Strategy of Developing Oral Vaccine Candidates Against Co-infection of Porcine Diarrhea Viruses Based on a Lactobacillus Delivery System.

The number of co-infections with multiple porcine diarrhea viruses has increased in recent years. Inducing mucosal immunity through oral immunization is an effective approach for controlling these pathogens. To generate a multi-pathogen vaccine against viral co-infection, we employed the Lactobacillus vector platform, which was previously used to generate potent candidate vaccines against various diseases. Two strategies were used to test the protective efficiency of recombinant Lactobacillus against multiple diarrhea viruses. First, we used a mixture of recombinant Lactobacillus separately expressing antigens of transmissible gastroenteritis virus (TGEV), porcine epidemic diarrhea virus (PEDV), and porcine rotavirus (PoRV). Next, we used a recombinant Lactobacillus expressing an antigen fusion protein of the above viruses. Twenty-four newborn piglets were divided into three groups and orally immunized with a mixture of recombinant Lactobacillus, recombinant Lactobacillus expressing the antigen fusion protein, or sterile phosphate-buffered saline daily for seven consecutive days after birth. After immunization, the piglets were randomly selected from each group for oral administration of PEDV, and these piglets were then cohabited with piglets without PEDV infection for 7 days. The protective effect against PEDV was evaluated based on clinical symptoms, viral shedding, and intestinal pathological damage. Piglets immunized with recombinant Lactobacillus showed specific mucosal and humoral immune responses to the three viruses and were protected against severe diarrhea and intestinal pathology. Our results highlight the potential of an oral multi-pathogen vaccine based on Lactobacillus to prevent transmission and limit the severity of viral co-infection.

Delivery of antigen to porcine dendritic cells by fusing antigen with porcine dendritic cells targeting peptide.

Dendritic cells (DCs) are professional antigen-presenting cells that can recognize, capture, and process antigens. Fusing molecules targeting DCs with antigens can effectively improve the efficiency with which antigens are recognized and captured by DCs. This targeting strategy can be used for vaccine development to effectively improve the efficiency of antigen recognition and capture by DCs. The targeting sequence of porcine cytotoxic T-lymphocyte associated protein 4 (CTLA4), which binds porcine DCs, was identified in this study. Recombinant Lactobacillus reuteri (L. reuteri) expressing CTLA4-6aa (LYPPPY) and CTLA4-87aa fused to the porcine epidemic diarrhea virus (PEDV) protective antigen core neutralizing epitope (COE) were used to evaluate the ability of the two targeting motifs to bind the B7 molecule on DCs. Our results demonstrate that CTLA4-6aa could bind porcine DCs, and recombinant Lactobacillus expressing the CTLA4-6aa captured by porcine DCs was more efficient than those expressing CTLA4-87aa. In addition, the expression of DC markers, toll-like receptors, and cytokines was significantly higher in the 6aa-COE/L. reuteri-stimulated porcine DCs compared to DCs treated with 87aa-COE/L. reuteri (p<0.01) and recombinant Lactobacillus expressing CTLA4-6aa enhanced the ability of porcine DCs to activate T-cell proliferation. Our analysis of the protein structure revealed that CTLA4-87aa contains intramolecular hydrogen bonds, which may have weakened the intermolecular force between the residues on porcine CTLA4 and that on B7. In conclusion, recombinant Lactobacillus expressing CTLA4-6aa were more efficiently captured by porcine DCs and had a stronger ability to promote DC maturation and enhance T-cell proliferation. The LYPPPY motif is the optimal sequence for binding to porcine DCs. Piglets immunized with recombinant Lactobacillus showed that recombinant Lactobacillus expressing CTLA4-6aa induced significant levels of anti-PEDV-specific IgG and IgA antibody responses. Our study may promote research on DC-targeting strategies to enhance the effectiveness of porcine vaccines.

Detecting QTL and Candidate Genes for Plant Height in Soybean via Linkage Analysis and GWAS.

Soybean is an important global crop for edible protein and oil, and plant height is a main breeding goal which is closely related to its plant shape and yield. In this research, a high-density genetic linkage map was constructed by 1996 SNP-bin markers on the basis of a recombinant inbred line population derived from Dongnong L13 × Henong 60. A total of 33 QTL related to plant height were identified, of which five were repeatedly detected in multiple environments. In addition, a 455-germplasm population with 63,306 SNP markers was used for multi-locus association analysis. A total of 62 plant height QTN were detected, of which 26 were detected repeatedly under multiple methods. Two candidate genes, Glyma.02G133000 and Glyma.05G240600, involving in plant height were predicted by pathway analysis in the regions identified by multiple environments and backgrounds, and validated by qRT-PCR. These results enriched the soybean plant height regulatory network and contributed to molecular selection-assisted breeding.

