Search Results (5,249)

This study aimed to investigate culture conditions for the efficient micropropagation of apple cultivars ‘Golden Delicious’ and ‘Royal Gala’ in liquid medium by temporary immersion. RITA^® bioreactors were used for the multiplication stage whereas RITA^® or Plantform™ were used for the rooting stage. Murashige and Skoog media (MS) with N⁶-benzyladenine (BA) was used for shoot multiplication and indole-3-butyric acid (IBA) for root induction. During the multiplication phase, we evaluated the mineral medium, BA concentration, immersion frequency, silver nitrate and activated charcoal supplementation and the use of physical supports to hold explants in an upright position. The results demonstrated that longer incubation periods (10 weeks) were better than shorter periods (6 weeks) for decreasing hyperhydricity and increasing the multiplication coefficient (MC). For ‘Golden Delicious’, the highest MC were obtained either with explants placed directly on the bioreactor basket and immersed six times per day for 60 s in MS with 2.2 µM BA or explants placed between rockwool cubes cultivated with 4.4 µM BA (both yielding MC of 8.9 and 5–10% hyperhydricity). These results were superior to ‘Royal Gala’, which showed a MC of 7.3 and 23% of hyperhydricity when cultivated in MS with half nitrates, 1.55 µM BA and rockwool cubes. Both varieties rooted efficiently (96–100%), and resulting plantlets were successfully acclimated. This is the first report in the micropropagation of these two commercial fruiting cultivars in temporary immersion, demonstrating the potential of this technology to enhance large-scale plant production for the apple nursery industry. Full article

Iris laevigata is an ornamental plant and so its wild genetic resources need to be protected. However, traditional inefficient propagation limits its landscape applications. In this study, we assessed the effects of phytohormones on growth of I. laevigata at various culture stages using roots and hypocotyls as explants and established an efficient micropropagation system. The highest callus induction of hypocotyl (75.0%) was obtained using Murashige and Skoog medium containing 6-benzylaminopurine (6-BA), 0.5 mg L⁻¹ + 2,4-dichlorophenoxyacetic acid (2,4-D), 1.0 mg L⁻¹ + 1-naphthylacetic acid (NAA), and 0.4 mg L⁻¹. Similarly, the highest callus induction (73.3%) of roots was achieved with 6-BA 0.5 mg L⁻¹ + 2,4-D 0.5 mg L⁻¹ + NAA 0.4 mg L⁻¹. The calli induced from hypocotyl and root tissues achieved 39.7% and 49.5% adventitious shoot induction on a medium containing indole-3-butyric acid (IBA) 0.5 mg L⁻¹ + 6-BA 1.5 mg L⁻¹ + NAA 1.0 mg L⁻¹ and 6-BA 2.0 mg L⁻¹ + NAA 0.4 mg L⁻¹ + kinetin (KT) 1.0 mg L⁻¹, respectively. The rooting of adventitious shoots reached 93.3% in the medium supplemented with NAA 0.2 mg L⁻¹. The survival of regenerated plants reached 90.0% after being transplanted into soil. This study provides an efficient and reliable propagation method for I. laevigata for landscape applications and the preservation of wild genetic material. Full article

Lactiplantibacillus plantarum is a widely studied probiotic species with significant strain-specific functional diversity, yet the molecular mechanisms underlying these variations remain largely unexplored. In this study, whole genome sequencing (WGS) and untargeted metabolomics were employed to comprehensively characterize the genetic architecture and extracellular metabolic profile of Lp. plantarum FRT4 (CGMCC 17955), a probiotic strain previously studied for its metabolic effects in animal models. WGS revealed a circular chromosome and five plasmids, encoding 3301 protein-coding genes enriched in amino acid biosynthesis, carbohydrate metabolism, and environmental response pathways. Carbohydrate-active enzymes (CAZy) annotation revealed 135 carbohydrate-active enzyme genes, dominated by glycoside hydrolases and glycosyl transferases. Untargeted metabolomic analysis comparing the fermentation supernatant of FRT4 with non-inoculated MRS medium revealed significant alterations in metabolite composition, including elevated levels of acetylcholine, nicotinamide adenine dinucleotide (NAD), and trans-3-coumarate, and reduced levels of uridine, inosine, and fructose-1-phosphate, indicating active modulation of neurotransmission, redox balance, and purine metabolism. KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment showed significant regulation of pathways related to amino acid metabolism, carbon metabolism, and cofactor biosynthesis. These findings highlight the metabolic versatility and functional potential of FRT4, offering mechanistic insights into its probiotic effects and providing a basis for its potential application in fermentation-based formulations. Full article

