Novel antiviral discoveries for Japanese encephalitis virus infections through reporter virus-based high-throughput screening.

Japanese encephalitis (JE) caused by JE virus (JEV), remains a global public health concern. Currently, there is no specific antiviral drug approved for the treatment of JE. While vaccines are available for prevention, they may not cover all at-risk populations. This underscores the urgent need for prophylaxis and potent anti-JEV drugs. In this context, a high-content JEV reporter system expressing Nanoluciferase (Nluc) was developed and utilized for a high-throughput screening (HTS) of a commercial antiviral library to identify potential JEV drug candidates. Remarkably, this screening process led to the discovery of five drugs with outstanding antiviral activity. Further mechanism of action analysis revealed that cepharanthine, an old clinically approved drug, directly inhibited virus replication by blocking GTP binding to the JEV RNA-dependent RNA polymerase. Additionally, treatment with cepharanthine in mice models alleviated JEV infection. These findings warrant further investigation into the potential anti-JEV activity of cepharanthine as a new therapeutic approach for the treatment of JEV infection. The HTS method employed here proves to be an accurate and convenient approach that facilitates the rapid development of antiviral drugs.

Clean and efficient synthesis of LiFePO4 cathode material using titanium white waste and calcium dihydrogen phosphate.

Large amounts of titanium white waste are generated in the production of titanium dioxide using sulphate method, which in turn can be used to prepare LiFePO4 cathode material, thereby reducing environmental risks and achieving resource recovery. However, a key challenge lies in the elimination of impurities. In this work, a cost-efficient and straightforward approach based on phase transformation during hydrothermal treatment was proposed to utilize titanium white waste with calcium dihydrogen phosphate for the preparation of LiFePO4 cathode material. The content of Fe in the leachate was enriched to 81.5 g/L after purification, while 99.9 % of Ti and 98.36 % of Al and were successfully removed. In the subsequent process for Fe/P mother liquor preparation, the losses of Fe and P were only 5.82 % and 2.81 %, respectively. The Fe and P contents of the synthesized FePO4 product were 29.47 % and 17.08 %, respectively, and the Fe/P molar ratio was 0.986. Crystal phase of the product matched well with standard iron phosphate, and the lamellar microstructure of FePO4 was uniform with the particle size ranging from 3 to 5 µm. Moreover, the contents of impurities in the product were far below the standard. The initial discharge of LiFePO4 synthesized by the iron phosphate was 160.6 mAh.g-1 at 0.1C and maintained good reversible capacity after 100 cycles. This work may provide new strategy for preparing LiFePO4 cathode material from industrial solid waste.

Both chimpanzee adenovirus-vectored and DNA vaccines induced long-term immunity against Nipah virus infection.

Nipah virus (NiV) is a highly lethal zoonotic paramyxovirus that poses a severe threat to humans due to its high morbidity and the lack of viable countermeasures. Vaccines are the most crucial defense against NiV infections. Here, a recombinant chimpanzee adenovirus-based vaccine (AdC68-G) and a DNA vaccine (DNA-G) were developed by expressing the codon-optimized full-length glycoprotein (G) of NiV. Strong and sustained neutralizing antibody production, accompanied by an effective T-cell response, was induced in BALB/c mice by intranasal or intramuscular administration of one or two doses of AdC68-G, as well as by priming with DNA-G and boosting with intramuscularly administered AdC68-G. Importantly, the neutralizing antibody titers were maintained for up to 68 weeks in the mice that received intramuscularly administered AdC68-G and the prime DNA-G/boost AdC68-G regimen, without a significant decline. Additionally, Syrian golden hamsters immunized with AdC68-G and DNA-G via homologous or heterologous prime/boost immunization were completely protected against a lethal NiV virus challenge, without any apparent weight loss, clinical signs, or pathological tissue damage. There was a significant reduction in but not a complete absence of the viral load and number of infectious particles in the lungs and spleen tissue following NiV challenge. These findings suggest that the AdC68-G and DNA-G vaccines against NiV infection are promising candidates for further development.

