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Unmanned Aerial Vehicles (UAVs) combined with target detection algorithms can enhance the detection of road cracks. In response to the challenges presented by complex crack shapes and textures, small sizes, and highly integrated backgrounds, this article developed a UAV-based road crack target detection algorithm using road crack you only look once (RCYOLO). RCYOLO was composed of a C2f_DySnakeConv (C2f_DSConv) module located in the ninth layer and a simulated attention mechanism (SimAM) module situated above the Spatial Pyramid Pooling--Fast (SPPF), along with a dyhead attention detection head that integrated three types of attention mechanisms. Initially, the C2f_DSConv was proposed to effectively extract tubular features of cracks. Subsequently, the SimAM addressed the issue of target-background fusion, enhancing feature recognition of the targets while suppressing background interference. Finally, the dyhead strategy incorporated three types of attention mechanisms, effectively resolving the issue of small target omissions. Our results showed that on the custom UAV dataset road crack image, which included close-range and long-range images, RCYOLO outperformed the baseline network YOLOv8 by 5.9% in mAP@0.5, 6.5% in recall, and 9.8% in precision. On the public dataset Detection of Objects in Aerial Images, mAP@0.5 also exceeded YOLOv8 by 5.8%, indicating that RCYOLO performed well in other remote sensing image tasks, making this target detection algorithm more suitable for high-altitude photography of crack targets than other mainstream algorithms.

Objective To evaluate the effect of modified nursing shift schedule based on SBAR communication model in perioperative management of patients with early gastric cancer. Methods Totally 100 patients who received endoscopic submucosal dissection (ESD) for early gastric cancer between January and May 2021 were included as the control group, another 100 patients who received ESD for early gastric cancer between August and December 2021 were included as the observation group. The routine nursing shift management was adopted in the control group, and the modified nursing shift schedule based on SBAR communication model was carried out in the observation group. The quality of nursing shift, patient’s satisfaction and nurse’s critical thinking ability were measured and compared between two groups. Results After implementation of modified nursing shift schedule, the quality of nursing shift was improved, and nurses achieved a higher score of The California Critical Thinking Disposition Inventory (CCTDI) compared with that before implementation of modified nursing shift schedule (259. 95±38. 60 vs. 228. 68±36. 44, P＜0. 05). The score of satisfaction assessment in the observation group was higher than that in the control group (21. 47±2. 96 vs. 7. 39±2. 90, P＜0. 05). Conclusion The modified nursing shift schedule based on SBAR communication model is an effective tool of improve quality of nursing shift, patient’s satisfaction and nurse’s critical thinking ability. (目的 探讨基于SBAR模式改良护理交班表在早期胃癌患者围手术期管理中的应用效果。方法 选取2021年1月—5月医院消化内科收治的100例行内镜黏膜下剥离术(ESD)治疗的早期胃癌患者作为对照组, 2021年8月—12月收治的100例行ESD治疗的早期胃癌患者作为观察组。对照组使用传统交班模式, 观察组采用基于SBAR模式改良的护理交班表完成护理交班管理。评估两组的交班质量、患者对护士的满意度及护士的评判性思维能力。结果 实施改良护理交班表后, 护士交班质量全面性明显提高, 护士的评判性思维得分(259. 95±38. 60)分也高于实施前的(228. 68±36. 44)分, 差异有统计学意义(P＜0. 05)。观察组患者对护士满意度评分(21. 47±2. 96)分, 高于对照组的(17. 39±2. 90)分, 差异有统计学意义(P＜0. 05)。结论 采用基于SBAR模式改良的护理交班表可以提高交班质量以及患者对护士的满意度, 还能提高护士的评判性思维能力。)

Microbiomes provide multiple life-support functions for plants, including nutrient acquisition and tolerance to abiotic and biotic stresses. Considering the importance of C4 cereal and biofuel crops for food security under climate change conditions, more attention has been given recently to C4 plant microbiome assembly and functions. Here, we review the current status of C4 cereal and biofuel crop microbiome research with a focus on beneficial microbial traits for crop growth and health. We highlight the importance of environmental factors and plant genetics in C4 crop microbiome assembly and pinpoint current knowledge gaps. Finally, we discuss the potential of foxtail millet as a C4 model species and outline future perspectives of C4 plant microbiome research., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Objectives: While the Partnership Long-Term Care Insurance (PLTC) Program was designed to mitigate the low uptake of long-term care insurance and reduce Medicaid's financial burden, research has predominantly focused on its direct impacts, leaving the effects on informal caregiving unexplored. This study aimed to investigate how the program alters the dynamics of family-provided care, leveraging nationally representative data to unveil the broader consequences on informal caregiving arrangements among older individuals., Methods: Data for this study were sourced from the U.S. Health and Retirement Study (1992-2018) and linked with the timing of the PLTC program implementation across states. The analysis compared individuals exposed to the program with those who were not, employing two-way-fixed-effects and dynamic models to assess its impact on long-term care insurance coverage and reliance on informal caregiving., Results: The program positively affected long-term care insurance coverage, increasing insurance uptake among older individuals in the long run. Conversely, a significant negative effect was observed on the receipt of assistance from any helper, indicating a reduced reliance on informal care. This reduction extended specifically to family helpers and children's assistance with activities of daily living. The analysis suggests that the program effectively reduced the necessity for informal caregiving across several domains., Discussion: These findings highlight the program's potential to reshape caregiving dynamics, suggesting the need for policies that balance promoting private insurance uptake with the implications for family caregiving roles. Policymakers should consider both the economic benefits and the social shifts induced by such programs in the long-term care landscape., (© The Author(s) 2024. Published by Oxford University Press on behalf of The Gerontological Society of America.)