This work develops a novel phosphate-functionalized extraction resin (HEHEHP + Cyanex272)/SiO₂-P via the vacuum impregnation method for efficient separation of light rare earth element impurities from lanthanum (La³⁺) in nitric medium through synergistic extraction. Batch experiments have demonstrated superior adsorption selectivity toward impurity ions over La³⁺ in a pH 4 nitric acid solution. Column studies confirmed exceptional performance under ambient conditions, achieving a lanthanum treatment capacity of 120.6 mg/g and over 98% impurity removal, which surpasses most reported values. Notably, this purification process enables direct production of purified La³⁺ solutions through a single-column system without desorption, significantly enhancing efficiency and reducing costs. Mechanistic insights revealed combined ion exchange and coordination interactions between metal ions and P-OH/P=O groups, corroborated by advanced characterization and density functional theory calculations. These findings indicate a higher binding affinity of light rare earth compared with La³⁺. This strategy provides a scalable approach for ultra-high-purity lanthanum compound production in advanced optical and electronic applications. Full article

Background/Objectives: Co-culture models of neurons and Schwann cells have been used to explore the mechanisms of myelination during development, axonal regeneration after injury, and the pathogenesis of various demyelinating neuropathies. A spontaneously immortalized Fischer rat Schwann cell line 1 (IFRS1), established from the primary culture of adult Fischer344 rat peripheral nerves, can myelinate neurites in co-cultures with primary cultured dorsal root ganglion neurons and neuronal cell lines, such as nerve growth factor (NGF)-primed PC12 cells and NSC-34 motor neuron-like cells. In this study, we aimed to establish a stable co-culture system using IFRS1 cells and ND7/23 sensory neuron-like cells. Methods: ND7/23 cells were seeded at a low density (2 × 10³/cm²) and maintained for 7 days in serum-containing medium supplemented with NGF (10 ng/mL) and the Rho kinase inhibitor Y27632 (5 μM) to promote neurite elongation. The cells were then treated with the anti-mitotic agent mitomycin C (1 μg/mL) for 12–16 h to suppress proliferative activity. Following this, the cells were co-cultured with IFRS1 cells (2 × 10⁴/cm²) and maintained at 37 °C in serum-containing medium supplemented with ascorbic acid (50 μg/mL), NGF (10 ng/mL), and ciliary neurotrophic factor (10 ng/mL). Results: Double-immunofluorescence staining performed on day 21 of the co-culture revealed myelin protein 22- or myelin basic protein-immunoreactive IFRS1 cells surrounding βIII tubulin-immunoreactive neurites emerging from ND7/23 cells. Myelin formation was further confirmed via Sudan Black B staining and electron microscopy. Conclusions: This co-culture system may provide a valuable tool for studying the processes of myelination in the peripheral nervous system, as well as the pathogenesis of various sensory neuropathies and potential novel therapeutic approaches for these conditions. Full article

The environmental problems brought about by factory-based aquaculture have become increasingly prominent. Reducing nitrogen and phosphorus concentrations in tailwater has become the key to tailwater management. In order to assess the potential of microalgae in removing nitrogen and phosphorus ions from aquaculture wastewater, four microalgae species, i.e., Chlorella sp., Dicrateria zhanjiangensis, Nitzschia closterium minutissima, and Platymonas subcordiformis, were used in this study, and their growth and nitrogen and phosphorus removal rates in four nutrient concentrations of simulated aquaculture wastewater were systematically evaluated. After 15 days of cultivation, the cell counts of all four types of microalgae increased. Three species, i.e., Chlorella sp., N. closterium minutissima, and P. subcordiformis, grew best in high PO₄³⁻ and low NH₄⁺ medium, whereas D. zhanjiangensis possessed best growth in low PO₄³⁻ and high NH₄⁺ medium. The removal rate of PO₄³⁻, NH₄⁺, NO₃⁻, and NO₂⁻ by four microalgae species exceeded 82.64%, 89.06%, 59.27%, and 42.15%, respectively, even though the four microalgae had different performance in the removal of nitrogen and phosphorus. All microalgae in the low-phosphorus groups removed PO₄³⁻ at significantly lower rates than those in the high–phosphorus groups, while high NH₄⁺ removal rates were observed in all four microalgae groups. Moreover, in phosphorus-limited conditions, four microalgae exhibit lower removal rates of NO₃⁻ when nitrogen content was high. The chlorophyll a contents of microalgae in four culture media strictly corresponded to their final cell densities. P. subcordiformis exhibited the highest intracellular polysaccharide accumulation in high PO₄³⁻ and low NH₄⁺ type medium, whereas D. zhanjiangensis demonstrated the strongest protein synthesis capacity in high PO₄³⁻ and low NH₄⁺ medium. The activities of acid phosphatase in all microalgae were higher under phosphorus–deficient conditions than phosphorus-sufficient conditions. Our results might provide useful references for microalgae selection in the treatment of different aquaculture wastewater conditions. Full article

