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Summary: Premature ovarian failure (POF) is a complex and heterogeneous disease that causes infertility and subfertility. However, the molecular mechanism of POF has not been fully elucidated. Here, we show that the loss of adenylyl cyclase III (Adcy3) in female mice leads to POF and a shortened reproductive lifespan. We found that Adcy3 is abundantly expressed in mouse oocytes. Adcy3 knockout mice exhibited the excessive activation of primordial follicles, progressive follicle loss, follicular atresia, and ultimately POF. Mechanistically, we found that mitochondrial oxidative stress in oocytes significantly increased with age in Adcy3-deficient mice and was accompanied by oocyte apoptosis and defective folliculogenesis. In contrast, compared with wild-type female mice, humanized ADCY3 knock-in female mice exhibited improved fertility with age. Collectively, these results reveal that the previously unrecognized Adcy3 signaling pathway is tightly linked to female ovarian aging, providing potential pharmaceutical targets for preventing and treating POF.

Mortar stands as one of the fundamental components in masonry construction, significantly enriching the global cultural heritage, particularly evident in ancient masonry structures. In this paper, the improvements of shear strength of mortar were studied, based on the original formulation material of sticky-rice lime mortar. The study considered the effect of different aggregates and additives on the shear strength of mortar, the shear tests and scanning electron microscopy were employed to analyze the shear strength index and microscopic shear basis of the mortar. The results showed that 2 wt% Na-SiO2 significantly enhanced the shear strength of the mortar. Furthermore, the adhesion and frictional resistance between fiber and lime played a crucial role in enhancing the mortar's shear resistance. Incorporating a 0.5 wt% fiber additive not only effectively improved the mortar's brittleness but also acted as a deterrent to crack formation. Compared to pure lime mortar, the mixture of sand and clay aggregate exhibited a larger capacity at the same volume. This mortar also showcased better plasticity and ductility, making it more suitable for structures with significant shear deformation. These analytical findings provide valuable insights for modifying mortars in the restoration of historical buildings.

Ebola disease is a lethal viral hemorrhagic fever caused by ebolaviruses within the Filoviridae family with mortality rates of up to 90%. Monoclonal antibody (mAb) based therapies have shown great potential for the treatment of EVD. However, the potential emerging ebolavirus isolates and the negative effect of decoy protein on the therapeutic efficacy of antibodies highlight the necessity of developing novel antibodies to counter the threat of Ebola. Here, 11 fully human mAbs were isolated from transgenic mice immunized with GP protein and recombinant vesicular stomatitis virus-bearing GP (rVSV-EBOV GP). These mAbs were divided into five groups according to their germline genes and exhibited differential binding activities and neutralization capabilities. In particular, mAbs 8G6, 2A4, and 5H4 were cross-reactive and bound at least three ebolavirus glycoproteins. mAb 4C1 not only exhibited neutralizing activity but no cross-reaction with sGP. mAb 7D8 exhibited the strongest neutralizing capacity. Further analysis on the critical residues for the bindings of 4C1 and 8G6 to GPs was conducted using antibodies complementarity-determining regions (CDRs) alanine scanning. It has been shown that light chain CDR3 played a crucial role in binding and neutralization and that any mutation in CDRs could not improve the binding of 4C1 to sGP. Importantly, mAbs 7D8, 8G6, and 4C1 provided complete protections against EBOV infection in a hamster lethal challenge model when administered 12 h post-infection. These results support mAbs 7D8, 8G6, and 4C1 as potent antibody candidates for further investigations and pave the way for further developments of therapies and vaccines.

Thin-walled concrete-filled steel tubes are efficient and economical with promising applications in civil and light industrial buildings. However, their local buckling resistance and deformation capacity are low, which adversely affects the seismic safety of structures. There are relatively few studies on thin-walled concrete-filled steel tubular columns with ultra-large diameter-to-thickness ratios, and there is also a lack of relevant experimental research on them. In this study, horizontal hysteresis tests were conducted on concrete columns with a large diameter-to-thickness ratio. The seismic performances of regular and straight-ribbed specimens were analyzed and compared, including the analyses of load-displacement hysteresis curves, strain distribution, skeleton curves, ductility, and energy dissipation capacity. Using these results, a restoring force model for concrete columns with a large diameter-to-thickness ratio was established. The findings indicate that under horizontal loading, the ductility of concrete columns with a regular thin-walled steel tube is 3.9, with an equivalent viscous damping coefficient of 1.65. Meanwhile, the ultimate bearing capacity is 201 kN. After adding stiffening ribs, the ultimate bearing capacity reaches 266 kN and the ductility coefficient reaches 4.4, resulting in the stiffeners increasing the ultimate bearing capacity and ductility by >30% and 12.8%, respectively. However, they have a less pronounced effect on deformation and energy dissipation. Building on these research outcomes, we propose a dimensionless three-line skeleton curve model and a restoring force model. The calculation results from these models align well with the test results, offering valuable insights for the seismic safety analysis of real-world engineering structures.

