Search Results (903)

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

The anaerobic co-digestion of agricultural residues emerges as a promising strategy for energy recovery and nutrient recycling within circular agricultural systems. This study aimed to optimize co-digestion parameters for vinegar residue (VR) and cattle manure (CM) using an orthogonal experimental design. Three key variables were investigated which are the co-substrate ratio (VR to CM), feedstock-to-inoculum (F/I) ratio, and total solids (TS) content. Nine experimental combinations were tested to evaluate methane yield, feedstock degradation, and digestate characteristics. Results showed that the optimal condition for methane yield comprised a 2:3 co-substrate ratio, 1:2 F/I ratio, and 20% TS, achieving the highest methane yield of 267.84 mL/g volatile solids (VS) and a vs. degradation rate of 58.65%. Digestate analysis indicated this condition generated the most nutrient-rich liquid digestate and solid digestate, featuring elevated N, P, and K concentrations, acceptable seed germination indices (GI), and moderate humification levels. While total nutrient content did not meet commercial organic fertilizer standards, the digestate is suitable for direct land application in rural settings. This study underscores the need to balance energy recovery and fertilizer quality in anaerobic co-digestion systems, providing practical guidance for decentralized biogas plants seeking to integrate waste treatment with agricultural productivity. Full article

Micromeria graeca (L.) Benth. ex Rchb. (Lamiaceae) is a promising medicinal plant valued for its antioxidant, anti-hyperglycemic, anti-hypertensive, antimicrobial, and anti-aflatoxigenic properties. It is rich in phenolic and flavonoid compounds, supporting its traditional use for digestive, respiratory, cardiovascular, and dermatological conditions. Plant tissue culture facilitates controlled in vitro propagation to study plant growth and bioactive properties. The effects of activated charcoal and varying subculture intervals on multiplication and biomass production in M. graeca shoot cultures were investigated. The phenolic composition of methanolic extracts from in vitro-grown plants was characterized using high-performance liquid chromatography (HPLC), identifying rosmarinic, caffeic, and syringic acids as the primary phenolic compounds. Antimicrobial activity against selected microbial strains was evaluated using a micro-well dilution assay. Anticancer activity of selected extracts was assessed in human hepatocellular carcinoma cell line HepG2, with flow cytometry (Annexin-V/PI staining) used to analyze cell death mechanisms, and compared to pure rosmarinic acid (RA). Activated charcoal showed no beneficial effects on multiplication or biomass production, but significantly increased phenolic acid content (up to 4-fold). RA dominated the phenolic profiles, with other phenolic acids present in lower amounts. Methanolic extracts exhibited negligible antimicrobial activity compared to reference antibiotics and fungicide. Extracts from 4-week-old shoot cultures displayed modest anti-hepatoma activity (IC50 values of CV assay ranging from 193 to 274 µg mL⁻¹), inducing HepG2 cell apoptosis via oxidative stress, independent of RA. Our results suggest that the metabolic output of M. graeca shoot cultures and consequently their biological activity can be modulated by varying in vitro culture conditions. These findings underscore the potential of their methanolic extracts for biotechnological production and therapeutic applications. Full article