Vaccines based on the fusion protein consensus sequence protect Syrian hamsters from Nipah virus infection.

Nipah virus (NiV), a bat-borne paramyxovirus, results in neurological and respiratory diseases with high mortality in humans and animals. Developing vaccines is crucial for fighting these diseases. Previously, only a few studies focused on the fusion (F) protein alone as the immunogen. Numerous NiV strains have been identified, including 2 representative strains from Malaysia (NiV-M) and Bangladesh (NiV-B), which differ significantly from each other. In this study, an F protein sequence with the potential to prevent different NiV strain infections was designed by bioinformatics analysis after an in-depth study of NiV sequences in GenBank. Then, a chimpanzee adenoviral vector vaccine and a DNA vaccine were developed. High levels of immune responses were detected after AdC68-F, pVAX1-F, and a prime-boost strategy (pVAX1-F/AdC68-F) in mice. After high titers of humoral responses were induced, the hamsters were challenged by the lethal NiV-M and NiV-B strains separately. The vaccinated hamsters did not show any clinical signs and survived 21 days after infection with either strain of NiV, and no virus was detected in different tissues. These results indicate that the vaccines provided complete protection against representative strains of NiV infection and have the potential to be developed as a broad-spectrum vaccine for human use.

Recovery of iron and aluminum from iron-rich bauxite residue by an integrated phase reconstruction approach.

The comprehensive recovery of iron and aluminum from iron-rich bauxite residue (IRBR) is of critical importance both in terms of resource utilization and environment protection, which, however, is challenging due to the intertwined phases between Iron and aluminum. In this study, an integrated phase reconstruction approach, consisting of alkali roasting, two-stage column leaching, and carbonation decomposition, was proposed for Fe/Al recovery from IRBR. The results demonstrated that aluminum and sodium were fused into soluble substances such as sodium aluminate (Na7Al3O8, NaAlO2, and Na2O (Al2O3)11) in the alkali roasting process, allowing for the separation of Al and Fe in the subsequent leaching process. Following water/FeCl3 solution leaching, the removal efficiency of aluminum reached 84.66%, and Fe content in the residue could be enriched to 55.56%. Fe can be recycled as iron concentrate, and Al in the leaching solution with 75.95 g/L can be recovered in the form of Al(OH)3 through carbonation decomposition. This work provides an alternative strategy for the recovery of resources from IRBR, with potential implications for the sustainable development of the aluminum industry.

Identification and characterization of gonadotropin-releasing hormone (GnRH) in Zhikong scallop Chlamys farreri during gonadal development.

Gonadotropin-releasing hormone (GnRH) controls synthesis of sex steroid hormones through hypothalamic-pituitary-gonadal (HPG) axis in vertebrates. But in mollusks, research on neuroendocrine control of gonadal function, such as the function of GnRH during gonadal development is limited. In this study, we investigated the morphology and structure of the nerve ganglia of Zhikong scallop Chlamys farreri by physiological and histological observations. We also cloned the ORF and studied the expression patterns of GnRH in the scallop. Tissue expression analysis showed that GnRH was highly expressed in parietovisceral ganglion (PVG). The in situ hybridization result further confirmed that GnRH mRNA only distributed in some good-sized neurons in the posterior lobe (PL) and some pint-sized neurons in the lateral lobe (LL). In addition, by examining the expression of GnRH during gonadal development in ganglia, we found GnRH displayed higher expression in the female scallops, and showed significant high expression at the growing stage of female scallops in PVG. This study would contribute to gaining insight into the mechanism underlying reproduction regulation by GnRH in the scallop and help to provide a better understanding of reproductive neuroendocrine in mollusks.

Evolution of Immune Evasion and Host Range Expansion by the SARS-CoV-2 B.1.1.529 (Omicron) Variant.