To address the issue of local buckling in thin-walled steel tube concrete columns, a form of helical stiffening ribs was proposed. Axial compression tests were conducted on five sections of square steel tube concrete column specimens. The research revealed that, compared to ordinary steel tube concrete columns, the axial compression bearing capacity and deformation capacity of steel tube concrete columns with helical rib constraints increased by 18.5% and 7.7%, respectively. The helical ribs effectively enhanced the buckling resistance of the thin-walled steel tube concrete components. The failure pattern of this new type of component was characterized by diagonal cracks in the encased concrete aligning with the direction of the helical ribs, and the buckling of the steel tube walls was concentrated between the helical stiffening ribs. Based on the experiments, an analysis of the buckling performance of thin-walled steel tubes with helical rib constraints was conducted, and this was incorporated into the bearing capacity calculation. The test, simulation, and theoretical calculations showed that the bearing capacity error of the composite columns for each specimen was within 10%. Ultimately, a formula for the critical buckling bearing capacity of the helical rib steel tubes was proposed. The research findings provide a foundation for the engineering application of this new type of component.

Abstract Primary cilia are conserved organelles in most mammalian cells, acting as “antennae” to sense external signals. Maintaining a physiological cilium length is required for cilium function. MicroRNAs (miRNAs) are potent gene expression regulators, and aberrant miRNA expression is closely associated with ciliopathies. However, how miRNAs modulate cilium length remains elusive. Here, using the calcium‐shock method and small RNA sequencing, a miRNA is identified, namely, miR‐669a‐5p, that is highly expressed in the cilia‐enriched noncellular fraction. It is shown that miR‐669a‐5p promotes cilium elongation but not cilium formation in cultured cells. Mechanistically, it is demonstrated that miR‐669a‐5p represses ras‐GTPase‐activating protein SH3‐domain‐binding protein (G3BP) expression to inhibit histone deacetylase 6 (HDAC6) expression, which further upregulates A‐kinase anchor protein 12 (AKAP12) expression. This effect ultimately blocks cilia disassembly and leads to greater cilium length, which can be restored to wild‐type lengths by either upregulating HDAC6 or downregulating AKAP12. Collectively, these results elucidate a previously unidentified miR‐669a‐5p/G3BP/HDAC6/AKAP12 signaling pathway that regulates cilium length, providing potential pharmaceutical targets for treating ciliopathies.

The deterioration of concrete materials in the sewage environment is prominent, significantly shortening the structure's service life. Based on the composite modification method, modified basalt fiber high-durability concrete is proposed to solve the problem of insufficient material durability in the sewage environment. The acid environment erosion test was carried out for the modified concrete materials. The macroscopic performance indexes were obtained, such as apparent corrosion characteristics, water absorption rate and compressive strength. The corrosion mechanism and deterioration law of concrete materials under an acid environment were revealed by combining the analysis of the fine structure and corrosion product composition. The results show that the hydration reaction of the modified concrete was adequate. The generation of C–S–H gel and Ca(OH)2 increased significantly. The porosity was reduced by 40%, and the initial strength was increased by 12%. After 240d corrosion, the strength of ordinary concrete was degraded by 41%. The degradation rate of modified concrete was 26%, its residual strength was 1.4 times that of ordinary concrete, and its corrosion-resistant ability was greatly improved. Based on the experiment, the corrosion resistance mechanism analysis was carried out, and it was found that the inorganic aluminium salt compound salt crystal expansion modification effect is better than the sodium methyl silicate reticulated resin film modification. However, after double doping, the corrosion resistance is reduced instead, and it is recommended that inorganic aluminium salts be preferred as anti-corrosion and anti-seepage adding materials. The research results of this paper can better enhance the acid environment erosion resistance of concrete, engineering application prospect is good.

