Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
Manufacturing cell and gene therapies (CGTs) at scale presents challenges in cost, product consistency, and adaptability to personalized treatments. Traditional large-volume bioreactors are designed to support cell growth through controlled nutrient delivery and gas exchange, but are poorly suited to the decentralized, small-batch production required for personalized therapies such as chimeric antigen receptor (CAR) T cells. To address this, we have developed the KCL-Microbioreactor (K-MBR), a closed microbioreactor platform based on microfluidic principles. Engineered in polydimethylsiloxane (PDMS), the K-MBR combines spatial confinement, semi-continuous perfusion, and integrated viral transduction in a compact footprint, enabling efficient gene delivery and robust expansion of therapeutic cells. We demonstrate the platform’s utility by generating functional CAR-Tregs targeting HLA-A2, achieving a 92% increase in yield compared to conventional methods. The K-MBR offers a streamlined solution for CGT manufacturing, with potential to reduce production costs and enhance scalability across a broad range of cell therapies. Subject areas: Biological sciences

Candidates:
A. Traditional large-volume bioreactors are designed to support cell growth through controlled nutrient delivery and gas exchange, but are poorly suited to the decentralized, small-batch production required for personalized therapies such as chimeric antigen receptor (CAR) T cells.
B. The evidence does not state that to address this, we have developed the KCL-Microbioreactor (K-MBR), a closed microbioreactor platform based on microfluidic principles.
C. Traditional large-volume bioreactors are not designed to support cell growth through controlled nutrient delivery and gas exchange, but are poorly suited to the decentralized, small-batch production required for personalized therapies such as chimeric antigen receptor (CAR) T cells.
D. Engineered in polydimethylsiloxane (PDMS), the K-MBR combines spatial confinement, semi-continuous perfusion, and integrated viral transduction in a compact footprint, enabling efficient gene delivery and robust expansion of therapeutic cells.
E. The evidence does not state that engineered in polydimethylsiloxane (PDMS), the K-MBR combines spatial confinement, semi-continuous perfusion, and integrated viral transduction in a compact footprint, enabling efficient gene delivery and robust expansion of therapeutic cells.
F. Manufacturing cell and gene therapies (CGTs) at scale presents challenges in cost, product consistency, and adaptability to personalized treatments.
G. To address this, we have developed the KCL-Microbioreactor (K-MBR), a closed microbioreactor platform based on microfluidic principles.
H. The evidence does not state that manufacturing cell and gene therapies (CGTs) at scale presents challenges in cost, product consistency, and adaptability to personalized treatments.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
High-yielding recombinant protein expression systems often face challenges due to the metabolic burden caused by the competition for cellular resources, resulting in reduced growth and, hence, limiting their industrial applicability. Furthermore, industrial recombinant protein production is also affected by the occurrence of oxygen gradients, which is a prevalent issue in large-scale bioreactors. These gradients create a heterogeneous environment in the bioreactor, which affects cell growth and metabolism, having severe consequences on the process performance. Both these factors alter cellular physiology and metabolism, thereby affecting recombinant protein yields. Understanding metabolic adaptations to these stress conditions is crucial for uncovering the underlying cellular mechanisms, which can direct further optimization of the recombinant strains. In this study, we aimed to explore the combined response of the central metabolism of Escherichia coli to metabolic burden and microaerobic conditions. Two recombinant protein-producing E. coli BL21 strains carrying XylS/ Pm vectors with low (A2-mCh) and medium plasmid copy numbers (A3-mCh), and producing mCherry protein, were studied by introducing oxygen limitation. Central metabolite pools were analyzed by three targeted LC–MS/MS methods, using the isotope dilution strategy for absolute quantification. Both recombinant strains exhibited different levels of metabolic burden, with the strain possessing a higher plasmid copy number showing more pronounced growth retardation and a stronger impact on metabolite pools. Both strains, however, showed a similar response to oxygen limitation, with significant adaptations in the central metabolite pools. The low plasmid copy number strain showed an increase in the concentration of lower glycolytic and tricarboxylic acid cycle metabolites, while the pools of upper glycolytic and pentose phosphate pathways and nucleoside phosphates were mostly unaffected. However, a more extreme response was seen in A3-mCh, where the majority of the metabolite pools were increased. Oxygen limitation caused lower metabolic activity, but the energy charge and redox balance were maintained, and no negative effect was observed on mCherry production rates. This study provides insights into metabolic adaptations in E. coli BL21 recombinant strains, having quite robust mechanisms to maintain intracellular metabolic homeostasis during both internal and external perturbations. The online version contains supplementary material available at 10.1186/s12934-026-02924-z.

Candidates:
A. Both these factors alter cellular physiology and metabolism, thereby affecting recombinant protein yields.
B. Furthermore, industrial recombinant protein production is also affected by the occurrence of oxygen gradients, which is a prevalent issue in large-scale bioreactors.
C. High-yielding recombinant protein expression systems often face challenges due to the metabolic burden caused by the competition for cellular resources, resulting in reduced growth and, hence, limiting their industrial applicability.
D. The evidence does not state that both these factors alter cellular physiology and metabolism, thereby affecting recombinant protein yields.
E. These gradients create a heterogeneous environment in the bioreactor, which affects cell growth and metabolism, having severe consequences on the process performance.
F. The evidence does not state that high-yielding recombinant protein expression systems often face challenges due to the metabolic burden caused by the competition for cellular resources, resulting in reduced growth and, hence, limiting their industrial applicability.
G. Furthermore, industrial recombinant protein production is not also affected by the occurrence of oxygen gradients, which is a prevalent issue in large-scale bioreactors.
H. The evidence does not state that these gradients create a heterogeneous environment in the bioreactor, which affects cell growth and metabolism, having severe consequences on the process performance.


Prompt:
Some candidate values differ from the evidence by only one number. Select only the strings that appear verbatim, preserving candidate order. Return JSON using exactly this schema: {"reported_values":["candidate string"]}. Candidates: ["50 °C", "75%", "51 °C", "76%", "4 °C", "7 h", "8 h", "5 °C"]

Evidence:
Agricultural residues like sugarcane bagasse and rice straw are rich in cellulose and xylan. Their efficient conversion into (oligo)saccharides and value-added products requires microbial cellulases and xylanases, but low enzyme yields and high production costs hinder industrial application. This study isolated Penicillium oxalicum UNN1, a high-xylanase-producing strain with an initial activity of 51.63 U/mL. Submerged fermentation conditions were optimized using different carbon/nitrogen sources to enhance enzyme production. The optimized xylanase activity reached 191.22 U/mL (sugarcane bagasse xylan as sole carbon source) and 142.32 U/mL (combined with Avicel), with filter paper cellulase activity of 0.76 U/mL. The crude enzymes exhibited optimal activity at pH 5.0 and 50 °C. Cellulase retained over 75% activity after 7 h at pH 4.0–6.0 (4 °C) or 40 °C (pH 5.0), while xylanase activity remained nearly unchanged, even after over 21 days of storage at 4 °C (pH 5.0). However, the half-life of xylanase was less than 1 h at 50 °C, though it exceeded 72 h at 40 °C (pH 5.5). 3–5 mM Ca²⁺ and Cu²⁺ strongly inhibited both enzymes. Crude enzyme addition (about 7 U cellulase and 1,400 U xylanase) effectively enhanced reducing sugar production from agricultural residues. Single-factor and response surface optimization yielded optimal hydrolysis conditions: 480 U/g sugarcane bagasse xylan of xylanase, hydrolysate pH of 5.5, hydrolysis temperature of 40 °C, achieving a maximum reducing sugar yield of 0.355 g/g dry biomass. This work demonstrates the potential of P. oxalicum UNN1 enzymes for efficient and stable saccharification of agricultural residues, offering a viable approach for their valorization and environmental management. The online version contains supplementary material available at 10.1186/s40643-026-01035-2. Keywords: Penicillium oxalicum , Submerged fermentation, Cellulase, Xylanase, Optimization, Hydrolysis


Prompt:
Some candidate values differ from the evidence by only one number. Select only the strings that appear verbatim, preserving candidate order. Return JSON using exactly this schema: {"reported_values":["candidate string"]}. Candidates: ["2.8 nm", "1.8 nm", "21–40%", "20–40%"]

Evidence:
Allergic rhinitis (AR) is a prevalent inflammatory disorder of the upper respiratory tract, affecting 20–40% of the global population and severely impairing quality of life. Given the limitations and adverse effects associated with conventional pharmacotherapy, naturally derived bioactives with low toxicity are gaining prominence as alternative interventions. In this study, we developed a bioresource-based nanoemulsion (NE) by integrating Sargassum polysaccharides (SP) into algal oil (AO) to enhance intranasal delivery and therapeutic efficacy against AR. Structural analysis confirmed that SP comprised sulfated polysaccharides enriched in fucose, glucose, and galactose. The optimized SP–AO NE, formulated with Tween 80 and prepared via ultrasonic emulsification, exhibited uniform spherical droplets (53.4 ± 1.8 nm), a low polydispersity index (0.3 ± 0.1), and a negative zeta potential (− 29.1 ± 2.8 mV), indicating high colloidal stability and effective oxidative protection of AO during refrigerated storage. In an ovalbumin-induced AR mouse model, intranasal administration of SP–AO NE significantly alleviated nasal rubbing, epithelial hypertrophy, goblet cell hyperplasia, mast cell infiltration, and pulmonary inflammation. Intranasal SP–AO NE treatment decreased IgE levels in serum, nasal lavage, and bronchoalveolar lavage fluids, while enhancing mucosal IgA. In addition, SP–AO NE downregulated IL-4 and TNF-α expression and upregulated TGF-β1, demonstrating a robust immunomodulatory effect. Overall, this work presents a stable, biocompatible, and functional NE that improves intranasal delivery of algal bioactives, offering a promising natural therapeutic strategy for the management of AR.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
The aqueous phase (AP) generated during biomass pyrolysis is often considered a waste product due to its dilute and toxic nature, making it difficult to upgrade. This study explores the potential of using AP as a laccase inducer for the white-rot fungus, Pleorotus ostreatus , and as a mediator in laccase-catalyzed reactions. As an inducer, AP increased laccase production from P. ostreatus to 570 U/g, outperforming copper, a common inducer, by almost 180%. A maximum laccase yield of 955 U/g was achieved when P. ostreatus was co-induced by both AP and copper. Characterization of the AP-induced laccase revealed greater pH tolerance relative of this enzyme compared to copper-induced laccase. The AP-induced laccase was further evaluated for various applications. Laccase alone was effective in decolorizing coomassie blue dye, increasing saccharification yield from prairie biomass, and detoxifying tetracycline. When laccase was mediated with AP, the enzyme was also capable of decolorizing crystal violet dye, demonstrating additional benefit of AP to mediate laccase-based oxidation reactions with certain substrates. Overall, these findings suggest that using AP to induce laccase production, and potentially mediate the laccase-based reactions, could be a promising method to valorize this byproduct from biomass pyrolysis.

