Establishment and application of a VP3 antigenic domain-based peptide ELISA for the detection of antibody against goose plague virus infection.

The detection of antibody against goose plague virus (GPV) infection has never had a commercialized test kit, which has posed challenges to the prevention and control of this disease. In this study, bioinformatics software was used to analyze and predict the dominant antigenic regions of the main protective antigen VP3 of GPV. Three segments of bovine serum albumin (BSA) vector-coupled peptides were synthesized as ELISA coating antigens. Experimental results showed that the VP3-1 (358-392aa) peptide had the best reactivity and specificity. By using the BSA-VP3-1 peptide, a detection method for antibody against GPV infection was established, demonstrating excellent specificity with no cross-reactivity with common infectious goose pathogen antibodies. The intra-batch coefficient of variation and inter-batch coefficient of variation were both less than 7%, indicating good stability and repeatability. The dynamic antibody detection results of gosling vaccines and the testing of 120 clinical immune goose serum samples collectively demonstrate that BSA-VP3-1 peptide ELISA can be used to detect antibody against GPV in the immunized goose population and has higher sensitivity than traditional agar gel precipitation methods. Taken together, the developed peptide-ELISA based on VP3 358-392aa could be useful in laboratory viral diagnosis, routine surveillance in goose farms. The main application of the peptide-ELISA is to monitor the antibody level and vaccine efficacy for GPV, which will help the prevention and control of gosling plague.

Comprehensive characterization of MUC16 mutations in lung adenocarcinoma for immunotherapies and prognosis: An observational study.

Lung adenocarcinoma (LUAD) is a non-small-cell lung cancer and is the leading cause of cancer-related deaths worldwide. Immunotherapy is a promising candidate for LUAD, and tumor mutation burden (TMB) could be a new biomarker to monitor the response of cancer patients to immunotherapy. It is known that the mucin 16 (MUC16) mutation is the most common and affects the progression and prognosis of several cancers. However, whether MUC16 mutations are associated with TMB and tumor-infiltrating immune cells in LUAD is not fully elucidated. All the data were obtained from the cancer genome atlas database to assess the prognostic value and potential mechanism of MUC16 in LUAD. An immune prognostic model (IPM) was developed based on immune-related genes that could be differentially expressed between MUC16MUT and MUC16WT LUAD patients. Later, the IPM effect on the prognosis and immunotherapy of LUAD was comprehensively evaluated. MUC16 was frequently mutated in LUAD, with a mutational frequency of 43.4%, significantly associated with higher TMB and better clinical prognosis. Based on 436 patients with LUAD, an IPM was established and validated to differentiate patients with a low or high risk of poor survival. The univariate and multivariate Cox regression analyses demonstrated that the IPM was an independent prognostic indicator for LUAD patients. Elevated expressions of PD-L1, LAG3, PDCD1, and SIGLEC15, and most of the T-effector and interferon-γ gene signatures, were depicted in the high-risk group. Moreover, the nomogram using the IPM and clinical prognostic factors also predicted the overall survival and clinical utility. Our project developed a robust risk signature depending on the MUC16 status and provided novel insights for individualized treatment options for LUAD patients.

Discovery of Novel, Selective Prostaglandin EP4 Receptor Antagonists with Efficacy in Cancer Models.

Prostaglandin E2 (PGE2) receptor 4 (EP4) is one of four EP receptors commonly upregulated in the tumor microenvironment and plays vital roles in stimulating cell proliferation, invasion, and metastasis. Biochemical blockade of the PGE2-EP4 signaling pathway is a promising strategy for controlling inflammatory and immune related disorders. Recently combination therapies of EP4 antagonists with anti-PD-1 or chemotherapy agents have emerged in clinical studies for lung, breast, colon, and pancreatic cancers. Herein, a novel series of indole-2-carboxamide derivatives were identified as selective EP4 antagonists, and SAR studies led to the discovery of the potent compound 36. Due to favorable pharmacokinetics properties and good oral bioavailability (F = 76%), compound 36 was chosen for in vivo efficacy studies. Compound 36 inhibited tumor growth in a CT-26 colon cancer xenograft better than E7046 and a combination of 36 with capecitabine significantly suppressed tumor growth (TGI up to 94.26%) in mouse models.

