Search Results (327)

The olive oil sector constitutes a fundamental pillar in the Mediterranean region from socio-economic and cultural perspectives. Nonetheless, it produces significant amounts of waste, leading to numerous environmental issues. These waste streams contain valuable compounds that can be recovered and utilized as inputs for various applications. This study introduces a novel value chain for olive wastes, focused on extracting lignin from olive pomace by ionic liquids and polyphenols from olive mill wastewater, which are then incorporated as hybrid nanoparticles in the formulation of an innovative starch-based biofertilizer. This biofertilizer, obtained by using residual wastewater as a source of soluble nitrogen, acting at the same time as a plasticizer for the biopolymer, was demonstrated to surpass traditional NPK biofertilizers’ efficiency, allowing for root growth and foliage in drought conditions. In order to recognize the environmental impact due to its production and align it with the technical output, the circularity and environmental performance of the proposed system were innovatively evaluated through a combination of Life Cycle Assessment (LCA) and the Material Circularity Indicator (MCI). LCA results indicated that the initial upcycling process was potentially characterized by significant hot spots, primarily related to energy consumption (>0.70 kWh/kg of water) during the early processing stages. As a result, the LCA score of this preliminary version of the biofertilizer may be higher than that of conventional commercial products, due to reliance on thermal processes for water removal and the substantial contribution (56%) of lignin/polyphenol precursors to the total LCA score. Replacing energy-intensive thermal treatments with more efficient alternatives represents a critical area for improvement. The MCI value of 0.84 indicates limited potential for further enhancement. Full article

Background/Objectives: Epilepsy affects approximately 50 million people worldwide, with antiepileptic drugs (AEDs) remaining the cornerstone of treatment. Due to their narrow therapeutic windows, AEDs are ideal candidates for therapeutic drug monitoring (TDM). Oral fluid is increasingly considered a viable alternative to blood and urine, as it reflects the free (active) concentration of many AEDs. Its non-invasive collection, which does not require trained personnel, makes it particularly suitable for TDM in paediatric and geriatric populations. However, as samples are often stored for extended periods before analysis, analyte stability becomes a critical concern. This study aimed to evaluate the stability of four commonly used AEDs in dried saliva spot (DSS) samples. Methods: Phenobarbital, phenytoin, carbamazepine, and carbamazepine-10,11-epoxide were analysed in oral fluid samples collected via spitting and stored as DSSs. Quantification was performed using high-performance liquid chromatography with diode array detection (HPLC-DAD). Design of experiments tools were used to assess the effects of preservatives, storage temperatures, light exposure, and storage durations on analyte stability. Results: Optimal conditions were refrigeration in the dark, with a low concentration of ascorbic acid as preservative. Samples at 10 µg/mL remained stable for 14 days longer than those without preservative or reported in previous studies. Unexpectedly, at 0.5 µg/mL, analytes in samples without preservative showed greater stability. Conclusions: To our knowledge, this is the first study combining DSS and HPLC-DAD to assess the stability of these AEDs in oral fluid, providing valuable insights for non-invasive TDM strategies and supporting the feasibility of saliva-based monitoring in clinical settings. Full article

During fruit ripening in Capsicum species, substantial amounts of carotenoids accumulate in the pericarp. While the carotenoid biosynthesis pathway in Capsicum species has been extensively investigated from various angles, the transcriptional regulation of genes encoding carotenoid biosynthetic enzymes remains less understood in this non-climacteric horticultural crop compared to tomato, a climacteric fruit. In the present study, we investigated the function of the NAM, ATAF1/2 or CUC2 81 (CaNAC81) transcription factor gene. This gene was selected through RNA-Seq co-expression analysis based on the correlation between expressed transcription factor gene profiles and those of carotenoid structural genes. To determine its role in regulating the expression of biosynthetic-related carotenogenic genes, we performed Virus-Induced Gene Silencing (VIGS) assays in the Serrano-type C. annuum ‘Tampiqueño 74’. Fruits from plants infected with a pTRV2:CaNAC81 construct (silenced fruits) exhibited altered carotenoid pigmentation accumulation, manifested as yellow-orange spots, in contrast to fruits from non-agroinfected controls (NTC) and fruits from plants infected with the empty TRV2 construct (red fruits). Quantitative real-time PCR (qPCR) assays confirmed decreased transcript levels of CaNAC81 in fruits displaying altered pigmentation, along with reduced transcription of the PSY gene, which encodes the carotenoid biosynthetic enzyme phytoene synthase (PSY). High-performance liquid chromatography (HPLC) analysis revealed a distinct carotenoid pigment accumulation pattern in fruits from plants showing silencing symptoms, characterized by low concentrations of capsanthin and zeaxanthin and trace amounts of capsorubin, compared to control plants (NTC). These findings suggest the involvement of CaNAC81 in the regulatory network of the carotenoid biosynthetic pathway in chili pepper fruits. Full article