In recent years, there has been growing interest in using wood biomass for energy production, which has led to an increase in post-processing waste in the form of wood biomass ash (WBA). Due to the rich composition of WBA, its fertilising potential should be considered. In the conducted studies, WBA was used both alone and in combination with digestate (DG). The WBA was obtained from the Municipal Heat Energy Company and the DG from the Agricultural Biogas Plant in the form of unseparated liquid digestate (ULD), separated solid digestate (SSD) and separated liquid digestate (SLD). The studies included four series: (1) WBA, (2) WBA + ULD, (3) WBA + SSD and (4) WBA + SLD. In each series, WBA was introduced in three increasing doses (0.5, 1.0 and 1.5, expressed in hydrolytic acidity units (HACs) and determined based on the general alkalinity of the material). The digestates (DGs) were applied in fixed doses, which were balanced with respect to the nitrogen introduced into the soil. The test plant was the maize (Zea mays L.) variety Garantio, which was grown in a vegetation hall. The obtained results indicate that the combined use of WBA and DGs (especially ULD and SLD) had a positive effect on the plant height, leaf greenness index (SPAD), and thus, maize yield and dry matter content. In the series with DG addition, the maize yield ranged from 615.5 g (WBA + SSD) to 729.6 g pot⁻¹ (WBA + SLD), which was 28–52% higher than in the series with WBA alone. In turn, the application of increasing doses of WBA alone did not significantly affect the biomass yield but significantly increased the content of N (34%), K (60%), Mg (56%), Ca (60%) and Na (4%). In the series with WBA and DGs, the increase in the content of the above-mentioned macronutrients depended on the type of DG and the dose of WBA. The exception among the macronutrients was P, whose content generally decreased (by 4–23%) with an increasing WBA dose, regardless of the test series. The most favourable results in terms of the chemical composition, excluding the P content, were observed following the combined application of WBA and liquid forms of DG (ULD and SLD). Full article

This study investigated the feasibility of using both the solid and the liquid fractions of waste from the anaerobic digestion process—the digestate—as a possible liquid growth medium for fungal production of chitosan. An enriched liquid phase (ELP), combining both fractions, and derived from mild acid hydrolysis treatment at 120 °C with 6% H₂SO₄ (w/v) for 70 min, was screened for its ability to support biomass and chitosan production by 17 fungal strains. The best results were obtained with Absidia blakesleeana NRRL 2696 and Rhizopus oryzae NRRL 1510 cultures, which yielded chitosan volumetric productions of 444 and 324 mg L⁻¹, respectively. The chitosan preparations of the former and the latter strain, characterized by infrared spectroscopy, elemental analysis, viscosimetry and thermogravimetric analysis, showed deacetylation degrees of 79% and 84.2%, respectively, and average viscosimetric molecular weights of around 20 and 5.4 kDa, respectively. Moreover, both fungal chitosan samples exerted significant antibacterial activity towards Gram-negative (i.e., Pseudomonas syringae and Escherichia coli) and Gram-positive (i.e., Bacillus subtilis) species. Full article

Lately, anaerobic digestion has become a promising method for producing bioenergy from organic waste and is considered a model of the circular economy. At the same time, the concept of circular economy has gained particular attention in environmental policy agendas supporting the transition towards climate neutrality and the promotion of clean energy sources. Although the main objective of anaerobic digestion is to produce biogas, a significant part of the used substrate is converted into digestate, a by-product. Digestate is composed of organic and inorganic matter, which are considered dangerous contaminants for the environment if not properly treated, but also potential renewable resources if properly recovered. Digestate has enormous potential as an organic fertilizer, soil improver and landfill cover soil, but its disposal and use present significant challenges. The main aim of this review paper is to present the current routes for solid and liquid anaerobic digestate valorization according to circular economy principles and to highlight the relation between anaerobic digestion processes and circular economy models. It further focuses on the aspects regarding anaerobic digestate processing technologies, standards and regulations for digestate use and environmental benefits of its use as soil fertilizer. Full article

Long non-coding RNAs (lncRNAs) are increasingly recognized as key regulators of gene expression and cellular signaling in cancer. Their functions are primarily mediated through interactions with specific protein partners that modulate chromatin structure, epigenetic remodeling, transcription, and signal transduction. In this review, we explore reports and strategies for the proteomic characterization of lncRNA-associated proteins, particularly emphasizing high-throughput liquid chromatography–mass spectrometry (LC-MS)-based techniques. Affinity-based methods such as RNA pull-down, ChIRP MS, RAP-MS, BioID-MS, and SILAC-MS enable sensitive and specific mapping of lncRNA and protein complexes. These approaches reveal cancer-specific proteomic signatures, post-translational modifications, and mechanistic insights into tumor biology. The use of label-free quantification, bituminization, and crosslinking strategies further enhances the resolution of dynamic RNA–protein networks. Validation tools following bioinformatic analyses, such as Western blotting, immunohistochemistry, immunofluorescence, and ELISA, are used to prioritize and confirm findings. Candidate biomarkers from hepatocellular carcinoma to colorectal and prostate cancers, profiling lncRNA-associated proteins, hold promise for identifying clinically actionable biomarkers and therapeutic targets. This review highlights the translational relevance of lncRNA protein studies and advocates for their broader adoption in oncological research. In LC-MS workflows, proteins bound to lncRNAs are enzymatically digested into peptides, separated via nano-LC, and analyzed using high-resolution tandem MS. Label-free or isotope-labeled methods quantify differential enrichment, followed by bioinformatics-driven pathway annotation. Full article