Pollen carries extensive genetic information, which may provide clues regarding the kinship of Malus, whose genetic relationships are complex. In this study, the phenotypic variation of pollen from 107 Malus taxa was investigated using combined methods of intraspecific/interspecific uniformity testing, cluster analysis, and Pearson correlation analysis. The family aggregation distributions in Malus sections, species, and cultivars were analyzed to infer their pedigree relationships. The results showed that (1) compared with pollen size and morphology, aberrant pollen rate and ornamentation were highly interspecifically differentiated, but ornamentation was also intraspecifically unstable, especially perforation densities (c.v.¯ > 15%). (2) The pollen alteration direction from the original to the evolutionary population of Malus was large to small, with elliptic to rectangular morphologies, large and compact to small and sparse ridges, and low to high perforation densities. However, there was no significant change in pollen size. (3) The 107 studied taxa were divided into four groups. Malus species were relatively clustered in the same section, while homologous cultivars showed evidence of family aggregation distribution characteristics (92.60% of cultivars were clustered with their parents). (4) M . baccata and M . pumilar var. neidzwetzkyana were high-frequency parents, participating in 38.7% and 20.7% of cross-breeding, respectively. Overall, this study provides a reference for identifying Malus ' pedigree relationship.

The application of nanoscale scaffolds has become a promising strategy in vaccine design, with protein-based nanoparticles offering desirable avenues for the biocompatible and efficient delivery of antigens. Here, we presented a novel endogenous capsid-forming protein, activated-regulated cytoskeleton-associated protein (ARC), which could be engineered through the plug-and-play strategy (SpyCatcher3/SpyTag3) for multivalent display of antigens. Combined with the self-assembly capacity and flexible modularity of ARC, ARC-based vaccines elicited robust immune responses against Mpox or SARS-CoV-2, comparable to those induced by ferritin-based vaccines. Additionally, ARC-based nanoparticles functioned as immunostimulants, efficiently stimulating dendritic cells and facilitating germinal center responses. Even without adjuvants, ARC-based vaccines generated protective immune responses in a lethal challenge model. Hence, this study showed the feasibility of ARC as a novel protein-based nanocarrier for multivalent surface display of pathogenic antigens and demonstrated the potential of exploiting recombinant mammalian retrovirus-like protein as a delivery vehicle for bioactive molecules., (© 2024. The Author(s).)

The Medicaid Aging Waiver program (MAW) subsidizes the cost of long-term care (LTC) at home or in communities to satisfy older people's increasing desire to age in place. The MAW program might be health improving for older people by allowing them to age at home. However, less quality and quantity of home-based care comparing to nursing home care could offset some of the potential benefits. I use policy expenditure across states over time linked with detailed health information from the Health and Retirement Study (HRS) to identify the associated effects of MAWs on health outcomes of older adults who are at risk of needing LTC and who are resources constrained to be potentially eligible for Medicaid. Overall, the findings suggest that the MAW program is beneficial to health: a $1,000 increase in MAW spending for each older person results is associated with a 1.4 percent improvement in self-reported health status, a 1.5 percent reduction in functional mobility limitations, a 1.6 percent decrease in Instrumental Activities of Daily Living (IADL) limitations, and a 1.7 percent improvement in negative psychological feelings. For older people who are most likely not eligible for MAWs, such as those who are wealthy or in good health and do not require LTC, these health-improving effects have not been observed., (© 2024. The Author(s).)

Human brucellosis caused by Brucella is a widespread zoonosis that is prevalent in many countries globally. The high homology between members of the Brucella genus and Ochrobactrum spp. often complicates the determination of disease etiology in patients. The efficient and reliable identification and distinction of Brucella are of primary interest for both medical surveillance and outbreak purposes. A large amount of genomic data for the Brucella genus was analyzed to uncover novel probes containing single-nucleotide polymorphisms (SNPs). GAMOSCE v1.0 software was developed based on the above novel eProbes. In conjunction with clinical requirements, an RPA-Cas12a detection method was developed for the on-site determination of B. abortus and B. melitensis by fluorescence and lateral flow dipsticks (LFDs). We demonstrated the potential of these probes for rapid and accurate detection of the Brucella genus and five significant Brucella species in silico using GAMOSCE. GAMOSCE was validated on different Brucella datasets and correctly identified all Brucella strains, demonstrating a strong discrimination ability. The RPA-Cas12a detection method showed good performance in detection in clinical blood samples and veterinary isolates. We provide both in silico and on-site methods that are convenient and reliable for use in local hospitals and public health programs for the detection of brucellosis.

This paper examines the long-standing and severe public health crisis, the opioid epidemic in the United States, which has been worsening since the mid-1990s. In contrast to previous research, it investigates the broader impacts of this epidemic, particularly on family members and healthcare systems. Using a comprehensive dataset spanning from 1998 to 2010, the study analyzes opioid use at the three-digit ZIP code level, utilizing data from the Drug Enforcement Agency (DEA) and individual-level data from the Health and Retirement Study (HRS) in a two-way fixed effect model. The findings reveal significant negative effects on family caregivers, notably adult children, due to the opioid epidemic. Additionally, opioid exposure is associated with increased healthcare utilization, including home health care and hospital use. This research contributes to a deeper understanding of the multifaceted consequences of the opioid epidemic., Competing Interests: Declaration of Competing Interest The authors declare that they have no relevant or material financial interests that relate to the research described in the paper., (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)

