Molecular Basis for Ligand Modulation of a Mammalian Voltage-Gated Ca2+ Channel.

The L-type voltage-gated Ca2+ (Cav) channels are modulated by various compounds exemplified by 1,4-dihydropyridines (DHP), benzothiazepines (BTZ), and phenylalkylamines (PAA), many of which have been used for characterizing channel properties and for treatment of hypertension and other disorders. Here, we report the cryoelectron microscopy (cryo-EM) structures of Cav1.1 in complex with archetypal antagonistic drugs, nifedipine, diltiazem, and verapamil, at resolutions of 2.9 Å, 3.0 Å, and 2.7 Å, respectively, and with a DHP agonist Bay K 8644 at 2.8 Å. Diltiazem and verapamil traverse the central cavity of the pore domain, directly blocking ion permeation. Although nifedipine and Bay K 8644 occupy the same fenestration site at the interface of repeats III and IV, the coordination details support previous functional observations that Bay K 8644 is less favored in the inactivated state. These structures elucidate the modes of action of different Cav ligands and establish a framework for structure-guided drug discovery.

Structure of the Nav1.4-ß1 Complex from Electric Eel.

Voltage-gated sodium (Nav) channels initiate and propagate action potentials. Here, we present the cryo-EM structure of EeNav1.4, the Nav channel from electric eel, in complex with the ß1 subunit at 4.0 Å resolution. The immunoglobulin domain of ß1 docks onto the extracellular L5I and L6IV loops of EeNav1.4 via extensive polar interactions, and the single transmembrane helix interacts with the third voltage-sensing domain (VSDIII). The VSDs exhibit "up" conformations, while the intracellular gate of the pore domain is kept open by a digitonin-like molecule. Structural comparison with closed NavPaS shows that the outward transfer of gating charges is coupled to the iris-like pore domain dilation through intricate force transmissions involving multiple channel segments. The IFM fast inactivation motif on the III-IV linker is plugged into the corner enclosed by the outer S4-S5 and inner S6 segments in repeats III and IV, suggesting a potential allosteric blocking mechanism for fast inactivation.

Impact of specific electromagnetic radiation on wakefulness in mice.

Electromagnetic radiation (EMR) in the environment, particularly in the microwave range, may constitute a public health concern. Exposure to 2.4 GHz EMR modulated by 100 Hz square pulses was recently reported to markedly increase wakefulness in mice. Here, we demonstrate that a similar wakefulness increase can be induced by the modulation frequency of 1,000 Hz, but not 10 Hz. In contrast to the carrier frequency of 2.4 GHz, 935 MHz EMR of the same power density has little impact on wakefulness irrespective of modulation frequency. Notably, the replacement of the 100 Hz square-pulsed modulation by sinusoidal-pulsed modulation of 2.4 GHz EMR still allows a marked increase of wakefulness. In contrast, continuous sinusoidal amplitude modulation of 100 Hz with the same time-averaged power output fails to trigger any detectable change of wakefulness. Therefore, alteration of sleep behavior by EMR depends upon not just carrier frequency but also frequency and mode of the modulation. These results implicate biological sensing mechanisms for specific EMR in animals.

Cryo-EM structures of apo and antagonist-bound human Cav3.1.

Among the ten subtypes of mammalian voltage-gated calcium (Cav) channels, Cav3.1-Cav3.3 constitute the T-type, or the low-voltage-activated, subfamily, the abnormal activities of which are associated with epilepsy, psychiatric disorders and pain1-5. Here we report the cryo-electron microscopy structures of human Cav3.1 alone and in complex with a highly Cav3-selective blocker, Z9446,7, at resolutions of 3.3 Å and 3.1 Å, respectively. The arch-shaped Z944 molecule reclines in the central cavity of the pore domain, with the wide end inserting into the fenestration on the interface between repeats II and III, and the narrow end hanging above the intracellular gate like a plug. The structures provide the framework for comparative investigation of the distinct channel properties of different Cav subfamilies.