Background/Objectives: Over the past few decades, controlled substance abuse in drug-facilitated sexual assaults (DFSAs) has significantly increased worldwide, leading to an urgency to develop rapid and selective drug detection methods for field use (i.e., on-spot detection). Currently, techniques for detecting DFSA drug-associated samples are laborious and require skilled personnel to analyze/interpret the results. Moreover, most DFSA-associated drugs have a short half-life, making them more challenging to detect promptly. For instance, the timely detection of gamma-hydroxybutyrate (GHB) has been of ultimate concern for decades due to its fast elimination from the body. This study describes the development of a fluorescent ionic liquid nanosensor that can be used to rapidly detect GHB in the field. Methods: Trihexyltetradecylphosphonium fluorescein (THP₂FL) ionic liquid was synthesized and evaluated for its potential application in detecting GHB. THP₂FL nanoparticles in deionized water were synthesized with a size of 199 nm by a reprecipitation method. Results: The addition of GHB to THP₂FL nanoparticles resulted in up to a 60% increase in fluorescence intensity and a 79% increase in absorbance. These results suggest potential applications for using the fluorescent THP₂FL nanoparticles to detect GHB. The sensor’s selectivity was tested on compounds structurally similar to GHB, and the results showed that 1,4-butanediol (a precursor of GHB) is a potentially interfering species. Conclusion: This fluorescent technique allows for field deployable sensors, which would benefit screening GHB onsite. Full article

This study evaluated the acaricidal effects of biosurfactants produced by Serratia ureilytica against the two-spotted spider mite Tetranychus urticae and their compatibility with the predatory mite Ambliseus swirski. The biosurfactants were obtained via liquid cultures of the bacterial strains. In the laboratory, T. urticae was exposed via acaricide-immersed leaves and A. swirskii via acaricide-coated glass vials. In the greenhouse, mite-infested plants were sprayed with the biosurfactants. In the laboratory, biosurfactants produced by S. ureilytica NOD-3 and UTS exhibited strong acaricidal activity, causing 95% mortality in adults and reducing egg viability by more than 60%. In the greenhouse trial, all biosurfactants significantly suppressed T. urticae populations at all evaluated periods (7, 14, and 21 days post-application). Gas chromatography–mass spectrometry (GC-MS) analysis of the biosurfactants identified several fatty acids, including hexadecanoic acid, pentanoic acid, octadecanoic acid, decanoic acid, and tetradecanoic acid, as well as the amino acids L-proline, L-lysine, L-valine, and glutamic acid. These fatty acids and amino acids are known structural components of lipopeptides. Furthermore, the bioinformatic analysis of the genomes of the three S. ureilytica strains revealed nonribosomal peptide synthetase (NRPS) gene clusters homologous to those involved in the biosynthesis of lipopeptides. These findings demonstrate that S. ureilytica biosurfactants are promising eco-friendly acaricides, reducing T. urticae populations by >95% while partially sparing A. swirskii. Full article

The pathways and mechanisms of primary hydrocarbon migration, which are still not well understood, are of great significance for evaluating both conventional and unconventional oil and gas resources, understanding the mechanisms of shale oil retention, and predicting sweet spots. To investigate the petrography, geochemistry, and pore systems of organic-rich mudstones and organic-lean sand-silt intervals in core samples from the Yanchang shale in the Ordos Basin, China, we conducted thin-section observation, X-ray diffraction, Rock-Eval pyrolysis, field emission scanning electron microscopy (FE-SEM), and porosity analysis. Sand-silt intervals are heterogeneously developed within the Yanchang shale. The petrology, mineral composition, geochemistry, type, and content of solid organic matter as well as the pore type, pore size, and porosity of these intervals differ significantly from those of mudstones. Compared with mudstones, sand-silt intervals typically have coarser detrital grain sizes, higher contents of quartz, feldspar, and migrated solid bitumen (MSB), larger pore sizes, higher porosity, and higher oil saturation index (OSI). In contrast, they have lower contents of clay minerals, total organic carbon (TOC), free liquid hydrocarbons (S₁), and total residual hydrocarbons (S₂). The sand-silt intervals in the Yanchang shale serve as both pathways for hydrocarbon primary migration and “micro reservoirs” for hydrocarbon storage. The interconnected inorganic and organic pore systems, organic matter networks, fractures, and sand-silt intervals form the hydrocarbons’ primary migration pathways within the Yanchang shale. A model for the primary migration of hydrocarbons within the Yanchang shale is proposed. Full article

