SY18ΔL60L: a new recombinant live attenuated African swine fever virus with protection against homologous challenge

IntroductionAfrican swine fever (ASF) is an acute and highly contagious disease and its pathogen, the African swine fever virus (ASFV), threatens the global pig industry. At present, management of ASF epidemic mainly relies on biological prevention and control methods. Moreover, due to the large genome of ASFV, only half of its genes have been characterized in terms of function.MethodsHere, we evaluated a previously uncharacterized viral gene, L60L. To assess the function of this gene, we constructed a deletion strain (SY18ΔL60L) by knocking out the L60L gene of the SY18 strain. To evaluate the growth characteristics and safety of the SY18ΔL60L, experiments were conducted on primary macrophages and pigs, respectively.ResultsThe results revealed that the growth trend of the recombinant strain was slower than that of the parent strain in vitro. Additionally, 3/5 (60%) pigs intramuscularly immunized with a 105 50% tissue culture infectious dose (TCID50) of SY18ΔL60L survived the 21-day observation period. The surviving pigs were able to protect against the homologous lethal strain SY18 and survive. Importantly, there were no obvious clinical symptoms or viremia.DiscussionThese results suggest that L60L could serve as a virulence- and replication-related gene. Moreover, the SY18ΔL60L strain represents a new recombinant live-attenuated ASFV that can be employed in the development of additional candidate vaccine strains and in the elucidation of the mechanisms associated with ASF infection

A Companion Cell–Dominant and Developmentally Regulated H3K4 Demethylase Controls Flowering Time in Arabidopsis via the Repression of FLC Expression

Flowering time relies on the integration of intrinsic developmental cues and environmental signals. FLC and its downstream target FT are key players in the floral transition in Arabidopsis. Here, we characterized the expression pattern and function of JMJ18, a novel JmjC domain-containing histone H3K4 demethylase gene in Arabidopsis. JMJ18 was dominantly expressed in companion cells; its temporal expression pattern was negatively and positively correlated with that of FLC and FT, respectively, during vegetative development. Mutations in JMJ18 resulted in a weak late-flowering phenotype, while JMJ18 overexpressors exhibited an obvious early-flowering phenotype. JMJ18 displayed demethylase activity toward H3K4me3 and H3K4me2, and bound FLC chromatin directly. The levels of H3K4me3 and H3K4me2 in chromatins of FLC clade genes and the expression of FLC clade genes were reduced, whereas FT expression was induced and the protein expression of FT increased in JMJ18 overexpressor lines. The early-flowering phenotype caused by the overexpression of JMJ18 was mainly dependent on the functional FT. Our findings suggest that the companion cell–dominant and developmentally regulated JMJ18 binds directly to the FLC locus, reducing the level of H3K4 methylation in FLC chromatin and repressing the expression of FLC, thereby promoting the expression of FT in companion cells to stimulate flowering

Distribution, Site-Specific Water Quality Criteria, and Ecological Risk Assessment of Heavy Metals in Surface Water in Fen River, China

Due to a lack of toxicity reference values that match the regional environmental characteristics, the ecological risk of metals in water bodies cannot be accurately assessed. The Fen River is the second-largest tributary of the Yellow River in China, and the sustainability of this area is threatened by heavy metal pollution caused by intensive industrial and agricultural activities. In this study, site-specific water quality criteria (WQCs) for heavy metals in the Fen River were derived considering toxicity data from native aquatic organisms and regional water quality factors (e.g., water hardness). Short-term WQCs for Mn, Cu, Cd, Zn, Cr, Pb, and Ni were 2026.15, 98.62, 10.02, 63.07, 6.06, 166.74, and 132.73 μg/L, respectively, and long-term WQCs were 166.53, 29.71, 2.18, 19.29, 4.15, 6.38, and 14.76 μg/L, respectively. The distribution characteristics of these metals during the wet season in 2020 were explored, and their average concentrations in the river water did not exceed the environmental quality standards for surface water in China but were higher than the world average levels. Cr was the main pollutant in the sampling sites of Yaodu region, Hongdong Shitan, Xiao River, and Duanchun River, as was Pb in Duanchun River. Based on the site-specific WQCs, using hazardous quotient (HQ) and margin of safety (MOS10) approaches, a high risk of Pb was identified in the Duanchun River, and a medium risk of Cr might occur at midstream and downstream of Yaodu and Xiaodian. The results will provide a reference basis for heavy metal pollution control and water quality management in the Fen River