Recently, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variant B.1.1.529 (Omicron) has rapidly become the dominant strain, with an unprecedented number of mutations within its spike gene. However, it remains unknown whether these variants have alterations in their entry efficiency, host tropism, and sensitivity to neutralizing antibodies and entry inhibitors. In this study, we found that Omicron spike has evolved to escape neutralization by three-dose inactivated-vaccine-elicited immunity but remains sensitive to an angiotensin-converting enzyme 2 (ACE2) decoy receptor. Moreover, Omicron spike could use human ACE2 with a slightly increased efficiency while gaining a significantly increased binding affinity for a mouse ACE2 ortholog, which exhibits limited binding with wild-type (WT) spike. Furthermore, Omicron could infect wild-type C57BL/6 mice and cause histopathological changes in the lungs. Collectively, our results reveal that evasion of neutralization by vaccine-elicited antibodies and enhanced human and mouse ACE2 receptor engagement may contribute to the expanded host range and rapid spread of the Omicron variant. IMPORTANCE The recently emerged SARS-CoV-2 Omicron variant with numerous mutations in the spike protein has rapidly become the dominant strain, thereby raising concerns about the effectiveness of vaccines. Here, we found that the Omicron variant exhibits a reduced sensitivity to serum neutralizing activity induced by a three-dose inactivated vaccine but remains sensitive to entry inhibitors or an ACE2-Ig decoy receptor. Compared with the ancestor strain isolated in early 2020, the spike protein of Omicron utilizes the human ACE2 receptor with enhanced efficiency while gaining the ability to utilize mouse ACE2 for cell entry. Moreover, Omicron could infect wild-type mice and cause pathological changes in the lungs. These results reveal that antibody evasion, enhanced human ACE2 utilization, and an expanded host range may contribute to its rapid spread.

Outer fold is sole effective tissue among three mantle folds with regard to oyster shell colour.

Molluscs constitute the second largest phylum of animals in the world, and shell colour is one of their most important phenotypic characteristics. In this study, we found among three folds on the mantle edge of oyster, only the outer fold had the same colour as the shell. Transcriptome and mantle cutting experiment indicated that the outer fold may be mainly reflected in chitin framework formation and biomineralisation. There were obvious differences in SEM structure and protein composition between the black and white shell periostraca. The black shell periostraca had more proteins related to melanin biosynthesis and chitin binding. Additionally, we identified an uncharacterized protein gene (named as CgCBP) ultra-highly expressed only in the black outer fold and confirmed its function of chitin-binding and CaCO3 precipitation promoting. RNAi also indicated that CgCBP knockdown could change the structure of shell periostracum and reduce shell pigmentation. All these results suggest that the mantle outer fold plays multiple key roles in shell periostraca bioprocessing, and shell periostracum structure affected by chitin-binding protein is functionally correlated with shell pigmentation. The investigation of oyster shell periostracum structure and shell colour will provide a better understanding in pigmentation during biological mineralisation in molluscs.

CfIRF8-like interacts with the TBK1/IKKε family protein and regulates host antiviral innate immunity.

The interferon regulatory factor (IRF) family, a class of transcription factors with key functions, are important in host innate immune defense and stress response. However, further research is required to determine the functions of IRFs in invertebrates. In this study, the coding sequence of an IRF gene was obtained from the Zhikong scallop (Chlamys farreri) and named CfIRF8-like. The open reading frame of CfIRF8-like was 1371 bp long and encoded 456 amino acids. Protein domain prediction revealed a typical IRF domain in the N-terminus of the CfIRF8-like protein and a typical IRF3 domain in the C-terminus. Multiple sequence alignment confirmed the conservation of the amino acid sequences of these two functional protein domains. Phylogenetic analysis showed that CfIRF8-like clustered with mollusk IRF8 proteins and then clustered with vertebrate IRF3, IRF4, and IRF5 subfamily proteins. Quantitative real-time PCR detected CfIRF8-like mRNA in all tested scallop tissues, with the highest expression in the gills. Simultaneously, the expression of CfIRF8-like transcripts in gills was significantly induced by polyinosinic-polycytidylic acid challenge. The results of protein interaction experiments showed that CfIRF8-like could directly bind the TBK1/IKKε family protein of scallop (CfIKK2) via its N-terminal IRF domain, revealing the presence of an ancient functional TBK1/IKKε-IRF signaling axis in scallops. Finally, dual-luciferase reporter assay results showed that the overexpression of CfIRF8-like in human embryonic kidney 293T cells could specifically activate the interferon ß promoter of mammals and the interferon-stimulated response element promoter in dose-dependent manners. The findings of this preliminary analysis of the signal transduction and immune functions of scallop CfIRF8-like protein lay a foundation for an in-depth understanding of the innate immune function of invertebrate IRFs and the development of comparative immunology. The experimental results also provide theoretical support for the breeding of scallop disease-resistant strains.