Pear black spot disease seriously threatens the pear industry. Currently, its control mainly relies on chemical fungicides while biological control using antagonistic microorganisms represents a promising alternative approach. This study identified and characterized Bacillus velezensis TRMB57782 as a biocontrol strain through whole-genome sequencing. AntiSMASH analysis predicted the strain’s potential to produce secondary metabolites such as surfactin, difficidin, and bacilysin. In vitro experiments demonstrated that TRMB57782 inhibited the growth of Alternaria gaisen. In vivo experiments using excised branches and pear fruits at two different stages also showed significant control effects. A preliminary exploration of the metabolic substances of TRMB57782 was carried out. The strain can produce siderophores and three biocontrol enzymes. Crude extracts obtained by the hydrochloric acid precipitation and ammonium sulfate saturation precipitation of the bacterial liquid exhibited significant activity and volatile organic compounds showed biocontrol activity. Meanwhile, the effects of strain TRMB57782 on the hyphae of pathogenic fungi were studied, leading to hyphal atrophy and spore shrinkage. This paper provides an effective biocontrol strategy for fragrant pear black spot disease, reveals the antibacterial mechanism of Bacillus velezensis TRMB57782, and offers a new option for the green control of pear black spot disease. Full article

A double-sided telecentric zoom optical system can ensure the measurement and detection accuracy for different workpiece sizes and plays a crucial role in industrial detection. The conventional double-sided telecentric mechanical zoom system is faced with the problem of complex structure and difficult focusing. To address these issues, a liquid lens is applied to design a non-displacement double-sided telecentric zoom system in this paper. Here, the design method of the conventional double telecentric zoom system is analyzed first; then, the liquid lenses are substituted for the mechanical motion groups. Finally, a double-sided telecentric zoom system with a detection range of 25~60 mm and a magnification of −0.44×~−0.183× has been designed and optimized by Zemax software. The design results show that in the process of magnification, the root mean square radius of the diffuse spot in the system is smaller than the pixel size during the process of zooming. The modulation transfer function values at the Nyquist frequency of 80 lp/mm are all above 0.4, distortion is controlled within 0.2, and telecentricity is less than 0.5°, indicating that the system has excellent imaging quality, low distortion, and high telecentric and other characteristics, which meet system requirements. The design method proposed in this paper can provide an effective solution for the rapid conversion of displacement zoom systems to non-displacement zoom systems. Full article

Background: Furosemide is a loop diuretic used extensively to treat adult and pediatric patients. In some hospitals, furosemide oral liquids are not available in stock, thus necessitating the extemporaneous preparation of the drug. This study evaluates the stability of on-the-spot formulations of furosemide oral suspensions from crushed tablets evaluated in various vehicles: Dextrose 50%, Dextrose 70%, Ora-Sweet, and Ora-Plus over 60 days. This examination was prompted by the frequent shortage of certain excipients in the hospital, leading to the need to switch to Dextrose 50% or Dextrose 70% when Ora-Sweet and Ora-Plus are out of stock. Methods: The extemporaneous furosemide oral suspensions were prepared following the same compounding method used in the pharmacy. The suspensions were maintained at 4 °C in the refrigerator and assessed immediately and later, on days 7, 14, 30, and 60. The assessed parameters included visual appearance, redispersion time, sedimentation volume, and pH levels for stability analysis. We also examined the drug content, dissolution of the suspension, and microbiological stability. Results: Initial examinations indicated that Dextrose 50% and Ora-Plus maintained pH levels and stable appearances, while significant changes, mainly in appearance and redispersion time, indicated the instability of Dextrose 70%. Ora-Sweet showed fluctuations but stabilized by day 30. Dissolution studies demonstrated that Ora-Plus had dissolution characteristics superior to the other formulations, while Dextrose 50% showed declining dissolution percentages over time. Overall, the Ora-Plus vehicle showed superior stability (60 days), followed by Ora-Sweet (30 days), while Dextrose 70% and Dextrose 50% showed shorter stability durations of 14 and 7 days, respectively. The microbiological test results showed no microbial growth. Conclusions: This study demonstrates that the vehicle used in extemporaneous furosemide suspensions critically affects their stability and performance. Ora-Plus emerged as the most suitable vehicle, maintaining physical, chemical, and microbiological stability over 60 days, with consistent pH, redispersion, and dissolution behavior. Ora-Sweet showed intermediate stability (30 days), while Dextrose 50% and 70% exhibited early instability—7 and 14 days, respectively—marked by sedimentation, poor redispersibility, and declining drug release. These findings underscore the importance of vehicle selection and regular stability monitoring in compounded formulations to ensure therapeutic reliability and patient safety. Full article