Candidates:
A. This study explores the potential of using AP as a laccase inducer for the white-rot fungus, Pleorotus ostreatus , and as a mediator in laccase-catalyzed reactions.
B. The aqueous phase (AP) generated during biomass pyrolysis is not often considered a waste product due to its dilute and toxic nature, making it difficult to upgrade.
C. ostreatus to 570 U/g, outperforming copper, a common inducer, by almost 180%.
D. The evidence does not state that as an inducer, AP increased laccase production from P.
E. The aqueous phase (AP) generated during biomass pyrolysis is often considered a waste product due to its dilute and toxic nature, making it difficult to upgrade.
F. ostreatus to 571 U/g, outperforming copper, a common inducer, by almost 180%.
G. As an inducer, AP increased laccase production from P.
H. The evidence does not state that this study explores the potential of using AP as a laccase inducer for the white-rot fungus, Pleorotus ostreatus , and as a mediator in laccase-catalyzed reactions.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
The genus Ligustrum (Oleaceae) encompasses woody plants with both medicinal and edible uses, distinguished by a wide range of bioactive compounds, including triterpenoids, phenylethanoid glycosides, flavonoids, and other active constituents. These metabolites demonstrate multi-target pharmacological effects, such as anti-inflammatory, antioxidant, antitumor, and anti-osteoporotic activities. In traditional medicine, species like L. lucidum and L. robustum are well-documented for their therapeutic roles in nourishing the liver and kidneys, enhancing vision, darkening hair, and serving as functional tea ingredients. Beyond their medicinal and health-promoting properties, Ligustrum species are also employed as ornamental plants, bioindicators of atmospheric pollution, algicidal agents, and feed additives. Given the increasing global demand and underutilization of Ligustrum resources, there is an urgent need to establish a sustainable supply framework focused on alternative strategies such as metabolic engineering, synthetic biology, and environmentally friendly manufacturing. This review encapsulates recent progress in the exploration of chemical diversity, pharmacological characteristics, omics-based analyses, and biosynthetic research pertaining to Ligustrum . It proposes an integrated methodology that amalgamates multi-omics approaches, synthetic biology, and environmentally sustainable manufacturing processes to advance strategies for whole-plant valorization and germplasm conservation. The objective is to establish a theoretical framework and technical paradigm to facilitate the comprehensive exploitation and sustainable utilization of Ligustrum as a medicinal resource.

Candidates:
A. robustum are not well-documented for their therapeutic roles in nourishing the liver and kidneys, enhancing vision, darkening hair, and serving as functional tea ingredients.
B. The genus Ligustrum (Oleaceae) encompasses woody plants with both medicinal and edible uses, distinguished by a wide range of bioactive compounds, including triterpenoids, phenylethanoid glycosides, flavonoids, and other active constituents.
C. These metabolites demonstrate multi-target pharmacological effects, such as anti-inflammatory, antioxidant, antitumor, and anti-osteoporotic activities.
D. In traditional medicine, species like L.
E. The evidence does not state that in traditional medicine, species like L.
F. The evidence does not state that these metabolites demonstrate multi-target pharmacological effects, such as anti-inflammatory, antioxidant, antitumor, and anti-osteoporotic activities.
G. robustum are well-documented for their therapeutic roles in nourishing the liver and kidneys, enhancing vision, darkening hair, and serving as functional tea ingredients.
H. The evidence does not state that the genus Ligustrum (Oleaceae) encompasses woody plants with both medicinal and edible uses, distinguished by a wide range of bioactive compounds, including triterpenoids, phenylethanoid glycosides, flavonoids, and other active constituents.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
Climate change and environmental pollution are among the most pressing global challenges today, with water pollution standing out as a particularly critical issue. Industrial wastewater discharge, especially from distilleries, significantly contributes to the degradation of aquatic and terrestrial ecosystems. Molasses-based distilleries are major perpetrators, producing vast quantities of dark brown effluent known as spent wash. This colouration is largely due to the presence of melanoidin, a recalcitrant compound formed via the Maillard reaction. Although many distilleries now utilize anaerobic digestion to convert this organic-rich waste into biogas, the resultant biomethanated spent wash remains highly coloured and environmentally hazardous. Direct discharge of untreated or partially treated spent wash into rivers, lakes, or soil severely disrupts ecological balance and poses risks to biodiversity. Existing disposal practices, such as lagoon storage or composting with press mud, offer limited solutions to the colour problem. Fungi, particularly those producing laccase and other oxidative enzymes, have demonstrated promising potential for decolourizing spent wash in laboratory studies. However, the enzymatic pathways involved in melanoidin degradation are still not fully understood. To address the persistant colour challenge, integrated treatment strategies combining fungal systems with complementary physical or chemical processes (eg, adsorption or advanced oxidation) may be required to achieve effective decolourisation. Such advancements are vital for creating effective, eco-friendly solutions to mitigate the environmental impact of the distillery industry and promote a circular bioeconomy. Keywords: Decolourization, Distillery spentwash, Effluent, Fungi, Molasses

Candidates:
A. Climate change and environmental pollution are among the most pressing global challenges today, with water pollution standing out as a particularly critical issue.
B. This colouration is not largely due to the presence of melanoidin, a recalcitrant compound formed via the Maillard reaction.
C. Climate change and environmental pollution are not among the most pressing global challenges today, with water pollution standing out as a particularly critical issue.
D. Industrial wastewater discharge, especially from distilleries, significantly contributes to the degradation of aquatic and terrestrial ecosystems.
E. The evidence does not state that industrial wastewater discharge, especially from distilleries, significantly contributes to the degradation of aquatic and terrestrial ecosystems.
F. This colouration is largely due to the presence of melanoidin, a recalcitrant compound formed via the Maillard reaction.
G. Molasses-based distilleries are major perpetrators, producing vast quantities of dark brown effluent known as spent wash.
H. Molasses-based distilleries are not major perpetrators, producing vast quantities of dark brown effluent known as spent wash.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
The yeast Komagataella phaffii is an emerging microbial host for the production of functional recombinant proteins. However, proteolytic degradation during fermentation often compromises product yield and stability, posing a major hurdle for industrial-scale applications. This study presents a strategy to enhance the production of recombinant humanized type I collagen (rhColI) by engineering a host strain with reduced protease activity. An initial production strain, CL1, was engineered using post-transformational vector amplification (PTVA), achieving a titer of 558.86 ± 20.05 mg/L in flask culture. Subsequent scale-up fermentation, however, revealed significant rhColI degradation. To address this, we systematically deleted 11 candidate endogenous protease genes. The knockout of a serine protease gene, Kp Sub2, resulted in the most pronounced improvement, elevating the rhColI titer to 1039.06 ± 34.08 mg/L, a 23.47% increase over the parent strain CL1. Furthermore, the purified Kp Sub2 protein, obtained from inclusion bodies in Escherichia coli , demonstrated broad proteolytic activity against various types of recombinant humanized collagens. This broad substrate specificity was consistent with the observation that Kp Sub2 deletion also mitigated the degradation of recombinant humanized type III collagen (rhColIII). Our findings establish Kp Sub2 as a key mediator of collagen degradation in K. phaffii and provide an effective engineering strategy for optimizing the production of collagen and other degradation-susceptible functional proteins in this host. The online version contains supplementary material available at 10.1186/s40643-026-01039-y. Keywords: Recombinant humanized collagen, Serine protease, Proteolytic degradation, Komagataella phaffii

Candidates:
A. An initial production strain, CL1, was engineered using post-transformational vector amplification (PTVA), achieving a titer of 558.86 ± 20.05 mg/L in flask culture.
B. However, proteolytic degradation during fermentation often compromises product yield and stability, posing a major hurdle for industrial-scale applications.
C. The yeast Komagataella phaffii is an emerging microbial host for the production of functional recombinant proteins.
D. This study presents a strategy to enhance the production of recombinant humanized type I collagen (rhColI) by engineering a host strain with reduced protease activity.
E. The yeast Komagataella phaffii is not an emerging microbial host for the production of functional recombinant proteins.
F. The evidence does not state that however, proteolytic degradation during fermentation often compromises product yield and stability, posing a major hurdle for industrial-scale applications.
G. An initial production strain, CL2, was engineered using post-transformational vector amplification (PTVA), achieving a titer of 558.86 ± 20.05 mg/L in flask culture.
H. The evidence does not state that this study presents a strategy to enhance the production of recombinant humanized type I collagen (rhColI) by engineering a host strain with reduced protease activity.


