C-FOS inhibition promotes pancreatic cancer cell ferroptosis by transcriptionally regulating the expression of SLC7A11.

Cellular proto-oncogene C-Fos forms the AP-1 transcription factor by dimerizing with proto-oncogene c-Jun; this factor upregulates the transcription of genes associated with different malignancies. However, its functions in pancreatic adenocarcinoma (PAAD) remain poorly understood. In this study, the c-Fos was increased in PAAD cells and tissues through bioinformatic analysis, RT-PCR, and WB. In two PAAD cell lines, PANC-1 and BxPC-3, we performed c-Fos knockdown studies using short hairpin RNA (shRNA). Functional analysis indicated that c-Fos depletion in PAAD cells inhibits cell proliferation and promotes ferroptosis. Chromatin Immunoprecipitation (ChIP) and Dual-luciferase experiments showed that c-Fos coupled to the promoter region of SLC7A11 stimulated its transcription, providing mechanistic insight into the process. Moreover, SLC7A11 blocked the decline of proliferation and ferroptosis by c-Fos knockdown in PAAD cells. Furthermore, a xenograft nude mouse model was established to study the impact of c-Fos on tumorigenesis in vivo. Depletion of c-Fos could suppress PC tumor growth and the expressions of SLC7A11, ki-67, and 4HNE, but overexpression of SLC7A11 reversed this process. In summary, our investigation has shown that c-Fos acts as a transcriptional regulator of SLC7A11, which may enhance tumour growth in pancreatic cancer by inhibiting ferroptosis. These results indicate that c-Fos might be a promising target for treating ferroptosis in PAAD.

Combining an Enhanced Polyphosphate Kinase-Driven UDP-Glucose Regeneration System with the Screening of Key Glycosyltransferases for Efficient In Vitro Synthesis of Nucleoside Disaccharides.

Nucleoside disaccharides are essential glycosides that naturally occur in specific living organisms. This study developed an enhanced UDP-glucose regeneration system to facilitate the in vitro multienzyme synthesis of nucleoside disaccharides by integrating it with nucleoside-specific glycosyltransferases. The system utilizes maltodextrin and polyphosphate as cost-effective substrates for UDP-glucose supply, catalyzed by α-glucan phosphorylase (αGP) and UDP-glucose pyrophosphorylase (UGP). To address the low activity of known polyphosphate kinases (PPKs) in the UDP phosphorylation reaction, a sequence-driven screening identified RhPPK with high activity against UDP (>1000 U/mg). Computational design further led to the creation of a double mutant with a 2566-fold increase in thermostability at 50 °C. The enhanced UDP-glucose regeneration system increased the production rate of nucleoside disaccharide synthesis by 25-fold. In addition, our UDP-glucose regeneration system is expected to be applied to other glycosyl transfer reactions.

Mining unique cysteine synthetases and computational study on thoroughly eliminating feedback inhibition through tunnel engineering.

L-cysteine is an essential component in pharmaceutical and agricultural industries, and synthetic biology has made strides in developing new metabolic pathways for its production, particularly in archaea with unique O-phosphoserine sulfhydrylases (OPSS) as key enzymes. In this study, we employed database mining to identify a highly catalytic activity OPSS from Acetobacterium sp. (AsOPSS). However, it was observed that the enzymatic activity of AsOPSS suffered significant feedback inhibition from the product L-cysteine, exhibiting an IC50 value of merely 1.2 mM. A semi-rational design combined with tunnel analysis strategy was conducted to engineer AsOPSS. The best variant, AsOPSSA218R was achieved, totally eliminating product inhibition without sacrificing catalytic efficiency. Molecular docking and molecular dynamic simulations indicated that the binding conformation of AsOPSSA218R with L-cys was altered, leading to a reduced affinity between L-cysteine and the active pocket. Tunnel analysis revealed that the AsOPSSA218R variant reshaped the landscape of the tunnel, resulting in the construction of a new tunnel. Furthermore, random acceleration molecular dynamics simulation and umbrella sampling simulation demonstrated that the novel tunnel improved the suitability for product release and effectively separated the interference between the product release and substrate binding processes. Finally, more than 45 mM of L-cysteine was produced in vitro within 2 h using the AsOPSSA218R variant. Our findings emphasize the potential for relieving feedback inhibition by artificially generating new product release channels, while also laying an enzymatic foundation for efficient L-cysteine production.