Background: Lassa fever is a hemorrhagic disease caused by Lassa virus (LASV), which has been classified by the World Health Organization as one of the top infectious diseases requiring prioritized research. Previous studies have provided insights into the classification and geographic characteristics of LASV lineages. However, the factor of the distribution and evolution characteristics and phylodynamics of the virus was still limited., Methods: To enhance comprehensive understanding of LASV, we employed phylogenetic analysis, reassortment and recombination detection, and variation evaluation utilizing publicly available viral genome sequences., Results: The results showed the estimated the root of time of the most recent common ancestor (TMRCA) for large (L) segment was approximately 634 (95% HPD: [385879]), whereas the TMRCA for small (S) segment was around 1224 (95% HPD: [10301401]). LASV primarily spread from east to west in West Africa through two routes, and in route 2, the virus independently spread to surrounding countries through Liberia, resulting in a wider spread of LASV. From 1969 to 2018, the effective population size experienced two significant increased, indicating the enhanced genetic diversity of LASV. We also found the evolution rate of L segment was faster than S segment, further results showed zinc-binding protein had the fastest evolution rate. Reassortment events were detected in multiple lineages including sub-lineage IIg, while recombination events were observed within lineage V. Significant amino acid changes in the glycoprotein precursor of LASV were identified, demonstrating sequence diversity among lineages in LASV., Conclusion: This study comprehensively elucidated the transmission and evolution of LASV in West Africa, providing detailed insights into reassortment events, recombination events, and amino acid variations., (© 2024. The Author(s).)

Brevibacillus laterosporus ( B. laterosporus ) is widely distributed in nature and demonstrates significant potential for applications in biological control, environmental protection, agricultural production, and clinical medicine. This review provides a comprehensive overview of the applications of B. laterosporus in crop cultivation and animal feeding, as well as an examination of the antimicrobial peptides produced by B. laterosporus and their antibacterial mechanisms. B. laterosporus enhances crop cultivation by secreting hydrolases to improve nutrient absorption capabilities, synthesizing hormones to promote crop growth, and producing proteins to inhibit the reproduction of harmful organisms. B. laterosporus has been used to improve animal production by regulating the structure of the intestinal microbiota and inhibiting the growth of pathogenic bacteria through the secretion of various antimicrobial peptides. The bactericidal activity of Brevilaterins secreted by B. laterosporus is attributed to their ability to bind to lipopolysaccharide/lipid II molecules on the cell membrane, thereby altering permeability. Brevilaterins also inhibit bacterial reproduction by affecting relevant gene pathways in the cell membranes of pathogenic bacteria. These pathways include ATP synthesis, peptidoglycan biosynthesis, membrane transport, and cellular metabolism. In conclusion, B. laterosporus exhibits substantial potential as a probiotic activity in crop and animal production. However, applications of B. laterosporus in animal production could be improved, necessitating further research to elucidate the underlying probiotic mechanisms.

The highly diverse microbial ecosystem of the human body colonizes the gastrointestinal tract has a profound impact on the host's immune, metabolic, endocrine, and other physiological processes, which are all interconnected. Specifically, gut microbiota has been found to play a crucial role in facilitating the adaptation and initiation of immune regulatory response through the gastrointestinal tract affecting the other distal mucosal sites such as lungs. A tightly regulated lung-gut axis during respiratory ailments may influence the various molecular patterns that instructs priming the disease severity to dysregulate the normal function. This review provides a comprehensive summary of current research on gut microbiota dysbiosis in respiratory diseases including asthma, pneumonia, bronchopneumonia, COPD during infections and cancer. A complex-interaction among gut microbiome, associated metabolites, cytokines, and chemokines regulates the protective immune response activating the mucosal humoral and cellular response. This potential mechanism bridges the regulation patterns through the gut-lung axis. This paper aims to advance the understanding of the crosstalk of gut-lung microbiome during infection, could lead to strategize to modulate the gut microbiome as a treatment plan to improve bad prognosis in various respiratory diseases., (© 2024. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

We investigate the effects of a significant health insurance expansion in rural China known as the New Cooperative Medical Scheme (NCMS). Our analysis is based on a nationwide dataset spanning from 2004 to 2011. We find that the NCMS effectively increases healthcare utilization, particularly inpatient admissions, and reduces the incidence for infectious diseases. In addition to the increased healthcare utilization, the reduction in the incidence for infectious diseases can be attributed to improved health knowledge and health behavior, both of which are associated with the expansion of insurance coverage. Our findings affirm the importance of insurance coverage in safeguarding low-income individuals from the adverse health consequences linked to infectious diseases., Competing Interests: The authors declare that they have no relevant or material financial interests that relate to the research described in the paper., (© 2024 The Authors.)

Brucellosis is a major threat to public health and animal husbandry. Several in vivo vertebrate models, such as mice, guinea pigs, and nonhuman primates, have been used to study Brucella pathogenesis, bacteria-host interactions, and vaccine efficacy. However, these models have limitations whereas the invertebrate Galleria mellonella model is a cost-effective and ethical alternative. The aim of the present study was to examine the invertebrate G. mellonella as an in vivo infection model for Brucella . Infection assays were employed to validate the fitness of the larval model for Brucella infection and virulence evaluation. The protective efficacy of immune sera was evaluated by pre-incubated with a lethal dose of bacteria before infection. The consistency between the mouse model and the larval model was confirmed by assessing the protective efficacy of two Brucella vaccine strains. The results show that G. mellonella could be infected by Brucella strains, in a dose- and temperature-dependent way. Moreover, this larval model can effectively evaluate the virulence of Brucella strains in a manner consistent with that of mammalian infection models. Importantly, this model can assess the protective efficacy of vaccine immune sera within a day. Further investigation implied that haemolymph played a crucial role in the protective efficacy of immune sera. In conclusion, G. mellonella could serve as a quick, efficient, and reliable model for evaluating the virulence of Brucella strains and efficacy of immune sera in an ethical manner.

