A novel transcription and replication-competent virus-like particles system modelling the Nipah virus life cycle.

AbstractNipah virus (NiV), a highly pathogenic Henipavirus in humans, has been responsible for annual outbreaks in recent years. Experiments revolving live NiV are highly restricted to biosafety level 4 (BSL-4) laboratories, which impedes NiV research. In this study, we developed a transcription and replication-competent NiV-like particles (trVLP-NiV) lacking N, P, and L genes. This trVLP-NiV exhibited the ability to infect and continuously passage in cells ectopically expressing N, P, and L proteins while maintaining stable genetic characteristics. Moreover, the trVLP-NiV displayed a favourable safety profile in hamsters. Using the system, we found the NiV nucleoprotein residues interacting with viral RNA backbone affected viral replication in opposite patterns. This engineered system was sensitive to well-established antiviral drugs, innate host antiviral factors, and neutralising antibodies. We then established a high-throughput screening platform utilizing the trVLP-NiV, leading to the identification of tunicamycin as a potential anti-NiV compound. Evidence showed that tunicamycin inhibited NiV replication by decreasing the infectivity of progeny virions. In conclusion, this trVLP-NiV system provided a convenient and versatile molecular tool for investigating NiV molecular biology and conducting antiviral drug screening under BSL-2 conditions. Its application will contribute to the development of medical countermeasures against NiV infections.

Applying improved ddPCR to reliable quantification of MPXV in clinical settings.

Monkeypox virus (MPXV) poses a global health threat. Droplet digital PCR (ddPCR) holds potential as an accurate diagnostic tool for clinical microbiology. However, there is limited literature on the applicability of ddPCR in clinical settings. In this study, the clinical features of patients with MPXV during the initial outbreak in China in June 2023 were reviewed, and an optimized ddPCR method with dilution and/or inhibitor removal was developed to enhance MPXV detection efficiency. Eighty-two MPXV samples were tested from nine different clinical specimen types, including feces, urine, pharyngeal swabs, anal swabs, saliva, herpes fluid, crust, and semen, and the viral load of each specimen was quantified. A comparative analysis was performed with qPCR to assess sensitivity and specificity and to investigate the characteristics of MPXV infection by analyzing viral loads in different clinical specimens. Consequently, common pharyngeal and gastrointestinal symptoms were observed in patients with MPXV. The optimized ddPCR method demonstrated relatively high sensitivity for MPXV quantification in the clinical materials, with a limit of detection of 0.1 copies/µL. This was particularly evident in low-concentration samples like whole blood, semen, and urine. The optimized ddPCR demonstrated greater detection accuracy compared with normal ddPCR and qPCR, with an area under the curve (AUC) of 0.939. Except for crust samples, viral loads in the specimens gradually decreased as the disease progressed. Virus levels in feces and anal swabs kept a high detection rate at each stage of post-symptom onset, and feces and anal swabs samples may be suitable for clinical diagnosis and continuous monitoring of MPXV. IMPORTANCE: The ddPCR technique proved to be a sensitive and valuable tool for accurately quantifying MPXV viral loads in various clinical specimen types. The findings provided valuable insights into the necessary pre-treatment protocols for MPXV diagnosis in ddPCR detection and the potentially suitable sample types for collection. Therefore, such results can aid in comprehending the potential characteristics of MPXV infection and the usage of ddPCR in clinical settings.

RNA binding motif 4 inhibits the replication of ebolavirus by directly targeting 3'-leader region of genomic RNA.

Ebola virus (EBOV) belongs to Filoviridae family possessing single-stranded negative-sense RNA genome, which is a serious threat to human health. Nowadays, no therapeutics have been proven to be successful in efficiently decreasing the mortality rate. RNA binding proteins (RBPs) are reported to participate in maintaining cell integrity and regulation of viral replication. However, little is known about whether and how RBPs participate in regulating the life cycle of EBOV. In our study, we found that RNA binding motif protein 4 (RBM4) inhibited the replication of EBOV in HEK293T and Huh-7 cells by suppressing viral mRNA production. Such inhibition resulted from the direct interaction between the RRM1 domain of RBM4 and the "CU" enrichment elements located in the PE1 and TSS of the 3'-leader region within the viral genome. Simultaneously, RBM4 could upregulate the expression of some cytokines involved in the host innate immune responses to synergistically exert its antiviral function. The findings therefore suggest that RBM4 might serve as a novel target of anti-EBOV strategy.

Endothelial BACE1 Impairs Cerebral Small Vessels via Tight Junctions and eNOS.