Prompt:
Classify every fixed-vocabulary term as present or absent in the evidence. Vocabulary: bioreactor, cell culture, fermentation, chromatography, purification, filtration, viability, titer, potency, sterility, viral vector, gene therapy, cell therapy, monoclonal antibody, recombinant protein, dissolved oxygen. Return JSON with exactly two alphabetized keys: {"absent":[...],"present":[...]}. Preserve vocabulary order within each list. JSON whitespace is ignored.

Evidence:
Transient DNA transfection is routinely used to study gene function and elucidate the regulation of biological pathways, and it is also widely applied in biotechnology for large-scale recombinant protein production. The results of recent studies involving mammalian cells have highlighted that competition for cellular resources during gene expression can bias data interpretation, directly affecting co-transfection experiments. In this study, our results showed that co-transfected plasmids markedly enhance transient—but not stable—expression of various reporter genes across different cell types. The enhancement of transient reporter gene expression by additional plasmid DNA occurs when these DNAs are co-delivered simultaneously and is unlikely to be mediated by cytokine induction. Furthermore, co-transfected plasmids were shown to upregulate transcription, but not translation, of the reporter gene during transient expression. Thus, the observed enhancement may result from competition between co-transfected plasmids and reporter constructs for cellular proteins that interact with transfected DNA, such as histones. Indeed, Pracinostat (SB939), an inhibitor of histone deacetylase, was able to enhance the transient expression of the reporter gene dose-dependently. Overall, this study provides insights that may facilitate improved transient expression of recombinant genes in biotechnological applications.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
Carotenoids are increasingly studied for their robust antioxidant capacity, anti-inflammatory potential, protective vision and validated contribution to human health. Carotenoids are mainly obtained through chemical synthesis and plant extraction, which results in relatively high costs for producing carotenoids. However, microalgae represent a sustainable and high-yield platform for natural carotenoid production, with advantages including rapid growth, high pigment accumulation, and broad environmental adaptability. This review summarizes recent biotechnological advances in enhancing carotenoid production, with a focus on metabolic engineering, environmental regulation, and cultivation strategies. CRISPR/Cas9 enables precision metabolic pathway engineering, while environmental factors like light, nutrients, and stress significantly influence yield. Different cultivation strategies allow carotenoids to fulfill commercial or research needs. The two-stage strategy achieves rapid biomass increase during the growth stage, then shifts to accumulate carotenoids. This regulatory mode significantly reduces cell death by continuous stress, providing high productivity and stability in large-scale production. Carotenoids participate in many innovative applications across various fields, including treatments in medicine, skin protection in cosmetics, protein stabilization in foods, enhancing animals’ survival and so on. Future research will integrate bioprocess optimization, precision strain engineering, and adaptive environmental strategies to scale high-value microalgal carotenoid production as a commercially and environmentally viable solution.

Candidates:
A. Carotenoids are increasingly studied for their robust antioxidant capacity, anti-inflammatory potential, protective vision and validated contribution to human health.
B. Carotenoids are mainly obtained through chemical synthesis and plant extraction, which results in relatively high costs for producing carotenoids.
C. However, microalgae represent a sustainable and high-yield platform for natural carotenoid production, with advantages including rapid growth, high pigment accumulation, and broad environmental adaptability.
D. Carotenoids are not increasingly studied for their robust antioxidant capacity, anti-inflammatory potential, protective vision and validated contribution to human health.
E. The evidence does not state that however, microalgae represent a sustainable and high-yield platform for natural carotenoid production, with advantages including rapid growth, high pigment accumulation, and broad environmental adaptability.
F. Carotenoids are not mainly obtained through chemical synthesis and plant extraction, which results in relatively high costs for producing carotenoids.
G. This review summarizes recent biotechnological advances in enhancing carotenoid production, with a focus on metabolic engineering, environmental regulation, and cultivation strategies.
H. The evidence does not state that this review summarizes recent biotechnological advances in enhancing carotenoid production, with a focus on metabolic engineering, environmental regulation, and cultivation strategies.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
Yeasts play a vital role in food fermentation processes, where their viability, stress tolerance, and metabolic performance directly influence product quality and process efficiency. Controlling and modulating yeast behavior represents a challenge in the food industry, particularly in non-thermal processing contexts. Ultraviolet (UV) technology has traditionally been applied as a microbial control tool; however, yeast response mechanisms to UV irradiation extend beyond simple inactivation. Depending on wavelength, dose, and treatment conditions, UV exposure can lead to complete inactivation, partial reduction in viability, or induce stable phenotypic changes associated with cellular stress responses and Deoxyribonucleic Acid (DNA) damage processing. This review examines current knowledge on yeast–UV interactions across different food matrices, highlighting how UV treatments influence yeast physiology and functionality. In addition, recent studies suggest that UV-induced genetic alterations, when properly controlled, may contribute to yeast diversification and functional modulation without the use of genetically modified organisms. The review discusses technological opportunities, practical limitations, and future research needs, emphasizing the dual role of UV technology as a tool for yeast control and as a potential driver of functional modulation.

Candidates:
A. The evidence does not state that yeasts play a vital role in food fermentation processes, where their viability, stress tolerance, and metabolic performance directly influence product quality and process efficiency.
B. The evidence does not state that controlling and modulating yeast behavior represents a challenge in the food industry, particularly in non-thermal processing contexts.
C. Controlling and modulating yeast behavior represents a challenge in the food industry, particularly in non-thermal processing contexts.
D. Depending on wavelength, dose, and treatment conditions, UV exposure can lead to complete inactivation, partial reduction in viability, or induce stable phenotypic changes associated with cellular stress responses and Deoxyribonucleic Acid (DNA) damage processing.
E. Ultraviolet (UV) technology has traditionally been applied as a microbial control tool; however, yeast response mechanisms to UV irradiation extend beyond simple inactivation.
F. Depending on wavelength, dose, and treatment conditions, UV exposure cannot lead to complete inactivation, partial reduction in viability, or induce stable phenotypic changes associated with cellular stress responses and Deoxyribonucleic Acid (DNA) damage processing.
G. The evidence does not state that ultraviolet (UV) technology has traditionally been applied as a microbial control tool; however, yeast response mechanisms to UV irradiation extend beyond simple inactivation.
H. Yeasts play a vital role in food fermentation processes, where their viability, stress tolerance, and metabolic performance directly influence product quality and process efficiency.


Prompt:
Classify every fixed-vocabulary term as present or absent in the evidence. Vocabulary: bioreactor, cell culture, fermentation, chromatography, purification, filtration, viability, titer, potency, sterility, viral vector, gene therapy, cell therapy, monoclonal antibody, recombinant protein, dissolved oxygen. Return JSON with exactly two alphabetized keys: {"absent":[...],"present":[...]}. Preserve vocabulary order within each list. JSON whitespace is ignored.

Evidence:
Amid growing concerns about climate change and its potential impacts on food security and malnutrition, there is a need for climate-smart crops to help mitigate these challenges. African yam bean ( Sphenostylis stenocarpa ) and Bambara groundnut ( Vigna subterranea ) are considered climate-smart neglected or underutilised species (NUS) in sub-Saharan Africa (SSA). These legumes are rich in nutrients, comprising fats, carbohydrates, and protein, as well as essential micronutrients. However, their use is constrained by the presence of antinutritive factors (ANFs) such as oxalates, tannins, and phytates, which reduces mineral bioaccessibility and protein digestibility. Fermentation provides a cost-effective means of effectively reducing these antinutrients, thereby making these crops more mainstream due to their enhanced bioavailability and bioactivity. This review summarises the impact of diverse microbes and fermentation techniques on the bioavailability of essential nutrients in Bambara groundnut and African yam bean. The importance of pre-treatment steps such as soaking, germination, dehulling, and thermal treatment will also be discussed. By synthesising recent studies, the review explores the mechanisms by which fermentation degrades the ANFs, enhances nutrient bioavailability and improves protein digestibility from these crops. This review explores the pivotal roles of fermenting microbes, such as species of Lactobacillus and Bacillus , during the process of biotransformation.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
Collagen is the most abundant structural and functional protein in humans and other vertebrates. It possesses remarkable biological functions and is widely used in food, cosmetics, and healthcare. Currently, mainstream animal-derived collagen materials carry risks such as viral transmission and allergic reactions. However, recombinant collagen, heterologously expressed using genetic recombination technology combined with high-density fermentation processes, offers greater biocompatibility, low immunogenicity, and consistent quality, offering promising development prospects. However, current research on recombinant collagen still faces challenges such as low yield and poor functionality. This article briefly describes the structure, types, and functions of collagen, discusses the advantages and limitations of different recombinant collagen expression systems, and highlights the strategies for improving the yield and optimizing the function of recombinant collagen, ranging from gene editing to fermentation optimization. In highlighting practical approaches to achieving high yield, we present a series of case examples to illustrate the successful application of these principles. This review aims to help researchers, engineers, and industry practitioners better understand research trends in the expression and production of recombinant collagen, and to promote its further development and commercialization across diverse application areas.

