Plant-Based Recombinant Vaccines for Foot-and-Mouth Disease: A Meta-Synthesis

Background: Transient and stable gene transformation systems play a crucial role in elucidating gene functions and driving genetic improvement in plants. However, their application in medicinal woody plants has been hampered by inefficient procedures for isolating protoplasts and regenerating plant in vitro. Results: Embryogenic callus protoplast isolation and transient transformation system were successfully established, the highest yield of protoplasts was achieved at approximately 1.88×106 cells per gram with a viability of 90% under the combination of 1.5% cellulase and 0.2% lysozyme, with enzymatic digestion for 6 h in darkness followed by centrifugation at 400×g for 5 min. The transient transfection rate of protoplast reached 45.56% at a PEG 4000 concentration of 40%, a transfection time of 40 min, 16 h of dark incubation, a plasmid concentration of 1.5 ng·μL-1, and a heat shock of 25 min at 45°C. Additionally, 15 Agrobacterium tumefaciens-mediated GUS-positive seedlings were obtained through the somatic embryogenetic pathway under the optimized conditions. Conclusion: This study successfully established both transient and stable genetic transformation systems, paving the way for future molecular biology research in A. senticosus.

BackgroundTransient and stable gene transformation systems play a crucial role in elucidating gene functions and driving genetic improvement in plants. However, their application in medicinal woody plants has been hampered by inefficient procedures for isolating protoplasts and regenerating plants in vitro.ResultsEmbryogenic callus protoplast isolation and transient transformation system were successfully established. The highest yield of protoplasts was approximately 1.88 × 106 cells per gram with a viability of 90% under the combination of 1.5% cellulase and 0.2% macerozyme, with enzymatic digestion for 6 h in darkness followed by centrifugation at 400×g for 5 min. The transient transfection rate of protoplast reached 45.56% at a PEG 4000 concentration of 40%, a transfection time of 40 min, 16 h of dark incubation, a plasmid concentration of 1.5 ng μL−1, and 25 min heat shock at 45 °C. In addition, 15 Agrobacterium tumefaciens-mediated GUS-positive seedlings were obtained through the somatic embryogenetic pathway under the optimized conditions.ConclusionThis study successfully established both transient and stable genetic transformation systems, paving the way for future molecular biology research on A. senticosus.Graphical

Vaccination as a means of control of foot-and-mouth disease in sub-saharan Africa

With the continuous improvement of people’s quality of life and tremendous changes in consumption concepts, people’s demand for fresh agricultural products is increasing, which has led to a huge gap in the supply of fresh agricultural products logistics. In recent years, frequent food quality and safety issues have attracted the attention of consumers and have been highly valued by the country, making enterprises deeply aware of the importance of the refrigeration supply chain. However, the traditional cold chain supply of agricultural products is far from being able to meet the strict quality requirements of food for consumers in terms of production, processing, storage, transportation and sales. Therefore, how to improve the efficiency of the product refrigeration supply chain, reduce logistics costs, and ensure product freshness and quality assurance has become the focus and difficulty of establishing a cold chain supply chain business. This article is based on the analysis and design of the intelligent cold chain logistics simulation model under the Internet of Things technology. Firstly, the literature research method is used to summarize the problems of the intelligent cold chain logistics under the Internet of Things technology and the advantages of the intelligent cold chain logistics under the Internet of Things technology. The feasibility of the intelligent cold chain logistics simulation model under the Internet of Things technology, and then through the analysis of the cold chain logistics process unit and organization, the modeling is carried out based on the analysis results.

With the development of the times and the improvement of people’s pursuit of quality of life, cold chain logistics has emerged quietly. Cold chain logistics can maximize the freshness of transported products, and it has become the first choice for transportation, especially for fresh products. From the perspective of public management, this paper explores the current situation of cold chain logistics, and puts forward reasonable suggestions combined with the problems existing in the development of cold chain logistics, aiming to improve the service quality of cold chain logistics, better highlight the advantages of cold chain logistics, and provide suggestions for the development of the entire logistics industry.