Agriculture is one of the most important sectors of the global economy, but it increasingly faces sustainability challenges in meeting rising food demands. The intensive use of mineral fertilizers not only improves yields, but also causes negative environmental impacts such as increasing greenhouse gas emissions, water eutrophication, and soil degradation. To develop more sustainable solutions, the focus is on organic fertilizers, which are produced using waste and biostimulants such as amino acids. The aim of this study was to develop and characterize liquid nitrogen–calcium–magnesium fertilizers produced by decomposing dolomite with nitric acid followed by further processing and to enrich them with a powdered amino acid concentrate Naturamin-WSP and liquid extracts from digestate, a by-product of biogas production. Nutrient-rich extracts were obtained using water and potassium hydroxide solutions, with the latter proving more effective by yielding a higher organic carbon content (4495 ± 0.52 mg/L) and humic substances, which can improve soil structure. The produced fertilizers demonstrated favourable physical properties, including appropriate viscosity and density, as well as low crystallization temperatures (eutectic points from –3 to –34 °C), which are essential for storage and application in cold climates. These properties were achieved by adjusting the content of nitrogenous compounds and bioactive extracts. The results of the study show that liquid fertilizers enriched with organic matter can be an effective and more environmentally friendly alternative to mineral fertilizers, contributing to the development of the circular economy and sustainable agriculture. Full article

Digestate valorization is essential for sustainable waste management and circular economy strategies, yet large-scale adoption faces technical, economic, and environmental challenges. Beyond waste-to-energy conversion, digestate is a valuable soil amendment, enhancing soil structure and reducing reliance on synthetic fertilizers. However, its agronomic benefits depend on feedstock characteristics, treatment processes, and application methods. This study reviews digestate composition, treatment technologies, regulatory frameworks, and environmental impact assessment through Life Cycle Assessment. It analyzes the influence of functional unit selection and system boundary definitions on Life Cycle Assessment outcomes and the effects of feedstock selection, pretreatment, and post-processing on its environmental footprint and fertilization efficiency. A review of 28 JCR-indexed articles (2018–present) analyzed LCA studies on digestate, focusing on methodologies, system boundaries, and impact categories. The findings indicate that Life Cycle Assessment methodologies vary widely, complicating direct comparisons. Transportation distances, nutrient stability, and post-processing strategies significantly impact greenhouse gas emissions and nutrient retention efficiency. Techniques like solid–liquid separation and composting enhance digestate stability and agronomic performance. Digestate remains a promising alternative to synthetic fertilizers despite market uncertainty and regulatory inconsistencies. Standardized Life Cycle Assessment methodologies and policy incentives are needed to promote its adoption as a sustainable soil amendment within circular economy frameworks. Full article