Nipah virus (NiV) and Hendra virus (HeV) are newly emerging dangerous zoonotic pathogens of the Henipavirus genus of the Paramyxoviridae family. NiV and HeV (HNVs) which are transmitted by bats cause acute respiratory disease and fatal encephalitis in humans. To date, as there is a lack of antiviral drugs or effective antiviral therapies, the development of vaccines against those two viruses is of primary importance, and the immunogen design is crucial to the success of vaccines. In this study, the full-length protein (G), the ectodomain (Ge) and the head domain (Gs) of NiV attachment glycoprotein were delivered by the replication-defective type 5 adenovirus vector (Ad5) respectively, and the recombinant Ad5-NiV vaccine candidates (Ad5-NiVG, Ad5-NiVGe and Ad5-NiVGs) were constructed and their immunogenicity were evaluated in mice. The results showed that all the vaccine candidates stimulated specific humoral and cellular immune responses efficiently and rapidly against both NiV and HeV, and the Ad5-NiVGe elicited the strongest immune responses after a single-dose immunization. Furthermore, the potent conserved T-cell epitope DTLYFPAVGFL shared by NiV and HeV was identified in the study, which may provide valid information on the mechanism of HNVs-specific cellular immunity. In summary, this study demonstrates that the Ad5-NiVGe could be a potent vaccine candidate against HNVs by inducing robust humoral and cellular immune responses., Competing Interests: Beijing Institute of Biotechnology has filed several patent applications for Ad5-NiV vaccines., (Copyright © 2023 Huang, Li, Li, Wang, Yang, Wang, Yin, Zai, Zhang and Xu.)

Aluminum (alum) adjuvant is the most extensively used protein subunit vaccine adjuvant, and its effectiveness and safety have been widely recognized. The surface charge of the antigen determines its electrostatic adsorption to alum adjuvant, which directly affects the immune efficacy of the protein vaccine. In our study, we precisely modified its surface charge by inserting charged amino acids into the flexible region of the SARS-CoV-2 receptor-binding domain (RBD), achieving electrostatic adsorption and a site-specific anchor between the immunogen and alum adjuvant. This innovative strategy extended the bioavailability of the RBD and directionally displayed the neutralizing epitopes, thereby significantly enhancing humoral and cellular immunity. Furthermore, the required dose of antigen and alum adjuvant was greatly reduced, which improved the safety and accessibility of the protein subunit vaccine. On this basis, the wide applicability of this novel strategy to a series of representative pathogen antigens such as SARS-RBD, MERS-RBD, Mpox-M1, MenB-fHbp, and Tularemia-Tul4 was further confirmed. Charge modification of antigens provides a straightforward approach for antigenicity optimization of alum-adjuvanted vaccines, which has great potential to be adopted as a global defense against infectious diseases., Competing Interests: C.W., J.X., X. Zai, C.Z., Z.Z., F.Z., Y.Z., J.Z., Y. Yin, Y. Yang, R.L., and Y.L. are listed as inventors on a pending patent application for charge-modification vaccines., (© 2023 The Authors.)

Tularemia is a highly contagious disease caused by infection with Francisella tularensis ( Ft ), a pathogenic intracellular gram-negative bacterium that infects a wide range of animals and causes severe disease and death in people, making it a public health concern. Vaccines are the most effective way to prevent tularemia. However, there are no Food and Drug Administration (FDA)-approved Ft vaccines thus far due to safety concerns. Herein, three membrane proteins of Ft , Tul4, OmpA, and FopA, and a molecular chaperone, DnaK, were identified as potential protective antigens using a multifactor protective antigen platform. Moreover, the recombinant DnaK, FopA, and Tul4 protein vaccines elicited a high level of IgG antibodies but did not protect against challenge. In contrast, protective immunity was elicited by a replication-defective human type 5 adenovirus (Ad5) encoding the Tul4, OmpA, FopA, and DnaK proteins (Ad5-Tul4, Ad5-OmpA, Ad5-FopA, and Ad5-DnaK) after a single immunization, and all Ad5-based vaccines stimulated a Th1-biased immune response. Moreover, intramuscular and intranasal vaccination with Ad5-Tul4 using the prime-boost strategy effectively eliminated Ft lung, spleen and liver colonization and provided nearly 80% protection against intranasal challenge with the Ft live vaccine strain (LVS). Only intramuscular, not intranasal vaccination, with Ad5-Tul4 protected mice from intraperitoneal challenge. This study provides a comprehensive comparison of protective immunity against Ft provided by subunit or adenovirus-vectored vaccines and suggests that mucosal vaccination with Ad5-Tul4 may yield desirable protective efficacy against mucosal infection, while intramuscular vaccination offers greater overall protection against intraperitoneal tularemia., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Zhao, Zhai, Zai, Mao, Hu, Wei, Li, Li, Liu, Xu, Yu and Chen.)

The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) had become a global concern because of its unexpectedly high pathogenicity and transmissibility. SARS-CoV-2 variants that reduce the immune protection elicited from previous vaccination or natural infection raise challenges in controlling the spread of the pandemic. The development of universal vaccines against these variants seems to be a practical solution to alleviate the physical and economic effects caused by this disease, but it is hard to achieve. In this review, we describe the high mutation rate of RNA viruses and dynamic molecular structures of SARS-CoV-2 variants in several major neutralizing epitopes, trying to answer the question of why universal vaccines are difficult to design. Understanding the biological basis of immune evasion is crucial for combating these obstacles. We then summarize several advancements worthy of further study, including heterologous prime-boost regimens, construction of chimeric immunogens, design of protein nanoparticle antigens, and utilization of conserved neutralizing epitopes. The fact that some immunogens can induce cross-reactive immune responses against heterologous coronaviruses provides hints for universal vaccine development. We hope this review can provide inspiration to current universal vaccine studies., (© 2022. The Author(s).)