Live-attenuated vaccines face a critical challenge in balancing immunogenicity with safety. To address this, we engineered programmable finite-replicated organisms (FROs) by depositing a limited number of indispensable components (such as noncanonical amino acids, ncAAs) within the cell, consuming the coenabling precise control of bacterial replication capability while preserving antigenic breadth. Two strategies were adopted to achieve the following purposes: (1) encoding ncAA in essential genes; (2) encoding ncAA in antitoxin of toxin–antitoxin (TA) systems. As noncanonical amino acids, 3,5-dichlorotyrosine (Cl2Y) was encoded by the amber codon (TAG) and inserted into the essential genes (e.g., serS, murG, and dnaA) or antitoxin genes. After optimizing expression and the number of amber codons in the storage genes, the FRO cells can grow up to six generations, achieving amplification approaching 100 times after depletion of the ncAA in the growth medium. The escape frequencies are 10⁻⁵ to 10⁻⁷, which need to be optimized by combining multiple storage genes in the same genome in the future. This work holds the potential to amplify the amounts of antigens for vaccines, potentially accelerating the development of next-generation vaccines against antibiotic-resistant threats. Full article

A continuous effort is needed to develop sustainable production methods for industrial platform chemicals. B. succiniciproducens, a natural succinic acid-producer, can metabolize five and six carbon atoms containing sugars in pure form as well as from agro-industrial wastes. In our work, we investigate the conversion of industrial by-products, apple pomace from apple juice production, and whey waste from milk processing to succinic acid and other organic acids (lactic, formic, and acetic acid). We obtained a succinic acid yield of 0.224 g/g total consumed fermentable sugars, lactic acid yield was 0.087 g/g, in turn, formic acid was produced at a 0.034 g/g yield, and acetic acid was obtained at 0.010 g/g total consumed fermentable sugars, using a thermal pretreated apple pomace-based medium. In the case of pretreated whey-based medium formulation, we obtained a succinic acid yield of 0.236 g/g consumed lactose, while formic acid and acetic acid were produced as well (0.09 g/g and 0.101 g/g, respectively). We demonstrate that lactose is a promising carbon source for organic acid production by B. succiniciproducens, while our study is the first to propose the use of a similarly available agro-industrial by-product, apple pomace, for the fermentative production of succinic acid by B. succiniciproducens. Full article

Protoplasts offer a promising alternative to embryogenic cell suspensions (ECS) for gene editing in banana, potentially overcoming several limitations associated with ECS-based transformation systems. This study aimed to optimize protoplast isolation and regeneration in Cavendish banana (cv. Williams) and to assess their suitability for transient gene expression. Enzymatic digestion of ECS using cellulase and macerozyme consistently yielded approximately 3 × 10⁶ protoplasts per milliliter of settled cell volume. Protoplast yield was further enhanced, by approximately threefold, through the addition of an antioxidant mixture (ascorbic acid, citric acid and L-cysteine) combined with 0.01% bovine serum albumin. Polyethylene glycol-mediated transfection with a green fluorescent protein reporter gene yielded transient expression in approximately 0.75% of protoplasts five days post-transfection. While phenotypically normal plants were regenerated from untransfected protoplasts after 12 weeks in agarose bead culture with conditioned liquid medium, no regeneration was observed from transfected cells. These findings establish a reproducible protocol for protoplast isolation and plant regeneration in Cavendish banana and provide insight into the barriers limiting successful regeneration following transfection. Full article