BACKGROUND: Cerebral small vessel injury, including loss of endothelial tight junctions, endothelial dysfunction, and blood-brain barrier breakdown, is an early and typical pathology for Alzheimer's disease, cerebral amyloid angiopathy, and hypertension-related cerebral small vessel disease. Whether there is a common mechanism contributing to these cerebrovascular alterations remains unclear. Studies have shown an elevation of BACE1 (ß-site amyloid precursor protein cleaving enzyme 1) in cerebral vessels from cerebral amyloid angiopathy or Alzheimer's disease patients, suggesting that vascular BACE1 may involve in cerebral small vessel injury. METHODS: To understand the contribution of vascular BACE1 to cerebrovascular impairments, we combined cellular and molecular techniques, mass spectrometry, immunostaining approaches, and functional testing to elucidate the potential pathological mechanisms. RESULTS: We observe a 3.71-fold increase in BACE1 expression in the cerebral microvessels from patients with hypertension. Importantly, we discover that an endothelial tight junction protein, occludin, is a completely new substrate for endothelial BACE1. BACE1 cleaves occludin with full-length occludin reductions and occludin fragment productions. An excessive cleavage by elevated BACE1 induces membranal accumulation of caveolin-1 and subsequent caveolin-1-mediated endocytosis, resulting in lysosomal degradation of other tight junction proteins. Meanwhile, membranal caveolin-1 increases the binding to eNOS (endothelial nitric oxide synthase), together with raised circulating Aß (ß-amyloid peptides) produced by elevated BACE1, leading to an attenuation of eNOS activity and resultant endothelial dysfunction. Furthermore, the initial endothelial damage provokes chronic reduction of cerebral blood flow, blood-brain barrier leakage, microbleeds, tau hyperphosphorylation, synaptic loss, and cognitive impairment in endothelial-specific BACE1 transgenic mice. Conversely, inhibition of aberrant BACE1 activity ameliorates tight junction loss, endothelial dysfunction, and memory deficits. CONCLUSIONS: Our findings establish a novel and direct relationship between endothelial BACE1 and cerebral small vessel damage, indicating that abnormal elevation of endothelial BACE1 is a new mechanism for cerebral small vessel disease pathogenesis.

Millimeter-wave SFCW SAR imaging system based on in-phase signal measurement with simplified transceiver.

A conventional millimeter-wave stepped frequency continuous wave (SFCW) synthetic aperture radar (SAR) imaging system generally utilizes IQ modulation technique to acquire the amplitude and phase of the waves scattered from a target object. Due to measure both in-phase signal and quadrature signal, the transceiver of the conventional system is complicate and costly, and the IQ imbalance problem makes the system difficult to calibrate. To reduce hardware complexity and enhance efficiency-cost ratio, a novel SFCW SAR imaging system only measuring in-phase signal is proposed and demonstrated. For lack of quadrature signal measurement, an algorithm based on Fourier transform is proposed to estimate the amplitude and phase. The ultimate images are obtained through an image-reconstruction algorithm, which uses the estimated amplitude and phase as input parameters. The proposed system is verified by both simulation and experiment, where the frequencies are set from 24GHz to 30GHz. The imaging results with high resolution and low noise are demonstrated. Compared to the conventional system, the image quality of the proposed system is almost identical, but the transceiver of the proposed system is greatly simplified.

Terahertz image super-resolution based on a deep convolutional neural network.

We propose an effective and robust method for terahertz (THz) image super-resolution based on a deep convolutional neural network (CNN). A deep CNN model is designed. It learns an end-to-end mapping between the low- and high-resolution images. Blur kernels with multiple width and noise with multiple levels are taken into the training set so that the network can handle THz images very well. Quantitative comparison of the proposed method and other super-resolution methods on the synthetic THz images indicates that the proposed method performs better than other methods in accuracy and visual improvements. Experimental results on real THz images show that the proposed method significantly improves the quality of THz images with increased resolution and decreased noise, which proves the practicability and exactitude of the proposed method.

Using sounds for making decisions: greater tube-nosed bats prefer antagonistic calls over non-communicative sounds when feeding.

Bats vocalize extensively within different social contexts. The type and extent of information conveyed via their vocalizations and their perceptual significance, however, remains controversial and difficult to assess. Greater tube-nosed bats, Murina leucogaster, emit calls consisting of long rectangular broadband noise burst (rBNBl) syllables during aggression between males. To experimentally test the behavioral impact of these sounds for feeding, we deployed an approach and place-preference paradigm. Two food trays were placed on opposite sides and within different acoustic microenvironments, created by sound playback, within a specially constructed tent. Specifically, we tested whether the presence of rBNBl sounds at a food source effectively deters the approach of male bats in comparison to echolocation sounds and white noise. In each case, contrary to our expectation, males preferred to feed at a location where rBNBl sounds were present. We propose that the species-specific rBNBl provides contextual information, not present within non-communicative sounds, to facilitate approach towards a food source.

Situational and Age-Dependent Decision Making during Life Threatening Distress in Myotis macrodactylus.

Echolocation and audiovocal communication have been studied extensively in bats. The manner in which these abilities are incorporated within escape behaviors during life-threatening distress is largely unknown. Here we tested the hypothesis that behavioral response profiles expressed during distress are relatively stereotypic given their evolutionary adaptations to avoid predators. We subjected juvenile and adult big-footed myotis (Myotis macrodactylus) to a sequence of three types of life threatening distress: 1) trapping them in a mist-net (environmental threat), 2) approaching them when trapped (predator threat), and 3) partially restraining their freedom to move (arrest), and recorded their escape behavior in each of the three conditions. Response profiles differed across individuals and with the context in which they were expressed. During environmental and predator threat, bats displayed significantly more biting and wing-flapping behaviors and emitted more echolocation pulses than during arrest. Response profiles also varied with age. During arrest, juveniles were more likely than adults to emit distress calls and vice-versa for biting and wing flapping during environmental and predator threat. Overall, individualized response profiles were classified into ten clusters that were aligned along two divergent response trajectories when viewed within two-dimensional, multifactorial decision space. Juvenile behaviors tended to follow a predominantly "social-dependence" trajectory, whereas adult behaviors were mostly aligned along a "self-reliance" trajectory. We conclude that bats modify their vocal behavior and make age-appropriate and contextually adaptive decisions when distressed. This decision-making ability is consistent with observations in other social species, including humans.