafusal fibers may play in the development of the central projections of these Ia sensory axons is unclear. In the present study, we assessed the influence of intrafusal fibers in muscle spindles on the formation of monosynaptic connections between Ia (muscle spindle) sensory axons and motoneurons (MNs) using two transgenic strains of mice. Deletion of the ErbB2 receptor from developing myotubes disrupts the formation of intrafusal muscle fibers and causes a nearly complete absence of functional synaptic connections between Ia axons and MNs. Monosynaptic connectivity can be fully restored by postnatal administration of neurotrophin-3 (NT-3), and the synaptic connections in NT-3-treated mice are as specific as in wild-type mice. Deletion of the Egr3 transcription factor also impairs the development of intrafusal muscle fibers and disrupts synaptic connectivity between Ia axons and MNs. Postnatal injections of NT-3 restore the normal strengths and specificity of Ia-motoneuronal connections in these mice as well. Severe deficits in intrafusal fiber development, therefore, do not disrupt the establishment of normal, selective patterns of connections between Ia axons and MNs, although these connections require the presence of NT-3, normally supplied by intrafusal fibers, to be functional.

3D printing lamellar Ti3C2T x MXene/graphene hybrid aerogels for enhanced electromagnetic interference shielding performance.

Two-dimensional (2D) transition-metal carbides and nitrides (MXenes), especially Ti3C2T x nanosheets, offer high conductivities comparable to metal, and are very promising for fabricating high performance electromagnetic interference (EMI) shielding materials. Due to the weak gelation capability of MXenes, MXene/graphene hybrid aerogels were mostly studied. Among those studied, anisotropic hybrid aerogels showed excellent electrical properties in certain direction due to the intrinsic anisotropic properties of 2D materials. However, the present preparation methods for anisotropic hybrid aerogels lack freedom of geometry, and their electrical performances still have room for improvement. In this study, based on our previous work, the lamellar Ti3C2T x MXene/graphene hybrid aerogels generated by 3D printing with Ti3C2T x MXene/graphene oxide (GO) water-TBA dispersions demonstrated enhanced conductivity and electromagnetic interference (EMI) shielding performance. The addition of MXene deeply influenced the lamellar structure of the hybrid aerogels, and made the structure more ordered than that in the 3D printed lamellar graphene aerogels. The printed lamellar MXene/graphene hybrid aerogels achieved a maximum electrical conductivity of 1236 S m-1. The highest EMI shielding efficiency (EMI SE) of the hybrid aerogels was up to 86.9 dB, while the absolute shielding effectiveness (SSE/t) was up to 25 078.1 dB cm2 g-1 at 12.4 GHz. These values are higher than those of most reported anisotropic MXene-based nanocomposite aerogels.

Self-Organizing, Environmentally Stable, and Low-Cost Copper-Nickel Complex Inks for Printed Flexible Electronics.

Cost-effective copper conductive inks are considered as the most promising alternative to expensive silver conductive inks for use in printed electronics. However, the low stability and high sintering temperature of copper inks hinder their practical application. Herein, we develop rapidly customizable and stable copper-nickel complex inks that can be transformed in situ into uniform copper@nickel core-shell nanostructures by a self-organized process during low-temperature annealing and immediately sintered under photon irradiation to form copper-nickel alloy patterns on flexible substrates. The complex inks are synthesized within 15 min via a simple mixing process and are particle-free, air-stable, and compatible with large-area screen printing. The manufactured patterns exhibit a high conductivity of 19-67 µΩ·cm, with the value depending on the nickel content, and can maintain high oxidation resistance at 180 °C even when the nickel content is as low as 6 wt %. In addition, the printed copper-nickel alloy patterns exhibit high flexibility as a consequence of the local softening and mechanical anchoring effect between the metal pattern and the flexible substrate, showing strong potential in the additive manufacturing of highly reliable flexible electronics, such as flexible radio-frequency identification (RFID) tags and various wearable sensors.

Preparation of graphene oxide liquid crystals with long-range highly-ordered flakes using a coat-hanger die.