For the first time, an alternative and sustainable approach is reported using phenyl-2-pyridyl ketoxime (PPKO) as a selective extracting agent for the recovery of Cu(II) from alkaline solutions in the presence of tartrate ions. The advantages relative to conventional processes carried out in acidic media are outlined. Through potentiometric and spectrophotometric analyses, the sequential formation of a 1:2 metal cation–ligand Cu(II)-(PPKO)₂ complex was identified as the predominant species in alkaline aqueous solutions. The high removal capacity of the extractant for Cu(II), as assessed from liquid–liquid extraction, and its efficient performance are comparable to widely used commercial extractants. Thermodynamic studies of the complexation between the copper(II) ion and PPKO demonstrated that the process exhibits an endothermic character. A progressive decrease in the performance of the extractant was observed after reuse without a regenerative treatment. This deterioration was partially reversed through a controlled reprotonation process using an acetate buffer solution. Overall, the results support the potential of PPKO as an effective and selective alternative ligand for hydrometallurgical applications in alkaline medium. Full article

The conversion from fast-growing tachyzoites to slow-growing bradyzoites is the key factor in establishing the chronic infection and long-term persistence of Toxoplasma gondii. Environmental stressors, such as amino acid starvation and alkaline medium, can trigger the transformation of tachyzoites into bradyzoites. Under such stress conditions, ribosomes slow down, potentially leading to stalling, and ribosomal collisions typically activate ribosome-associated quality control (RQC) pathways. In this study, we investigated the role of T. gondii ribosome quality control complex subunit 2 (TgRqc2), which contains both NFACT and coiled-coil domains, in the parasite’s survival and stage conversion. NFACT represents the “domain” found in the central players involved in RQC, human NEMF and its orthologs FbpA (known as RqcH), Caliban, and Tae2 (known as Rqc2). Phylogenetic analyses revealed that TgRqc2 formed a distinct clade with its orthologs in apicomplexan parasites. The deletion of TgRqc2 impaired T. gondii’s invasion and replication. The Rqc2-knockout strain showed defects in plaque formation and bradyzoite development. Our findings demonstrate that TgRqc2 is essential for T. gondii’s lytic cycle and the conversion of tachyzoites into bradyzoites. RNA-seq analysis further showed that the depletion of TgRqc2 significantly disrupted global transcriptional activity. However, the detailed molecular mechanisms involved remain to be elucidated. In conclusion, our results proved valuable insights that may aid in the development of therapeutic strategies to prevent chronic infection. Full article

This two-part study assesses the impact of biogas inhibition on large-scale waste-to-energy anaerobic digestion (WtE-AD) plants through techno-economic and life cycle assessment approaches. The first part addresses technical and economic aspects. An anaerobic co-digestion system using vegetable waste (FVW) and meat waste (MW) was operated at laboratory scale in a semi-continuous regime with daily feeding to establish a stable process and induce programmed failures causing methanogenic inhibition, achieved by removing MW from the reactor feed and drastically reducing the protein content. Experimental data, combined with bioprocess scale-up models and cost engineering methods, were then used to evaluate the effect of inhibition periods on the profitability of large-scale WtE-AD processes. In the experimental stage, the stable process achieved a yield of 521.5 ± 21 mL CH₄ g⁻¹ volatile solids (VS) and a biogas productivity of 0.965 ± 0.04 L L⁻¹ d⁻¹ (volume of biogas generated per reactor volume per day), with no failure risk detected, as indicated by the volatile fatty acids/total alkalinity ratio (VFA/TA, mg VFA L⁻¹/mg L⁻¹) and the VFA/productivity ratio (mg VFA L⁻¹/L L⁻¹ d⁻¹), both recognized as effective early warning indicators. However, during the inhibition period, productivity decreased by 64.26 ± 11.81% due to VFA accumulation and gradual TA loss. With the progressive reintroduction of the FVW:MW management and the addition of fresh inoculum to the reaction medium, productivity recovered to 96.7 ± 1.70% of its pre-inhibition level. In WtE-AD plants processing 60 t d⁻¹ of waste, inhibition events can reduce net present value (NPV) by up to 40.2% (from 0.98 M USD to 0.55 M USD) if occurring once per year. Increasing plant capacity (200 t d⁻¹), combined with higher revenues from waste management fees (99.5 USD t⁻¹) and favorable electricity markets allowing higher selling prices (up to 0.23 USD kWh⁻¹), can enhance resilience and offset inhibition impacts without significantly compromising profitability. These findings provide policymakers and industry stakeholders with key insights into the economic drivers influencing the competitiveness and sustainability of WtE-AD systems. Full article