Evaluation of waterlogging tolerance and responses of protective enzymes to waterlogging stress in pumpkin.

Waterlogging caused by short and severe, or prolonged precipitation can be attributed to global warming. Pumpkin plants are drought-tolerant but not tolerate to waterlogging stress. Under frequent rain and waterlogging conditions, the production of pumpkins is of lower quality, sometimes rotten, and harvest failure occurs in severe cases. Therefore, it is of great significance to assess the waterlogging tolerance mechanism of pumpkin plants. In this study, 10 novel pumpkin varieties from Baimi series were used. The waterlogging tolerance level of pumpkin plants was evaluated by measuring waterlogging tolerance coefficient of biomass and physiological indices using waterlogging stress simulation method. The criteria to evaluate the waterlogging tolerance capacities of pumpkin plants were also explored. Using principal component and membership function analysis, waterlogging tolerance levels of the pumpkin varieties were ranked as follows: Baimi No. 10>; Baimi No. 5>; Baimi No. 1>; Baimi No. 2>; Baimi No. 3>; Baimi No. 7>; Baimi No. 9>; Baimi No. 6>; Baimi No. 4>; Baimi No. 8. Based on the results, Baimi No. 10 was identified with strong waterlogging tolerance and Baimi No. 8 with weak waterlogging tolerance. The responses of malondialdehyde (MDA), proline, key enzymes responsible for anaerobic respiration, and antioxidant enzymes to waterlogging stress were studied in pumpkin plants. The relative expression levels of related genes were determined using real-time fluorescence quantitative PCR technique. The aim of our study was to assess the waterlogging tolerance mechanism of pumpkin plants, thus laying a theoretical foundation for breeding waterlogging-tolerant varieties in the future. After flooding stress treatment, the antioxidant enzyme activities, contents of proline and alcohol dehydrogenases of Baimi No. 10 and Baimi No. 8 displayed an increase followed by a decrease. All indices of Baimi No. 10 were higher than Baimi No. 8. MDA contents gradually increased, with the content being higher in Baimi No. 8 than Baimi No. 10. The activities of pyruvate decarboxylases (PDCs) in Baimi No. 8 and Baimi No. 10 exhibited a decrease initially, followed by an increase, and then a decrease again. The PDC activity in Baimi No. 8 was generally higher than Baimi No. 10. The relative expression levels of genes encoding superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase were consistent with their corresponding enzyme activities. During the early stage of flooding stress, pumpkin plants waterlogging tolerance was improved by enhancing the expression levels of antioxidant enzyme encoding genes and increasing the antioxidant enzyme activities.

Distracted driving behavior in patients with insomnia.

Insomnia is one of the most common sleep disorders and is characterized by a subjective perception of difficulty falling asleep. Drivers with insomnia are vulnerable to distraction and exhibit higher levels of risk while driving. This study investigated the effect of two sources of in-vehicle distractions on the driving performance of drivers with insomnia and good sleepers by analyzing different driving behavior measures. Twenty-one drivers with insomnia and twenty-one healthy volunteers were recruited to complete simulated driving dual tasks. The primary task required the participants to perform: (a) a lane-keeping task, and (b) a lane-change task. The secondary task required the participants to deal with: (a) baseline (non-task), (b) internal distraction task, and (c) external distraction task. The internal distraction task required participants to complete quantitative reasoning tasks, while the external distraction task was a 0-back test. The relationship between distracted driving ability and cognitive function was also investigated. The results demonstrate that for lane-keeping tasks, drivers with insomnia had significantly higher standard deviations (SD) for speed, throttle position, acceleration, and lateral position than healthy drivers under internal distraction, but the driving performance did not differ significantly between groups under internal distraction or baseline. In the lane-change task, drivers with insomnia had higher SDs for steering wheel angle, steer angular velocity, lateral acceleration, and lateral speed than healthy drivers under external distraction. Moreover, external distraction impaired driving behavior in the healthy group, while internal distraction impaired driving ability in both groups. Healthy drivers with cognitive impairment displayed impaired lane-keeping abilities under internal distractions and impaired lane-changing abilities under external distractions. Driving performance in the insomnia group was not significantly associated with cognitive function. The results demonstrate that insomnia and distraction impair driving ability, and driver performance is affected differently by the distraction source (internal or external). The driving ability of healthy drivers with decreased cognition was impaired, but not that of insomniacs.The findings of this study provide new insights for preventing and estimating the potential influence of distracted driving behavior in individuals with insomnia.