Turkey herpesvirus (HVT) is widely used as an effective recombinant vaccine vector for expressing protective antigens of multiple avian pathogens from different loci of the HVT genome. These include the HVT029/031 (UL22-23) locus for the insertion of IBDV VP2 and the recently identified HVT005/006 locus as a novel site for expressing heterologous proteins. In order to compare the efficacy of recombinant vaccines with the HA gene at different sites, the growth curves and the HA expression levels of HVT-005/006-hCMV-HA, HVT-005/006-MLV-HA, and HVT-029/031-MLV-HA were first examined in vitro. While the growth kinetics of three recombinant viruses were not significantly different from those of parent HVT, higher expression of the HA gene was achieved from the HVT005/006 site than that from the HVT029/031 site. The efficacy of the three recombinant viruses against avian influenza H9N2 virus was also evaluated using one-day-old SPF chickens. Chickens immunized with HVT-005/006-MLV-HA or HVT-005/006-hCMV-HA displayed reduced virus shedding compared to HVT-029/031-MLV-HA vaccinated chickens. Moreover, the overall hemagglutination inhibition (HI) antibody titers of HVT-005/006-HA-vaccinated chickens were higher than that of HVT-029/031-HA-vaccinated chickens. However, HVT-005/006-MLV-HA and HVT-005/006-hCMV-HA did not result in a significant difference in the level of HA expression in vitro and provided the same protective efficacy (100%) at 5 days after challenge. In the current study, the results suggested that recombinant HVT005/006 vaccines caused better expression of HA than recombinant HVT029/031 vaccine, and that HVT-005/006-MLV-HA or HVT-005/006-hCMV-HA could be a candidate vaccine for the protection of chickens against H9N2 influenza.

Adjuvants are crucial components of vaccines that can enhance and modulate antigen-specific immune responses. Herein, we reported for the first time that human metallothionein-3 (MT3), a low molecular weight cysteine-rich metal-binding protein, was a novel promising adjuvant candidate that could help protein antigens to induce rapid, effective, and durable antigen-specific immune responses. In the present study, MT3 was fused to outer membrane protein 19 (Omp19) of Brucella abortus (MT3-Omp19, MO) and C fragment heavy chain (Hc) of tetanus neurotoxin (MT3-Hc, MH), respectively. The results showed that MT3 as a built-in adjuvant increased the Omp19- or Hc-specific antibody responses by 100-1000 folds in seven days after primary immunization. Compared to other commercially available adjuvants, MT3 could stimulate earlier (4 days after primary injection) and stronger (10-100 folds) antibody response with lower antigen dose, and its adjuvanticity relied on fusion to antigen. Although the mechanism was not clear yet, the fusion protein MO was observed to directly activate DCs, promote germinal center formation and improve the speed of Ig class switching. Interestingly, our subsequent study found that other members of the mammalian MT family (human MT1 or murine MT3 for examples) also had potential adjuvant effects, but their effects were lower than human MT3. Overall, this study explored a new function of human MT3 as a novel built-in adjuvant, which may have important clinical application potential in vaccine development against global pandemics., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. Beijing Institute of Biotechnology has submitted patent applications related to use of MTs as vaccine adjuvants., (Copyright © 2022 Yin, Gu, Zai, Li, Zhu, Yu, Zhang, Wang, Zhang, Lin, Xu and Chen.)

Highly divergent SARS-CoV-2 variants have continuously emerged and spread around the world, and updated vaccines and innovative vaccination strategies are urgently needed to address the global SARS-COV2 pandemic. Here, we established a series of Ad5-vectored SARS-CoV-2 variant vaccines encoding multiple spike proteins derived from the Alpha, Beta, Gamma, Epsilon, Kappa, Delta and Omicron lineages and analyzed the antibody immune responses induced by single-dose and prime-boost vaccination strategies against emerging SARS-CoV-2 variants of concern (VOCs). Single-dose vaccination with SARS-CoV-2 variant vaccines tended to elicit the optimal self-matched neutralizing effects, and Ad5-B.1.351 produced more broad-spectrum cross-neutralizing antibodies against diverse variants. In contrast, prime-boost vaccination further strengthened and broadened the neutralizing antibody responses against highly divergent SARS-CoV-2 variants. The heterologous administration of Ad5-B.1.617.2 and Ad5-B.1.429 to Ad5-WT-primed mice resulted in superior antibody responses against most VOCs. In particular, the Omicron spike could only stimulate self-matched neutralizing antibodies with infrequent cross-reactivities to other variants used in single-dose vaccination strategies; moreover, with prime-boost regimens, this vaccine elicited an optimal specific neutralizing antibody response to Omicron, and prompted cross-antibody responses against other VOCs that were very similar to those obtained with Ad5-WT booster. Overall, this study delineated the unique characteristics of antibody responses to the SARS-CoV-2 VOC spikes with the single-dose or prime-boost vaccination strategies and provided insight into the vaccine development of next SARS-CoV-2 VOCs., (© 2022. The Author(s).)

Turkey herpesvirus (HVT) has been widely used as a successful live virus vaccine against Marek's disease (MD) in chickens for more than five decades. Increasingly, HVT is also used as a highly effective recombinant vaccine vector against multiple avian pathogens. Conventional recombination, or recombineering, techniques that involve the cloning of viral genomes and, more recently, gene editing methods have been used for the generation of recombinant HVT-based vaccines. In this study, we used NHEJ-dependent CRISPR/Cas9-based approaches to insert the mCherry cassette for the screening of the HVT genome and identifying new potential sites for the insertion of foreign genes. A novel intergenic site HVT-005/006 in the unique long (UL) region of the HVT genome was identified, and mCherry was found to be stably expressed when inserted at this site. To confirm whether this site was suitable for the insertion of other exogenous genes, haemagglutinin (HA) of the H9N2 virus was inserted into this site, and a recombinant HVT-005/006-HA was rescued. The recombinant HVT-HA can grow well and express HA protein stably, which demonstrated that HVT-005/006 is a promising site for the insertion of foreign genes., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Zai, Shi, Shao, Qian, Ye, Yao, Nair and Qin.)