This study evaluated the effects of liquid oregano oil, chitosan nanoparticles with encapsulated liquid oregano oil, and a negative control of empty chitosan nanoparticles on in vitro ruminal fermentation. Three Boer goats were used as ruminal fluid donors, fed with a formulated ration for 21 d for inoculum adaptation. Treatments tested on in vitro assays were diet without oregano oil or nanoparticles (CON); diet with 100 ppm of oregano oil in nanoparticles (100N); diet with 300 ppm of liquid oregano oil (300L); diet with 300 ppm of oregano oil in nanoparticles (300N); and diet with 300 ppm of empty nanoparticles (300CHN). The variables studied were in vitro dry matter digestibility (ivDMD), in vitro neutral detergent fiber digestibility (ivNDFD_om), total gas production (TGP), ammonia nitrogen concentration (NH₃), and pH. The experimental design was a randomized complete block design. Linear and quadratic regressions were used to identify dependence and inflection points. The ivDMD increased at 12, 36, and 48 h, with 300N and with 300L exhibiting increased ivNDFD_om at 36 h. Ruminal pH was highest (p < 0.05) with 300CHN at 36 h. For first-order regression analysis of TGP, coefficients (β) were highest (p < 0.05) for 300N. In conclusion, 300N increased ruminal fermentation in vitro, as reflected by increases in ivDMD, ivNDFD_om, and TGP. Full article

Devices that meet the needs of agricultural biogas plants represent a significant share of the energy balance of the source. The digester mixer is a crucial component installed in the fermentation chamber. Energy consumption during mixing depends on the regime and intensity, as well as the rheological properties of the carrier liquid, the dry matter content, and the dimensions of the fibers. Bioreactor operators often oversize mixers and extend mixing duration to avoid disruptions in biogas production. This paper analyzed the influence of digester mixer operations on selected electrical power quality parameters. For this purpose, two agricultural biogas plants with a capacity of 40 kW, connected to the low-voltage grid, were studied (one located approximately 120 m from the transformer station and the second 430 m away). As shown by the correlations presented in the article, the connection point of the biogas plant significantly impacted the magnitude of the influence of mixer operations on the analyzed voltage parameters. In the second biogas plant, switching on the mixers (in the absence of generation) caused the grid voltage to drop to the lower value permitted by regulations. (Switching on the mixers caused a change in voltage by about 30 V.) The most disturbances were introduced into the grid when the power generated by the biogas plant was equal to the power consumed by its internal equipment. (THD_I then reached as high as 63.2%, while in other cases, it did not exceed 17%.) Furthermore, the operation of the mixers alone resulted in a reduction of approximately 1 MWh of energy exported to the power grid per month. Full article

Ginger (Zingiber officinale Roscoe) has been widely recognized for its antioxidant properties, primarily attributed to its phenolic compounds such as gingerols and shogaols. However, limited data exist regarding how different pharmaceutical forms influence the bioaccessibility and antioxidant efficacy of these compounds. This study aimed to evaluate the antioxidant capacity and bioaccessibility of ginger in different pharmaceutical forms—capsules (20 mg, 40 mg, and 80 mg), a pure powdered extract, and a liquid formulation—standardized to ≥6% gingerols. The phenolic profile of each formulation was characterized using HPLC-DAD (High-Performance Liquid Chromatography with Diode Array Detection), followed by the evaluation of antioxidant capacity through DPPH (2,2-Diphenyl-1-picrylhydrazyl) and ORAC (Oxygen Radical Absorbance Capacity) assays, and the assessment of bioaccessibility via an in vitro digestion model. The results demonstrated that antioxidant activity was positively correlated with extract concentration and was highest in the liquid formulation (426.0 ± 0.05 µmol Trolox equivalents (TE) and 11,336.7 ± 0.20 µmol TE in the DPPH and ORAC assays, respectively). The bioaccessibility of 6-gingerol and 6-shogaol significantly increased in the liquid form, reaching 23.44% and 11.31%, respectively, compared to ≤4% in the pure extract. These findings highlight the influence of the formulation matrix on compound release and support the use of liquid preparations to enhance the functional efficacy of ginger-derived nutraceuticals. This standardized comparative approach, using formulations derived from the same extract, offers new insights into how the delivery matrix influences the functional performance of ginger compounds, providing guidance for the development of more effective nutraceutical strategies. Full article