Protoplast culture and transformation technology offer a novel method for developing new plant varieties. Nonetheless, the effective preparation of protoplasts and transformation technology specific to celery has yet to be achieved. This study utilized celery seedling leaves as the primary materials to examine the key factors influencing protoplast isolation. The aim was to prepare leaf protoplasts with a high yield and of high quality and subsequently conduct transient gene transformation and expression. The findings indicated that the most effective procedure for isolating and purifying protoplasts was enzymatic digestion using an enzyme solution consisting of 2.0% cellulase, 0.1% pectolase, and 0.6 M mannitol for a duration of 8 h. Subsequently, the protoplasts were filtered through a 400-mesh sieve and purified through centrifugation at 200× g. Within this system, the overall protoplast yield was exceptionally high, reaching a viability rate of up to 95%. The transient transformation system yielded a maximum transformation efficiency of approximately 53%, as evaluated using the green fluorescent protein (GFP) as a reporter gene. The parameters of the transient transformation system were as follows: a protoplast concentration of 5 × 105 cells·mL−1, exogenous DNA concentration of 500 μg·mL−1, final concentration of PEG4000 at 40%, and transformation duration of 15 min. The transient transformation system was also utilized to further analyze the protein localization characteristics of the celery transcription factor AgMYB80. The findings indicated that AgMYB80 predominantly localizes in the nucleus, thereby confirming the reliability and effectiveness of the transient transformation system. This study successfully established an efficient system for isolating, purifying, and transforming celery protoplasts, and will serve as a basis for future studies on molecular biology and gene function.

The recent COVID-19 pandemic has highlighted the value of technologies that allow a fast setup and production of biopharmaceuticals in emergency situations. The plant factory system can provide a fast response to epidemics/pandemics. Thanks to their scalability and genome plasticity, plants represent advantageous platforms to produce vaccines. Plant systems imply less complicated production processes and quality controls with respect to mammalian and bacterial cells. The expression of vaccines in plants is based on transient or stable transformation systems and the recent progresses in genome editing techniques, based on the CRISPR/Cas method, allow the manipulation of DNA in an efficient, fast, and easy way by introducing specific modifications in specific sites of a genome. Nonetheless, CRISPR/Cas is far away from being fully exploited for vaccine expression in plants. In this review, an overview of the potential conjugation of the renewed vaccine technologies (i.e., virus-like particles—VLPs, and industrialization of the production process) with genome editing to produce vaccines in plants is reported, illustrating the potential advantages in the standardization of the plant platforms, with the overtaking of constancy of large-scale production challenges, facilitating regulatory requirements and expediting the release and commercialization of the vaccine products of genome edited plants.

The nopaline synthase (nos) promoter is expressed in a wide range of plant cell types and regulated by various developmental and environmental factors. The nos upstream control region essential for this regulation was studied by means of synthetic oligomers using transient and stable transformation systems. Insertion of a 20 nucleotide sequence containing two hexamer motifs and a spacer region into deletion mutants lacking the upstream control region was essential for promoter activity. Mutation of one or more nucleotides of either hexamer sequence significantly altered the strength of expression of the nos promoter. Point mutations within the spacer region also strongly influenced promoter strength. Insertion of multiple copies of the 20 nucleotide sequence into the nonfunctional deletion mutants proportionally increased the promoter activity. These results suggest that this twenty nucleotide sequence is essential for the nos promoter to function. Substitution of the nos element with the ocs or 35S as-1 which contain similar hexamer motifs restored not only promoter activity but also responses to wounding, auxin, methyl jasmonate, and salicylic acid.

Efficient scar-free knock-ins of several kilobases in plants by engineered CRISPR-Cas endonucleases

The decentralized clinical trials make clinical research accessible by using digital tools such as e-consent, telehealth, and remote monitoring. Decentralized clinical trials in India have the potential to reduce participant burden and increase trial participation. However, the nation's disparate healthcare system, digital divide, and changing regulatory environment make broad adoption difficult. The current status of decentralized clinical trials in India is examined in this review, including significant operational, regulatory gaps, and potential implementation strategies. The aim is to offer information which will help in scaling decentralized clinical trials in India. This review compiles findings from journals, regulatory guidelines, white papers, blogs and case studies from Indian decentralized clinical trial platforms. It focuses on trends, logistical challenges, regulatory readiness in decentralized clinical trials implementation in the Indian context. To support key details tables and figures are used. India's pilot decentralized clinical trials demonstrate increased cost-effectiveness and reach, particularly when it comes to extending access outside of cities. However, region-specific infrastructure deficiencies, untrained staff for decentralization operations, and absence of e-consent provisions in New Drugs and Clinical Trial Rules 2019, restricts scalability. The 2024 draft guidance of United States’s Food and Drug Administration provides framework for decentralized clinical trials which can be implemented in India. In order to ensure ethical, and successful implementation of decentralized clinical trials for India's diverse healthcare system investment in digital health infrastructure, regulatory revisions, and site personnel training for decentralized clinical trial operations is necessary.