Fatigue performance in patients with chronic insomnia.

Insomnia is associated with fatigue and poor driving performance, thus increasing the risk of traffic accidents. This study aimed to evaluate the effect of fatigue on driving in patients with chronic insomnia in a free-flow traffic scenario and car-following scenario, and to investigate the relationships between driving performance, cognitive function, and insomnia. The Trail Making Test (TMT), Stroop Color and Word Test (SCWT), Symbol Digit Modalities Test (SDMT), and Digit Span Test (DST) of 15 participants with mild-to-moderate chronic insomnia and 16 healthy participants were assessed. During the fatigue driving task, drivers completed simulated driving tasks under free-flow traffic and car-following scenarios. The mean speed (MS), mean acceleration (MA), mean lateral position (MLP), and standard deviation of lateral position (SDLP) were measured to assess driving performance. During fatigued tasks, the MA and MLP in the free-driving scenario were higher than those in the car-following scenario (P < 0.01), the SDLP was higher in the insomnia group than in the healthy group (P = 0.02), and the interaction effect was significantly different for MLP between the groups (P = 0.03). MS was negatively correlated with TMT score, SDMT score, and DST score, and positively correlated with time to complete TMT, errors in SCWT, and time to complete SCWT. SDLP was negatively correlated with DST score and positively correlated with time to complete SCWT. Furthermore, the insomnia group had poorer lateral vehicle control ability than the healthy group. The insomnia group had a more impaired driving performance in the free-driving scenario than in the car-following scenario. Drivers with impaired cognitive function exhibited impaired driving performance.

Measuring the effects of mind wandering in people with insomnia: A driving simulator study.

Purpose: Studies have shown that individuals with insomnia experience more frequent and longer episodes of mind wandering (MW) while driving. However, the effect of the interaction between insomnia and MW on driving behavior is not fully understood. This study aimed to gain deeper insights into the relationships among insomnia, MW, and driving behavior. Patients and methods: Forty-two participants (21 diagnosed with insomnia and 21 controls) were recruited, and subjective sleep quality and cognitive function were assessed. A driving simulator experiment with a within-subject design was performed, involving two distraction tasks (no-distraction task versus MW task) and two driving scenarios (lane-keeping versus lane-changing). Results: In the lane-keeping scenario, there was no significant between-group difference (people with insomnia and controls) in longitudinal driving performance for the no-distraction task, although the interaction between MW and insomnia significantly increased drivers' longitudinal control variation. Correlation analysis confirmed that longitudinal driving performance was positively correlated with sleep quality and the cognitive level. Unlike longitudinal driving performance, lateral driving performance was significantly weaker in people with insomnia than in controls under both distraction tasks. In the lane-changing scenario, although there was no between-group difference in driving performance, the MW task led to significant changes in driving performance within each group compared with the no-distraction task, and these findings were associated with cognitive function, but not with sleep quality. Conclusion: These findings show that insomnia and MW combined can lead to reduced driving performance. Further research is needed to elucidate the factors that influence this phenomenon.

Effects of ischemia-hypoperfusion on neuro-vascular units in dual transgenic mice with Alzheimer's disease.