Pseudomonas aeruginosa (P. aeruginosa) is a common antibiotic-resistant pathogen, posing significant public health threats worldwide. It is a major cause of ocular infections, mostly linked to contact lens wear. P. aeruginosa often produces biofilm during infections, and these are also associated with antibiotic resistance. Bacteriophage (phage) therapy is emerging as a promising approach for treating multidrug-resistant P. aeruginosa. Objective: This study aimed to assess the antibiofilm effects of six phages against P. aeruginosa biofilms isolated from patients with corneal infections. Method: This study examined P. aeruginosa strains for their ability to form biofilms using crystal violet assay. Six P. aeruginosa bacteriophages (DiSu1 to DiSu6) were used, which were isolated from sewage water in Melbourne, Australia. Spot tests were used to assess phage sensitivity. The effect of phages against P. aeruginosa strains was determined using time–kill assay and efficiency of plating. The ability of phage to inhibit biofilm formation over 24 h or reduce preformed biofilms was also studied and confirmed using confocal laser scanning microscopy with Live/Dead staining. Result: After 24 h of incubation, all tested P. aeruginosa strains formed moderate to strong biofilms. All P. aeruginosa strains were sensitive to at least four of the six phages. The highest level of bacterial growth inhibition in the liquid infection model was observed when phages were applied at a multiplicity of infection (MOI) of 100. Certain bacteria/phage combinations were able to inhibit biofilm formation over 24 h, with the combination of strain PA235 and phage DiSu3 producing the highest inhibition (83%) at a MOI of 100. This was followed by the combinations of PA223/DiSu3 (56%), and PA225/DiSu5 (52%). For the reduction in preformed biofilms, the best combinations were PA235 (90%), PA221 (61%), and PA213 and PA225 (57% each), all with DiSu3 after 3 h. However, exposing the biofilm with phages for over 24 h appeared to promote phage resistance as there was evidence of biofilm growth, with the only combination still showing a significant reduction being PA221/DiSu3 (58%) at MOI of 100. Conclusions: This study showed that the effect of phages against P. aeruginosa is concentration (MOI) dependent. Phages at higher MOI have the ability to disrupt, inhibit, and reduce P. aeruginosa biofilms. However, prolonged exposure of the biofilm with phages appeared to promote phage resistance. To enhance phage efficacy and address this form of resistance, further studies utilizing phage cocktails or a combination of phages and antibiotics is warranted. Full article