The SARS-CoV-2 Omicron variant shows substantial resistance to neutralization by infection- and vaccination-induced antibodies, highlighting the demands for research on the continuing discovery of broadly neutralizing antibodies (bnAbs). Here, we developed a panel of bnAbs against Omicron and other variants of concern (VOCs) elicited by vaccination of adenovirus-vectored COVID-19 vaccine (Ad5-nCoV). We also investigated the human longitudinal antibody responses following vaccination and demonstrated how the bnAbs evolved over time. A monoclonal antibody (mAb), named ZWD12, exhibited potent and broad neutralization against SARS-CoV-2 variants Alpha, Beta, Gamma, Kappa, Delta, and Omicron by blocking the spike protein binding to the angiotensin-converting enzyme 2 (ACE2) and provided complete protection in the challenged prophylactic and therapeutic K18-hACE2 transgenic mouse model. We defined the ZWD12 epitope by determining its structure in complex with the spike (S) protein via cryo-electron microscopy. This study affords the potential to develop broadly therapeutic mAb drugs and suggests that the RBD epitope bound by ZWD12 is a rational target for the design of a broad spectrum of vaccines., (© 2022. The Author(s).)

Root diameter and rooting depth lead to morphological and architectural heterogeneity of plant roots; however, little is known about their effects on root-associated microbial communities. Bacterial community assembly was explored across 156 samples from three rhizocompartments (the rhizosphere, rhizoplane, and endosphere) for different diameters (0.0-0.5 mm, 0.5-1.0 mm, 1.0-2.0 mm, and>2.0 mm) and depths (0-5 cm, 5-10 cm, 10-15 cm, and 15-20 cm) of soybean [ Glycine max (L.) Merrill] root systems. The microbial communities of all samples were analyzed using amplicon sequencing of bacterial 16S rRNA genes. The results showed that root diameter significantly affected the rhizosphere and endosphere bacterial communities, while rooting depth significantly influenced the rhizosphere and rhizoplane bacterial communities. The bacterial alpha diversity decreased with increasing root diameter in all three rhizocompartments, and the diversity increased with increasing rooting depth only in the rhizoplane. Clearly, the hierarchical enrichment process of the bacterial community showed a change from the rhizosphere to the rhizoplane to the endosphere, and the bacterial enrichment was higher in thinner or deeper roots (except for the roots at a depth of 15-20 cm). Network analysis indicated that thinner or deeper roots led to higher bacterial network complexity. The core and keystone taxa associated with the specific root diameter class and rooting depth class harbored specific adaptation or selection strategies. Root diameter and rooting depth together affected the root-associated bacterial assembly and network complexity in the root system. Linking root traits to microbiota may enhance our understanding of plant root-microbe interactions and their role in developing environmentally resilient root ecosystems., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Luo, Zai, Sun, Li, Li, Li, Wei and Chen.)

Eggplant is a popular vegetable in Asia; however, it has a short storage life and considerable economic losses have resulted from eggplant browning. Calcium has been reported to play a key role in the postharvest storage of plants. Here, we found that exogenous calcium application could delay eggplant fruit browning and maintain higher storage quality. The increased browning index (BI), relative electrolytic leakage (REL), and water loss were suppressed by calcium treatment during storage. Delayed browning with calcium treatment might result from a higher phenolic level and suppressed the activity of polyphenol oxidase (PPO). Less H 2 O 2 and O 2 - but more activated reactive oxygen species (ROS) scavenging enzymes accumulated in calcium-treated fruits than in H 2 O-treated fruits. Moreover, the nonenzymatic antioxidant, ascorbic acid (AsA), was accumulated more in calcium-treated eggplant fruits. Taken together, our data demonstrated that exogenous calcium application delayed eggplant fruit browning by regulating phenol metabolism and enhancing antioxidant systems., Competing Interests: The authors have no conflicts of interest to declare., (© 2021 The Authors. Food Science & Nutrition published by Wiley Periodicals LLC.)

The high plasticity of root morphology, physiology, and function influences root-associated microbiomes. However, the variation in root-associated microbiome diversity and structures in response to root diameter at different root depths remains poorly understood. Here, we selected black locust (Robinia pseudoacacia L.) as a model plant to investigate the selection and network interactions of rhizospheric and root endophytic bacterial microbiomes associated with roots of different diameters (1, 1-2, and > 2 mm) among root depths of 0-100 cm via the Illumina sequencing of the 16S rRNA gene. The results showed that the alpha diversity of the root-associated bacterial communities decreased with increasing root diameters among different root depths; fewer orders with higher relative abundance, especially in the endosphere, were enriched in association with coarse roots (> 2 mm) than fine roots among root depths. Furthermore, the variation in the enriched bacterial orders associated with different root diameters was explained by bulk soil properties. Higher co-occurrence network complexity and stability emerged in the rhizosphere microbiomes of fine roots than those of coarse roots, in contrast to the situation in the endosphere microbiomes. In particular, the endosphere of roots with a diameter of 1-2 mm exhibited the lowest network complexity and stability and a high proportion of keystone taxa (e.g., Cytophagia, Flavobacteriia, Sphingobacteriia, β-Proteobacteria, and γ-Proteobacteria), suggesting a keystone taxon-reliant strategy in this transitional stage. In summary, this study indicated that root diameter at different root depths differentially affects rhizospheric and endophytic bacterial communities, which implies a close relationship between the bacterial microbiome, root function, and soil properties., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC part of Springer Nature.)