How do Chinese and Egyptian science textbooks differ? A cross-country comparative research.

Textbooks have a crucial role in shaping students' knowledge, behaviors, and attitudes in different school subjects. This study compares the structure and content of science textbooks of grade nine in Egypt and China to reveal the common and different features in the textbook design. It opts for a horizontal analysis of four science textbooks in the associated countries. The results revealed that the distribution of science subjects has partial similarities to some extent among the preparatory stage between the Chinese and Egyptian science textbooks besides the overlapping in the associated topics, presenting Biology as a common subject of interest. Moreover, the number of activities distributed within units and subjects have the highest shares in the Chinese textbooks, and most of the activities in the Egyptian textbooks focused on Chemistry and Physics subjects. In addition to the structure analysis, this study also explored the textbooks content in both countries, covering three dimensions: (1) cognitive expectations, (2) learning goals, and (3) efficiency of illustration. The results provide valuable insights for textbook designers and curriculum developers to enhance the quality of science curricula and textbooks. Therefore, the study recommends considering instructional design and lesson plans when distributing the learning activities and developing international standards for designing school science textbooks.

CTAC Self-Assembled Alkylated ß-Cyclodextrin Loaded onto Functionalized MWCNTs Electrochemical Sensor for NP Detection.

Nonylphenol (NP) is an important fine chemical raw material and intermediate that is widely utilized in industry and may be distributed in aquatic ecosystems. Following its entry into the food and water cycles, it can subsequently enter the human body and potentially harm the human reproductive system. For the purpose of monitoring NP in water, it is thus essential to build a straightforward, affordable, and robust electrochemical sensor. Based on a two-step chemical modification proceeding and an electrostatic self-assembly effect, a double-modified ß-cyclodextrin functionalized multiwalled carbon nanotube sensor (HE-ß-CD-CTAC/F-MWCNTs) has been successfully constructed. It incorporates the excellent host-guest interaction ability of ß-cyclodextrin and the high chemical activity of cetyltrimethylammonium chloride (CTAC), and the carbon nanotubes have an enormous particular surface area and strong electrical conductivity. The electrochemical oxidation reaction of NP with the sensor is controlled by a surface adsorption process of equal numbers of protons and electrons. In accordance with the optimized experimental parameters, the limit of detection (LOD) for the sensor is 0.13 µM, and it responds linearly to NP in the concentration range of 1-200 µM. Meanwhile, the sensor has excellent repeatability, stability, and immunity to interference. For the detection of NP in real water samples, the sensor also showed an excellent recovery rate (92.8%-98.5%) and relative standard deviation (1.16%-3.26%).

Research on comprehensive evaluation & development of aesthetic education based on PCA and CEM model.

Aesthetic education, conveyed through public art courses, serves as a vital form of humanistic literacy education. It represents an effective approach to fostering innovative and creative thinking among college students. In order to effectively analyze the aesthetic education work of 46 universities, an aesthetic education index evaluation system is constructed, involving indicators including faculty strength, curriculum setting, teaching management, artistic practice, and teaching support. The secondary indicators corresponding to the five indicators are statistically analyzed, and a comprehensive evaluation analysis of the current development status of aesthetic education in 46 universities in Anhui Province is conducted by combining theoretical analysis with empirical analysis. Based on principal component analysis, an integrated evaluation model for the development of aesthetic education in universities in Anhui Province is further constructed. The model designed quantifies the influence weight of each aesthetic education index on the development of aesthetic education in Anhui Province, and forges a theoretical basis for determining the precursors of rapid development of aesthetic education in Anhui Province. Additionally, a novel approach is introduced to gauge the progression of aesthetic education within universities in Anhui Province, considering the dispersion of aesthetic education index data across the province. The comprehensive evaluation model for the development of aesthetic education in Anhui Province exhibits an overall declining trend. Hence, it is suggested to utilize the maximum value of the first derivative of the comprehensive evaluation model as an indicator of the imminent rapid development of aesthetic education in Anhui Province. On this basis, the probability equation of sustainable development of aesthetic education in Anhui Province is defined. Overall, the research results lay a theoretical foundation for the development of aesthetic education in Anhui Province.