Among grapevine trunk diseases, Eutypa dieback, caused by the fungus Eutypa lata, is one of the most critical ones, due to its widespread infection in vineyards and the lack of effective treatments. This fungus is a vascular pathogen that enters grapevines through pruning wounds. The infection process is associated with phytotoxic metabolites produced by the fungus, and as such, the identification of new metabolites from different culture conditions and broths could provide valuable insights into the fungus’s enzymatic system and help its control. For the purposes of this study, the OSMAC (one strain, many compounds) approach was applied to investigate the secondary metabolism of E. lata strain 311 isolated from Vitis vinifera plants in Spain. A total of twenty metabolites were isolated, including five reported for the first time from E. lata and four that are newly identified compounds in the literature: eulatagalactoside A, (R)-2-(4′-hydroxy-3′-methylbut-1′-yn-1′-yl)-4-(hydroxymethyl)phenol, (S)-7-hydroxymethyl-3-methyl-2,3-dihydro-1-benzoxepin-3-ol, and (3aR,4S,5R,7aS)-4,5-dihydroxy-6-((R)-3′-methylbuta-1′,3′-dien-1′-ylidene)hexahydrobenzo[d][1,3]dioxol-2-one. These compounds were extracted from fermentation broths using silica gel column chromatography and high-performance liquid chromatography (HPLC). Their structures were elucidated through extensive 1D and 2D NMR spectroscopy, along with high-resolution electrospray ionization mass spectrometry (HRESIMS). Compounds were evaluated for phytotoxicity against Phaseolus vulgaris, with only eulatagalactoside A producing white spots after 48 h. Additionally, the antibacterial activity against Escherichia coli, Staphylococcus aureus, and Klebsiella pneumoniae of selected compounds was tested. The compounds (R)-2-(4′-hydroxy-3′-methylbut-1′-yn-1′-yl)-4-(hydroxymethyl)phenol and (S)-7-(hydroxymethyl)-3-methyl-2,3-dihydrobenzo[b]oxepin-3-ol showed the most significant antimicrobial activity against Gram-positive bacteria, inhibiting S. aureus by over 75%, with IC₅₀ values of 511.4 µg/mL and 617.9 µg/mL, respectively. Full article

Skin aging is a complex process influenced by several factors, including UV exposure, environmental stressors, and lifestyle choices. The demand for effective, natural skincare products has driven research into plant-based oils rich in bioactive compounds. Rosehip oil has garnered attention for its high content of carotenoids, phenolics, and antioxidants, which are known for their anti-aging, photoprotective, and skin-rejuvenating properties. Despite the growing interest in rosehip oil, limited studies have investigated its efficacy on human skin using advanced imaging technologies. This study aims to fill this gap by evaluating the efficacy of cold-pressed Rosa canina seed oil on facial skin characteristics, specifically wrinkles, ultraviolet (UV) spot reduction, and erythema mitigation, using imaging technologies (the VISIA analysis system). Seed oil pressed from R. canina collected from the Băișoara area of Cluj County has been selected for this study due to its high carotenoid, phenolic, and antioxidant contents. The oil has also been analyzed for the content of individual carotenoids (i.e., lutein, lycopene, β Carotene, and zeaxanthin) using HPLC-DAD (High-Performance Liquid Chromatography—Diode Array Detector), along with lutein and zeaxanthin esters and diesters. After the preliminary screening of multiple Rosa species for carotenoid, phenolic, and antioxidant contents, the R. canina sample with the highest therapeutic potential was selected. A cohort of 27 volunteers (aged 30–65) underwent a five-week treatment protocol, wherein three drops of the selected rosehip oil were topically applied to the face daily. The VISIA imaging was conducted before and after the treatment to evaluate changes in skin parameters, including the wrinkle depth, UV-induced spots, porphyrins, and texture. Regarding the bioactivities, rosehip oil showed a significant total carotenoids content (28.398 μg/mL), with the highest levels in the case of the β-carotene (4.49 μg/mL), lutein (4.33 μg/mL), and zexanthin (10.88 μg/mL) contents. Results indicated a significant reduction in mean wrinkle scores across several age groups, with notable improvements in individuals with deeper baseline wrinkles. UV spots also showed visible declines, suggesting ideal photoprotective and anti-pigmentary effects attributable to the oil’s high vitamin A and carotenoid content. Porphyrin levels, often correlated with bacterial activity, decreased in most subjects, hinting at an additional antimicrobial or microbiome-modulatory property. However, skin responses varied, possibly due to individual differences in skin sensitivity, environmental factors, or compliance with sun protection. Overall, the topical application of R. canina oil appeared to improve the facial skin quality, reduce the appearance of age-related markers, and support skin health. These findings reinforce the potential use of rosehip oil in anti-aging skincare formulations. Further long-term, large-scale studies are warranted to refine dosing regimens, investigate mechanisms of action, and explore synergistic effects with other bioactive compounds. Full article