Specific electromagnetic radiation in the wireless signal range increases wakefulness in mice.

Electromagnetic radiation (EMR) in the environment has increased sharply in recent decades. The effect of environmental EMR on living organisms remains poorly characterized. Here, we report the impact of wireless-range EMR on the sleep architecture of mouse. Prolonged exposure to 2.4-GHz EMR modulated by 100-Hz square pulses at a nonthermal output level results in markedly increased time of wakefulness in mice. These mice display corresponding decreased time of nonrapid eye movement (NREM) and rapid eye movement (REM). In contrast, prolonged exposure to unmodulated 2.4-GHz EMR at the same time-averaged output level has little impact on mouse sleep. These observations identify alteration of sleep architecture in mice as a specific physiological response to prolonged wireless-range EMR exposure.

Comparative structural analysis of human Nav1.1 and Nav1.5 reveals mutational hotspots for sodium channelopathies.

Among the nine subtypes of human voltage-gated sodium (Nav) channels, the brain and cardiac isoforms, Nav1.1 and Nav1.5, each carry more than 400 missense mutations respectively associated with epilepsy and cardiac disorders. High-resolution structures are required for structure-function relationship dissection of the disease variants. We report the cryo-EM structures of the full-length human Nav1.1-ß4 complex at 3.3 Å resolution here and the Nav1.5-E1784K variant in the accompanying paper. Up to 341 and 261 disease-related missense mutations in Nav1.1 and Nav1.5, respectively, are resolved. Comparative structural analysis reveals several clusters of disease mutations that are common to both Nav1.1 and Nav1.5. Among these, the majority of mutations on the extracellular loops above the pore domain and the supporting segments for the selectivity filter may impair structural integrity, while those on the pore domain and the voltage-sensing domains mostly interfere with electromechanical coupling and fast inactivation. Our systematic structural delineation of these mutations provides important insight into their pathogenic mechanism, which will facilitate the development of precise therapeutic interventions against various sodium channelopathies.

Visible Light-Responsive Crystalline B←N Host Adducts with Solvent-Induced Allosteric Effect for Guest Release.

Photo-responsive organic crystals, capable of converting light energy into chemical energy to initiate conformational transitions, present an emerging strategy for developing lightweight and versatile smart materials. However, visible light-triggered tailored guests capture and release behaviors in all-organic solids are rarely reported. Here, we introduce a photoreactive crystalline boron-nitrogen (B←N) host adduct with the ability to undergo [2+2] photocycloaddition upon 447 nm light exposure. This process facilitates single-crystal-to-single-crystal (SCSC) photodimerization in the mother liquor, maintaining the original B←N host structure. Weakened intermolecular interactions within the photodimer host contribute to fast guest release in air under irradiation. Furthermore, the dynamic B←N bonds enable reversible transformations between organic host adducts and adduct cocrystals under the solvent-induced allosteric effect. As a result, four B←N host adduct crystals containing individual alkane guest are easily obtained and exhibited the ability of photo-controlled alkane release. Therefore, the integration of photo reactivity and structural transformation within B←N host adduct enables customized capture and release of guest molecules.

Expression and Activity of the Transcription Factor CCAAT/Enhancer-Binding Protein ß (C/EBPß) Is Regulated by Specific Pulse-Modulated Radio Frequencies in Oligodendroglial Cells.

The rapid growth of wireless electronic devices has raised concerns about the harmful effects of leaked electromagnetic radiation (EMR) on human health. Even though numerous studies have been carried out to explore the biological effects of EMR, no clear conclusions have been drawn about the effect of radio frequency (RF) EMR on oligodendrocytes. To this end, we exposed oligodendroglia and three other types of brain cells to 2.4 GHz EMR for 6 or 48 h at an average input power of 1 W in either a continuous wave (CW-RF) or a pulse-modulated wave (PW-RF, 50 Hz pulse frequency, 1/3 duty cycle) pattern. RNA sequencing, RT-qPCR, and Western blot were used to examine the expression of C/EBPß and its related genes. Multiple reaction monitoring (MRM) was used to examine the levels of expression of C/EBPß-interacting proteins. Our results showed that PW-RF EMR significantly increased the mRNA level of C/EBPß in oligodendroglia but not in other types of cells. In addition, the expression of three isoforms and several interacting proteins and targeted genes of C/EBPß were markedly changed after 6-h PW-RF but not CW-RF. Our results indicated that RF EMR regulated the expression and functions of C/EBPß in a waveform- and cell-type-dependent manner.