Paeonia lactiflora Pall. is known as the king of herbaceous flowers with high ornamental and precious medicinal value. However, the lack of a stable genetic transformation system has greatly affected the research of gene function in P. lactiflora. The Agrobacterium-mediated transient gene expression is a powerful tool for the characterization of gene function in plants. In this study, the seedlings of P. lactiflora were used as the transformation receptor materials, and the efficient transient transformation system with a GUS reporter gene was successfully established by Agrobacterium harboring pCAMBIA1301. To optimize the system, we investigated the effects of germination time, Agrobacterium cell density, infection time, acetosyringone (AS) concentration, co-culture time, negative pressure intensity, Tween-20 concentration and different receptor materials on the transient transformation efficiency of P. lactiflora. The results showed that the highest transient transformation efficiency (93.3%) could be obtained when seedlings in 2-3 cm bud length were subjected to 12 h infection of resuspension solution comprising 1.2 OD600 Agrobacterium, 200 μM AS and 0.01% Tween-20 under 10 of negative pressure intensity followed by 3 days of co-culture in darkness condition. This method is more suitable for the study of gene function in P. lactiflora. Subsequently, stress resistance genes PlGPAT, PlDHN2 and PlHD-Zip were used to verify the effectiveness of this transformation system. These results can provide critical information for identification of key genes in non-model plants, such as P. lactiflora, and promote the development of molecular biology research for P. lactiflora.

This mixed-methods study investigates the cognitive and methodological structure of AI-mediated recursive dialogue as a generative process for scholarly inquiry—and as the analytic engine behind this very study. Using nine full-length transcripts from an unsupervised 56-day research sprint, I tracked how dialogic engagement with ChatGPT catalyzed theory emergence, narrative coherence, and publication-ready academic work. ChatGPT autonomously parsed, coded, and sequenced the transcripts in real time, while I provided the conceptual framework and interpretive synthesis. Through thematic coding and quantitative analysis, I identified patterns of reflection, synthesis, and conceptual pivot points, emphasizing the recursive rhythm of: Prompt → Reflection → Clarification → Synthesis. This study demonstrates how recursive dialogue with AI can produce replicable cognitive insights and contribute to emergent models of methodological co-construction. I argue that when intentionally scaffolded, AI dialogue is not only a valid method of inquiry, but one capable of generating new epistemologies and expediting their study from within.

Targeted protein degradation plays a wide range of important roles in plant growth and development, but analyzing protein turnover in vivo is technically challenging. Until recently, there has been no straightforward methodology for quantifying protein dynamics at subcellular resolution during cellular transitions in plants. A tandem fluorescent protein timer (tFT) is a fusion of two different fluorescent proteins with distinct fluorophore maturation kinetics, which allows estimation of relative protein age from the ratio of fluorescence intensities of the two fluorescent proteins. Here, we describe approaches to use this technology to report relative protein lifetime in both transient and stable plant transformation systems. tFTs enable in vivo, real-time protein lifetime assessment within subcellular compartments and across tissues, permitting the analysis of protein degradation dynamics in response to stresses or developmental cues and in different genetic backgrounds.

Toxic work culture has become a major concern in organizational psychology due to its impact on mental health and productivity. This study explores how power imbalances, weak organizational systems, and the absence of protective mechanisms create psychosocial stress in the workplace. Using a qualitative case study in the Contact Center Industry in Central Java and Yogyakarta, 10 participants from various job levels were interviewed through semi-structured interviews, observations, and document analysis. Data were analyzed using thematic coding. Findings revealed three dominant themes: psychological insecurity (63%), reflected in withdrawal, fear of speaking up, and loss of initiative; toxic behaviors (18%), such as sarcasm and verbal abuse tolerated due to high performance; and structural dysfunction (23%), including the absence of conflict SOPs and weak HR intervention. Employees responded with self-protection strategies like social withdrawal. This study contributes to developing ethical and psychologically safe organizational systems through ethics-based evaluation and stronger HR functions.

Stagnant Perceptions of Nursing Among High School Students: Results of a Shadowing Intervention Study