INTRODUCTION: The study aimed to investigate the effects of ischemia on neuro-vascular units in transgenic mice, and to investigate the role of ischemia-hypoperfusion in the model of dual transgenic mice with dementia. MATERIAL AND METHODS: In this study, the ischemic model was generated by operating a bilateral common carotid artery micro-embolism. Mice were divided into four groups, including group 1: C57BL sham surgery group (control), group 2: C57BL ischemic group, group 3: amyloid precursor protein/presenilin-1 (APP/PS1) group, and group 4: APP/PS1 ischemic group. Each group comprised 20 mice. Spatial behavior and memory ability of mice were detected by Morris water maze and jumping platform test. Mouse hippocampus was observed by HE staining and Congo red staining. Ultrastructure of each group of neuro-cyclic units was observed by electron microscopy. Various biochemical indicators were detected by ELISA. Western blot detected the amount of protein expression. qRT-PCR identified mRNA expression. RESULTS: The results indicated that learning and memory functions of C57 ischemic mice were lower than those of control group. Positive expression area of APP in APP/PS1 ischemic group was higher than in APP/PS1 group. In APP/PS1 group and APP/PS1 ischemic group, the content of Ab was significantly higher than in C57 ischemic group. Electron microscopic observation revealed that there were more mitochondrial vacuoles in hippocampal neurons of APP/PS1 mice, and the structure was relatively intact. Mitochondrial vacuoles in hippocampus increased significantly, and vascular wall proliferated in APP/PS1 ischemic group. Compared with C57 control group, the content of vascular endothelial growth factor (VEGF) increased significantly in C57 ischemic group. CONCLUSIONS: Ischemia deteriorates the learning and memory function of transgenic mice, aggravates the damage of neuro-vascular units, and impairs the blood-brain barrier transport of Ab, leading to an increase in the concentration of Ab cerebrospinal fluid, and further deterioration of neuro-vascular units. At the same time, ischemia is an effective stimulating factor in the release of VEGF.

Antiviral Effect of Lithium Chloride on Replication of Marek's Disease Virus in Chicken Embryonic Fibroblasts.

To investigate the antiviral effect of lithium chloride (LiCl) on the replication of Marek's disease virus (MDV) in chicken embryonic fibroblast (CEF) cells, real-time PCR, Western blotting, plaque counting, and indirect immunofluorescence experiments were performed at different time points of LiCl treated CEF cells with virus infection. The results demonstrated that LiCl could affect multiple steps of virus replication and inhibit viral gene expression and protein synthesis in a dose- and time-dependent manner. Moreover, LiCl could directly affect viral infectivity as well. In addition, LiCl significantly affected the gene expression of IFN-ß related genes in virus-infected cells. These results indicate that LiCl significantly inhibits MDV replication and proliferation in CEF cells and it has the potential to be used as an antiviral agent against MDV.

Regulation of Avian Leukosis Virus Subgroup J Replication by Wnt/ß-Catenin Signaling Pathway.

Wnt/ß-catenin signaling is a highly conserved pathway related to a variety of biological processes in different cells. The regulation of replication of various viruses by Wnt/ß-catenin signaling pathway has been reported. However, the interaction between the Wnt/ß-catenin pathway and avian leukosis virus is unknown. In the present study, we investigated the effect of modulating the Wnt/ß-catenin pathway during avian leukosis virus subgroup J (ALV-J) infection. The activation of the Wnt/ß-catenin pathway by GSK-3 inhibitor increased ALV-J mRNA, viral protein expression, and virus production in CEF cells. This increase was suppressed by iCRT14, one of the specific inhibitors of the Wnt/ß-catenin signaling pathway. Moreover, treatment with iCRT14 reduced virus titer and viral gene expression significantly in CEF and LMH cells in a dose-dependent manner. Inhibition Wnt/ß-catenin signaling pathway by knockdown of ß-catenin reduced virus proliferation in CEF cells also. Collectively, these results suggested that the status of Wnt/ß-catenin signaling pathway modulated ALV-J replication. These studies extend our understanding of the role of Wnt/ß-catenin signaling pathway in ALV-J replication and make a new contribution to understanding the virus-host interactions of avian leukosis virus.