A three-year optimization study was conducted at a mechanical biological treatment plant with the aim of enhancing organic fractions recovery from mechanically separated fine fractions (MSFF) of residual waste using a screw press. The study aimed to optimize key operating parameters for the employed screw press, such as pressure, liquid-to-MSFF, feeding quantity per hour, and press basket mesh size, to enhance volatile solids and biogas recovery in the generated press water for anaerobic digestion. Experiments were performed at the full-scale facility to evaluate the efficiency of screw press extraction with other pretreatment methods, like press extrusion, wet pulping, and hydrothermal treatment. The results indicated that hydrolysis of the organic fractions in MSFF was the most important factor for improving organic extraction from the MSFF to press water for fermentation. Optimal hydrolysis efficiency was achieved with a digestate and process water-to-MSFF of approximately 1000 L/ton, with a feeding rate between 8.8 and 14 tons per hour. Increasing pressure from 2.5 to 4.0 bar had minimal impact on press water properties or biogas production, regardless of the press basket size. The highest volatile solids (29%) and biogas (50%) recovery occurred at 4.0 bar pressure with a 1000 L/ton liquid-to-MSFF. Further improvements could be achieved with longer mixing times before pressing. These findings demonstrate the technical feasibility of the pressing system for preparing an appropriate substrate for the fermentation process, underscoring the potential for optimizing the system. However, further research is required to assess the cost–benefit balance. Full article

Adequate treatment of the organic fraction of municipal solid waste (OFMSW) in co-digestion with sewage sludge (SS) through dark fermentation (DF) technologies has been widely studied and recognized. However, there is little experience with a high-solids approach, where practical and scalable conditions are established to lay the groundwork for further development of feasible industrial-scale projects. In this study, the biochemical hydrogen potential of OFMSW using a 7 L batch reactor at mesophilic conditions was evaluated. Parameters such as pH, redox potential, temperature, alkalinity, total solids, and substrate/inoculum ratio were adjusted and monitored. Biogas composition was analyzed by gas chromatography. The microbial characterization of SS and post-reaction percolate liquids was determined through metagenomics analyses. Results show a biohydrogen yield of 38.4 NmLH₂/gVS OFMSW, which forms ~60% of the produced biogas. Aeration was proven to be an efficient inoculum pretreatment method, mainly to decrease the levels of methanogenic archaea and metabolic competition, and at the same time maintain the required total solid (TS) contents for high-solids conditions. The microbial community analysis reveals that biohydrogen production was carried out by specific anaerobic and aerobic bacteria, predominantly dominated by the phylum Firmicutes, including the genus Bacillus (44.63% of the total microbial community), Clostridium, Romboutsia, and the phylum Proteobacteria, with the genus Proteus. Full article

Mechanical–biological treatment plants face challenges in effectively separating organic fractions from residual municipal solid waste for biological treatment. This study investigates the optimization measures carried out at the Erbenschwang MBT facility, which transitioned from solely aerobic treatment to integrated anaerobic digestion using a screw press. This study focused on evaluating the efficiency of each mechanical pretreatment step by investigating the composition of the residual waste, organic fraction recovery rate, and screw press performance in recovering organic material and biogas to press water. The results showed that 92% of the organic material from the residual waste was recovered into fine fractions after shredding and trommel screening. The pressing experiments produced high-quality press water with less than 3% inert material (0.063–4 mm size). Mass balance analysis revealed that 47% of the input fresh mass was separated into press water, corresponding to 24% of the volatile solids recovered. Biogas yield tests showed that the press water had a biogas potential of 416 m³/ton VS, recovering 38% of the total biogas potential. In simple terms, the screw press produced 32 m³ of biogas per ton of mechanically separated fine fractions and 20 m³ per ton of input residual waste. This low-pressure, single-step screw press efficiently and cost-effectively prepares anaerobic digestion feedstock, making it a promising optimization for both existing and new facilities. The operational configuration of the screw press remains an underexplored area in current research. Therefore, further studies are needed to systematically evaluate key parameters such as screw press pressure (bar), liquid-to-waste (L/ton), and feed rate (ton/h). Full article