Mandarin fish von Hippel-Lindau protein regulates the NF-κB signaling pathway via interaction with IκB to promote fish ranavirus replication.

The von Hippel-Lindau tumor suppressor protein (VHL), an E3 ubiquitin ligase, functions as a critical regulator of the oxygen-sensing pathway for targeting hypoxia-inducible factors. Recent evidence suggests that mammalian VHL may also be critical to the NF-κB signaling pathway, although the specific molecular mechanisms remain unclear. Herein, the roles of mandarin fish ( Siniperca chuatsi) VHL ( scVHL) in the NF-κB signaling pathway and mandarin fish ranavirus (MRV) replication were explored. The transcription of scVHL was induced by immune stimulation and MRV infection, indicating a potential role in innate immunity. Dual-luciferase reporter gene assays and reverse transcription quantitative PCR (RT-qPCR) results demonstrated that scVHL evoked and positively regulated the NF-κB signaling pathway. Treatment with NF-κB signaling pathway inhibitors indicated that the role of scVHL may be mediated through scIKKα, scIKKß, scIκBα, or scp65. Co-immunoprecipitation (Co-IP) analysis identified scIκBα as a novel target protein of scVHL. Moreover, scVHL targeted scIκBα to catalyze the formation of K63-linked polyubiquitin chains to activate the NF-κB signaling pathway. Following MRV infection, NF-κB signaling remained activated, which, in turn, promoted MRV replication. These findings suggest that scVHL not only positively regulates NF-κB but also significantly enhances MRV replication. This study reveals a novel function of scVHL in NF-κB signaling and viral infection in fish.

Adaptive Navigation Performance Evaluation Method for Civil Aircraft Navigation Systems with Unknown Time-Varying Sensor Noise.

During civil aviation flights, the aircraft needs to accurately monitor the real-time navigation capability and determine whether the onboard navigation system performance meets the required navigation performance (RNP). The airborne flight management system (FMS) uses actual navigation performance (ANP) to quantitatively calculate the uncertainty of aircraft position estimation, and its evaluation accuracy is highly dependent on the position estimation covariance matrix (PECM) provided by the airborne integrated navigation system. This paper proposed an adaptive PECM estimation method based on variational Bayes (VB) to solve the problem of ANP misevaluation, which is caused by the traditional simple ANP model failing to accurately estimate PECM under unknown time-varying noise. Combined with the 3D ANP model proposed in this paper, the accuracy of ANP evaluation can be significantly improved. This enhancement contributes to ensured navigation integrity and operational safety during civil flight.

Constructing an artificial in vitro multi-enzyme cascade pathway to convert glycerol and CO2 into L-aspartic acid.

Developing utilization technologies for biomass resources, exploring their applications in the fields of energy and chemical engineering, holds significant importance for promoting sustainable development and constructing a green, low-carbon society. In this study, we designed a non-natural in vitro multi-enzyme system for converting glycerol and CO2 into L-aspartic acid (L-Asp). The coupled system utilized eight enzymes, including alditol oxidase (ALDO), catalase-peroxidase (CAT), lactaldehyde dehydrogenase (ALDH), glycerate 2-kinase (GK), phosphopyruvate hydratase (PPH), phosphoenolpyruvate carboxylase (PPC), L-aspartate dehydrogenase (ASPD), and polyphosphate kinase (PPK), to convert the raw materials into L-Asp in one-pot coupled with NADH and ATP regeneration. Under optimal reaction conditions, 18.6 mM of L-Asp could be produced within 2.0 h at a total enzyme addition of 4.85 mg/mL, demonstrating the high efficiency and productivity characteristics of the designed system. Our technological application provides new insights and methods for the development of biomass resource utilization technologies.