Candidates:
A. It possesses remarkable biological functions and is widely used in food, cosmetics, and healthcare.
B. Collagen is the most abundant structural and functional protein in humans and other vertebrates.
C. It possesses remarkable biological functions and is not widely used in food, cosmetics, and healthcare.
D. Collagen is not the most abundant structural and functional protein in humans and other vertebrates.
E. However, recombinant collagen, heterologously expressed using genetic recombination technology combined with high-density fermentation processes, offers greater biocompatibility, low immunogenicity, and consistent quality, offering promising development prospects.
F. Currently, mainstream animal-derived collagen materials carry risks such as viral transmission and allergic reactions.
G. The evidence does not state that currently, mainstream animal-derived collagen materials carry risks such as viral transmission and allergic reactions.
H. The evidence does not state that however, recombinant collagen, heterologously expressed using genetic recombination technology combined with high-density fermentation processes, offers greater biocompatibility, low immunogenicity, and consistent quality, offering promising development prospects.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
Due to their exceptional nutritional benefits, microalgae and cyanobacteria are recognized as sustainable food sources and key contributors to the circular bioeconomy. Arthrospira sp. has garnered significant attention as one of the most promising cyanobacteria for a wide range of applications. The purpose of this study is to systematically analyze and synthesize global research trends in Arthrospira sp. applications. In this context, a “systematic review” refers to an integrated bibliometric and thematic analysis encompassing publication trends, geographical distribution of research outputs, leading journals, key application sectors, market development, and associated challenges and future prospects. Consequently, extensive research has been conducted on this species, leading to diverse areas of interest and application. This review article is the first of its kind, offering a comprehensive summary of trends in Arthrospira sp. applications over the past 15 years. It presents a bibliometric analysis of publications from 2010 to 2024 in journals indexed by Scopus. The analysis revealed that Bioresource Technology is the leading journal in publishing related research, with China producing the highest number of studies. Furthermore, phycocyanin extraction emerged as the most frequently studied application. Recently explored applications include its use as a biofertilizer, in bioplastic production, and in cosmetics. The Arthrospira sp. market is currently valued at an estimated $619 million in 2024, positioning it as a dominant player in the global industry. However, challenges persist, including safety concerns related to potential allergies and toxicity, as well as regulatory hurdles that may affect commercialization and market expansion.

Candidates:
A. The purpose of this study is to systematically analyze and synthesize global research trends in Arthrospira sp.
B. The purpose of this study is not to systematically analyze and synthesize global research trends in Arthrospira sp.
C. In this context, a “systematic review” refers to an integrated bibliometric and thematic analysis encompassing publication trends, geographical distribution of research outputs, leading journals, key application sectors, market development, and associated challenges and future prospects.
D. The evidence does not state that has garnered significant attention as one of the most promising cyanobacteria for a wide range of applications.
E. Due to their exceptional nutritional benefits, microalgae and cyanobacteria are recognized as sustainable food sources and key contributors to the circular bioeconomy.
F. The evidence does not state that in this context, a “systematic review” refers to an integrated bibliometric and thematic analysis encompassing publication trends, geographical distribution of research outputs, leading journals, key application sectors, market development, and associated challenges and future prospects.
G. Due to their exceptional nutritional benefits, microalgae and cyanobacteria are not recognized as sustainable food sources and key contributors to the circular bioeconomy.
H. has garnered significant attention as one of the most promising cyanobacteria for a wide range of applications.


Prompt:
Some candidate values differ from the evidence by only one number. Select only the strings that appear verbatim, preserving candidate order. Return JSON using exactly this schema: {"reported_values":["candidate string"]}. Candidates: ["91%", "90%"]

Evidence:
The removal of cadmium from contaminated water remains a critical challenge due to its high toxicity, persistence, and limited treatability at low concentrations. In this study, we propose a novel algal–nanoparticle system that integrates cadmium adsorption by Chlorella vulgaris with zinc ferrite (ZnFe 2 O 4 ) nanoparticle-assisted sedimentation, with the aim of addressing a significant operational challenge in algal remediation. The microalgal biomass demonstrated the capacity to remove cadmium with efficiencies exceeding 90%, facilitated by adsorption through surface functional groups. The incorporation of ZnFe 2 O 4 nanoparticles promoted the formation of dense, magnetically responsive aggregates, significantly accelerating biomass settling without the necessity for additional chemical flocculants. The strategy’s efficacy is evidenced by its enhancement of metal removal and solid–liquid separation processes, which renders it a potentially scalable and environmentally sustainable approach for the treatment of cadmium-contaminated wastewater. The strategy holds relevance for effluents derived from mining, electroplating, fertilizer production and battery manufacturing.


Prompt:
Some candidate values differ from the evidence by only one number. Select only the strings that appear verbatim, preserving candidate order. Return JSON using exactly this schema: {"reported_values":["candidate string"]}. Candidates: ["75%", "51%", "73.00%", "72.00%", "50%", "71.37%", "70.37%", "76%"]

Evidence:
Nitrogen supplements such as yeast extract and peptone/tryptone are the main cost drivers in bacterial cellulose (BC) fermentation. This study evaluated fourteen cereal, pseudo-cereal and legume flours as media substitutes for Komagataeibacter xylinus DSMZ 2325 using two strategies: (i) constant total nitrogen (CTN; 0.6 g·L −1 ) and (ii) constant nitrogen-source mass (CNSM; 5.0 g·L −1 ). BC yield (dry g·L −1 ) was determined under static cultivation and analyzed by ANOVA, correlation statistics and techno-economic assessment. Flour type and substitution level significantly influenced BC production ( p < 0.05). CTN substitution enhanced production, with the highest peak yields obtained for W-BC, C-BC, M-BC and SP-BC (6.68–8.97 g·L −1 ). CNSM substitution limited production, with O-BC and T-BC performing best (4.24–5.14 g·L −1 ). Techno-economic analysis further showed that the CTN regime substantially improved cost efficiency and reduced BC unit production cost, with the maximum reduction observed for TR-BC at 75% substitution (from 0.27 to 0.08 €/g; 70.37%) relative to the corresponding CTN HS control. Under the CNSM regime, the maximum reduction was observed for BY-BC at 50% substitution (from 0.25 to 0.07 €/g; 72.00%) relative to the corresponding CNSM HS control. These findings demonstrate that graded nitrogen substitution is an effective strategy for economically sustainable and scalable BC production.


Prompt:
Classify every fixed-vocabulary term as present or absent in the evidence. Vocabulary: bioreactor, cell culture, fermentation, chromatography, purification, filtration, viability, titer, potency, sterility, viral vector, gene therapy, cell therapy, monoclonal antibody, recombinant protein, dissolved oxygen. Return JSON with exactly two alphabetized keys: {"absent":[...],"present":[...]}. Preserve vocabulary order within each list. JSON whitespace is ignored.

Evidence:
Despite well-known benefits, the increasing use of single-use technology (SUT) in biopharmaceutical processes has raised concerns about the environmental impact of plastic waste. This paper provides the first bioprocessing industry example of a “closing the loop” proof of concept by implementing a circular economy model for a polycarbonate (PC) bioreactor vessel used in process development applications. Through a collaborative effort between an end user, a SUT supplier, and a resin supplier, a lab-scale study was initiated to collect, decontaminate, and mechanically recycle material from vessels after use in mammalian cell culture experiments to produce new vessels. The study demonstrates that using recycled PC reduces vessels' environmental footprint and does not adversely impact vessel extractables. Even in direct contact with cells and media, recycled PC left cell culture performance and monoclonal antibody production largely unaffected. This work paves the way for broader adoption of circular practices in the industry. Life Cycle Assessment (LCA) was used to evaluate closed-loop recycling of PC across multiple environmental indicators (e.g., climate change, resource use, water use, etc.). Benefits typically favor the closed-loop system, but sensitivity analysis indicates that variations in parameters such as recovery yield, contamination rate, means of transport, and electricity mix can erode these advantages in non-ideal settings. The paper outlines the logistics, challenges, and learnings from this program, emphasizing the need for standardized procedures and collaboration across teams to achieve sustainable SUT circularity. • Recycling bioreactor polycarbonate vessels reduces environmental footprint. • Equivalent extractables observed in virgin and recycled polycarbonate vessels. • Cell culture performance is comparable across recycled and virgin vessels. The online version contains supplementary material available at 10.1007/s00253-026-13796-z.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
The rising demand for renewable and sustainable oleochemicals underscores the necessity of producing yeast oil from lignocellulosic hydrolysates. We isolated two oleaginous yeast strains of Candida maltosa (UFV-1 and UFV-2) able to assimilate xylose and tolerate hemicellulosic hydrolysate inhibitors (acetic acid, furfural, hydroxymethylfurfural and formic acid). To optimize growth and lipid production by C. maltosa in malt bagasse hemicellulosic hydrolysate, we applied a central composite rotational design with response surface methodology, evaluating initial pH and yeast extract:peptone ratio (P:YE). The maximum lipid responses were recorded in: (P:YE = 1.0:0.0 g/L, pH = 6.0) for UFV-1 and (P:YE = 0.58:0.42 g/L, pH = 6.5) for UFV-2. Biomass and lipid production were higher in the detoxified hydrolysate than in the non-detoxified hydrolysate. Biomass titers increased from 9.12 to 17.0 g/L and from 10.5 to 12.9 g/L for UFV-1 and UFV-2, respectively; whilst the lipid titers increased from 0.84 to 3.14 g/L and 0.91 to 1.58 g/L for UFV-1 and UFV-2, respectively. Importantly, the levels of 16:0, 18:0, 10:0 fatty acids increased in the detoxified medium. Cultivation of the UFV-1 strain from detoxified hydrolysate in a benchtop bioreactor improved biomass formation, achieving a lipid productivity of 0.29 ± 0.02 g/L h, highlighting its potential to produce oil from hemicellulosic hydrolysates in biorefineries. The online version contains supplementary material available at 10.1007/s11274-026-04919-9. Keywords: Sustainable oleochemicals, Yeast, Response surface methodology, Biorefineries

Candidates:
A. We isolated two oleaginous yeast strains of Candida maltosa (UFV-2 and UFV-2) able to assimilate xylose and tolerate hemicellulosic hydrolysate inhibitors (acetic acid, furfural, hydroxymethylfurfural and formic acid).
B. maltosa in malt bagasse hemicellulosic hydrolysate, we applied a central composite rotational design with response surface methodology, evaluating initial pH and yeast extract:peptone ratio (P:YE).
C. The evidence does not state that the rising demand for renewable and sustainable oleochemicals underscores the necessity of producing yeast oil from lignocellulosic hydrolysates.
D. To optimize growth and lipid production by C.
E. The evidence does not state that to optimize growth and lipid production by C.
F. The rising demand for renewable and sustainable oleochemicals underscores the necessity of producing yeast oil from lignocellulosic hydrolysates.
G. The evidence does not state that maltosa in malt bagasse hemicellulosic hydrolysate, we applied a central composite rotational design with response surface methodology, evaluating initial pH and yeast extract:peptone ratio (P:YE).
H. We isolated two oleaginous yeast strains of Candida maltosa (UFV-1 and UFV-2) able to assimilate xylose and tolerate hemicellulosic hydrolysate inhibitors (acetic acid, furfural, hydroxymethylfurfural and formic acid).