Brucella spp. are Gram-negative, facultative intracellular bacteria that cause brucellosis in humans and various animals. The threat of brucellosis has increased, yet currently available live attenuated vaccines still have drawbacks. Therefore, subunit vaccines, produced using protein antigens and having the advantage of being safe, cost-effective and efficacious, are urgently needed. In this study, we used core proteome analysis and a compositive RV methodology to screen potential broad-spectrum antigens against 213 pathogenic strains of Brucella spp. with worldwide geographic distribution. Candidate proteins were scored according to six biological features: subcellular localization, antigen similarity, antigenicity, mature epitope density, virulence, and adhesion probability. In the RV analysis, a total 32 candidate antigens were picked out. Of these, three proteins were selected for assessment of immunogenicity and preliminary protection in a mouse model: outer membrane protein Omp19 (used as a positive control), type IV secretion system (T4SS) protein VirB8, and type I secretion system (T1SS) protein HlyD. These three antigens with a high degree of conservation could induce specific humoral and cellular immune responses. Omp19, VirB8 and HlyD could substantially reduce the organ bacterial load of B. abortus S19 in mice and provide varying degrees of protection. In this study, we demonstrated the effectiveness of this unique strategy for the screening of potential broad-spectrum antigens against Brucella. Further evaluation is needed to identify the levels of protection conferred by the vaccine antigens against wild-type pathogenic Brucella species challenge.

Polysaccharides isolated from natural plants may represent a novel source of vaccine adjuvants. In this research, we focused on a natural plant polysaccharide, PCP-I, which is derived from Poria cocos , a Chinese traditional herbal medicine. We chose the anthrax protective antigen (PA) as a model to evaluate the adjuvant ability of PCP-I in enhancing the immunogenicity and protection of a PA-based anthrax vaccine. According to our results, PCP-I could significantly enhance anthrax specific anti-PA antibodies, toxin-neutralizing antibodies, anti-PA antibody affinity, as well as IgG1 and IgG2a levels. Besides, PCP-I increased the frequency of PA-specific memory B cells, increased the proliferation of PA-specific splenocytes, significantly stimulated the secretion of IL-4, and enhanced the activation of Dendritic cells (DCs) in vitro . The combination of PCP-I and CpG significantly enhanced the level of anti-PA antibodies and neutralizing antibodies, particularly PA-specific IgG2a, and shifted the Th2-bias to a Th1/Th2 balanced response. In addition, PCP-I with or without CpG could significantly improve the survival rate of immunized mice following challenge with the anthrax lethal toxin. These findings suggest that PCP-I may be a promising vaccine adjuvant that warrants further study.

The genus Henipavirus (HNVs) includes two fatal viruses, namely Nipah virus (NiV) and Hendra virus (HeV). Since 1994, NiV and HeV have been endemic to the Asia-Pacific region and responsible for more than 600 cases of infections. Two emerging HNVs, Ghana virus (GhV) and Mojiang virus (MojV), are speculated to be associated with unrecognized human diseases in Africa and China, respectively. Despite many efforts to develop vaccines against henipaviral diseases, there is presently no licensed human vaccine. As HNVs are highly pathogenic and diverse, it is necessary to develop universal vaccines to prevent future outbreaks. The attachment enveloped glycoprotein (G protein) of HNVs mediates HNV attachment to the host cell's surface receptors. G proteins have been used as a protective antigen in many vaccine candidates for HNVs. We performed quantitative studies on the antibody responses elicited by the G proteins of NiV, HeV, GhV, and MojV. We found that the G proteins of NiV and HeV elicited only a limited cross-reactive antibody response. Further, there was no cross-protection between MojV, GhV, and highly pathogenic HNVs. We then constructed a bivalent vaccine where the G proteins of NiV and HeV were fused with the human IgG1 Fc domain. The immunogenicity of the bivalent vaccine was compared with that of monovalent vaccines. Our results revealed that the Fc-based bivalent vaccine elicited a potent antibody response against both NiV and HeV. We also constructed a tetravalent Fc heterodimer fusion protein that contains the G protein domains of four HNVs. Immunization with the tetravalent vaccine elicited broad antibody responses against NiV, HeV, GhV, and MojV in mice, indicating compatibility among the four antigens in the Fc-fusion protein. These data suggest that our novel bivalent and tetravalent Fc-fusion proteins may be efficient candidates to prevent HNV infection.

The virus-encoded microRNAs (miRNAs) have been demonstrated to have important regulatory roles in herpesvirus biology, including virus replication, latency, pathogenesis and/or tumorigenesis. As an emerging efficient tool for gene editing, the clustered regularly interspaced short palindromic repeat (CRISPR)/Cas9 system has been successfully applied in manipulating the genomes of large DNA viruses. Herein, utilizing the CRISPR/Cas9 system with a double-guide RNAs transfection/virus infection strategy, we have established a new platform for mutagenesis of viral miRNAs encoded by the Marek's disease virus serotype 1 (MDV-1), an oncogenic alphaherpesvirus that can induce rapid-onset T-cell lymphomas in chickens. A series of miRNA-knocked out (miR-KO) mutants with deletions of the Meq- or the mid-clustered miRNAs, namely RB-1B∆Meq-miRs, RB-1B∆M9-M2, RB-1B∆M4, RB-1B∆M9 and RB-1B∆M11, were generated from vvMDV strain RB-1B virus. Interestingly, mutagenesis of the targeted miRNAs showed changes in the in vitro virus growth kinetics, which is consistent with that of the in vivo proliferation curves of our previously reported GX0101 mutants produced by the bacterial artificial chromosome (BAC) clone and Rec E/T homologous recombination techniques. Our data demonstrate that the CRISPR/Cas9-based gene editing is a simple, efficient and relatively nondisruptive approach for manipulating the small non-coding genes from the genome of herpesvirus and will undoubtedly contribute significantly to the future progress in herpesvirus biology.