Cervical spondylosis is a widespread medical condition that significantly impacts patients' quality of life. Treatment options include surgical and conservative approaches, with conservative treatment often being the preferred choice. Rehabilitation therapy is an essential component of conservative treatment, and advancements in technology have the way to the development of new physiotherapy techniques. The effectiveness of treatment largely hinges on the patient's ability to improve their dysfunction. This study aims to provide valuable insights into the use of new physical therapy techniques, such as Sling Exercises Training (SET), fascia manipulation, muscle energy technique (MET), and proprioceptive neuromuscular facilitation (PNF), that aid the rehabilitation of cervical spondylosis. By scrutinizing the current research status of these techniques, this study aims to present innovative ideas enhancing the rehabilitation process and outcomes for patients suffering from cervical spondylosis.

BackgroundScatter-hoarding animals store food in multiple locations within their home range and rely on spatial memory for subsequent localization and retrieval. The relationship between memory and scatter-hoarding behavior has been widely demonstrated, but the association of gut microbiota with spatial memory and seed-hoarding behavior of animals remains unclear.MethodsIn this study, by using enclosure behavior tests, memory tests including an object location test (OLT) and a novel object recognition test (NORT), and fecal microbiota transplantation (FMT) experiment, we evaluated the role of gut microbiota in affecting the memory and seed-hoarding behavior of rodents. According to their scatter-hoarding intensity, South China field mice (Apodemus draco) were divided into scatter-hoarding group (SG) and non-scatter-hoarding group (NG).ResultsWe found that the SG performed better than the NG in the NORT. FMT from SG donor mice altered the NG recipient mice’s gut microbiota structure. Further tests demonstrated FMT from SG donor mice increased memory of NG recipient mice in laboratory tests and seed larder hoarding intensity of NG recipient mice in enclosures.ConclusionOur results suggest gut microbiota could modulate the memory and seed-hoarding behavior of animals.

To study the dynamic characteristics and seismic damage of the Bayun Pagoda,the dynamic characteristics are determined via an in-situ dynamic test, and the flexibility curvature method is used to identify the damage. Moreover, the numerical calculation results of the dynamic characteristics are compared with the test results. The mechanical parameters of the materials are determined in the consistency of the dynamic characteristics, and a plastic damage model is also introduced. From the vertical compressive stress the initial damage is defined, and the seismic damage is analyzed by inputting El-Centro, Taft, and Lanzhou artificial wave. As results, the first-order natural frequencies are close to each other in the horizontal East-West and North-South directions, and the initial damage rates of the 6th, 8th, 10th floors are higher. The distributions of the principal tensile stress excited by different seismic are similarbut the distributions of seismic damage are slightly different; the area with serious seismic damage is in the middle and bottom floors, and the damage accumulation of the first floor is more obvious than those of the other floors. Moreover, the outer edge of each floor, the pagoda eaves, and the periphery of the archway are found to be easily damaged.

The concrete structure will be seriously eroded in a sewage environment, causing substantial economic losses. Therefore, it is of great significance to repair the existing corrosion structure. In order to quickly restore the mechanical properties and enhance the durability performance of eroded concrete structures, this paper develops a new composite high-durability mortar by adding polypropylene fibers, basalt fibers and impermeable agents, which can achieve the dual requirements of structural repair and protection. For the new material, this test set up a total of five groups of mixing ratios. This test analyzed the water absorption, mass loss rate, compressive strength and other performance indicators of the test block in the corrosive environment, and scanned and studied the microstructure of each mortar test block after corrosion. It was found that the new anti-corrosion mortar has relatively less crystallization, dense internal structure, and significantly lighter erosion. From the mechanical properties, the strength of ordinary polymer mortar after erosion decreased by 6%; high-durability mortar instead improved the strength by nearly 20%, showing better resistance to erosion. Taken together, the use of mineral dopants and water repellents can effectively enhance the strength and corrosion resistance of mortar, and the cost is low, with good prospects for engineering applications.