Objectives: Although 3D-printing has been identified as a promising technique for personalised medicine manufacturing, developing complex formulations that are suitable for the process can be challenging. This study evaluates the use of a mixture design for the targeted development of an optimised formulation designed for the 3D-printing of oral dosage forms containing the drug sertraline hydrochloride featuring immediate-release drug dissolution. Methods: The polymers Eudragit E PO, Kollidon 17 PF and hydroxypropyl cellulose were compared in simple screening experiments regarding their extrudability, printability and disintegration. A combination of Eudragit E PO and Kollidon 17 PF proved superior and therefore served as the basis for the mixture design. The resulting blends were processed via hot melt extrusion to produce filaments, which were then measured for bending stress using a 3-point-bending-test, and 3D-printed sample plates were used to determine the crystallinity index of sertraline hydrochloride using X-ray diffraction in a previously identified range with low interference from the other components. The formulation was optimised using statistically based models with the aim of minimising the bending stress to obtain flexible, process-robust filaments and simultaneously minimising the crystallinity index with the intention of improving the solubility of the drug by maximising its amorphous content. Results: The filaments made from the optimised formulation could be reliably printed, and the amorphous state of the active ingredient therein was confirmed. The oral dosage forms produced from these showed immediate release characteristics in an acidic medium. Conclusions: This study demonstrates the advantages of a mixture design for optimising complex formulations in a time- and resource-efficient way and could serve as a basis for other research groups to develop innovative, customisable drug delivery systems more effectively. Full article

Microbial proteins offer a sustainable alternative for animal nutrition. Rossellomorea marisflavi NDS, a bacterium isolated from seawater, was previously identified as a promising candidate due to its high protein content. This study aimed to enhance its single cell protein production through systemic fermentation optimization. Single-factor optimization in shake flask determined the optimal conditions to be: a salinity of 20‰ NaCl, a temperature of 32 °C, and an initial pH of 7.3, and a medium composed of 1% (w/v) corn flour, 1% peptone, 0.3% beef extract, and 0.2% KCl. Scaling up to a 10 L bioreactor demonstrated that a two-stage agitation strategy (150 rpm for the first 20 h followed by 180 rpm for the remaining 12 h) enhanced single cell protein yield. Furthermore, allowing the pH to fluctuate freely was more beneficial for protein production than maintaining a constant pH of 7.3 ± 0.02. Under these optimized conditions, the biomass composition (wet weight) was determined to be 2.3767 ± 0.0205% crude ash, 15.6013 ± 0.0082% crude protein, 0.1023 ± 0.0026% crude lipid, and 2.6997 ± 0.0021% carbohydrates. Amino acid analysis revealed a rich profile, with lysine and glutamic acid being the predominant essential and non-essential amino acids, respectively. Fatty acids analysis indicated that C14:1n5 was the most dominant. These findings underscore the potential of R. marisflavi NDS as a high-quality dietary protein supplement and provide a solid foundation for its industrial-scale production. Full article

Background: Potential advantages for improving plant growth, stress tolerance, and valuable metabolites generation are provided by the implementation of nanotechnology into plant biotechnology. A recently discovered technique with significant promise for agricultural practices is the use of biopolymer-based nanomaterials, like peptidomimetics, as insecticides, growth regulators, and nutrient carriers. This study explores the impact of biopolymer-based organic nanofibers—specifically peptidomimetics formed through the self-assembly of L-valine and nicotinic acid (NA) (denoted as M6) on Stevia rebaudiana in vitro propagation and specialized metabolite production. The central hypothesis was that such nanofibers, particularly when used as hormone carriers, can beneficially influence plant morphology, physiology, and biochemistry, thereby promoting the synthesis of antioxidant compounds with therapeutic potential. Methods: The nanofibers were tested either alone (M6) or as carriers of the plant hormone indole-3-acetic acid (IAA) (M6+IAA), supplemented to the cultivation MS medium at variable concentrations (0, 1, 10, and 50 mg L⁻¹). Results: The results revealed that treatment with 10 mg L⁻¹ M6 significantly enhanced shoot growth parameters, including the highest fresh weight (0.249 g), mean shoot height (9.538 cm), shoot number (1.95), and micropropagation rate. Plants treated with M6 alone outperformed those treated with M6+IAA in terms of shoot growth, total soluble sugars, and steviol glycoside content. Conversely, M6+IAA treatment more effectively promoted root initiation, the increased accumulation of mono- and dicaffeoylquinic acids, and boosted antioxidant enzyme activity. Conclusions: These findings highlight the potential of organic nanofibers, both with and without hormone loading, as novel tools for optimizing micropropagation and metabolite enhancement in Stevia rebaudiana. Full article