Impairment of the Cell Wall Ligase, LytR-CpsA-Psr Protein (LcpC), in Methicillin Resistant Staphylococcus aureus Reduces Its Resistance to Antibiotics and Infection in a Mouse Model of Sepsis.

Staphylococcus aureus is a major opportunistic pathogen, infecting animals, and human beings. The bacterial cell wall plays a crucial role in antimicrobial resistance and its infection to host cells. Peptidoglycans (PGs) are a major component of the cell wall in S. aureus, which is heavily decorated with wall teichoic acids (WTAs) and capsular polysaccharides (CPs). The ligation of WTAs and CPs to PGs is catalyzed by LytR-CpsA-Psr (LCP) family proteins, including LcpA, LcpB, and LcpC. However, the involvement of LcpC in antimicrobial resistance of S. aureus and its infection to host cells remains unknown. By creating the LcpC-knockout strains, we showed that the deficiency in LcpC decreased the antimicrobial resistance to ß-lactams and glycopeptides and impeded the binding to various epithelial cells. These changes were accompanied by the morphological changes in bacterial cell wall. More importantly, the knockout of LcpC significantly reduced the pathogenicity of methicillin-resistant S. aureus (MRSA) in mice. Our results suggest that LcpC might be an appealing target for developing a therapeutic approach against MRSA infections.

Transcriptome analysis on responses of orchardgrass (Dactylis glomerata L.) leaves to a short term flooding.

BACKGROUND: Orchardgrass (Dactylis glomerata L.) is a popular cool-season perennial grass with a high production value, and orchardgrass seed is the fourth top-selling forage grass seed in the world. However, its yield and quality are often affected by flooding. To date, the molecular responses of orchardgrass to flooding were poorly understood. RESULTS: Here, we performed mRNA-seq to explore the transcriptomic responses of orchardgrass to a short term flooding (8 h and 24 h). There were 1454 and 565 differentially expressed genes identified in the 8 h and 24 h of flooding, respectively, compared to well control. GO functional enrichment analysis showed that oxidoreductase activity and oxidation-reduction process were highly present, suggesting that flooding induced the response to oxygen stress. Pathways enrichment analysis highlights the importance of glutathione metabolism, peroxidase, glycolysis and plant hormone signal transduction in response to flooding acclimation. Besides, the ROS clearance system is activated by significantly expressed glutathione S-transferase and genes encoding SOD and CAT (CAT1 and CDS2). The significant positive correlation between RNA sequencing data and a qPCR analysis indicated that the identified genes were credible. CONCLUSION: In the process of orchardgrass response to flooding stress, multiple differential genes and biological processes have participated in its acclimation to flooding, especially the biological processes involved in the removal of ROS. These results provide a basis for further research on the adaptation mechanism of orchardgrass to flood tolerance.

Transcriptome profiling of two Dactylis glomerata L. cultivars with different tolerance in response to submergence stress.

Submergence is one of the environmental stresses that limit plant growth and development. Dactylis glomerata L. is an important cool-season forage grass globally. To investigate the genes related to submergence response and the molecular mechanism associated with submergence tolerance, the transcriptome of D. glomerata in response to waterlogging treatment was analyzed. RNA-sequencing was performed in two D. glomerata cultivars, submergence tolerant 'Dianbei' and submergence sensitive 'Anba'. A total of 50,045 unique genes matched the known proteins in the NCBI nr database by BLAST searches and 60.8% (30,418) of these genes were annotated with GO terms. Among these, 1395 genes only differentially expressed in 'Dianbei' and 18 genes shown different expression all the time were detected between the submergence tolerant 'Dianbei' and sensitive 'Anba'. Gene ontology (GO) and KEGG pathway enrichment analyses demonstrated that the DEGs were mainly implicated in oxidation-reduction system, nucleic acid binding transcription factor activity, and glycerol kinase activity. The D. glomerata assembled transcriptome provided substantial molecular resource for further genomic analysis of forage grasses in response to submergence stress. The significant difference in expression of specific unigenes may account for waterlogging tolerance or acclimation in the two different D. glomerata cultivars. This study provided new insights into the molecular basis of submergence tolerance in D. glomerata.