The coronavirus disease 2019 pandemic has caused more than 532 million infections and 6.3 million deaths to date. The reactive and neutralizing fully human antibodies of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are effective detection tools and therapeutic measures. During SARS-CoV-2 infection, a large number of SARS-CoV-2 reactive and neutralizing antibodies will be produced. Most SARS-CoV-2 reactive and neutralizing fully human antibodies are isolated from human and frequently encoded by convergent heavy-chain variable genes. However, SARS-CoV-2 viruses can mutate rapidly during replication and the resistant variants of neutralizing antibodies easily survive and evade the immune response, especially in the face of such focused antibody responses in humans. Therefore, additional tools are needed to develop different kinds of fully human antibodies to compensate for current deficiency. In this study, we utilized antibody humanized CAMouseHG mice to develop a rapid antibody discovery method and examine the antibody repertoire of SARS-CoV-2 RBD-reactive hybridoma cells derived from CAMouseHG mice by using high-throughput single-cell V(D)J sequencing analysis. CAMouseHG mice were immunized by 28-day rapid immunization method. After electrofusion and semi-solid medium screening on day 12 post-electrofusion, 171 hybridoma clones were generated based on the results of SARS-CoV-2 RBD binding activity assay. A rather obvious preferential usage of IGHV6-1 family was found in these hybridoma clones derived from CAMouseHG mice, which was significantly different from the antibodies found in patients with COVID-19. After further virus neutralization screening and antibody competition assays, we generated a noncompeting two-antibody cocktail, which showed a potent prophylactic protective efficacy against SARS-CoV-2 in cynomolgus macaques. These results indicate that humanized CAMouseHG mice not only provide a valuable platform to obtain fully human reactive and neutralizing antibodies but also have a different antibody repertoire from humans. Thus, humanized CAMouseHG mice can be used as a good complementary tool in discovery of fully human therapeutic and diagnostic antibodies.

Marburg virus (MARV) is one of the principal members of the filovirus family, which can cause fatal hemorrhagic fever in humans. There are currently no prophylactic and therapeutic drugs on the market, and the high pathogenicity and infectivity of MARV make its research highly dependent on biosafety level 4 conditions, severely hindering the development of vaccines and therapies. Therefore, the development of medicines, such as MARV serological diagnosis, vaccines, and therapeutic antibody drugs, urgently needs a safe, convenient, and biosafety level 2 detection method to measure the neutralizing activity of MARV antibodies. To this end, we report a neutralization assay relying on a Rabies virus (RABV) reverse genetic operating system. We constructed infectious clones carrying the eGFP reporter gene and the full length of the original unmodified MARV GP gene. Based on the critical parameters of phylogenetic analysis, recombinant viruses targeting representative strains in the two major MARV lineages were successfully rescued. These pseudoviruses are safe in mice, and their inability to infect cells after being neutralized by antibodies can be visualized under a fluorescence microscope. We tested the system using the neutralizing antibody MR191. MR191 can significantly block the infection of BSR cells with pseudovirus. We compared it with the traditional lentivirus-type pseudovirus system to verify the system’s credibility and obtained the same results as reported in the literature. In general, we have established a safe and visualized method for evaluating the neutralizing activity of MARV antibodies. Compared with traditional methods, it has the advantages of convenient operation, short cycle, and low cost. It is a candidate method that can replace actual viruses for a neutralization assay.

Under corrosive environments, concrete material properties can deteriorate significantly, which can seriously affect structural safety. Therefore, it has important engineering applications to improve the durability performance at a lower economic cost. This paper proposes a new, highly durable concrete using inexpensive construction materials such as basalt fiber, sodium methyl silicate, and inorganic aluminum salt waterproofing agent. With the massive application of sewage treatment projects, the problem of concrete durability degradation is becoming more and more serious. In this paper, five types of concrete are developed for the sewage environment, and the apparent morphology and fine structure of the specimens after corrosion in sewage were analyzed. The density, water absorption, and compressive strength were measured to investigate the deterioration pattern of concrete properties. It was found that ordinary concrete was subject to significant corrosion, generating large deposits of algae on the surface and accompanied by sanding. The new concrete showed superior corrosion resistance compared to conventional concrete. Among other factors, the inorganic aluminum salt waterproofing agent effect was the most prominent. The study found that the strength of ordinary concrete decreased by about 15% in the test environment, while the new concrete had a slight increase. Comprehensive evaluation showed that the combination of basalt fiber and inorganic aluminum salt waterproofing agent had the best effect. Its use is recommended.

Abstract Neuronal primary cilia are crucial for body weight maintenance. Type III adenylyl cyclase (AC3) is abundantly enriched in neuronal cilia, and mice with global AC3 ablation are obese. However, whether AC3 regulates body weight through its ciliary expression and the mechanism underlying this potential regulation are not clear. In this study, humanized AC3 knock‐in mice that are resistant to high‐fat diet (HFD)‐induced obesity are generated, and increases in the number and length of cilia in the ventromedial hypothalamus (VMH) are shown. It is demonstrated that mice with specifically knocked down ciliary AC3 expression in the VMH show pronounced HFD‐induced obesity. In addition, in vitro and in vivo analyses of the VMH show that ciliary AC3 regulates autophagy by binding an autophagy‐related gene, gamma‐aminobutyric acid A receptor‐associated protein (GABARAP). Mice with GABARAP knockdown in the VMH exhibit exacerbated HFD‐induced obesity. Overall, the findings may reveal a potential mechanism by which ciliary AC3 expression regulates body weight in the mouse VMH.

