Search Results (99)

Porcine circovirus type 2 (PCV2) is a globally prevalent swine pathogen that induces immunosuppression, predisposing pigs to subclinical infections. In intensive farming systems, PCV2 persistently impairs growth performance and vaccine efficacy, leading to substantial economic losses in the swine industry. Emerging evidence suggests that certain viruses exploit Suppressor of Cytokine Signaling 3 (SOCS3), a key immune checkpoint protein, to subvert host innate immunity by suppressing cytokine signaling. While SOCS3 has been implicated in various viral infections, its regulatory role in PCV2 replication remains undefined. This study aims to elucidate the mechanisms underlying the interplay between SOCS3 and PCV2 during viral pathogenesis. Porcine SOCS3 was amplified using RT-PCR and stably overexpressed in PK-15 cells through lentiviral delivery. Bioinformatics analysis facilitated the design of three siRNA candidates targeting SOCS3. We systematically investigated the effects of SOCS3 overexpression and knockdown on PCV2 replication kinetics and host antiviral responses by quantifying the viral DNA load and the mRNA levels of cytokines. PCV2 infection upregulated SOCS3 expression at both transcriptional and translational levels in PK-15 cells. Functional studies revealed that SOCS3 overexpression markedly enhanced viral replication, whereas its knockdown suppressed viral proliferation. Intriguingly, SOCS3-mediated immune modulation exhibited a divergent regulation of antiviral cytokines: PCV2-infected SOCS3-overexpressing cells showed elevated IFN-β but suppressed TNF-α expressions, whereas SOCS3 silencing conversely downregulated IFN-β while amplifying TNF-α responses. This study unveils a dual role of SOCS3 during subclinical porcine circovirus type 2 (PCV2) infection: it functions as a host-derived pro-viral factor that facilitates viral replication while simultaneously reshaping the cytokine milieu to suppress overt inflammatory responses. These findings provide novel insights into the mechanisms underlying PCV2 immune evasion and persistence and establish a theoretical framework for the development of host-targeted control strategies. Although our results identify SOCS3 as a key host determinant of PCV2 persistence, the precise molecular pathways involved require rigorous experimental validation. Full article

Microalgae, with their unparalleled capabilities for sunlight-driven growth, CO₂ fixation, and synthesis of diverse high-value compounds, represent sustainable cell factories for a circular bioeconomy. However, industrial deployment has been hindered by biological constraints and the inadequacy of conventional genetic tools. The advent of CRISPR-Cas systems initially provided precise gene editing via targeted DNA cleavage. This review argues that the true transformative potential lies in moving decisively beyond cutting to harness CRISPR as a versatile synthetic biology “Swiss Army Knife”. We synthesize the rapid evolution of CRISPR-derived tools—including transcriptional modulators (CRISPRa/i), epigenome editors, base/prime editors, multiplexed systems, and biosensor-integrated logic gates—and their revolutionary applications in microalgal engineering. These tools enable tunable gene expression, stable epigenetic reprogramming, DSB-free nucleotide-level precision editing, coordinated rewiring of complex metabolic networks, and dynamic, autonomous control in response to environmental cues. We critically evaluate their deployment to enhance photosynthesis, boost lipid/biofuel production, engineer high-value compound pathways (carotenoids, PUFAs, proteins), improve stress resilience, and optimize carbon utilization. Persistent challenges—species-specific tool optimization, delivery efficiency, genetic stability, scalability, and biosafety—are analyzed, alongside emerging solutions and future directions integrating AI, automation, and multi-omics. The strategic integration of this CRISPR toolkit unlocks the potential to engineer robust, high-productivity microalgal cell factories, finally realizing their promise as sustainable platforms for next-generation biomanufacturing. Full article