Graphene oxide (GO) was discovered as a liquid crystalline (LC) phase formation in its water dispersion and expanded to a large number of applications, such as highly ordered GO sheets papers, films, and foams. However, there are still few efficient ways to prepare graphene oxide liquid crystals (GOLCs) with long-range highly ordered flakes. In this work, after carefully studying the rheological properties of GO aqueous dispersions at different concentrations, we have provided a new method to prepare holistically-oriented GOLCs through a designed coat-hanger die. Further, by simulating the extrusion process in the slot of the coat-hanger die, the die's dimensional sizes were optimized to apply efficient shear force on GO dispersions. Then, GOLCs with long-range highly ordered flakes of different GO concentrations were prepared using this method. Finally, a GO foam with a highly ordered structure was prepared using a layer-by-layer method, which exhibited improved conductivity compared to that of normal disordered GO foams after chemical reduction.

Layer-By-Layer Printing Strategy for High-Performance Flexible Electronic Devices with Low-Temperature Catalyzed Solution-Processed SiO2.

Additive printing techniques have been widely investigated for fabricating multilayered electronic devices. In this work, a layer-by-layer printing strategy is developed to fabricate multilayered electronics including 3D conductive circuits and thin-film transistors (TFTs) with low-temperature catalyzed, solution-processed SiO2 (LCSS) as the dielectric. Ultrafine, ultrasmooth LCSS films can be facilely formed at 90 °C on a wide variety of organic and inorganic substrates, offering a versatile platform to construct complex heterojunction structures with layer-by-layer fashion at microscale. The high-resolution 3D conductive circuits formed with gold nanoparticles inside the LCSS dielectric demonstrate a high-speed response to the transient voltage in less than 1 µs. The TFTs with semiconducting single-wall carbon nanotubes can be operated with the accumulation mode at a low voltage of 1 V and exhibit average field-effect mobility of 70 cm2  V-1  s-1 , on/off ratio of 107 , small average hysteresis of 0.1 V, and high yield up to 100% as well as long-term stability, high negative-gate bias stability, and good mechanical stability. Therefore, the layer-by-layer printing strategy with the LCSS film is promising to assemble large-scale, high-resolution, and high-performance flexible electronics and to provide a fundamental understanding for correlating dielectric properties with device performance.

Three-Dimensional Stretchable and Transparent Conductors with Controllable Strain-Distribution Based on Template-Assisted Transfer Printing.

Although stretchable transparent conductors, stemmed from the strategies of both conductive composite and structural design of nonstretchable conductors, have been extensively studied, these conductors either suffer from low stretchability or require a complex fabrication process, which drastically limits their practical applications. Here, we propose a novel strategy combining the design of substrates and a simple template-assisted transfer printing process to fabricate three-dimensional (3D) transparent conductors. The strategy not only eliminates the complex and costly fabrication processes but it also endows conductors with high stretchability and long-term stability, thanks to the controllable strain distribution as well as the seamless connection between the conductor layer and the substrate. These newly designed 3D conductors achieve a low sheet resistance of 1.0 Ω/sq with a high transmittance of above 85% and remain stable without obvious resistance change during 1000 stretching-relaxation cycles until 60% strain, which are superior to most reported conductors. A large-area stretchable heater based on the 3D conductor realizes the temperature fluctuation below 10% even under a large strain, thus showing huge application prospects in the field of wearable healthcare electronics. The simple solution-processed fabrication method and high performance such as stretchability and low resistance change over a large strain range promote the practical applications of these newly designed 3D conductors.

Highly Conductive Ag Paste for Recoverable Wiring and Reliable Bonding Used in Stretchable Electronics.

Stretchable wiring and stretchable bonding between a rigid chip/component and a stretchable substrate are two key factors for stretchable electronics. In this study, a highly conductive stretchable paste has been developed with commercial Ag microflakes and poly(dimethylsiloxane), which can be used to fabricate stretchable wirings and bondings under a low curing temperature of 100 °C with printing method. Herein, recoverabilities as to recovery time and recovery resistance of the wirings are defined and discussed. The effect of Ag composition and the tensile strain rate on the recoverability of the wirings are also examined. The wiring with a low resistivity of 8.7 × 10-5 Ω cm shows much better recoverability than nanowire-based wirings due to the flake nature of the Ag particles. When stretched to 50 and 100% of strain, the resistance of the patterned wiring increases by only 10 and 110%, respectively. Moreover, the resistance of the wiring during 20% tensile cyclic test remains within 1.1 times even after 1000 cycles, thus demonstrating significant durability. The paste was utilized to fabricate conductive tracks and stretchable bondings to assemble a rigid chip to fabricate a stretchable demo. When stretched to 50% of strain, resistance of the wiring was increased by 90%. It is anticipated that the newly developed paste will be used to fabricate various stretchable wirings, bondings, and packaging structures by a simple printing process, thus enabling mass production of stretchable electronic devices.