Gradient-assisted focusing light through scattering media.

Focusing light through a scattering medium is a longstanding challenge in biomedical optics, to which wavefront shaping is a powerful solution. The state-of-the-art feedback-based approach is the widely used genetic algorithm method. However, it can only achieve relatively low enhancement of the focus, and the genetic algorithm is known to be time-consuming. To tackle those issues, we propose a gradient-assisted strategy for wavefront shaping. The proposed method conducts optimization in the function distribution space. Specifically, when optimizing the parameters along each iteration, the consequent function distribution changes within a distance as measured by the Kullback-Leibler divergence. Taking advantage of the gradient information, the proposed method is over 60× faster to obtain the same peak-to-background ratio (PBR) level. Compared with the genetic algorithm that is able to optimize a number of 64×64 phase segments, the proposed gradient strategy is able to optimize 256×256 phase segments, and gives 20× higher focus enhancement as quantified by the PBR.

Laser curve extraction of a train wheelset based on an encoder-decoder network.

An algorithm of laser curve segmentation for a train wheelset based on an encoder- decoder network is proposed. Aiming at the rich local features and simple semantic features of the train wheelset laser curve image, a neural network with shallow depth, high resolution, and good detail performance was designed. The proposed neural network makes full use of the dense connection mechanism and the upsampling module to enhance feature reuse and feature propagation. It can extract context semantic information at multiple scales with fewer parameters. Experimental results show that the encoder-decoder network has better performance than other neural networks in laser curve extraction of train wheelset. Based on the encoder-decoder neural network, mIOU, Recall, Accuracy, and F1_score of the laser curve dataset of the train wheelset, the score index reached 86.5%, 89.2%, 99.9%, and 85.0%, which can accurately extract the laser stripe of the train wheelset. Additionally, the encoder-decoder network can diminish the influence of noise on the extraction of laser fringes of a train wheelset to a certain extent. Therefore, it has good application in railway safety.

Probabilistic natural mapping of gene-level tests for genome-wide association studies.

Genome-wide association studies (GWASs) generally focus on a single marker, which limits the elucidation of the genetic architecture of complex traits. Herein, we present a new computational framework, termed probabilistic natural mapping (PALM), for performing gene-level association tests. PALM robustly reveals the inherent genomic structures of genes and generates feature representations that can be seamlessly incorporated into conventional statistic tests. Our approach substantially improves the effectiveness of uncovering associations derived from a subgroup of variants with weak effects, which represents a known challenge associated with existing methods. We applied PALM in a gastric cancer GWAS and identified two additional gastric cancer-associated susceptibility genes, NOC3L and RUNDC2A. The robust susceptibility discoveries of PALM are widely supported by existing studies from other biological perspectives. PALM will be useful for further GWAS analytical strategies that use gene-level analyses.

One-Dimensional Face-Shared Perovskites with Broad-Band Bluish White-Light Emissions.

In recent years, two-dimensional (2D) hybrid lead halide perovskites based on corner-shared [PbX6] octahedrons have received extensive attention with important potentials in single-component white-light emitting diodes (WLEDs) due to the soft and distorted crystal lattices. However, limited research focused on the one-dimensional (1D) perovskites although they possess similar structural superiorities to achieve this performance. Herein, by using different types of organic amine cations as structural direction reagents, we report one new type of hybrid 1D perovskites of APbCl3 (A = (DTHPE)0.5, DMTHP, DBN) based on the same 1D face-shared octahedral [PbCl3]- chains. Upon UV light excitation, these 1D APbCl3 perovskites exhibit intrinsic broad-band bluish white-light emissions covering entire visible light spectra with the highest photoluminescence quantum yield (PLQY) of 6.99%, which catches up with the values of previously reported 2D perovskites. Through the systematical studies of time-resolved, temperature-dependent PL emissions, theoretical calculations, and so on, these broad-band light emissions can be ascribed to the radiative transition within conjugated organic cations. The facile assembly process, intrinsic broad-band light emissions, and high PLQYs enable these 1D APbCl3 perovskites as new types of promising candidates in fabricating single-component WLEDs.