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
Vero cells are extensively used in viral vaccine production, but their adaptation to serum-free suspension culture is hindered by excessive autophagy and strong anchorage dependence. In this study, we identified Connective Tissue Growth Factor (CTGF/CCN2) as significantly upregulated under starvation—an inducer of autophagy—via RNA-seq screening. A stable CTGF knockdown Vero cell line (knockdown efficiency >50%) was established using lentiviral shRNA. Functional characterization demonstrated that CTGF depletion concurrently attenuated autophagic flux (evidenced by reduced LC3-II/I ratio and lysosomal activity) and impaired cell adhesion (with adhesion rates decreased by 50%–60% on extracellular matrix proteins), while maintaining normal cell proliferation. Our findings reveal a new role for CTGF in regulating the environmental adaptation of Vero cells by coordinating autophagy and adhesion functions. The engineered cell line provides a novel strategy to overcome suspension adaptation bottlenecks, offering significant potential for improving vaccine production scalability.

Candidates:
A. Functional characterization demonstrated that CTGF depletion concurrently attenuated autophagic flux (evidenced by reduced LC3-II/I ratio and lysosomal activity) and impaired cell adhesion (with adhesion rates decreased by 50%–60% on extracellular matrix proteins), while maintaining normal cell proliferation.
B. A stable CTGF knockdown Vero cell line (knockdown efficiency >51%) was established using lentiviral shRNA.
C. A stable CTGF knockdown Vero cell line (knockdown efficiency >50%) was established using lentiviral shRNA.
D. In this study, we identified Connective Tissue Growth Factor (CTGF/CCN2) as significantly upregulated under starvation—an inducer of autophagy—via RNA-seq screening.
E. Vero cells are extensively used in viral vaccine production, but their adaptation to serum-free suspension culture is hindered by excessive autophagy and strong anchorage dependence.
F. Vero cells are extensively used in viral vaccine production, but their adaptation to serum-free suspension culture is not hindered by excessive autophagy and strong anchorage dependence.
G. In this study, we identified Connective Tissue Growth Factor (CTGF/CCN3) as significantly upregulated under starvation—an inducer of autophagy—via RNA-seq screening.
H. Functional characterization demonstrated that CTGF depletion concurrently attenuated autophagic flux (evidenced by reduced LC4-II/I ratio and lysosomal activity) and impaired cell adhesion (with adhesion rates decreased by 50%–60% on extracellular matrix proteins), while maintaining normal cell proliferation.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
Osteosarcoma (OS) remains a challenging malignancy with a high propensity for metastasis and poor survival outcomes. Bone marrow mesenchymal stem cell-derived extracellular vesicles (BMSC-EVs) have emerged as key mediators in the tumor microenvironment, promoting OS progression. This study identifies a novel molecular axis centered on circRNA-0010220 within BMSC-EVs that drives OS aggressiveness. We demonstrate that BMSC-EVs are internalized by OS cells, enhancing their proliferation, migration, and invasion. High-throughput sequencing revealed circRNA-0010220 as the most significantly upregulated circRNA in EV-treated OS cells. Functional studies showed that circRNA-0010220 knockdown in BMSCs attenuated the oncogenic effects of their EVs both in vitro and in vivo. Mechanistically, circRNA-0010220 recruits the histone methyltransferase EZH2 to the CTNNBIP1 promoter, facilitating H3K27me3-mediated epigenetic silencing. The subsequent downregulation of CTNNBIP1 leads to activation of the Wnt/β-catenin signaling pathway. This cascade was consistently observed across gain-of-function and loss-of-function experiments, and pharmacologic inhibition of β-catenin reversed the pro-tumorigenic effects. Our findings elucidate a complete signaling axis from BMSC-EVs to Wnt/β-catenin activation via circRNA-0010220/EZH2/CTNNBIP1, providing new insights into the epigenetic regulation of OS progression and suggesting potential therapeutic targets.

Candidates:
A. This study identifies a novel molecular axis centered on circRNA-0010220 within BMSC-EVs that drives OS aggressiveness.
B. This study identifies a novel molecular axis centered on circRNA-10221 within BMSC-EVs that drives OS aggressiveness.
C. Bone marrow mesenchymal stem cell-derived extracellular vesicles (BMSC-EVs) have emerged as key mediators in the tumor microenvironment, promoting OS progression.
D. We demonstrate that BMSC-EVs are not internalized by OS cells, enhancing their proliferation, migration, and invasion.
E. Osteosarcoma (OS) remains a challenging malignancy with a high propensity for metastasis and poor survival outcomes.
F. The evidence does not state that osteosarcoma (OS) remains a challenging malignancy with a high propensity for metastasis and poor survival outcomes.
G. We demonstrate that BMSC-EVs are internalized by OS cells, enhancing their proliferation, migration, and invasion.
H. The evidence does not state that bone marrow mesenchymal stem cell-derived extracellular vesicles (BMSC-EVs) have emerged as key mediators in the tumor microenvironment, promoting OS progression.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
In this review, we systematically categorize diverse organoid engineering strategies—including cellular programming, material engineering, and platform- or system-level innovations—according to their impact on reproducibility and scalability, and highlight representative applications and emerging directions. By reframing organoid generation as a manufacturing process, these technological advances pave the way toward standardized and high-fidelity organoid production for both fundamental research and translational applications. Subject terms: Biological techniques, Biotechnology, Engineering, Medical research, Stem cells

Candidates:
A. Subject terms: Biological techniques, Biotechnology, Engineering, Medical research, Stem cells
B. In this review, we systematically categorize diverse organoid engineering strategies—including cellular programming, material engineering, and platform- or system-level innovations—according to their impact on reproducibility and scalability, and highlight representative applications and emerging directions.
C. The evidence does not state that in this review, we systematically categorize diverse organoid engineering strategies—including cellular programming, material engineering, and platform- or system-level innovations—according to their impact on reproducibility and scalability, and highlight representative applications and emerging directions.
D. The evidence does not state that subject terms: Biological techniques, Biotechnology, Engineering, Medical research, Stem cells
E. By reframing organoid generation as a manufacturing process, these technological advances pave the way toward standardized and high-fidelity organoid production for both fundamental research and translational applications.
F. The evidence does not state that by reframing organoid generation as a manufacturing process, these technological advances pave the way toward standardized and high-fidelity organoid production for both fundamental research and translational applications.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
The rapid escalation of global plastic consumption, coupled with the environmental impacts of petrochemical polymers, has sparked a surge of interest in bioplastics, particularly those derived from microbial and enzymatic processes. This review provides a comprehensive overview of the metabolic pathways, structural properties and emerging technological innovations shaping the next generation of bioplastics, with a particular focus on polyhydroxyalkanoates (PHA). The following sections outline the conceptual distinctions between bio‐based and biodegradable plastics, the key bacterial pathways responsible for the biosynthesis of PHA, PLA precursors, bacterial cellulose, microbial polyamides and other bio‐derived polymers. The physicochemical and morphological features of PHA‐based materials are analysed as well. These features include monomer composition, crystallinity, copolymer architecture and molecular weight. The relationship between these features and the mechanical and thermal performance of the materials is then investigated. A dedicated section is allocated to recent advances in in vitro enzymatic PHA synthesis, covering PHA synthase (PhaC) classes, engineered variants, cell‐free metabolic engineering platforms, enzyme immobilisation and surface‐display strategies that enable fully programmable and modular polymerisation. Finally, we discuss future perspectives, with particular emphasis on sustainable feedstocks, process intensification through synthetic biology, techno‐economic challenges and the regulatory landscape required for large‐scale adoption. The present review integrates biochemical, structural and bioprocessing insights to map current progress and identify strategic directions for enabling enzymatic bioplastics as scalable, customisable and environmentally sound alternatives within a circular bioeconomy framework. This review highlights recent advances in microbial and enzymatic routes for producing polyhydroxyalkanoate‐based bioplastics, with emphasis on engineered enzymes and cell‐free systems. By integrating biochemical and bioprocess insights, it outlines strategies to enable scalable and sustainable biopolymer production within a circular bioeconomy. Keywords: bioplastic copolymers, enzymatic polymerisation, metabolic engineering, PHA synthase

Candidates:
A. The evidence does not state that the rapid escalation of global plastic consumption, coupled with the environmental impacts of petrochemical polymers, has sparked a surge of interest in bioplastics, particularly those derived from microbial and enzymatic processes.
B. The evidence does not state that the following sections outline the conceptual distinctions between bio‐based and biodegradable plastics, the key bacterial pathways responsible for the biosynthesis of PHA, PLA precursors, bacterial cellulose, microbial polyamides and other bio‐derived polymers.
C. The rapid escalation of global plastic consumption, coupled with the environmental impacts of petrochemical polymers, has sparked a surge of interest in bioplastics, particularly those derived from microbial and enzymatic processes.
D. The physicochemical and morphological features of PHA‐based materials are analysed as well.
E. The physicochemical and morphological features of PHA‐based materials are not analysed as well.
F. The following sections outline the conceptual distinctions between bio‐based and biodegradable plastics, the key bacterial pathways responsible for the biosynthesis of PHA, PLA precursors, bacterial cellulose, microbial polyamides and other bio‐derived polymers.
G. This review provides a comprehensive overview of the metabolic pathways, structural properties and emerging technological innovations shaping the next generation of bioplastics, with a particular focus on polyhydroxyalkanoates (PHA).
H. The evidence does not state that this review provides a comprehensive overview of the metabolic pathways, structural properties and emerging technological innovations shaping the next generation of bioplastics, with a particular focus on polyhydroxyalkanoates (PHA).