Cry2A is widely used in transgenic crops in combination with Cry1A toxins. The sensitive and robust detection of Cry2A toxin in food and the environment is necessary to monitor the safety of biopesticides. Here, we describe an approach that involves the use of phage-displayed peptide for the detection of Cry2Ad2-3-the main area of Cry2Ad2 insecticidal activity. After four rounds of panning, six positive monoclonal phage particles were obtained. Pep5 with a sequence of ACSYNHNSKCGGG displayed low cross-reactivity with other Cry toxins. The working range of detection for Cry2Ad2-3 toxin standards in the brush border membrane vesicle (BBMV)-peptide sandwich ELISA was 10-50.625 ng mL -1 and the detection limit (LOD) was 8 ng mL -1 . Molecular insight into the interaction of pep5 with Cry2Ad2-3 was gleaned using homology modeling and docking. Molecular docking results showed that high-affinity peptide tended to dock in the groove between the two domains of Cry2Ad2-3. The interactions within the toxin-pep5 complex were due to hydrogen bond and hydrophobic interaction. Pep5 also lead us to trap the binding region. Therefore, peptides may be a cost-efficient alternative for detecting Cry toxins and studying their mechanisms., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Anthrax is an acute and highly lethal disease caused by Bacillus anthracis. Protective antigen (PA) is the primary candidate antigen for the anthrax vaccines. However, PA suffers from poor immunogenicity with short-term anti-PA antibody response. High effectiveness, durable immunity, and minimal risk are required for development of an effective anthrax vaccine. In the present study, PA was self-conjugated by 8-arm polyethylene glycol (PEG) and further by thioester chemistry. As a result, 3-5 PA molecules were covalently conjugated and functioned as an antigen delivery system. The conjugate (PA-PEG) could maintain the structural properties of PA and increase the thermal stability of PA. PA-PEG could elicit a robust anti-PA IgG and neutralization antibody response in the magnitude and quality. The antibodies could be largely maintained for 180 days after three immunizations of PA-PEG. PA-PEG effectively stimulated the maturation of dendritic cell and rapidly induced the germinal center (GC) reaction. The percentages of the GC B-cells and T follicular helper (Tfh) cells were thus significantly augmented. The inflammatory response elicited by PA-PEG was comparable to those by PBS and PA. Therefore, PA-PEG is expected as an effective anthrax vaccine candidate with durable immunoprotection against anthrax., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The frequent occurrence of algal blooms in surface water has attracted more and more attention, which caused many water quality problems, including disinfection byproducts (DBPs). Algal organic matter (AOM) including intracellular organic matter (IOM) and extracellular organic matter (EOM), was a well-known precursor to DBPs formation in drinking water. This study evaluated the effect of ultraviolet (UV) irradiation on the cell integrity, IOM release and DBPs formation during subsequent chlorination of Chlorella sp. Results showed the damage rates of algal cells increased to 40.1% after the high UV irradiation of 528 mJ/cm 2 , which contributed to the release of IOM. In addition, UV irradiation was effective in reducing the formation of haloacetic acids (HAAs) both in AOM and IOM, but promoted the formation of nitrogenous DBPs (N-DBPs) from AOM in subsequent chlorination. Furthermore, neutral pH exerted a positive effect on the formation of DBPs. UV irradiation decreased the bromine substitution factor (BSF) value of AOM at a high bromide level. The BSF values increased with increasing of the concentration of bromide. Moreover, more amino acids and low molecular weight precursors were produced after UV irradiation in filtered supernatant, which contributed to the formation of N-DBPs with algal chlorination. Overall, this information demonstrated pre-oxidation of UV irradiation could be used to treat the algal-rich drinking water., (Copyright © 2019. Published by Elsevier B.V.)

Coagulase (Coa) secreted by Staphylococcus aureus is associated with the establishment of staphylococcal disease, which activates host prothrombin and generates fibrin shields. The R domain of Coa, consisting of several conserved repeats, is important in immune evasion during S. aureus infection. However, previous research showed that the Coa R domain induced very weak specific antibody responses. In this study, we constructed a new R domain, CoaR6, consisting of 6 repeats that occur most frequently in clinical isolates. By fusing CoaR6 with Hc, the C-terminal fragment of the heavy chain of tetanus neurotoxin, we successfully increased anti-CoaR6 IgG levels in immunized mice which were hardly detected in mice immunized with CoaR6 plus alum. To further improve anti-CoaR6 responses, the combination adjuvants alum plus CpG were formulated with the antigen and exhibited a significantly higher specific antibody response. Moreover, active Th1/Th17 immune responses were observed in Hc-CoaR6 immunized group rather than CoaR6. Active immunization of Hc-CoaR6 with alum plus CpG showed protective effects in a peritonitis model induced by two S. aureus strains with different coagulase types. Our results provided strategies to improve the immunogenicity of R domain and supporting evidences for R domain to be an S. aureus vaccine candidate., (Copyright © 2018 Institut Pasteur. Published by Elsevier Masson SAS. All rights reserved.)

Adenoviral vectors (AdV) are considered promising candidates for vaccine applications. A prominent group of Toll-like receptors (TLRs) participate in the adenovirus-induced adaptive immune response, yet there is little information regarding the role of TLR4 in AdV-induced immune responses in recent literature. We investigated the function of TLR4 in both adaptive and innate immune responses to an AdV-based anthrax vaccine. By immunizing wild-type and TLR4 knockout (TLR4-KO) mice, we revealed the requirement of TLR4 in AdV-induced innate responses. We also showed that TLR4 functions are required for germinal centre responses in immunized mice, as expression of the apoptosis-related marker Fas was down-regulated on germinal centre B cells from TLR4-KO mice. Likewise, decreased expression of inducible costimulator on follicular T helper cells was observed in immunized TLR4-KO mice. Moreover, a potent protective antigen-specific humoral immune response was mimicked using an adjuvant system containing the TLR4 agonist monophosphoryl lipid A. Overall, our findings showed that very rapid antigen-specific antibody production is correlated with the TLR4-imprinted germinal centre response to AdV-based vaccine. These results provide additional evidence for the use of the AdV and a TLR agonist to induce humoral responses. Our findings offer new insights into rational vaccine design., (© 2018 John Wiley & Sons Ltd.)