Three-dimensional structure of human γ-secretase.

The γ-secretase complex, comprising presenilin 1 (PS1), PEN-2, APH-1 and nicastrin, is a membrane-embedded protease that controls a number of important cellular functions through substrate cleavage. Aberrant cleavage of the amyloid precursor protein (APP) results in aggregation of amyloid-ß, which accumulates in the brain and consequently causes Alzheimer's disease. Here we report the three-dimensional structure of an intact human γ-secretase complex at 4.5 Å resolution, determined by cryo-electron-microscopy single-particle analysis. The γ-secretase complex comprises a horseshoe-shaped transmembrane domain, which contains 19 transmembrane segments (TMs), and a large extracellular domain (ECD) from nicastrin, which sits immediately above the hollow space formed by the TM horseshoe. Intriguingly, nicastrin ECD is structurally similar to a large family of peptidases exemplified by the glutamate carboxypeptidase PSMA. This structure serves as an important basis for understanding the functional mechanisms of the γ-secretase complex.

Deep learning model for ultrafast multifrequency optical property extractions for spatial frequency domain imaging.

Spatial frequency domain imaging (SFDI) is emerging as an important new method in biomedical imaging due to its ability to provide label-free, wide-field tissue optical property maps. Most prior SFDI studies have utilized two spatial frequencies (2-fx) for optical property extractions. The use of more than two frequencies (multi-fx) can vastly improve the accuracy and reduce uncertainties in optical property estimates for some tissue types, but it has been limited in practice due to the slow speed of available inversion algorithms. We present a deep learning solution that eliminates this bottleneck by solving the multi-fx inverse problem 300× to 100,000× faster, with equivalent or improved accuracy compared to competing methods. The proposed deep learning inverse model will help to enable real-time and highly accurate tissue measurements with SFDI.

Structural basis of human γ-secretase assembly.

The four-component intramembrane protease γ-secretase is intricately linked to the development of Alzheimer's disease. Despite recent structural advances, the transmembrane segments (TMs) of γ-secretase remain to be specifically assigned. Here we report a 3D structure of human γ-secretase at 4.32-Šresolution, determined by single-particle, electron cryomicroscopy in the presence of digitonin and with a T4 lysozyme fused to the amino terminus of presenilin 1 (PS1). The overall structure of this human γ-secretase is very similar to that of wild-type γ-secretase determined in the presence of amphipols. The 20 TMs are unambiguously assigned to the four components, revealing principles of subunit assembly. Within the transmembrane region, PS1 is centrally located, with its amino-terminal fragment (NTF) packing against Pen-2 and its carboxyl-terminal fragment (CTF) interacting with Aph-1. The only TM of nicastrin associates with Aph-1 at the thick end of the TM horseshoe, and the extracellular domain of nicastrin directly binds Pen-2 at the thin end. TM6 and TM7 in PS1, which harbor the catalytic aspartate residues, are located on the convex side of the TM horseshoe. This structure serves as an important framework for understanding the function and mechanism of γ-secretase.

Crystal structure of the γ-secretase component nicastrin.