Genome-Wide Identification of the Whirly Gene Family and Its Potential Function in Low Phosphate Stress in Soybean (Glycine max).

The Whirly (WHY) gene family, functioning as transcription factors, plays an essential role in the regulation of plant metabolic responses, which has been demonstrated across multiple species. However, the WHY gene family and its functions in soybean remains unclear. In this paper, we conducted genome-wide screening and identification to characterize the WHY gene family. Seven WHY members were identified and randomly distributed across six chromosomes. The phylogenetic evolutionary tree of WHY genes in soybean and other species was divided into five clades. An in-depth analysis revealed that segmental duplications significantly contributed to the expansion of GmWHYs, and the GmWHY gene members may have experienced evolutionary pressure for purifying selection in soybeans. The analysis of promoter Cis-elements in GmWHYs suggested their potential significance in addressing diverse stress conditions. The expression patterns of GmWHYs exhibited tissue-specific variations throughout the different stages of soybean development. Additionally, six GmWHY genes exhibited different responses to low phosphate stress. These findings will provide a theoretical basis and valuable reference for the future exploration of WHY gene function.

Instant catapult steam explosion combined with ammonia water: A complex technology for detoxification of aflatoxin-contaminated peanut cake with the aim of producing a toxicity-free and nutrients retention of animal feed.

Aflatoxin is one of the most toxic biotoxins found in contaminated agricultural products. It has strong mutagenicity, carcinogenesis and teratogenicity to humans and animals. In this study, instant catapult steam explosion combined with ammonia water was examined for its potential to degrade aflatoxin B1 in peanut cake in order to improve its utilization as a toxic-free animal feed. Incubation of AFB1-containing peanut cake followed by processing with Instant Catapult Steam Explosion (ICSE) led to approximately 79.03 % degradation of AFB1, while the degradation of AFB1 was up to 91.48 % under the treatment of ICSE combined with 4 % NH3·H2O at 1.2 MPa in 200 s of process time. After treatment, nutrients in peanut cake were not significantly changed. The toxicity of AFB1 degradation products was evaluated and the results showed that the toxicity of these products were found to be substantially less than that possessed by AFB1. A low chemical pollution, efficient and toxic-free technology system of AFB1 degradation was established, which detoxify aflatoxin-contaminated biomass for sustainable and safe utilization of agricultural biomass as animal feed.

Magnetic Resonance Imaging-Related Anatomic and Functional Parameters for the Diagnosis and Prognosis of Chiari Malformation Type I: A Systematic Review and Meta-analysis.

OBJECTIVE: Imaging parameters of Chiari malformation type I (CMI) development are not well established. This study aimed to collect evidence of general or specific imaging measurements in patients with CMI, analyze indicators that may assist in determining the severity of CMI, and guide its diagnosis and treatment. METHODS: A comprehensive search was conducted across various databases including the Cochrane Library, PubMed, MEDLINE, Scopus, and Embase, covering the period from January 2002 to October 2023, following predefined inclusion criteria. Meta-analyses were performed using RevMan (ver. 5.4). We performed a quantitative summary and systematic analysis of the included studies. This study was registered in the PROSPERO (International Prospective Register of Systematic Reviews) prior to initiation (CRD42023415454). RESULTS: Thirty-three studies met our inclusion criteria. The findings indicated that out of the 14 parameters examined, 6 (clivus length, basal angle, Boogard's angle, supraocciput lengths, posterior cranial fossa [PCF] height, and volume) exhibited significant differences between the CMI group and the control group. Furthermore, apart from certain anatomical parameters that hold prognostic value for CMI, functional parameters like tonsillar movement, obex displacement, and cerebrospinal fluid dynamics serve as valuable indicators for guiding the clinical management of the disease. CONCLUSION: We collated and established a set of linear, angular, and area measurements deemed essential for diagnosing CMI. However, more indicators can only be analyzed descriptively for various reasons, particularly in prognostic prediction. We posit that the systematic assessment of patients' PCF morphology, volume, and other parameters at a 3-dimensional level holds promising clinical application prospects.