Recovery from prolonged disorders of consciousness: A dual-center prospective cohort study in China.

BACKGROUND: Recent innovations in intensive care have improved the prognosis of patients with severe brain injuries and brought more patients with disorders of consciousness (DoC). Data are lacking regarding the long-term outcomes of those patients in China. It is necessary to study the long-term outcomes of patients with prolonged DoC in light of many factors likely to influence crucial decisions about their care and their life. AIM: To present the preliminary results of a DoC cohort. METHODS: This was a two-center prospective cohort study of inpatients with vegetative state (VS)/unresponsive wakefulness syndrome (UWS). The study outcomes were the recovery from VS/UWS to minimally conscious state (MCS) and the long-term status of patients with prolonged DoC considered in VS/UWS or MCS for up to 6 years. The patients were evaluated using the Glasgow coma scale, coma recovery scale-revised, and Glasgow outcome scale. The endpoint of follow-up was recovery of full consciousness or death. The changes in the primary clinical outcome improvement in clinical diagnosis were evaluated at 12 mo compared with baseline. RESULTS: The study population included 93 patients (62 VS/UWS and 31 MCS). The post-injury interval range was 28-634 d. Median follow-up was 20 mo (interquartile range, 12-37 mo). At the endpoint, 33 transitioned to an emergence from MCS or full consciousness, eight had a locked-in syndrome, and there were 35 patients remaining in a VS/UWS and 11 in an MCS. Seven (including one locked-in syndrome) patients (7.5%) died within 12 mo of injury. Compared with the unresponsive group (n = 52) at 12 mo, the responsive group (n = 41) had a higher proportion of males (87.8% vs 63.5%, P = 0.008), shorter time from injury (median, 40.0 d vs 65.5 d, P = 0.006), higher frequency of vascular etiology (68.3% vs 38.5%, P = 0.007), higher Glasgow coma scale score at admission (median, 9 vs 6, P < 0.001), higher coma recovery scale-revised score at admission (median, 9 vs 2.5, P < 0.001), at 1 mo (median, 14 vs 5, P < 0.001), and at 3 mo (median, 20 vs 6, P < 0.001), lower frequency of VS/UWS (36.6% vs 90.0%, P < 0.001), and more favorable Glasgow outcome scale outcome (P < 0.001). CONCLUSION: Patients with severe DoC, despite having strong predictors of poor prognosis, might recover consciousness after a prolonged time of rehabilitation. An accurate initial diagnosis of patients with DoC is critical for predicting outcome and a long-term regular follow-up is also important.

Coherence in P300 as a predictor for the recovery from disorders of consciousness.

Accurate and reliable prognostic judgments for disorders of consciousness (DOCs) can provide useful information for clinicians in the establishment of appropriate care plan, and may affect end-of-life decisions of caregivers. But until now no certain or standardized prognostic indicator has been constructed to predict the probability of awareness recovery. This study aims to assess higher-order cortical information processing in DOCs by event-related potential (ERP) and determine the value of P300 to predict the long-term prognosis. Two locked-in (LIS) patients and eighteen DOCs were evaluated with a hierarchical cognitive assessment by ERP. We used subject's own name (SON) as a deviant stimulus, 1000Hz tone and subject's derived name (SDN) as a standard stimulus in two paradigms respectively. P300 elicited by SON was used to assess the information processing. The patients' clinical outcomes were followed up at 2, 6 and 12 months after ERP recordings. The results showed that a P300 component was observed in both paradigms in two LIS patients. All of MCS and four out of nine UWS/VS showed an intact P300 in either paradigm. All of the five patients with P300 in both paradigms were finally awake after 12 months, while none of the eight patients without P300 regained consciousness. A highly significant relationship between P300 and subsequent recovery was found. The results provide evidence that P300 in the hierarchical bedside neurophysiologic oddball procedure can accurately characterize the level of cognitive preservation, and may serve as an alternative tool to predict the likelihood of recovery of DOCs.