Background: Fowl typhoid (FT), a septicemic infection caused by Salmonella Gallinarum (SG), and H9N2 avian influenza are two economically important diseases that significantly affect the global poultry industry. Methods: We exploited the live attenuated Salmonella Gallinarum (SG) mutant JOL3062 (SG: ∆lon ∆pagL ∆asd) as a delivery system for H9N2 antigens to induce an immunoprotective response against both H9N2 and FT. To enhance immune protection against H9N2, a prokaryotic and eukaryotic dual expression plasmid, pJHL270, was employed. The hemagglutinin (HA) consensus sequence from South Korean avian influenza A virus (AIV) was cloned under the Ptrc promoter for prokaryotic expression, and the B cell epitope of neuraminidase (NA) linked with matrix protein 2 (M2e) was placed for eukaryotic expression. In vitro and in vivo expressions of the H9N2 antigens were validated by qRT-PCR and Western blot, respectively. Results: Oral immunization with JOL3121 induced a significant increase in SG and H9N2-specific serum IgY and cloacal swab IgA antibodies, confirming humoral and mucosal immune responses. Furthermore, FACS analysis showed increased CD4+ and CD8+ T cell populations. On day 28 post-immunization, there was a substantial rise in the hemagglutination inhibition titer in the immunized birds, demonstrating neutralization capabilities of immunization. Both IFN-γ and IL-4 demonstrated a significant increase, indicating a balance of Th1 and Th2 responses. Intranasal challenge with the H9N2 Y280 strain resulted in minimal to no clinical signs with significantly lower lung viral titer in the JOL3121 group. Upon SG wildtype challenge, the immunized birds in the JOL3121 group yielded 20% mortality, while 80% mortality was recorded in the PBS control group. Additionally, bacterial load in the spleen and liver was significantly lower in the immunized birds. Conclusions: The current vaccine model, designed with a host-specific pathogen, SG, delivers a robust immune boost that could enhance dual protection against FT and H9N2 infection, both being significant diseases in poultry, as well as ensure public health. Full article

Epigenome engineering, particularly utilizing CRISPR/dCas-based systems, is a powerful strategy to modulate gene expression and genome functioning without altering the DNA sequence. In this review we summarized current achievements and prospects in dCas-mediated epigenome editing, primarily focusing on its applications in biomedicine, but also providing a wider context for its applications in biotechnology. The diversity of CRISPR/dCas architectures is outlined, recent innovations in the design of epigenetic editors and delivery methods are highlighted, and the therapeutic potential across a wide range of diseases, including hereditary, neurodegenerative, and metabolic disorders, is examined. Opportunities for the application of dCas-based tools in animal, agricultural, and industrial biotechnology are also discussed. Despite substantial progress, challenges, such as delivery efficiency, specificity, stability of induced epigenetic modifications, and clinical translation, are emphasized. Future directions aimed at enhancing the efficacy, safety, and practical applicability of epigenome engineering technologies are proposed. Full article

Antioxidant peptides derived from woody oil resource by-products exhibit strong free radical scavenging abilities and offer potential applications in functional foods, nutraceuticals, and cosmetics. This review summarizes the latest advances in preparation technologies, including enzymatic hydrolysis, microbial fermentation, chemical synthesis, recombinant expression, and molecular imprinting, each with distinct advantages in yield, selectivity, and scalability. The structure–activity relationships of antioxidant peptides are explored with respect to amino acid composition, molecular weight, and 3D conformation, which collectively determine their bioactivity and stability. Additionally, emerging delivery systems—such as nanoliposomes, microencapsulation, and cell-penetrating peptides—are discussed for their role in enhancing peptide stability, absorption, and targeted release. Mechanistic studies reveal that antioxidant peptides from woody oil resources act through network pharmacology, engaging core signaling pathways, including Nrf2/ARE, PI3K/Akt, AMPK, and JAK/STAT, to regulate oxidative stress, mitochondrial health, and inflammation. Preliminary safety data from in vitro, animal, and early clinical studies suggest low toxicity and favorable tolerability. The integration of omics technologies, molecular docking, and bioinformatics is accelerating the mechanism-driven design and functional validation of peptides. In conclusion, antioxidant peptides derived from woody oil resources represent a sustainable, multifunctional, and scalable solution for improving human health and promoting a circular bioeconomy. Future research should focus on structural optimization, delivery enhancement, and clinical validation to facilitate their industrial translation. Full article

The architecture, engineering, and construction (AEC) industry is experiencing significant digital transformation, creating a critical need to understand how future professionals perceive and accept emerging technologies. This study applies the Technology Acceptance Model (TAM) to investigate undergraduate construction students’ perceptions of Building Information Modelling (BIM) and examines how these factors influence their views on BIM applications. Using an exploratory mixed-methods approach, we analysed 773 responses from students at an Australian university across AEC disciplines, with 607 providing substantive qualitative feedback. Qualitative thematic analysis provided rich contextual understanding of student perspectives, while quantitative analysis revealed pattern frequencies across disciplines. Findings showed that perceived usefulness (PU) (37.7%) and attitude toward using (ATU) (68.4%) dominated student responses, while perceived ease of use (PEOU) (6.9%) received less attention. Productivity benefits (15.3%) and increased accuracy (7.9%) emerged as primary usefulness drivers. Disciplinary differences were significant, with Civil Engineering students emphasising design validation aspects and Construction Management students focusing on project delivery benefits of BIM. Notably, students exhibited sophisticated ambivalence, recognising BIM’s professional value while expressing concerns regarding the steep learning curve, especially when its adoption is coupled with the integration of emerging technologies such as artificial intelligence. This study contributes to the existing knowledge by: (1) documenting the current state of student perceptions in BIM education; and (2) revealing the complex interplay between technological enthusiasm and socio-professional concerns across both educational and industry settings. These findings provide evidence-based guidance for developing BIM curricula that address both socio-technical competencies and student perceptions, helping bridge the gap between educational outcomes and students’ understanding of industry requirements. Full article