Fatty Infiltration in Paraspinal Muscles: Predicting the Outcome of Lumbar Surgery and Postoperative Complications.

Lumbar spine disorders often cause lower back pain, lower limb radiating pain, restricted movement, and neurological dysfunction, which seriously affect the quality of life of middle-aged and older people. It has been found that pathological changes in the spine often cause changes in the morphology and function of the paraspinal muscles (PSMs). Fatty infiltration (FI) in PSMs is closely associated with disc degeneration and Modic changes. And FI causes inflammatory responses that exacerbate the progression of lumbar spine disease and disrupt postoperative recovery. Magnetic resonance imaging can better distinguish between fat and muscle tissue with the threshold technique. Three-dimensional magnetic resonance imaging multi-echo imaging techniques such as water-fat separation and proton density are currently popular for studying FI. Muscle fat content obtained based on these imaging sequences has greater accuracy, visualization, acquisition speed, and utility. The proton density fat fraction calculated from these techniques has been shown to evaluate more subtle changes in PSMs. Magnetic resonance spectroscopy can accurately reflect the relationship between FI and the degeneration of PSMs by measuring intracellular and extracellular lipid values to quantify muscle fat. We have pooled and analyzed published studies and found that patients with spinal disorders often exhibit FI in PSMs. Some studies suggest an association between FI and adverse surgical outcomes, although conflicting results exist. These suggest that clinicians should consider FI when assessing surgical risks and outcomes. Future studies should focus on understanding the biological mechanisms underlying FI and its predictive value in spinal surgery, providing valuable insights for clinical decision-making.

Identification of ferroptosis-related gene signature for tuberculosis diagnosis and therapy efficacy.

Diagnosis of tuberculosis remains a challenge when microbiological tests are negative. Immune cell atlas of patients with tuberculosis and healthy controls were established by single-cell transcriptome. Through integrated analysis of scRNA-seq with microarray and bulk RNA sequencing data, a ferroptosis-related gene signature containing ACSL4, CTSB, and TLR4 genes that were associated with tuberculosis disease was identified. Four gene expression datasets from blood samples of patients with tuberculosis, latent tuberculosis infection, and healthy controls were used to assess the diagnostic value of the gene signature. The areas under the ROC curve for the combined gene signature were 1.000, 0.866, 0.912, and 0.786, respectively, in differentiating active tuberculosis from latent infection. During anti-tuberculosis treatment, the expression of the gene signature decreased significantly in cured patients with tuberculosis. In conclusion, the ferroptosis-related gene signature was associated with tuberculosis treatment efficacy and was a promising biomarker for differentiating active tuberculosis from latent infection.

Haplotype-resolved assembly of the mule duck genome using high-fidelity sequencing technology.

Mule duck is vitally important to the production of global duck meat. Here, we present two high-quality haplotypes of a female mule duck (haplotype 1 (H1):1.28 Gb, haplotype 2 (H2): 1.40 Gb). The continuity (H1: contig N50 = 14.90 Mb, H2: contig N50 = 15.70 Mb) and completeness (BUSCO: H1 = 96.9%, H2 = 97.3%) are substantially better than those of other duck genomes. We detected the structural variations (SVs) in H1 and H2. We observed a positive correlation between autosome length and the number of SVs. Z chromosome was some deficient in deletions and insertions, but W chromosome was some excessive. A total of 1,451 genes were haplotype specific expression (HSEs). Among them, 737 specifically expressed in H1, and 714 specifically expressed in H2. We found that H1 and H2 HSEs tended to be involved in similar biological processes, such as myometrial relaxation and contraction pathways, muscle structure development and phosphorylation. Our haplotype-resolved genome assembly provides a powerful platform for future functional genomics, molecular breeding, and genome editing in mule duck.

Combination of total glucosides of paeony, narrow-band ultraviolet B, and oral corticosteroid mini-pulse therapy for nonsegmental vitiligo: A retrospective study.