The production of gas and condensate from liquid-rich shale reservoirs, particularly within heterogeneous lacustrine systems, remains a critical challenge in unconventional hydrocarbon exploration due to intricate multiphase hydrocarbon partitioning, including gases (C₁–C₂), volatile liquids (C₃–C₇), and heavier liquids (C₇₊). This study investigates a 120-meter-thick interval dominated by lacustrine deposits from the Lower Cretaceous Shahezi Formation (K₁sh) in the Songliao Basin. This interval, characterized by high clay mineral content and silicate–pyrite laminations, was examined to identify the factors controlling hybrid shale gas condensate systems. We proposed the Hybrid Shale Condensate Index (HSCI), defined as the molar ratios of (C₁–C₇)/C₇₊, to categorize fluid phases and address shortcomings in traditional GOR/API ratios. Over 1000 samples were treated by geochemical pyrolysis logging, X-ray fluorescence (XRF) spectrum element logging, SEM-based automated mineralogy, and in situ gas desorption, revealing four primary controls: (1) Thermal maturity thresholds. Mature to highly mature shales exhibit peak condensate production and the highest total gas content (TGC), with maximum gaseous and liquid hydrocarbons at T_max = 490 °C. (2) Lithofacies assemblage. Argillaceous shales rich in mixed carbonate and clay minerals exhibit an intergranular porosity of 4.8 ± 1.2% and store 83 ± 7% of gas in intercrystalline pore spaces. (3) Paleoenvironmental settings. Conditions such as humid climate, saline water geochemistry, anoxic bottom waters, and significant input of volcanic materials promoted organic carbon accumulation (TOC reaching up to 5.2 wt%) and the preservation of organic-rich lamination. (4) Laminae and fracture systems. Silicate laminae account for 78% of total pore space, and pyrite laminations form interconnected pore networks conducive to gas storage. These findings delineate the “sweet spots” for unconventional hydrocarbon reservoirs, thereby enhancing exploration for gas condensate in lacustrine shale systems. Full article

The fungus Alternaria alternata, which causes tobacco brown spot disease, poses a serious threat to the tobacco industry. Beneficial microorganisms and their secondary metabolites have emerged as a promising green strategy for disease management. This study recovered 16 endophytic bacterial strains from Mentha haplocalyx Briq., a therapeutic herb. The study revealed that strain B5, with an inhibition rate of 82.76%, exhibited the highest antifungal activity against A. alternata. This strain exhibited broad-spectrum antifungal activity, with inhibition rates ranging from 66.34% to 87.23%. Phylogenetic analysis of 16S rDNA and gyrA gene sequences identified it as Bacillus velezensis (GenBank: PV168970 and PV173738). Further characterization revealed that strain B5 can secrete cell wall-degrading enzymes, produce IAA, and synthesize siderophores. The growth of mycelium in A. alternata was greatly reduced by both the ethyl acetate extract and the filtered liquid from the sterile fermentation, resulting in marked morphological abnormalities. Multiple antifungal active substances were identified through liquid LC-MS analysis. Greenhouse experiments demonstrated that the B5 fermentation broth effectively suppressed the occurrence of tobacco brown spot disease, achieving a relative control efficacy of 60.66%, comparable to that of 10% difenoconazole water dispersible granule (WDG). Additionally, strain B5 enhances plant disease resistance by activating the activities of key defense enzymes. B. velezensis B5 serves as a safe alternative to chemical fungicides and is highly effective at controlling tobacco brown spot disease. Full article

The areca palm (Areca catechu L.), a medicinal tropical crop, hosts three novel viruses, areca palm necrotic ringspot virus (ANRSV), areca palm necrotic spindle-spot virus (ANSSV), and ANRSV2, which form a new genus Arepavirus in the family Potyviridae. Both viruses feature a unique tandem leader protease arrangement (HCPro1-HCPro2). To elucidate HCPro2’s role, this study identified its interaction partners in infected cells using affinity purification coupled with liquid chromatography-tandem mass spectrometry, a yeast two-hybrid system, and co-immunoprecipitation. Thirteen host proteins and five viral factors (HCPro1, 6K2, VPg, NIa-Pro, NIb) were validated as HCPro2 interactors. Among the host proteins interacting with HCPro2, the expression of five genes (NbeIF4A, NbSAMS1α, NbTEF1α, NbUEP1, and NbRan2) was upregulated under the condition of viral infection, while the expression of another five genes (NbpsbS1, NbPGK, NbchIP, NbClpC1A, and NbCysPrx) was downregulated. Functional assays showed that silencing NbeIF4A or NbPGK significantly reduced viral accumulation in Nicotiana benthamiana. These findings reveal HCPro2’s network of virus-host interaction, highlighting its critical role in viral pathogenesis. Further exploration of these interactions may clarify the evolutionary significance of tandem leader proteases and inform novel plant antiviral strategies. Full article