Porcine reproductive and respiratory syndrome virus (PRRSV) continues to be a major threat to the global swine industry, causing significant economic losses. To address this, we developed a scalable recombinant adeno-associated virus (rAAV)-based strategy for the delivery of soluble viral receptors (SVRs) to treat and potentially eliminate PRRSV infections. This strategy involves fusing the virus-binding domains of two key cellular receptors, sialoadhesin (Sn4D) and CD163 (SRCR5-9), with an Fc fragment. We then used an insect cell–baculovirus expression vector system to produce the rAAV-SRCR59-Fc/Sn4D-Fc vector. Through a series of optimizations, we determined the best conditions for rAAV production, including a baculovirus co-infection ratio of 0.5:1.0, an initial insect cell density of 2.0 × 10⁶ cells/mL, a fetal bovine serum concentration of 2%, and a culture temperature of 30 °C. Under these optimized conditions, we achieved a high titer of rAAV-SRCR59-Fc/Sn4D-Fc in a 2 L bioreactor, reaching 5.4 ± 0.9 × 10⁹ infectious viral particles (IVPs)/mL. Notably, in vitro neutralization assays using a Transwell co-culture system demonstrated a 4.3 log reduction in viral titers across genetically diverse PRRSV-2 strains, including VR2332, JXA1, JS07, and SH1705. Collectively, this study provides a robust platform for large-scale rAAV production and highlights the potential of SVR-based gene therapy to address the antigenic diversity of PRRSV-2. Full article

Astaxanthin, a xanthophyll carotenoid, exhibits potent biological functions, including antioxidant, immune regulation, growth promotion, improved reproductive capacity, and enhancement of the body color of aquatic animals. In recent years, with the rapid development of the aquaculture industry, the application of astaxanthin in aquaculture has garnered increasing attention. Studies have demonstrated that astaxanthin significantly enhances the antioxidant capacity of aquatic animals, reduces oxidative damage, and regulates the expression of immune-related genes, thereby improving immunity and disease resistance. Moreover, astaxanthin promotes growth and reproductive performance, particularly in high-value aquaculture species, where it also serves as a natural pigment to increase market competitiveness. However, the low bioavailability and high production costs of astaxanthin remain major constraints to its widespread use in aquaculture. To address these limitations, various strategies—such as microencapsulation, liposomal delivery, and nanotechnology—have been explored to improve its stability and water solubility. Additionally, expanding astaxanthin sources and optimizing production processes are effective approaches to reducing costs. This review summarizes recent advances in astaxanthin research within aquaculture, highlights its multifunctional roles in promoting the health and production efficiency of aquatic animals, and discusses the current challenges and future research directions. Full article

Available interventions for the management of chronic hepatitis B (hepB) exhibit limited efficacy and barriers to vaccination against the hepatitis B virus (HBV) have hampered prophylaxis programmes. Development of potent therapeutics capable of functional cure of chronic hepB thus remains a relevant medical objective. RNA interference (RNAi) can be exploited to effect potent and specific silencing of target genes through the introduction of RNA sequences that mimic the natural activators of the pathway. To achieve a therapeutic effect, artificial primary microRNAs (pri-miRNAs) have been used extensively to target various viruses, including HBV. To date artificial pri-miRNAs have exclusively been produced from DNA expression cassettes. Although this achieves impressive silencing, eventual translation of this platform to the clinic is complicated by the requirement for viral vectors to deliver DNA. Consequently, clinical translation has been slow. Recently, the use of in vitro transcribed RNA, specifically to produce mRNA vaccines at industrial scale, has gained significant interest. We therefore sought to evaluate the feasibility of using in vitro transcribed artificial pri-miRNAs for the inhibition of HBV gene expression. Artificial HBV-targeting pri-miR-31 sequences, which are highly effective when expressed in cells from a DNA template, demonstrated modest silencing of viral replication when incorporated into mRNA that was transcribed in vitro. Off-target effects were also observed. Characterisation revealed that intracellular processing of the artificial pri-miRNAs was inefficient and non-specific effects were caused by stimulation of the interferon response. Nevertheless, optimised nuclear delivery of the artificial pri-miRNAs should improve their processing and achieve better anti-hepB efficacy. Full article