BACKGROUND: The total glucoside of paeony (TGP) is recognized for its immunomodulatory properties and anti-inflammatory effects. This study evaluates the efficacy of TGP combined with oral mini-pulse therapy (OMP) and narrow-band ultraviolet B (NB-UVB) in treating active nonsegmental vitiligo (NSV). MATERIALS AND METHODS: The combination therapy was contrasted against those from a group treated solely with OMP and NB-UVB. Data from 62 patients undergoing TGP combination treatment and 55 without were analyzed over a 3-month period. After 6 months, the differences in recurrence rate were investigated by follow-up. RESULTS: The findings indicate that integrating TGP may yield superior outcomes compared to OMP + NB-UVB alone. Moreover, the patient's oxidative stress makers were significantly reduced after the treatment. The majority of patients in the TGP cohort exhibited enhanced skin pigmentation over the duration. Notably, no increase in side effects or recurrence was observed in this group. Especially, patients with vitiligo on their head and neck experienced pronounced improvements. CONCLUSION: The efficacy of the combination treatment group was better than that of the control group at 2 and 3 months, and there was no difference in recurrence rate and side effects, suggesting that TGP may continue to show efficacy in NSV for a longer period of time by reducing the level of oxidative stress, and is especially suitable for patients with head and neck lesions.

De novo biosynthesis of ß-arbutin in Corynebacterium glutamicum via pathway engineering and process optimization.

BACKGROUND: ß-Arbutin, a hydroquinone glucoside found in pears, bearberry leaves, and various plants, exhibits antioxidant, anti-inflammatory, antimicrobial, and anticancer effects. ß-Arbutin has wide applications in the pharmaceutical and cosmetic industries. However, the limited availability of high-performance strains limits the biobased production of ß-arbutin. RESULTS: This study established the ß-arbutin biosynthetic pathway in C. glutamicum ATCC13032 by introducing codon-optimized ubiC, MNX1, and AS. Additionally, the production titer of ß-arbutin was increased by further inactivation of csm and trpE to impede the competitive metabolic pathway. Further modification of the upstream metabolic pathway and supplementation of UDP-glucose resulted in the final engineered strain, C. glutamicum AR11, which achieved a ß-arbutin production titer of 7.94 g/L in the optimized fermentation medium. CONCLUSIONS: This study represents the first successful instance of de novo ß-arbutin production in C. glutamicum, offering a chassis cell for ß-arbutin biosynthesis.

Ecosystem engineers can regulate resource allocation strategies in associated plant species.

Balancing the biomass requirements of different functions for the purpose of population reproduction and persistence can be challenging for alpine plants due to extreme environmental stresses from both above- and below-ground sources. The presence of ecosystem engineers in alpine ecosystems effectively alleviates microenvironmental stresses, hence promoting the survival and growth of other less stress-tolerant species. However, the influence of ecosystem engineers on plant resource allocation strategies remains highly unexplored. In this study, we compared resource allocation strategies, including biomass accumulation, reproductive effort (RE), root fraction (RF), as well as relationships between different functions, among four alpine plant species belonging to Gentianaceae across bare ground, tussock grass-, cushion-, and shrub-engineered microhabitats. Shrub-engineered microhabitats exerted the strongest effects on regulating plant resource allocation patterns, followed by tussock grass- and cushion-engineered microhabitats. Additionally, apart from microhabitats, population background and plant life history also significantly influenced resource allocation strategies. Generally, plants established within engineered microhabitats exhibited higher biomass accumulation, as well as increased flower, leaf and stem production. Furthermore, individuals within engineered microhabitats commonly displayed lower RF, indicating a greater allocation of resources to above-ground functions while reducing allocation to root development. RE of annual plants was significantly higher than that of perennial plants. However, individuals of annual plants within engineered microhabitats showed lower RE compared to their counterparts in bare ground habitats; whereas perennial species demonstrated similar RE between microhabitat types. Moreover, RE was generally independent of plant size in bare-ground habitats but exhibited size-dependency in certain populations for some species within specific engineered microhabitat types. However, size-dependency did exist for absolute reproductive and root biomass allocation in most of the cases examined here. No trade-offs were observed between flower mass and flower number, nor between leaf mass and leaf number. The capacity of ecosystem engineers to regulate resource allocation strategies in associated plants was confirmed. However, the resource allocation patterns resulted synergistically from the ecosystem engineering effects, population environmental backgrounds, and plant life history strategies. In general, such regulations can improve individual survival and reproductive potential, potentially promoting population persistence in challenging alpine environments.