Association between Alanine Aminotransferase/Aspartate Aminotransferase Ratio (AST/ALT Ratio) and Coronary Artery Injury in Children with Kawasaki Disease

Objective. To investigate the association between the aspartate aminotransferase (AST)/alanine aminotransferase (ALT) ratio (AST/ALT ratio, AAR) and intravenous immunoglobulin (IVIG) resistance, coronary artery lesions (CAL), and coronary artery aneurysms (CAA) in children with Kawasaki disease (KD). Design. We retrospectively studied 2678 children with KD and divided them into two groups: a low-AAR group and a high-AAR group with a median AAR of 1.13 as the cut-off point. The differences in laboratory data, clinical manifestations, and coronary artery damage rates were compared between the two groups. Results. The incidence of CAL was higher in the low-AAR group than in the high-AAR group at 2 and 3-4 weeks after illness onset (p<0.001, respectively). The IVIG resistance rate was significantly higher in the low-AAR group than in the high-AAR group (29.94% vs 21.71%, p<0.001). The levels of C-reactive protein, erythrocyte sedimentation rate, white blood cell count, bilirubin, fibrinogen, thrombin time, D-dimer, and brain natriuretic peptide were also significantly higher in the low-AAR group compared with the high-AAR group. The levels of albumin and IgG were significantly lower in the low-AAR group compared with those of the high-AAR group. The proportion of typical KD cases in the low-AAR group was significantly higher than that in the high-AAR group. Low-AAR correlated with the risk of coronary artery damage and IVIG resistance. Conclusion. Children with KD who had low-AAR value were more likely to develop coronary artery damage and IVIG resistance. Low AAR is a risk factor for CAL, CAA, and IVIG resistance in KD

Evaluation of African Swine Fever Virus <i>E111R</i> Gene on Viral Replication and Porcine Virulence

African swine fever (ASF) is an acute infectious disease of domestic pigs and wild boars caused by the African swine fever virus (ASFV), with up to a 100% case fatality rate. The development of a vaccine for ASFV is hampered by the fact that the function of many genes in the ASFV genome still needs to be discovered. In this study, the previously unreported E111R gene was analyzed and identified as an early-expressed gene that is highly conserved across the different genotypes of ASFV. To further explore the function of the E111R gene, a recombinant strain, SY18ΔE111R, was constructed by deleting the E111R gene of the lethal ASFV SY18 strain. In vitro, the replication kinetics of SY18ΔE111R with deletion of the E111R gene were consistent with those of the parental strain. In vivo, high-dose SY18ΔE111R (105.0 TCID50), administered intramuscularly to pigs, caused the same clinical signs and viremia as the parental strain (102.0 TCID50), with all pigs dying on days 8–11. After being infected with a low dose of SY18ΔE111R (102.0 TCID50) intramuscularly, pigs showed a later onset of disease and 60% mortality, changing from acute to subacute infection. In summary, deletion of the E111R gene has a negligible effect on the lethality of ASFV and does not affect the viruses’ ability to replicate, suggesting that E111R could not be the priority target of ASFV live-attenuated vaccine candidates

Automated Machine Learning in Predicting 30-Day Mortality in Patients with Non-Cholestatic Cirrhosis