Middle East respiratory syndrome coronavirus (MERS-CoV) is an emergent coronavirus that has caused frequent zoonotic events through camel-to-human spillover. An effective camelid vaccination strategy is probably the best way to reduce human exposure risk. Here, we constructed and evaluated an inactivated rabies virus-vectored MERS-CoV vaccine in mice, camels, and alpacas. Potent antigen-specific antibody and CD8+ T-cell responses were generated in mice; moreover, the vaccination reduced viral replication and accelerated virus clearance in MERS-CoV-infected mice. Besides, protective antibody responses against both MERS-CoV and rabies virus were induced in camels and alpacas. Satisfyingly, the immune sera showed broad cross-neutralizing activity against the three main MERS-CoV clades. For further characterization of the antibody response induced in camelids, MERS-CoV-specific variable domains of heavy-chain-only antibody (VHHs) were isolated from immunized alpacas and showed potent prophylactic and therapeutic efficacies in the Ad5-hDPP4-transduced mouse model. These results highlight the inactivated rabies virus-vectored MERS-CoV vaccine as a promising camelid candidate vaccine.

Thin-walled steel pipe concrete has better economic performance, but the problem of local buckling is more prominent with a thin-walled steel pipe; meanwhile, thin-walled steel pipe is more sensitive to the environment and the influence of rusting is more prominent. To solve the above problems, this paper proposes new spiral stiffened rib thin-walled steel pipe concrete laminated members to obtain better force and economic performance. Based on axial compression tests on five forms of composite members, this paper studies the nonlinear behavior of the axial compression of this new type of laminated member and the factors influencing it. The following conclusions are obtained. Under the constraint of the spiral ribs, the new composite member has good integrity and each part can ensure cooperative stress; the buckling of the steel pipe is well limited and the mechanical performance is significantly improved. Compared with ordinary thin-walled concrete-filled steel tubular members, the bearing capacity is increased by about 20% and the deformation ability is increased by more than 30%. The nonlinear behavior of the member in compression can be better achieved through finite element analysis. The parametric analysis shows that the pitch and the steel tube width-to-thickness ratio greatly influence the force behavior of the member. In contrast, the spiral rib width-to-thickness ratio and the external reinforcement only need to meet the structural requirements. Finally, based on the superposition theory, the proposed method of calculating the member’s axial compressive load-bearing capacity is given and design suggestions are made. The results of this paper can provide some basis for the engineering application of this new combination member.

To enhance the local buckling resistance of thin-walled steel pipes and enhance their fire and corrosion resistance, a new spirally stiffened thin-walled square concrete-filled steel tubular laminated composite member with transverse ribs is proposed. Through the four forms of combined members for eccentric pressure testing, it was found that: ordinary thin-walled steel pipe concrete drum buckling is more severe; with spiral ribs, the buckling is limited between the stiffening ribs; and the deformation is significantly reduced. By addressing the problem of cooperative work between the inner and outer structural layers of new components, it was found that, after setting constraints such as steel bars, the integrity of the two can be ensured, and the stress performance is significantly improved; compared to ordinary steel pipe concrete, the load-carrying capacity is 17.9% higher, and the deformation capacity is roughly equivalent. Spiral ribs as a new form of spatial restraint, in addition to increasing the local bending stiffness, manifest an overall restraint role in limiting lateral deformation of the steel pipe, whereas the role of vertical stiffness is insignificant. Based on test evidence, the influences of the width to thickness ratio of spiral ribs and pitch were determined, and reasonable structural measures for the members were given. Through the N–M relationship curve, the limit of damage in compression and tension under eccentricity was obtained at an eccentricity of about 0.9. Finally, a method for calculating the eccentric compressive ultimate load capacity of this new composite member was proposed.

The PBL (Perfobond Leiste) shear connector has the advantages of high bearing capacity and strong constraint ability; however, the traditional PBL shear connector has strong and weak axis problems, and its stiffness is large, resulting in weak deformation ability. To this end, this paper proposes a new type of flange triple-web shear key and obtains the new shear key’s mechanical properties and failure mechanism through the push-out test. The results show that the failure mode of the new shear key is the deformation of the steel plate on the web and the edge of the opening, which has a high bearing capacity, outstanding deformation ability, and good integrity with concrete, showing obvious semi-rigid characteristics. Through numerical analysis, the effects of flange width, web height, and steel plate thickness on the mechanical properties of shear keys are obtained. Based on the fitting analysis method, the calculation formula of shear key bearing capacity is proposed. Finally, the horizontal seismic performance of the shear key is numerically simulated. It is found that the hysteretic curve of the shear key is full and shows good energy dissipation capacity.