Regulation of Tyrosinase Gene Expression by Retinoic Acid Pathway in the Pacific Oyster Crassostrea gigas.

Retinoic acid (RA) plays important roles in various biological processes in animals. RA signaling is mediated by two types of nuclear receptors, namely retinoic acid receptor (RAR) and retinoid x receptor (RXR), which regulate gene expression by binding to retinoic acid response elements (RAREs) in the promoters of target genes. Here, we explored the effect of all-trans retinoic acid (ATRA) on the Pacific oyster Crassostera gigas at the transcriptome level. A total of 586 differentially expressed genes (DEGs) were identified in C. gigas upon ATRA treatment, with 309 upregulated and 277 downregulated genes. Bioinformatic analysis revealed that ATRA affects the development, metabolism, reproduction, and immunity of C. gigas. Four tyrosinase genes, including Tyr-6 (LOC105331209), Tyr-9 (LOC105346503), Tyr-20 (LOC105330910), and Tyr-12 (LOC105320007), were upregulated by ATRA according to the transcriptome data and these results were verified by real-time quantitative polymerase chain reaction (RT-qPCR) analysis. In addition, increased expression of Tyr (a melanin-related TYR gene in C. gigas) and Tyr-2 were detected after ATRA treatment. The yeast one-hybrid assay revealed the DNA-binding activity of the RA receptors CgRAR and CgRXR, and the interaction of CgRAR with RARE present in the Tyr-2 promoter. These results provide evidence for the further studies on the role of ATRA and the mechanism of RA receptors in mollusks.

Perirenal hematoma and delayed contrast metabolism after cerebral intravascular therapy: A case report.

RATIONALE: Hematomas after percutaneous angiography often occur in the thigh, retroperitoneal, intraperitoneal, or abdominal wall. Renal hematoma after percutaneous angiography is very rare. DIAGNOSES: Herein, we present a case of perirenal hematoma and delayed contrast metabolism after cerebral angiograph, which may be caused by improper operation. INTERVENTIONS: Conservative treatments which development by multi-disciplinary collaboration. OUTCOMES: After treatment, the clinical symptoms of the patients gradually disappeared and the imaging results became negative. CONCLUSION: Though the patient missed timely diagnosis and treatment, fortunately no catastrophic events occurred. Meanwhile, the potential causes, diagnosis, and therapeutic management were all discussed.

Coordination-driven Cu-based Fenton-like process for humic acid treatment in wastewater.

Fenton oxidation process is effective in organic pollutant degradation during wastewater treatment, but subject to narrow pH range and secondary pollution. In this work, an application-promising alternative, i.e., coordination-driven Cu-based Fenton-like process, was proposed for wastewater treatment using humic-acid (HA) as the target contaminant. The results showed that the removal of HA through Cu-based Fenton-like process can reach 70% under the condition of pH 8.0, 146.8 mmol/L H2O2, 146.8 µmol/L Cu (II), 50 °C, and 4 h. Addition of Cl- could significantly accelerate the reaction process through coordination with copper ions, while HCO3- and P2O74- exhibited opposite effects. Increasing temperature is also beneficial for advancing the reaction, and the removal of HA followed pseudo-first-order kinetics. Fluorescence spectroscopic analyses showed that the removal of HA experienced a two-stage process, i.e., oxidation followed by degradation, which is dependent of the presence of coordination ions. Parallel factor analysis was used to characterize the change of fluorescence components. Three fluorescent components, i.e., terrestrial humic-like, UV/visible terrestrial humic-like and protein-like component were identified, all of which were effectively removed. This study deepens our understanding on Cu-based Fenton-like process, and may provide a promising technology for refractory wastewater treatment.