Due to the barriers imposed by the restriction–modification (RM) system, Nisin-producing industrial strains of Lactococcus lactis often encounter low transformation efficiency, which seriously hinders the widespread application of genetic engineering in non-model L. lactis. Herein, we present a novel pre-modification strategy (PMS) coupled with optimized plasmid delivery systems designed to systematically evade RM barriers and substantially improve Nisin biosynthesis in L. lactis. Through the use of engineered Escherichia coli strains with methylation profiles specifically optimized for L. lactis C20, we have effectively evaded RM barriers, thereby facilitating the efficient introduction of large Nisin biosynthetic gene clusters into L. lactis. The PMS tools, which significantly improve the transformation efficiency (~10³ transformants per microgram of DNA), have been further improved in combination with a Rolling Circle Amplification, resulting in a higher enhancement in transformation efficiency (~10⁴ transformants per microgram of DNA). Using this strategy, large Nisin biosynthetic gene clusters and the expression regulation of all genes within the cluster were introduced and analyzed in L. lactis, leading to a highest Nisin titer of 11,052.9 IU/mL through a fed-batch fermentation in a 5 L bioreactor. This is the first systematic report on the expression regulation and application of a complete Nisin biosynthesis gene cluster in L. lactis. Taken together, our studies provide a versatile and efficient strategy for systematic evasion and enhancement of RM barriers and Nisin biosynthesis, thereby paving the way for genetic modification and metabolic engineering in L. lactis. Full article

A number of viral diseases have significantly impeded the growth of the aquaculture industry. Antiviral drugs represent an effective means of controlling infection. However, the efficacy of the entire therapeutic process is contingent upon the availability of an efficient delivery system. This study selected three common antiviral drugs and constructed corresponding drug delivery systems utilising single-walled carbon nanotubes (SWCNTs) as carriers. The reliability of carbon nanotubes as delivery carriers was evaluated by detecting the therapeutic effect on infectious splenic and renal necrosis virus (ISKNV). The findings demonstrated that SWCNTs can effectively enhance the absorption of the three drugs into the body, prolong their metabolic half-life, and improve the survival rate of fish infected with ISKNV. The Ribavirin-SWCNTs (RBV-SWCNTs) group exhibited the most pronounced protective effect, with a mortality rate of less than 25%. It was observed that SWCNTs facilitated the rapid transportation of ribavirin, with the drug content in the RBV-SWCNTs group being approximately double that of the free ribavirin group. Furthermore, this system markedly diminished the viral load, augmented enzyme activities, and elevated antiviral gene expression. This study indicated that carbon nanotubes are optimal carriers for antiviral drugs, which have considerable potential as a delivery vehicle for antiviral drugs to prevent viral infections in aquaculture. Full article

Using plants as bioreactors, molecular farming has emerged as a versatile and sustainable platform for producing recombinant vaccines, therapeutic proteins, industrial enzymes, and nutraceuticals. This innovative approach leverages the unique advantages of plants, including scalability, cost-effectiveness, and reduced risk of contamination with human pathogens. Recent advancements in gene editing, transient expression systems, and nanoparticle-based delivery technologies have significantly enhanced the efficiency and versatility of plant-based systems. Particularly in vaccine development, molecular farming has demonstrated its potential with notable successes such as Medicago’s Covifenz for COVID-19, illustrating the capacity of plant-based platforms to address global health emergencies rapidly. Furthermore, edible vaccines have opened new avenues in the delivery of vaccines, mainly in settings with low resources where the cold chain used for conventional logistics is a challenge. However, optimization of protein yield and stability, the complexity of purification processes, and regulatory hurdles are some of the challenges that still remain. This review discusses the current status of vaccine development using plant-based expression systems, operational mechanisms for plant expression platforms, major applications in the prevention of infectious diseases, and new developments, such as nanoparticle-mediated delivery and cancer vaccines. The discussion will also touch on ethical considerations, the regulatory framework, and future trends with respect to the transformative capacity of plant-derived vaccines in ensuring greater global accessibility and cost-effectiveness of the vaccination. This field holds great promise for the infectious disease area and, indeed, for applications in personalized medicine and biopharmaceuticals in the near future. Full article