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
Bacteria‐based cancer therapy is a promising cancer treatment strategy that utilizes genetically engineered attenuated bacteria to specifically target tumor tissues, directly kill cancer cells, or activate the host's immune response. Engineering modifications of attenuated bacteria can achieve more effective anti‐tumor effects, and consequently, a number of clinical trials have been carried out. However, the clinical translation of attenuated bacteria faces multiple challenges, including safety risks, fluctuating therapeutic efficacy, difficulties in vivo monitoring, complex production quality control, and lack of regulatory approval standards. This review mainly summarizes five aspects of bacterial cancer therapy: the safety risks and attenuation strategies, the regulation of therapeutic stability, bacterial in vivo imaging technologies, the optimization of bacterial production processes, and market approval pathways. It aims to precisely control bacterial behavior through genetic circuits and enable real‐time monitoring of the treatment process by means of multimodal imaging, ultimately promoting the clinical translation of attenuated bacterial therapy.

Candidates:
A. The evidence does not state that this review mainly summarizes five aspects of bacterial cancer therapy: the safety risks and attenuation strategies, the regulation of therapeutic stability, bacterial in vivo imaging technologies, the optimization of bacterial production processes, and market approval pathways.
B. However, the clinical translation of attenuated bacteria faces multiple challenges, including safety risks, fluctuating therapeutic efficacy, difficulties in vivo monitoring, complex production quality control, and lack of regulatory approval standards.
C. This review mainly summarizes five aspects of bacterial cancer therapy: the safety risks and attenuation strategies, the regulation of therapeutic stability, bacterial in vivo imaging technologies, the optimization of bacterial production processes, and market approval pathways.
D. Bacteria‐based cancer therapy is not a promising cancer treatment strategy that utilizes genetically engineered attenuated bacteria to specifically target tumor tissues, directly kill cancer cells, or activate the host's immune response.
E. The evidence does not state that however, the clinical translation of attenuated bacteria faces multiple challenges, including safety risks, fluctuating therapeutic efficacy, difficulties in vivo monitoring, complex production quality control, and lack of regulatory approval standards.
F. Bacteria‐based cancer therapy is a promising cancer treatment strategy that utilizes genetically engineered attenuated bacteria to specifically target tumor tissues, directly kill cancer cells, or activate the host's immune response.
G. Engineering modifications of attenuated bacteria cannot achieve more effective anti‐tumor effects, and consequently, a number of clinical trials have been carried out.
H. Engineering modifications of attenuated bacteria can achieve more effective anti‐tumor effects, and consequently, a number of clinical trials have been carried out.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
Next-generation prebiotics are increasingly recognized for their ability to modulate gut microbiota and promote host health. Xylo-oligosaccharides (XOS) and malto-oligosaccharides (MOS), derived from cereal biomass, are particularly promising due to their structural stability, low effective dosages, and selective stimulation of beneficial gut microorganisms. This review synthesizes current knowledge on XOS and MOS, with emphasis on enzyme-enabled production technologies, microbial fermentation behavior, and health outcomes in humans and animals. Recent advances in tailored enzyme cocktails and sequential bioconversion strategies enable precise control over oligosaccharide yield and degree of polymerization, facilitating targeted production of functionally distinct fractions. Comparative analyses demonstrate that XOS and MOS offer greater microbial selectivity than conventional prebiotics, such as fructo- and galacto-oligosaccharides, and consistently enhance short-chain fatty acid production. These effects contribute to improved metabolic regulation, immune function, and resistance to enteric pathogens. A distinctive contribution of this review is the valorization of Kazakhstan’s underutilized cereal residues, including wheat, rice, barley, and millet straw and bran, as sustainable feedstocks for prebiotic production. Integrating regional biomass with engineered enzyme systems provides a scalable and environmentally sound platform. Furthermore, combined XOS–MOS formulations may exert synergistic microbiota-modulating effects through complementary fermentation dynamics. Collectively, these insights position enzyme-based conversion of cereal biomass as a novel, regionally relevant strategy for advancing next-generation prebiotics.

Candidates:
A. This review synthesizes current knowledge on XOS and MOS, with emphasis on enzyme-enabled production technologies, microbial fermentation behavior, and health outcomes in humans and animals.
B. The evidence does not state that recent advances in tailored enzyme cocktails and sequential bioconversion strategies enable precise control over oligosaccharide yield and degree of polymerization, facilitating targeted production of functionally distinct fractions.
C. The evidence does not state that this review synthesizes current knowledge on XOS and MOS, with emphasis on enzyme-enabled production technologies, microbial fermentation behavior, and health outcomes in humans and animals.
D. Next-generation prebiotics are not increasingly recognized for their ability to modulate gut microbiota and promote host health.
E. Xylo-oligosaccharides (XOS) and malto-oligosaccharides (MOS), derived from cereal biomass, are particularly promising due to their structural stability, low effective dosages, and selective stimulation of beneficial gut microorganisms.
F. Recent advances in tailored enzyme cocktails and sequential bioconversion strategies enable precise control over oligosaccharide yield and degree of polymerization, facilitating targeted production of functionally distinct fractions.
G. Xylo-oligosaccharides (XOS) and malto-oligosaccharides (MOS), derived from cereal biomass, are not particularly promising due to their structural stability, low effective dosages, and selective stimulation of beneficial gut microorganisms.
H. Next-generation prebiotics are increasingly recognized for their ability to modulate gut microbiota and promote host health.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
In biotechnological processes, cell density and physiology are critical parameters for controlling the feed rate, harvest time, and process performance. We developed an automated flow cytometry approach that enables continuous, real-time (fully automated, hourly) monitoring of bacterial populations in continuous bioreactors. The method employed a double-staining protocol that combined DAPI to assess total DNA content and Alexa Fluor 488-EdU via Click-iT technology to identify the proportions of cells undergoing active DNA replication through EdU incorporation. The integrated workflow included fixation, permeabilization, staining, and measurement steps and was applied to three Gram-negative strains: Bradyrhizobium sp., Escherichia coli , and Stenotrophomonas rhizophila . Automated analysis captured growth dynamics and cell cycle progression, providing insights into population behavior under different dilution rates. In this study, automated on-line sampling enabled hourly flow cytometry measurements of cell concentration and physiological indicators during continuous cultivation, supporting real-time monitoring and control in industrial biotechnology.

Candidates:
A. The method employed a double-staining protocol that combined DAPI to assess total DNA content and Alexa Fluor 489-EdU via Click-iT technology to identify the proportions of cells undergoing active DNA replication through EdU incorporation.
B. The evidence does not state that we developed an automated flow cytometry approach that enables continuous, real-time (fully automated, hourly) monitoring of bacterial populations in continuous bioreactors.
C. The integrated workflow included fixation, permeabilization, staining, and measurement steps and was not applied to three Gram-negative strains: Bradyrhizobium sp., Escherichia coli , and Stenotrophomonas rhizophila .
D. The integrated workflow included fixation, permeabilization, staining, and measurement steps and was applied to three Gram-negative strains: Bradyrhizobium sp., Escherichia coli , and Stenotrophomonas rhizophila .
E. The method employed a double-staining protocol that combined DAPI to assess total DNA content and Alexa Fluor 488-EdU via Click-iT technology to identify the proportions of cells undergoing active DNA replication through EdU incorporation.
F. In biotechnological processes, cell density and physiology are critical parameters for controlling the feed rate, harvest time, and process performance.
G. We developed an automated flow cytometry approach that enables continuous, real-time (fully automated, hourly) monitoring of bacterial populations in continuous bioreactors.
H. In biotechnological processes, cell density and physiology are not critical parameters for controlling the feed rate, harvest time, and process performance.