Adenylyl Cyclase 3 (AC3) plays an important role in the olfactory sensation-signaling pathway in mice. AC3 deficiency leads to defects in olfaction. However, it is still unknown whether AC3 deficiency affects gene expression or olfactory signal transduction pathways within the main olfactory epithelium (MOE). In this study, gene microarrays were used to screen differentially expressed genes in MOE from AC3 knockout (AC3−/−) and wild-type (AC3+/+) mice. The differentially expressed genes identified were subjected to bioinformatic analysis and verified by qRT-PCR. Gene expression in the MOE from AC3−/− mice was significantly altered, compared to AC3+/+ mice. Of the 41266 gene probes, 3379 had greater than 2-fold fold change in expression levels between AC3−/− and AC3+/+ mice, accounting for 8% of the total gene probes. Of these genes, 1391 were up regulated, and 1988 were down regulated, including 425 olfactory receptor genes, 99 genes that are specifically expressed in the immature olfactory neurons, 305 genes that are specifically expressed in the mature olfactory neurons, and 155 genes that are involved in epigenetic regulation. Quantitative RT-PCR verification of the differentially expressed epigenetic regulation related genes, olfactory receptors, ion transporter related genes, neuron development and differentiation related genes, lipid metabolism and membrane protein transport etc. related genes showed that P75NTR, Hinfp, Gadd45b, and Tet3 were significantly up-regulated, while Olfr370, Olfr1414, Olfr1208, Golf, Faim2, Tsg101, Mapk10, Actl6b, H2BE, ATF5, Kirrrel2, OMP, Drd2 etc. were significantly down-regulated. In summary, AC3 may play a role in proximal olfactory signaling and play a role in the regulation of differentially expressed genes in mouse MOE.

In this paper, the durability and mechanical properties of basalt fiber reinforced concrete under sodium sulfate erosion were studied. The concrete with 0%, 0.05%, 0.1%, 0.2% and 0.3% basalt fiber were respectively soaked in 5.0% sodium sulfate solution and clean water for 90 days, and the observations and records were used to analyse the changes in the water absorption rate of the test pieces and conduct compression tests. The main conclusions are as follows: The basalt fiber has little effect on the bearing capacity of concrete, but can improve the deformation capacity of concrete to certain extent. Under the erosion of sodium sulfate, basalt fiber can effectively lower the porosity growth rate, while excessive fiber adversely affects the durability, with the fiber content of about 0.1% leading to the best result. In view of the difficulty in basalt fiber dispersion, this paper put forward “Powder Particle Basalt Fiber Dispersion Method”. Therefore, this research will provide some theoretical foundation for engineering application of basalt fiber.

Middle East respiratory syndrome coronavirus (MERS-CoV), a new coronavirus that has been causing severe and fatal acute respiratory illnesses in humans since its outbreak in 2012, has raised public fear worldwide. The development of prophylactics and therapeutics is urgently needed to prevent and control MERS-CoV infections. In this study, a bacterium (Lactococcus lactis)-like particle (BLP) vaccine displaying the MERS-CoV receptor-binding domain (RBD) was developed, and gram-positive enhancer matrix (GEM) particles were used as substrates to externally bind to the MERS-CoV RBD through a protein anchor (PA). The designs included different numbers of lysin motif (LysM) repeats in the PAs linked by linkers (RBD-linker-PA2 (RLP2), RBD-linker-PA3 (RLP3) and RBD-PA3 (RP3)), and three LysM repeats and a linker in the fusion proteins increased the binding activity to the RBD. The specific immune responses were tested by intranasally immunizing mice with RLP3-GEM with or without the adjuvant GEL01. The results showed that GEL01-adjuvanted RLP3-GEM increased the systemic humoral, cellular and local mucosal immune responses in the mouse model, especially in the intestinal tract. The above results indicate that the MERS-CoV BLP product has the potential to be developed into a promising mucosal candidate vaccine to protect against MERS-CoV infections.