Counteracting health risks by Modulating Homeostatic Signaling.

Homeostasis was initially conceptualized by Bernard and Cannon around a century ago as a steady state of physiological parameters that vary within a certain range, such as blood pH, body temperature, and heart rate [1,2]. The underlying mechanisms that maintain homeostasis are explained by negative feedbacks that are executed by the neuronal, endocrine, and immune systems. At the cellular level, homeostasis, such as that of redox and energy steady state, also exists and is regulated by various cell signaling pathways. The induction of homeostatic mechanism is critical for human to adapt to various disruptive insults (stressors); while on the other hand, adaptation occurs at the expense of other physiological processes and thus runs the risk of collateral damages, particularly under conditions of chronic stress. Conceivably, anti-stress protection can be achieved by stressor-mimicking medicinals that elicit adaptive responses prior to an insult and thereby serve as health risk countermeasures; and in situations where maladaptation may occur, downregulating medicinals could be used to suppress the responses and prevent subsequent pathogenesis. Both strategies are preemptive interventions particularly suited for individuals who carry certain lifestyle, environmental, or genetic risk factors. In this article, we will define and characterize a new modality of prophylactic intervention that forestalls diseases via modulating homeostatic signaling. Moreover, we will provide evidence from the literature that support this concept and distinguish it from other homeostasis-related interventions such as adaptogen, hormesis, and xenohormesis.

Aluminum separation by sulfuric acid leaching-solvent extraction from Al-bearing LiFePO4/C powder for recycling of Fe/P.

Recovery of battery-grade FePO4 from Al-bearing spent LiFePO4 batteries (LFPs) is important for both prevention of environmental pollution and recycling of resources for LFPs industries. The premise for FePO4 recovery from spent LFPs is the separation of Al, because Al readily co-precipitates with FePO4 and lowers the electrochemical performance of the regenerated LiFePO4. In this work, an efficient approach involving sulfuric acid leaching followed by solvent extraction was developed to separate Al from spent LiFePO4/C powder. Di-(2-ethylhexyl) phosphoric acid (D2EHPA) in sulfonated kerosene was used as the extractant. The results showed that 96.4% of aluminum was extracted while the loss of iron was only 1.1% under the optimal conditions. The mass fraction of Al in the iron phosphate obtained from the extraction raffinate was only 0.007%, meeting the standard for preparing battery-grade FePO4. The extracted Al can be easily stripped by diluted H2SO4 solution and the extractants can be reused. Additionally, slope analysis method, FTIR spectroscopy, and ESI-MS analysis revealed that the extraction of Al in D2EHPA can be ascribed to the ion exchange between hydrogen ion of -PO(OH) and Al3+. This work may provide an economically feasible method for the recycling of valuable components from spent Al-bearing LiFePO4/C powder.

Preparation of Battery-Grade FePO4·2H2O Using the Stripping Solution Generated from Resource Recycling of Bauxite Residue.

A novel process for the high-value-use of iron from bauxite residue was proposed in this work. The process was trying to use the iron-containing stripping solution generated during resource recycling of bauxite residue to produce battery-grade FePO4·2H2O product. Thermodynamics calculation indicates that Fe and P in the stripping solution mainly existed in the form of FeHPO4+, and the theoretical pH for the conversion reaction from FePO4·2H2O to Fe(OH)3 was 1.72. The optimal condition for the synthesis of FePO4·2H2O using the stripping solution was determined as: reaction pH of 0.8, reaction temperature of 90°C, Fe/P ratio of 1, and reaction time of 24 h. XRD result showed that the synthesized FePO4·2H2O was well-crystallized and perfectly matched with the characteristic peaks of FePO4·2H2O. Moreover, all the parameters of the synthesized iron phosphate meet the quality requirements of battery precursor.

Colossal and reversible barocaloric effect in liquid-solid-transition materials n-alkanes.