Prompt:
Some candidate values differ from the evidence by only one number. Select only the strings that appear verbatim, preserving candidate order. Return JSON using exactly this schema: {"reported_values":["candidate string"]}. Candidates: ["65%", "3%", "37 °C", "64%", "101 rpm", "4%", "38 °C", "100 rpm"]

Evidence:
This study investigates the optimisation of poly(3-hydroxybutyrate) (PHB) production by Mycolicibacterium smegmatis using sugarcane bagasse (SCB) hydrolysate as a low-cost, renewable carbon source. Key fermentation parameters including temperature, pH, agitation, inoculum size and nitrogen supplementation were optimised to enhance biomass growth and PHB accumulation. Under optimised conditions (37 °C, pH 7, 100 rpm, 3% inoculum), M. smegmatis achieved a maximum PHB content of 64% of DCW from SCB hydrolysate, corresponding to a PHB titre of ~ 1.0 g L −1 and a volumetric productivity of 0.014 g L −1 h −1 after 72 h of cultivation, with yeast extract identified as the most effective nitrogen source. Thin-layer chromatography (TLC) analysis demonstrated that M. smegmatis could not only co-utilise glucose and xylose simultaneously but preferentially consumed xylose, which is a major advantage when processing lignocellulosic biomasses, where xylose is abundant and typically underutilised by many microorganisms. Whole-genome sequencing with Oxford Nanopore Technologies (ONT) confirmed the presence of PHB production genes ( phbA , phbB , phbC ) and a xylose-utilisation pathway, supporting its metabolic capability. These findings establish M. smegmatis as a promising candidate for converting agricultural waste into biodegradable bioplastics, contributing to circular bioeconomy strategies. The online version contains supplementary material available at 10.1186/s40643-026-01047-y.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
Membrane fouling during ultrafiltration/diafiltration (UF/DF) remains a major challenge for extending membrane lifetime in biologics manufacturing. Developing cleaning strategies that combine high efficiency with scalability is essential for robust downstream processing. In this study, different UF/DF membrane cleaning methods were assessed under bench-scale conditions with consideration for large-scale implementation. Permeate-closed cleaning (PCC), which temporarily closes the permeate port to enhance shear, effectively mitigated normalized water permeability (NWP) decline. Furthermore, forward and reverse PCC at elevated feed flux achieved comparable fouling control with fewer steps by leveraging hydrodynamic shear and localized backwashing. To simplify large-scale adoption, high-flux PCC was developed and demonstrated the ability to restore NWP and maintain stability over extended cycles. Quantitatively, high‑flux PCC reduced early NWP decay by ~ 18% over the first three cycles and recovered > 15% NWP in previously fouled membranes, stabilizing permeability over extended reuse. Backwashing under controlled negative transmembrane pressure (TMP) further improved pore-level cleaning and extended membrane usability. Additionally, combining sodium hypochlorite (NaClO) with sodium hydroxide (NaOH) provided strong oxidative and alkaline action; within the evaluated range (25–150 ppm NaClO), 50 ppm was identified as a practical lower limit for effective recovery, while 150 ppm restored NWP from ~ 70 to ~ 100% with residual NaClO < 0.02 ppm after final rinsing. Collectively, these findings present a portfolio of cleaning strategies—hydrodynamic, chemical, and combined approaches—that enhance cleaning efficiency, reduce chemical exposure, and support sustainable UF/DF operations in biologics manufacturing. The online version contains supplementary material available at 10.1186/s40643-026-01045-0. Keywords: Ultrafiltration/diafiltration (UF/DF), Membrane fouling, Cleaning strategies, Permeate-closed cleaning, Backwashing, Biologics downstream processing

Candidates:
A. The evidence does not state that membrane fouling during ultrafiltration/diafiltration (UF/DF) remains a major challenge for extending membrane lifetime in biologics manufacturing.
B. In this study, different UF/DF membrane cleaning methods were not assessed under bench-scale conditions with consideration for large-scale implementation.
C. Membrane fouling during ultrafiltration/diafiltration (UF/DF) remains a major challenge for extending membrane lifetime in biologics manufacturing.
D. Developing cleaning strategies that combine high efficiency with scalability is not essential for robust downstream processing.
E. Permeate-closed cleaning (PCC), which temporarily closes the permeate port to enhance shear, effectively mitigated normalized water permeability (NWP) decline.
F. The evidence does not state that permeate-closed cleaning (PCC), which temporarily closes the permeate port to enhance shear, effectively mitigated normalized water permeability (NWP) decline.
G. In this study, different UF/DF membrane cleaning methods were assessed under bench-scale conditions with consideration for large-scale implementation.
H. Developing cleaning strategies that combine high efficiency with scalability is essential for robust downstream processing.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
Acacia seyal is a medicinal plant rich in bioactive compounds known for their antimicrobial, antioxidant, and anticancer properties. Conventional plant extracts often suffer from poor stability and limited bioavailability, reducing their therapeutic effectiveness. The utilization of chitosan nanoparticles (CS-NPs) offers a promising strategy to enhance biological activity of plant extracts. Encapsulation of A. seyal seeds extract (ASSE) in CS-NPs (ASSE@CS-NPs) was the aim of the present investigation to compete Helicobacter pylori and HCT116 cancer cells. The phytochemical profile of ASSE revealed diverse phenolic acids and flavonoids via HPLC analysis. Methyl gallate was dominated (20.98 mg/g), followed by gallic acid, and catechin. FTIR analysis confirmed characteristic functional groups in ASSE and CS-NPs. Peak shifts and intensity variations in ASSE@CS-NPs indicated successful encapsulation through hydrogen bonding and electrostatic interactions. ASSE@CS-NPs showed the strongest anti- H. pylori effect with inhibition zone (23.7 mm) and lowest MIC/MBC (15.62 µg/mL). Compared to CS-NPs (20.7 mm, 31.25 µg/mL) and ASSE (19.7 mm, 31.25 µg/mL). ASSE@CS-NPs showed notable antioxidant activity, achieving 96% scavenging at 1000 µg/mL with IC 50 of 3µg/mL compared to ASSE (5 µg/mL) and CS-NPs (74 µg/mL). ASSE@CS-NPs exhibited potent cytotoxicity against HCT116 cells with IC 50 of 23 µg/mL, outperforming ASSE (588 µg/mL) and CS-NPs (120 µg/mL). The formulation achieved > 80% inhibition at 62.5 µg/mL. Molecular docking was performed to evaluate the binding affinity and interaction patterns of methyl gallate (a main constituent of ASSE) and chitosan against H. pylori urease (PDB ID: 6ZJA). Docking results revealed favorable binding scores for both ligands, with chitosan showing higher affinity (S score: − 6.42 kcal/mol) compared to methyl gallate (S score: − 5.45 kcal/mol). Key hydrogen bond interactions were observed with active site residues ASP362 and ALA169 for methyl gallate, and ASP223 and HIS323 for chitosan. These results suggest that ASSE@CS-NPs possess inhibitory potential against H. pylori and HCT116 cells. The online version contains supplementary material available at 10.1186/s40643-026-01031-6.

Candidates:
A. The evidence does not state that conventional plant extracts often suffer from poor stability and limited bioavailability, reducing their therapeutic effectiveness.
B. seyal seeds extract (ASSE) in CS-NPs (ASSE@CS-NPs) was the aim of the present investigation to compete Helicobacter pylori and HCT116 cancer cells.
C. The evidence does not state that the utilization of chitosan nanoparticles (CS-NPs) offers a promising strategy to enhance biological activity of plant extracts.
D. Acacia seyal is a medicinal plant rich in bioactive compounds known for their antimicrobial, antioxidant, and anticancer properties.
E. seyal seeds extract (ASSE) in CS-NPs (ASSE@CS-NPs) was the aim of the present investigation to compete Helicobacter pylori and HCT117 cancer cells.
F. Conventional plant extracts often suffer from poor stability and limited bioavailability, reducing their therapeutic effectiveness.
G. Acacia seyal is not a medicinal plant rich in bioactive compounds known for their antimicrobial, antioxidant, and anticancer properties.
H. The utilization of chitosan nanoparticles (CS-NPs) offers a promising strategy to enhance biological activity of plant extracts.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
The use of animal-derived reagents in biomedical research poses challenges for reproducibility due to batch-to-batch variability and inter-species differences, along with ethical concerns related to their origin. In pursuing a human-relevant in vitro model, an animal-free and defined cell culture process is preferred to improve relevance and reproducibility. We investigated the use of serum replacement (SR) consisting of human hepatocyte-derived proteins in cell culture and recombinant antibodies with a plant-derived blocking solution (animal-free blocker, AFB) in immunocytochemical staining of cells. Human serum (HS) instead of animal-derived serum was used in this study for comparison with SR. We showed that bone marrow stem/stromal cells (BMSCs) maintain their proliferation capacity and cell-specific morphology in SR-supplemented medium, whereas human umbilical vein endothelial cells (HUVECs) show compromised growth under similar conditions. In a more complex co-culture, BMSCs + HUVECs formed a stable vascular network in SR-supplemented medium. In immunocytochemical staining, we compared the performance of recombinant antibodies with animal-derived antibodies and an AFB solution with a bovine serum albumin (BSA)-based blocking solution. Adipose stem/stromal cells (ASCs) showed their typical spindle-shaped morphology when stained with recombinant antibodies against alpha-smooth muscle actin (αSMA) in both AFB and BSA-based blocking solutions. We detected partial non-specific binding of recombinant antibodies and animal-derived antibodies against β-tubulin III in ASC. In contrast, we did not observe non-specific binding on these neuronal antibodies in HUVECs in any tested condition. While protocol optimization depends on the cell type used, our findings indicate that animal-derived materials can reliably be replaced.

Candidates:
A. The evidence does not state that the use of animal-derived reagents in biomedical research poses challenges for reproducibility due to batch-to-batch variability and inter-species differences, along with ethical concerns related to their origin.
B. In pursuing a human-relevant in vitro model, an animal-free and defined cell culture process is preferred to improve relevance and reproducibility.
C. We investigated the use of serum replacement (SR) consisting of human hepatocyte-derived proteins in cell culture and recombinant antibodies with a plant-derived blocking solution (animal-free blocker, AFB) in immunocytochemical staining of cells.
D. Human serum (HS) instead of animal-derived serum was used in this study for comparison with SR.
E. Human serum (HS) instead of animal-derived serum was not used in this study for comparison with SR.
F. In pursuing a human-relevant in vitro model, an animal-free and defined cell culture process is not preferred to improve relevance and reproducibility.
G. The use of animal-derived reagents in biomedical research poses challenges for reproducibility due to batch-to-batch variability and inter-species differences, along with ethical concerns related to their origin.
H. The evidence does not state that we investigated the use of serum replacement (SR) consisting of human hepatocyte-derived proteins in cell culture and recombinant antibodies with a plant-derived blocking solution (animal-free blocker, AFB) in immunocytochemical staining of cells.