Discriminating a real underwater target echo from a synthetic echo is a key challenge to identifying an underwater target. The structure of an echo envelope contains information which closely relates to the physical parameters of the underwater target, and the characterization and extraction of echo features are problematic issues for active sonar target classification. In this study, firstly, the high-frequency envelope fluctuation of a complex underwater target echo was analyzed, the envelope fluctuation was characterized by the envelope fluctuation intensity, and a characterization model was established. The features of a benchmark model echo were extracted and analyzed by theoretical simulation and sea testing of a scaled model, and the result shows that the envelope fluctuation intensity varies with carrier frequency and azimuth of incident signal, but the echo envelope fluctuation of the synthetic target echo does not present these features. Then, based on the characteristics of echo envelope fluctuation, a novel method was developed for active sonar discrimination of a real underwater target echo from the synthetic echo. Through a sea experiment, the real target echo and synthetic echo were classified by their different echo envelope fluctuations, and the feasibility of the method was verified.

In this paper, beamforming and normal modes method are combined to calculate the underwater acoustical field. Firstly, beamforming technology is used to form directional sources by controlling the phase and relative amplitude. Secondly, considering the ocean speed, surface and bottom boundary, the point source sound field is calculated based on the normal modes model. Then underwater acoustical field with directional sources is achieved by adding all point source field. Taken the 11 elements vertical linear array as an example, the special directional source is formed by chebyshev amplitude weighting, and then underwater acoustical field is calculated in the Munk profile of different grazing angles. The results indicate that beam grazing angle is very essential to its sound field, and changing the grazing angle can enhance the sound intensity of the shadow zone correspondingly.

Intranasal medications are used to treat various nasal disorders. However, their effects on olfaction remain unknown. Zicam (zinc gluconate; Matrixx Initiatives, Inc), a homeopathic substance marketed to alleviate cold symptoms, has been implicated in olfactory dysfunction. Here, we investigated Zicam and several common intranasal agents for their effects on olfactory function. Zicam was the only substance that showed significant cytotoxicity in both mouse and human nasal tissue. Specifically, Zicam-treated mice had disrupted sensitivity of olfactory sensory neurons to odorant stimulation and were unable to detect novel odorants in behavioral testing. These findings were long-term as no recovery of function was observed after two months. Finally, human nasal explants treated with Zicam displayed significantly elevated extracellular lactate dehydrogenase levels compared to saline-treated controls, suggesting severe necrosis that was confirmed on histology. Our results demonstrate that Zicam use could irreversibly damage mouse and human nasal tissue and may lead to significant smell dysfunction.

BACKGROUND:A recent study of obesity in Swedish men found that polymorphisms in the type 3 adenylyl cyclase (AC3) are associated with obesity, suggesting the interesting possibility that AC3 may play a role in weight control. Therefore, we examined the weight of AC3 mice over an extended period of time. METHODOLOGY/PRINCIPAL FINDINGS:We discovered that AC3(-/-) mice become obese as they age. Adult male AC3(-/-) mice are about 40% heavier than wild type male mice while female AC3(-/-) are 70% heavier. The additional weight of AC3(-/-) mice is due to increased fat mass and larger adipocytes. Before the onset of obesity, young AC3(-/-) mice exhibit reduced physical activity, increased food consumption, and leptin insensitivity. Surprisingly, the obesity of AC3(-/-) mice is not due to a loss of AC3 from white adipose and a decrease in lipolysis. CONCLUSIONS/SIGNIFICANCE:We conclude that mice lacking AC3 exhibit obesity that is apparently caused by low locomotor activity, hyperphagia, and leptin insensitivity. The presence of AC3 in primary cilia of neurons of the hypothalamus suggests that cAMP signals generated by AC3 in the hypothalamus may play a critical role in regulation of body weight.

The reoccurrence of successive waves of SARS-CoV-2 variants suggests the exploration of more vaccine alternatives is imperative. Modified vaccinia virus Ankara (MVA) is a virus vector exhibiting excellent safety as well as efficacy for vaccine development. Here, a series of recombinant MVAs (rMVAs) expressing monomerized or trimerized S proteins from different SARS-CoV-2 variants are engineered. Trimerized S expressed from rMVAs is found predominantly as trimers on the surface of infected cells. Remarkably, immunization of mice with rMVAs demonstrates that S expressed in trimer elicits higher levels of binding IgG and IgA, as well as neutralizing antibodies for matched and mismatched S proteins than S in the monomer. In addition, trimerized S expressed by rMVA induces enhanced cytotoxic T-cell responses than S in the monomer. Importantly, the rMVA vaccines expressing trimerized S exhibit superior protection against a lethal SARS-CoV-2 challenge as the immunized animals all survive without displaying any pathological conditions. This study suggests that opting for trimerized S may represent a more effective approach and highlights that the MVA platform serves as an ideal foundation to continuously advance SARS-CoV-2 vaccine development. IMPORTANCE MVA is a promising vaccine vector and has been approved as a vaccine for smallpox and mpox. Our analyses suggested that recombinant MVA expressing S in trimer (rMVA-ST) elicited robust cellular and humoral immunity and was more effective than MVA-S-monomer. Importantly, the rMVA-ST vaccine was able to stimulate decent cross-reactive neutralization against pseudoviruses packaged using S from different sublineages, including Wuhan, Delta, and Omicron. Remarkably, mice immunized with rMVA-ST were completely protected from a lethal challenge of SARS-CoV-2 without displaying any pathological conditions. Our results demonstrated that an MVA vectored vaccine expressing trimerized S is a promising vaccine candidate for SARS-CoV-2 and the strategy might be adapted for future vaccine development for coronaviruses. [ABSTRACT FROM AUTHOR]