Engineering a Robust UDP-Glucose Pyrophosphorylase for Enhanced Biocatalytic Synthesis via ProteinMPNN and Ancestral Sequence Reconstruction.

UDP-glucose is a key metabolite in carbohydrate metabolism and plays a vital role in glycosyl transfer reactions. Its significance spans across the food and agricultural industries. This study focuses on UDP-glucose synthesis via multienzyme catalysis using dextrin, incorporating UTP production and ATP regeneration modules to reduce costs. To address thermal stability limitations of the key UDP-glucose pyrophosphorylase (UGP), a deep learning-based protein sequence design approach and ancestral sequence reconstruction are employed to engineer a thermally stable UGP variant. The engineered UGP variant is significantly 500-fold more thermally stable at 60 °C and has a half-life of 49.8 h compared to the wild-type enzyme. MD simulations and umbrella sampling calculations provide insights into the mechanism behind the enhanced thermal stability. Experimental validation demonstrates that the engineered UGP variant can produce 52.6 mM UDP-glucose within 6 h in an in vitro cascade reaction. This study offers practical insights for efficient UDP-glucose synthesis methods.

Recombinant Art v4.01 protein produces immunological tolerance by subcutaneous immunotherapy in a wormwood pollen-driven allergic asthma female mouse model.

Art v4.01 is a well-known profilin protein belonging to the pan-allergens group and is commonly involved in triggering allergic asthma, polyallergy, and cross-sensitization. It is also referred to as Wormwood due to its origin. Crude wormwood extracts are applied for allergen-specific immunotherapy (AIT). Whether the recombinant Art v4.01 (rArt v4.01) can produce in vivo immunological tolerance by subcutaneous immunotherapy (SCIT) remains elusive. In this study, to investigate the in vivo immunological response of rArt v4.01, Th2, Th1, Treg, Th17 type-related cytokines and phenotypes of immune cells were tested, facilitating the exploration of the underlying mechanisms. The expression and purification of Art v4.01 were carried out using recombinant techniques. Allergic asthma female BALB/c mice were induced by subcutaneous sensitization of wormwood pollen extract and intranasal challenges. SCIT without adjuvant was performed using the rArt v4.01 and wormwood pollen extract for 2 weeks. Following exposure to challenges, the levels of immunoglobulin E (IgE), cytokines, and inflammatory cells were assessed through enzyme-linked immunosorbent assay (ELISA) and histological examination of sera, bronchoalveolar lavage fluid (BALF), and lung tissue. These parameters were subsequently compared between treatment groups receiving rArt v4.01 and wormwood pollen extract. The rArt v4.01 protein was expressed, which had a high purity (>90%) and an allergenic potency. Compared with the pollen extract, rArt v4.01 was superior in terms of reducing the number of white blood cells (WBCs), total nucleated cells (TNCs), and monocytes (MNs) in BALF and the degree of lung inflammation (1.77±0.99 vs. 2.31±0.80, P > 0.05). Compared with the model group, only rArt v4.01 reduced serum IgE level (1.19±0.25 vs. 1.61±0.17 µg/ml, P = 0.062), as well as the levels of Th2 type-related cytokines (interleukin-4 (IL-4) (107.18±16.17 vs. 132.47±20.85 pg/ml, P < 0.05) and IL-2 (19.52±1.19 vs. 24.02±2.14 pg/ml, P < 0.05)). The study suggested that rArt v4.01 was superior to pollen extract in reducing the number of inflammatory cells in BALF, pneumonitis, levels of pro-inflammatory cytokines, and serum IgE level. These findings confirmed that Art v4.01 could be a potential candidate protein for allergen-specific immunotherapy.