Objective: To evaluate the feasibility of automated machine learning (AutoML) in predicting 30-day mortality in non-cholestatic cirrhosis. Methods: A total of 932 cirrhotic patients were included from the First Affiliated Hospital of Soochow University between 2014 and 2020. Participants were divided into training and validation datasets at a ratio of 8.5:1.5. Models were developed on the H2O AutoML platform in the training dataset, and then were evaluated in the validation dataset by area under receiver operating characteristic curves (AUC). The best AutoML model was interpreted by SHapley Additive exPlanation (SHAP) Plot, Partial Dependence Plots (PDP), and Local Interpretable Model Agnostic Explanation (LIME). Results: The model, based on the extreme gradient boosting (XGBoost) algorithm, performed better (AUC 0.888) than the other AutoML models (logistic regression 0.673, gradient boost machine 0.886, random forest 0.866, deep learning 0.830, stacking 0.850), as well as the existing scorings (the model of end-stage liver disease [MELD] score 0.778, MELD-Na score 0.782, and albumin-bilirubin [ALBI] score 0.662). The most key variable in the XGBoost model was high-density lipoprotein cholesterol, followed by creatinine, white blood cell count, international normalized ratio, etc. Conclusion: The AutoML model based on the XGBoost algorithm presented better performance than the existing scoring systems for predicting 30-day mortality in patients with non-cholestatic cirrhosis. It shows the promise of AutoML in its future medical application

Deleting the C84L Gene from the Virulent African Swine Fever Virus SY18 Does Not Affect Its Replication in Porcine Primary Macrophages but Reduces Its Virulence in Swine

(1) Background: African swine fever (ASF) is a highly contagious disease that causes high pig mortality. Due to the absence of vaccines, prevention and control are relatively challenging. The pathogenic African swine fever virus (ASFV) has a complex structure and encodes over 160 proteins, many of which still need to be studied and verified for their functions. In this study, we identified one of the unknown functional genes, C84L. (2) Methods: A gene deficient strain was obtained through homologous recombination and several rounds of purification, and its replication characteristics and virulence were studied through in vitro and in vivo experiments, respectively. (3) Results: Deleting this gene from the wild-type virulent strain SY18 did not affect its replication in porcine primary macrophages but reduced its virulence in pigs. In animal experiments, we injected pigs with a 102 TCID50, 105 TCID50 deletion virus, and a 102 TCID50 wild-type strain SY18 intramuscularly. The control group pigs reached the humane endpoint on the ninth day (0/5) and were euthanized. Two pigs in the 102 TCID50(2/5) deletion virus group survived on the twenty-first day, and one in the 105 TCID50(1/5) deletion virus group survived. On the twenty-first day, the surviving pigs were euthanized, which was the end of the experiment. The necropsies of the survival group and control groups’ necropsies showed that the surviving pigs’ liver, spleen, lungs, kidneys, and submaxillary lymph nodes did not show significant lesions associated with the ASFV. ASFV-specific antibodies were first detected on the seventh day after immunization; (4) Conclusions: This is the first study to complete the replication and virulence functional exploration of the C84L gene of SY18. In this study, C84L gene was preliminarily found not a necessary gene for replication, gene deletion strain SY18ΔC84L has similar growth characteristics to SY18 in porcine primary alveolar macrophages. The C84L gene affects the virulence of the SY18 strain

Low-Intensity Pulsed Ultrasound Attenuates Postoperative Neurocognitive Impairment and Salvages Hippocampal Synaptogenesis in Aged Mice

Postoperative neurocognitive impairment is an urgent problem with global aging accelerating. The prevention and treatment of postoperative neurocognitive impairment have been widely investigated but lack effective strategies. Low-intensity pulsed ultrasound (LIPUS), a non-invasive tool, has shown an effect on neuroprotection, but whether it could attenuate the postoperative neurocognitive impairment and the underlying mechanisms remains unknown. An experimental setup for LIPUS stimulation of the hippocampus was well established. A laparotomy model in aged mice was applied, and a Morris water maze was used to assess cognitive function. RT-qPCR and western blotting were used to detect levels of Piezo1, synapse-associated proteins in the hippocampus, respectively. Immunofluorescent staining was also used to determine the neural activation and Piezo1 expression. The results showed that LIPUS increased synapse-related proteins of the hippocampus and attenuated cognitive impairment in aged mice. Meanwhile, LIPUS suppressed the overexpression of Piezo1 in the hippocampus. We further found that LIPUS promoted Calpain1 activity and increased extracellular regulated protein kinases (Erk) phosphorylation. Our results suggested that LIPUS could improve cognitive impairment and increase hippocampal synaptogenesis through the Piezo1-mediated Calpain1/ Erk pathway. LIPUS could be used as an effective physical intervention to alleviate postoperative cognitive dysfunction in the aged population