Emerging caloric cooling technology provides a green alternative to conventional vapor-compression technology which brings about serious environmental problems. However, the reported caloric materials are much inferior to their traditional counterparts in cooling capability. Here we report the barocaloric (BC) effect associated with the liquid-solid-transition (L-S-T) in n-alkanes. A low-pressure of ~50 MPa reversibly triggers an entropy change of ~700 J kg-1 K-1, comparable to those of the commercial refrigerants in vapor-based compression systems. The Raman study and theoretical calculations reveal that applying pressure to the liquid state suppresses the twisting and random thermal motions of molecular chains, resulting in a lower configurational entropy. When the pressure is strong enough to drive the L-S-T, the configurational entropy will be fully suppressed and induce the colossal BC effect. This work could open a new avenue for exploring the colossal BC effect by evoking L-S-T materials.

A novel TBK1/IKKϵ is involved in immune response and interacts with MyD88 and MAVS in the scallop Chlamys farreri.

Inhibitor of κB kinase (IKK) family proteins are key signaling molecules in the animal innate immune system and are considered master regulators of inflammation and innate immunity that act by controlling the activation of transcription factors such as NF-κB. However, few functional studies on IKK in invertebrates have been conducted, especially in marine mollusks. In this study, we cloned the IKK gene in the Zhikong scallop Chlamys farreri and named it CfIKK3. CfIKK3 encodes a 773-amino acid-long protein, and phylogenetic analysis showed that CfIKK3 belongs to the invertebrate TBK1/IKKϵ protein family. Quantitative real-time PCR analysis showed that CfIKK3 mRNA is ubiquitously expressed in all tested scallop tissues. The expression of CfIKK3 transcripts was significantly induced after challenge with lipopolysaccharide, peptidoglycan, or poly(I:C). Co-immunoprecipitation (co-IP) assays confirmed the direct interaction of CfIKK3 with MyD88 (the key adaptor in the TLR pathway) and MAVS (the key adaptor in the RLR pathway), suggesting that this IKK protein plays a crucial role in scallop innate immune signal transduction. In addition, the CfIKK3 protein formed homodimers and bound to CfIKK2, which may be a key step in the activation of its own and downstream transcription factors. Finally, in HEK293T cells, dual-luciferase reporter gene experiments showed that overexpression of CfIKK3 protein activated the NF-κB reporter gene in a dose-dependent manner. In conclusion, our experimental results confirmed that CfIKK3 could respond to PAMPs challenge and participate in scallop TLR and RLR pathway signaling, ultimately activating NF-κB. Therefore, as a key signaling molecule and modulator of immune activity, CfIKK3 plays an important role in the innate immune system of scallops.

Separation and recovery of arsenic from As, Cu, and Zn rich leaching liquor using a reduction-crystallization approach.

As, Cu, and Zn rich leaching liquor is generated in the leaching process of copper dust, which contains various metals with high recovery value. Herein, an approach for the direct separation and recovery of arsenic from As, Cu, and Zn rich leaching liquor was proposed. The approach includes two steps, namely SO2 reduction and arsenic crystallization. The factors affecting the reduction of As(v) to As(iii) were investigated, including the pH, SO2 dosage, and reduction temperature. In the crystallization stage, the impacts of sulfuric acid consumption and temperature on the crystallization of arsenic (As2O3) were studied. The results show that the optimal H+ concentration, temperature, and SO2 input for the arsenic reduction were 3.95 mol L-1, 45 °C, and 1.14 L g-1 As(v), respectively. While the optimal temperature and sulfuric acid dosage in As recovery process were 5 °C and 0.1 L L-1 leaching liquor, respectively. Under these conditions, the As2O3 recovery percentage reached 96.53%, and the losses of Cu and Zn were only 3.12% and 0.75%, respectively. The precipitate contained 96.72% of As2O3, 0.83% of Cu, and 0.13% Zn. Compared with the traditional technologies, this new method can improve the recovery efficiency of As2O3 and reduce the loss percentage of other valuable metals (Cu and Zn).