γ-Secretase is an intramembrane protease responsible for the generation of amyloid-ß (Aß) peptides. Aberrant accumulation of Aß leads to the formation of amyloid plaques in the brain of patients with Alzheimer's disease. Nicastrin is the putative substrate-recruiting component of the γ-secretase complex. No atomic-resolution structure had been identified on γ-secretase or any of its four components, hindering mechanistic understanding of γ-secretase function. Here we report the crystal structure of nicastrin from Dictyostelium purpureum at 1.95-Å resolution. The extracellular domain of nicastrin contains a large lobe and a small lobe. The large lobe of nicastrin, thought to be responsible for substrate recognition, associates with the small lobe through a hydrophobic pivot at the center. The putative substrate-binding pocket is shielded from the small lobe by a lid, which blocks substrate entry. These structural features suggest a working model of nicastrin function. Analysis of nicastrin structure provides insights into the assembly and architecture of the γ-secretase complex.

Structural insights into the T6SS effector protein Tse3 and the Tse3-Tsi3 complex from Pseudomonas aeruginosa reveal a calcium-dependent membrane-binding mechanism.

The opportunistic pathogen Pseudomonas aeruginosa uses the type VI secretion system (T6SS) to deliver the muramidase Tse3 into the periplasm of rival bacteria to degrade their peptidoglycan (PG). Concomitantly, P. aeruginosa uses the periplasm-localized immunity protein Tsi3 to prevent potential self-intoxication caused by Tse3, and thus gains an edge over rival bacteria in fierce niche competition. Here, we report the crystal structures of Tse3 and the Tse3-Tsi3 complex. Tse3 contains an annexin repeat-like fold at the N-terminus and a G-type lysozyme fold at the C-terminus. One loop in the N-terminal domain (Loop 12) and one helix (α9) from the C-terminal domain together anchor Tse3 and the Tse3-Tsi3 complex to membrane in a calcium-dependent manner in vitro, and this membrane-binding ability is essential for Tse3's activity. In the C-terminal domain, a Y-shaped groove present on the surface likely serves as the PG binding site. Two calcium-binding motifs are also observed in the groove and these are necessary for Tse3 activity. In the Tse3-Tsi3 structure, three loops of Tsi3 insert into the substrate-binding groove of Tse3, and three calcium ions present at the interface of the complex are indispensable for the formation of the Tse3-Tsi3 complex.

Interactions between electromagnetic radiation and biological systems.

Even though the bioeffects of electromagnetic radiation (EMR) have been extensively investigated during the past several decades, our understandings of the bioeffects of EMR and the mechanisms of the interactions between the biological systems and the EMRs are still far from satisfactory. In this article, we introduce and summarize the consensus, controversy, limitations, and unsolved issues. The published works have investigated the EMR effects on different biological systems including humans, animals, cells, and biochemical reactions. Alternative methodologies also include dielectric spectroscopy, detection of bioelectromagnetic emissions, and theoretical predictions. In many studies, the thermal effects of the EMR are not properly controlled or considered. The frequency of the EMR investigated is limited to the commonly used bands, particularly the frequencies of the power line and the wireless communications; far fewer studies were performed for other EMR frequencies. In addition, the bioeffects of the complex EM environment were rarely discussed. In summary, our understanding of the bioeffects of the EMR is quite restrictive and further investigations are needed to answer the unsolved questions.

Pharmacological actions of the bioactive compounds of Epimedium on the male reproductive system: current status and future perspective.

ABSTRACT: Compounds isolated from Epimedium include the total flavonoids of Epimedium, icariin, and its metabolites (icaritin, icariside I, and icariside II), which have similar molecular structures. Modern pharmacological research and clinical practice have proved that Epimedium and its active components have a wide range of pharmacological effects, especially in improving sexual function, hormone regulation, anti-osteoporosis, immune function regulation, anti-oxidation, and anti-tumor activity. To date, we still need a comprehensive source of knowledge about the pharmacological effects of Epimedium and its bioactive compounds on the male reproductive system. However, their actions in other tissues have been reviewed in recent years. This review critically focuses on the Epimedium, its bioactive compounds, and the biochemical and molecular mechanisms that modulate vital pathways associated with the male reproductive system. Such intrinsic knowledge will significantly further studies on the Epimedium and its bioactive compounds that protect the male reproductive system and provide some guidances for clinical treatment of related male reproductive disorders.