The Ebola virus (EBOV) is a member of the Orthoebolavirus genus, Filoviridae family, which causes severe hemorrhagic diseases in humans and non-human primates (NHPs), with a case fatality rate of up to 90%. The development of countermeasures against EBOV has been hindered by the lack of ideal animal models, as EBOV requires handling in biosafety level (BSL)-4 facilities. Therefore, accessible and convenient animal models are urgently needed to promote prophylactic and therapeutic approaches against EBOV. In this study, a recombinant vesicular stomatitis virus expressing Ebola virus glycoprotein (VSV-EBOV/GP) was constructed and applied as a surrogate virus, establishing a lethal infection in hamsters. Following infection with VSV-EBOV/GP, 3-week-old female Syrian hamsters exhibited disease signs such as weight loss, multi-organ failure, severe uveitis, high viral loads, and developed severe systemic diseases similar to those observed in human EBOV patients. All animals succumbed at 2-3 days post-infection (dpi). Histopathological changes indicated that VSV-EBOV/GP targeted liver cells, suggesting that the tissue tropism of VSV-EBOV/GP was comparable to wild-type EBOV (WT EBOV). Notably, the pathogenicity of the VSV-EBOV/GP was found to be species-specific, age-related, gender-associated, and challenge route-dependent. Subsequently, equine anti-EBOV immunoglobulins and a subunit vaccine were validated using this model. Overall, this surrogate model represents a safe, effective, and economical tool for rapid preclinical evaluation of medical countermeasures against EBOV under BSL-2 conditions, which would accelerate technological advances and breakthroughs in confronting Ebola virus disease. [ABSTRACT FROM AUTHOR]

The coronavirus disease 2019 pandemic has caused more than 532 million infections and 6.3 million deaths to date. The reactive and neutralizing fully human antibodies of severe acute respiratory syndrome coronavirus 2 (SARSCoV-2) are effective detection tools and therapeutic measures. During SARSCoV-2 infection, a large number of SARS-CoV-2 reactive and neutralizing antibodies will be produced. Most SARS-CoV-2 reactive and neutralizing fully human antibodies are isolated from human and frequently encoded by convergent heavy-chain variable genes. However, SARS-CoV-2 viruses can mutate rapidly during replication and the resistant variants of neutralizing antibodies easily survive and evade the immune response, especially in the face of such focused antibody responses in humans. Therefore, additional tools are needed to develop different kinds of fully human antibodies to compensate for current deficiency. In this study, we utilized antibody humanized CAMouseHG mice to develop a rapid antibody discovery method and examine the antibody repertoire of SARS-CoV-2 RBD-reactive hybridoma cells derived from CAMouseHG mice by using high-throughput single-cell V (D)J sequencing analysis. CAMouseHG mice were immunized by 28-day rapid immunization method. After electrofusion and semi-solid medium screening on day 12 post-electrofusion, 171 hybridoma clones were generated based on the results of SARS-CoV-2 RBD binding activity assay. A rather obvious preferential usage of IGHV6-1 family was found in these hybridoma clones derived from CAMouseHG mice, which was significantly different from the antibodies found in patients with COVID-19. After further virus neutralization screening and antibody competition assays, we generated a noncompeting two-antibody cocktail, which showed a potent prophylactic protective efficacy against SARS-CoV-2 in cynomolgus macaques. These results indicate that humanized CAMouseHG mice not only provide a valuable platform to obtain fully human reactive and neutralizing antibodies but also have a different antibody repertoire from humans. Thus, humanized CAMouseHG mice can be used as a good complementary tool in discovery of fully human therapeutic and diagnostic antibodies. [ABSTRACT FROM AUTHOR]