Immunization against Gonadotropin-Releasing Hormone (GnRH) has been successfully explored and developed for the parenteral inoculation of animals, aimed at controlling fertility, reducing male aggressiveness, and preventing boar taint. Although effective, these vaccines may cause adverse reactions at the injection site, including immunosuppression and inflammation, as well as the involvement of laborious and time-consuming procedures. Oral vaccines represent an advancement in antigen delivery technology in the vaccine industry. In this study, a Salmonella enterica serovar Typhimurium (S. Typhimurium) mutant lacking the pathogenicity island 2 (S. Typhimurium ΔSPI2) was used as a vehicle and mucosal adjuvant to deliver two genetic constructs in an attempt to develop an oral immunological preparation against gonadotropin hormone-releasing hormone (GnRH). S. Typhimurium ΔSPI2 was transformed to carry two plasmids containing a modified GnRH gene repeated in tandem (GnRXG/Q), one under eukaryotic expression control (pDNA::GnRXG/Q) and another under prokaryotic expression control (pJexpress::GnRXG/Q). A group of three male BALB/c mice were orally immunized and vaccination-boosted 30 days later. The oral administration of S. Typhimurium ΔSPI2 transformed with both plasmids was effective in producing antibodies against GnRXG/Q, leading to a decrease in serum testosterone levels and testicular tissue atrophy, evidenced by a reduction in the transverse tubular diameter of the seminiferous tubules and a decrease in the number of layers of the seminiferous epithelium in the testes of the inoculated mice. These results suggest that S. Typhimurium ΔSPI2 can be used as a safe and simple system to produce an oral formulation against GnRH and that Salmonella-mediated oral antigen delivery is a novel, yet effective, alternative to induce an immune response against GnRH in a murine model, warranting further research in other animal species. Full article

In recent years, parcel volumes have reached record highs, prompting the logistics industry to explore innovative solutions to meet growing demand. In densely populated areas, delivery robots offer a promising alternative to traditional truck-based delivery systems. These autonomous electric robots operate on sidewalks and deliver time-sensitive goods, such as express parcels, medicine and meals. However, their limited cargo capacity and battery life require a return to a depot after each delivery. This challenge can be modeled as an electric vehicle-routing problem with soft time windows and single-unit capacity constraints. The objective is to serve all customers while minimizing the quadratic sum of delivery delays and ensuring each vehicle operates within its battery limitations. To address this problem, we propose a mixed-integer quadratic programming model and introduce an enhanced formulation using a layered graph structure. For this layered graph, we present two solution approaches based on relaxations that reduce the number of nodes and arcs compared to the expanded formulation. The first approach, Iterative Refinement, solves the current relaxation to optimality and refines the graph when the solution is infeasible for the expanded formulation. This process continues until a proven optimal solution is obtained. The second approach, Branch and Refine, integrates graph refinement into a branch-and-bound framework, eliminating the need for restarts. Computational experiments on modified Solomon instances demonstrate the effectiveness of our solution approaches, with Branch and Refine consistently outperforming Iterative Refinement across all tested parameter configurations. Full article

The popularity of online shopping in China has increased significantly, creating new development opportunities for the express delivery industry. However, the rapid expansion of the express industry has also created challenges in the parcel sorting process. The demanding nature of parcel sorting work, which is characterized by intense and prolonged repetitive tasks, makes individuals particularly vulnerable to the effects of fatigue. Fatigue is a complex condition that encompasses both physiological and psychological exhaustion. It often results in reduced energy levels and diminished functionality, significantly impacting an individual’s performance at work and their overall well-being. This study aimed to investigate how physiological and psychological fatigue affects sorting efficiency and to identify appropriate rest periods that will allow employees to maintain their performance levels. The research involved fifteen participants who took part in a 60 min continuous sorting experiment and a similar experiment with scheduled breaks. During both trials, we collected data on participants’ electromyography (EMG) and electrodermal activity (EDA), as well as subjective fatigue ratings (RPE). Signal features such as the median frequency (MF) of EMG and the skin conductance level (SCL) were analyzed to assess physiological and psychological fatigue, respectively. The results show that physiological fatigue mainly affects sorting efficiency in the first 30 min, while psychological fatigue becomes more influential in the following half-hour period. In addition, subjective fatigue levels during the first 30 min are primarily determined by psychological factors, while beyond that point, both physiological and psychological fatigue contribute to subjective fatigue. Rest periods of 415–460 s, based on EDA recovery times, effectively support sorting efficiency and participants’ recovery. This study highlights the complex ways in which fatigue affects parcel sorting performance and provides valuable theoretical and practical insights for establishing labor quotas and optimizing work schedules in the parcel sorting industry. Full article