Functional Analyses of Cassette Chromosome Recombinase C2 (CcrC2) and Its Use in Eliminating Methicillin Resistance by Combining CRISPR-Cas9.

Worldwide occurrence of methicillin-resistant Staphylococcus aureus (MRSA) poses enormous challenges for both communities and health care settings. Cassette chromosome recombinases (Ccr) specifically perform excision and acquisition of a staphylococcal cassette chromosome mec (SCC mec) in staphylococci and are responsible for the spread of methicillin resistance. This study explored the roles of CcrC2, a recently discovered Ccr, in the horizontal transfer of SCC mec and developed a potential means to control the spread of methicillin resistance. Knockout of CcrC2 completely aborted the excision of SCC mec, while overexpression of CcrC2 partially removed the SCC mec from the genome and transformed methicillin-resistant Staphylococcus aureus (MRSA) into methicillin-susceptible Staphylococcus aureus (MSSA). Moreover, two nucleotide residues (G5C6) in the direct repeat sequence within an att site were found to be critical for excision and acquisition efficiencies. To block the horizontal transfer of methicillin resistance, a SCC mec killer system was developed by combining the CcrC2-mediated SCC mec excision and the mecA-targeting CRISPR-Cas9 machinery. The SCC mec killer transformed MRSA to MSSA and disrupted the mecA-carrying SCC mec intermediate, thereby eliminating methicillin resistance determinant mecA gene inside a MRSA cell and blocking the horizontal transfer of SCC mec. The SCC mec killer was versatile for efficiently removing multiple types of SCC mec elements. It is envisioned that this approach could offer a new means to control the spread of methicillin resistance.

Juvenile myoclonic epilepsy and sleep.

Juvenile myoclonic epilepsy (JME) is a sleep-related epilepsy syndrome, and only a few studies have addressed the relationship between JME and sleep disorders. In this review, the sleep characteristics of patients with JME were summarized based on the features of circadian rhythm, the possible cause of the early morning seizures, the common subjective and objective sleep disorders, the alterations in sleep architecture, and the effect of sleep deprivation and sodium valproate (VPA). The aims of this study were to summarize the interaction between JME and sleep, to reveal JME sleep characteristics, to encourage clinicians to focus on JME and sleep, to heighten the positive diagnosis rate, to guide the treatment, to improve the prognosis, and to enhance the daily life quality of patients with JME. At the same time, this study aimed to present existing controversies, in order to necessitate further studies.

Global acquisition of genetic material from different bacteria into the staphylococcal cassette chromosome elements of a Staphylococcus epidermidis isolate.

Staphylococcus epidermidis has been suggested as a main reservoir of methicillin resistance and virulence genes facilitating the evolution of Staphylococcus aureus as a successful pathogen. However, it remains a mystery where and how S. epidermidis obtains these numerous genes to serve as the reservoir. In this study, methicillin-resistant S. epidermidis isolate NW32 from a mastitic milk sample was sequenced and its staphylococcal cassette chromosome (SCC) elements were characterised. The SCC composite island covered 3.5% of the genome and consisted of three intact SCC elements carrying resistance genes against ß-lactam antibiotics, several heavy metals and polyamines as well as genes for utilisation of sorbitol as a carbon source. Analysis of the postulated evolutionary route suggested that the three SCC elements were assembled from genetic material from various bacterial species (staphylococci, streptococci, salinicocci and Lysinibacillus) from three habitats (human, soil and cow) in different countries (Asia, North America, South America and Europe). We propose that the hsdS restriction-modification profile and the lack of CRISPR (clustered regularly interspaced short palindromic repeat) sequences in this bacterium may facilitate the genetic exchange of SCC elements among different staphylococcal species.