Printable and Flexible Copper-Silver Alloy Electrodes with High Conductivity and Ultrahigh Oxidation Resistance.

Printable and flexible Cu-Ag alloy electrodes with high conductivity and ultrahigh oxidation resistance have been successfully fabricated by using a newly developed Cu-Ag hybrid ink and a simple fabrication process consisting of low-temperature precuring followed by rapid photonic sintering (LTRS). A special Ag nanoparticle shell on a Cu core structure is first created in situ by low-temperature precuring. An instantaneous photonic sintering can induce rapid mutual dissolution between the Cu core and the Ag nanoparticle shell so that core-shell structures consisting of a Cu-rich phase in the core and a Ag-rich phase in the shell (Cu-Ag alloy) can be obtained on flexible substrates. The resulting Cu-Ag alloy electrode has high conductivity (3.4 µΩ·cm) and ultrahigh oxidation resistance even up to 180 °C in an air atmosphere; this approach shows huge potential and is a tempting prospect for the fabrication of highly reliable and cost-effective printed electronic devices.

Early postnatal development of reciprocal Ia inhibition in the murine spinal cord.

The pathway mediating reciprocal inhibition from muscle spindle afferents (Ia axons) to motoneurons (MNs) supplying antagonist muscles has been well studied in adult cats, but little is known about how this disynaptic pathway develops. As a basis for studying its development, we characterized this pathway in mice during the first postnatal week, focusing on the projection of quadriceps (Q) Ia axons to posterior biceps-semitendinosis (PBSt) MNs via Ia inhibitory interneurons. Synaptic potentials in PBSt MNs evoked by Q nerve stimulation are mediated disynaptically and are blocked by strychnine, implying that glycine is the major inhibitory transmitter as in adult cats. The specificity of neuronal connections in this reflex pathway is already high at birth; Q afferents evoke inhibitory synaptic potentials in PBSt MNs, but afferents supplying the adductor muscle do not. Similar to this disynaptic pathway in cats, Renshaw cells inhibit the interposed Ia interneurons, as they reduce the disynaptic input from Q axons but do not inhibit PBSt MNs directly. Reciprocal inhibition functionally inhibits the monosynaptic excitatory reflex in PBSt MNs by P3, but this functional inhibition is weak at P1. Finally, deletion of the transcription factor Pax6, which is required for the development of V1-derived Renshaw cells, does not block development of this pathway. This suggests either that Pax6 is not required for the phenotypic development of all V1-derived spinal interneurons or that these inhibitory interneurons are not derived from V1 precursors.

Combinatorial ShcA docking interactions support diversity in tissue morphogenesis.

Changes in protein-protein interactions may allow polypeptides to perform unexpected regulatory functions. Mammalian ShcA docking proteins have amino-terminal phosphotyrosine (pTyr) binding (PTB) and carboxyl-terminal Src homology 2 (SH2) domains, which recognize specific pTyr sites on activated receptors, and a central region with two phosphorylated tyrosine-X-asparagine (pYXN) motifs (where X represents any amino acid) that each bind the growth factor receptor-bound protein 2 (Grb2) adaptor. Phylogenetic analysis indicates that ShcA may signal through both pYXN-dependent and -independent pathways. We show that, in mice, cardiomyocyte-expressed ShcA directs mid-gestational heart development by a PTB-dependent mechanism that does not require the pYXN motifs. In contrast, the pYXN motifs are required with PTB and SH2 domains in the same ShcA molecule for the formation of muscle spindles, skeletal muscle sensory organs that regulate motor behavior. Thus, combinatorial differences in ShcA docking interactions may yield multiple signaling mechanisms to support diversity in tissue morphogenesis.