Prompt:
Some candidate values differ from the evidence by only one number. Select only the strings that appear verbatim, preserving candidate order. Return JSON using exactly this schema: {"reported_values":["candidate string"]}. Candidates: ["4 h", "93%", "18 h", "19 h", "2 h", "94%", "5 h", "3 h"]

Evidence:
Corynebacterium glutamicum is a major industrial cell factory for amino acid production. Acidic byproduct accumulation can lower broth pH, disrupt cytoplasmic pH homeostasis, induce oxidative stress, and ultimately compromise productivity. As low-pH stress is dynamic and process-dependent, distinguishing responses to acute acid shock versus sustained acidification is important for improving strain robustness. Here, we investigated low-pH adaptation under two regimes: acute, unbuffered pH 4.0 shock and sustained pH 5.5 stress with pH control, by integrating viability assays, time-series RNA-seq, and genetic validation. Acute shock caused 93% viability loss within 2 h, followed by broth neutralisation and regrowth, whereas sustained stress led to a 4.1-log 10 decline over 18 h. Transcriptomics identified 905/847 differentially expressed genes at 1/4 h under acute shock and 643/608/1857 genes at 1/8/18 h under sustained stress. Shared responses included repression of central metabolism and induction of β-ketoadipate catabolism, potassium uptake, sodium/proton antiport, urease-mediated ammonium release, amino acid biosynthesis, and oxidative and membrane-stress defences. Acute shock showed rapid global reprogramming dominated by oxidative protection, chaperone induction, and ion-flux control, whereas sustained stress induced progressive metabolic rewiring, cell envelope reinforcement, and redox buffering. Functional validation using both overexpression and knockout mutants confirmed the contribution of ion transport, iron regulation, β-ketoadipate metabolism, urease, and respiratory modules to low-pH tolerance in a regime-dependent manner. This study provides the first time-resolved, regime-specific transcriptomic dissection of low-pH adaptation in C. glutamicum and identifies key modules for engineering acid-resilient strains.


Prompt:
Classify every fixed-vocabulary term as present or absent in the evidence. Vocabulary: bioreactor, cell culture, fermentation, chromatography, purification, filtration, viability, titer, potency, sterility, viral vector, gene therapy, cell therapy, monoclonal antibody, recombinant protein, dissolved oxygen. Return JSON with exactly two alphabetized keys: {"absent":[...],"present":[...]}. Preserve vocabulary order within each list. JSON whitespace is ignored.

Evidence:
Thermophilic methanogens of the genus Methanothermobacter are established biocatalysts in power-to-gas applications, converting H 2 and CO 2 into CH 4 through the process of methanogenesis. Further expanding this platform for the bioproduction of value-added compounds (power-to-x) has the potential to increase the economic viability of such processes. This requires a genetic toolset that enables the controlled expression of recombinant pathways. Here, we report the fully autotrophic inducible recombinant bioproduction of acetoin from H 2 and CO 2 in Methanothermobacter thermautotrophicus ΔH. To facilitate inducible gene expression, we implemented an anhydrotetracycline (aTc)-inducible promoter system, expanding our available set of promoters. The aTc-inducible system enabled controlled expression of a codon-optimized acetoin-production operon comprising the acetolactate synthase- and acetolactate decarboxylase-encoding genes from Streptococcus thermophilus . Batch cultivation at 42 °C demonstrated aTc-dependent acetoin formation, yielding up to 0.45 ± 0.08 mM acetoin. Fed-batch bioreactor experiments confirmed growth-coupled, recombinant acetoin production, while eliminating the non-specific acetoin accumulation that we observed during non-growth phases in batch cultivation. Continuous cultivation in a chemostat resulted in stable acetoin production rates of 1.28 ± 0.07 μmol L −1 h −1 (0.11 ± 0.01 mg L −1 h −1 ) at 42 °C. Elevated temperatures led to reduced acetoin production, suggesting diminished activity or thermal instability of the heterologous enzymes. This study demonstrates the feasibility of value-added bioproduction in Methanothermobacter and establishes an inducible expression system suitable for pathway engineering in thermophilic methanogens. Together with genome-scale modeling and emerging enzyme engineering strategies, these results lay the foundation for developing robust, CH 4 -co-producing power-to-x bioprocesses with Methanothermobacter species. Keywords: Methanogenic archaea, Genetic engineering, Inducible promoter system, Bioreactors, Chemostat, Recombinant acetoin production


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
The immortalized NK-92 cell line is widely used to study natural killer (NK) cell biology and develop immunotherapies. NK-92 cells exhibit strong cytotoxic activity against tumor and virus-infected cells and are often used as a functional surrogate for primary NK cells. Beyond research applications, NK-92 cells are currently being evaluated in clinical trials as an allogeneic, off-the-shelf cell therapy. NK cells are typically cultured in static systems, which limits scalability. For clinical and commercial applications, large-scale cell expansion requires scalable platforms, such as bioreactors. However, conventional bubble-aerated bioreactors generate shear stress and foam, which can impair proliferation during prolonged culture and compromise cell quality. To address these limitations, a membrane-based stirring and aeration system was compared to a conventional pitched-blade impeller with microsparger aeration in 2 L stirred-tank bioreactors. NK-92 cells were expanded from static pre-cultures into shake flasks and subsequently inoculated into each bioreactor with continuous feeding to maintain ideal nutrient supply. Both systems supported comparable growth, viability, and metabolic profiles. Cells showed comparable growth, viability and metabolism profile in both systems. However, cells expanded in the membrane-based system exhibited markedly higher cytotoxicity and cytotoxic capacity. Computational fluid dynamic simulations of both systems suggest that this observation is likely attributable to the more homogeneous shear distribution in the membrane-stirred setup compared to the pitched-blade configuration. Overall, this work presents a novel cultivation method to produce highly cytotoxic NK-92 cells in a well scalable stirred-tank bioreactor platform for allogeneic off-the-shelf cell therapy.

Candidates:
A. The immortalized NK-92 cell line is widely used to study natural killer (NK) cell biology and develop immunotherapies.
B. Beyond research applications, NK-92 cells are currently being evaluated in clinical trials as an allogeneic, off-the-shelf cell therapy.
C. NK cells are not typically cultured in static systems, which limits scalability.
D. NK cells are typically cultured in static systems, which limits scalability.
E. NK-93 cells exhibit strong cytotoxic activity against tumor and virus-infected cells and are often used as a functional surrogate for primary NK cells.
F. The immortalized NK-93 cell line is widely used to study natural killer (NK) cell biology and develop immunotherapies.
G. NK-92 cells exhibit strong cytotoxic activity against tumor and virus-infected cells and are often used as a functional surrogate for primary NK cells.
H. Beyond research applications, NK-93 cells are currently being evaluated in clinical trials as an allogeneic, off-the-shelf cell therapy.


Prompt:
Some candidate values differ from the evidence by only one number. Select only the strings that appear verbatim, preserving candidate order. Return JSON using exactly this schema: {"reported_values":["candidate string"]}. Candidates: ["2 L", "81%", "80%", "2.7%", "100%", "3 L", "3.7%", "99%"]

Evidence:
Wastewater treatment faces increasing pressure to transition from energy-intensive technology to sustainable alternatives aligned with global resource efficiency and climate goals. Microalgae-based processes have emerged as promising solutions in environmental remediation applications; however, their large-scale deployment remains constrained by contamination risks, stringent operational requirements, and high downstream costs. These challenges are particularly evident in the treatment of nutrient-rich industrial influents such as dairy wastewater, which represents an environmental concern. Addressing this gap is important for strengthening overall climate action efforts and safeguarding ecosystems by reducing greenhouse gas emissions and transforming nutrient loads from pollution sources into potential resources. In this study, a two-stage biological treatment of raw dairy wastewater was tested as an alternative to conventional technology. The process relied on activated algae biomass, consisting of microalgae-bacteria consortia, operated in sequencing batch mode. The treatment stages were strategically designed to address elevated organic and ammonium loads while maintaining aerobic conditions exclusively through photosynthesis. The first stage operated at high COD loadings (>1 g O 2 L) and achieved organic matter removal above 80%, while the second stage, adapted to lower COD (<0.5 g O 2 L), ensured residual ammonium below the detection limit and overall COD removal up to 99%. Optimization of operational conditions further improved microalgae harvesting efficiency (from 88.6 ± 2.7% to 94.4 ± 1.8%) and enhanced floc stability through diversification of microalgae communities. Complementary, microfauna analysis outlined the presence of protozoan and metazoan populations confirming process stability and ecological balance comparable to traditional activated sludge system. The findings demonstrate potential of the activated algae system as a resilient and resource-efficient alternative to conventional wastewater treatment technology. By avoiding energy demand for mechanical aeration and ensuring nutrient recovery in line with environmental regulatory frameworks, the developed process supports sustainable wastewater treatment management while aligning with international goals on climate change mitigation and aquatic ecosystems protection.


Prompt:
Identify every candidate statement reproduced exactly from the evidence. Return JSON using exactly this schema: {"supported":["A"]}. Preserve candidate order. JSON whitespace is ignored.

Evidence:
Current academic research and industrial processes for cultivated meat production primarily utilize cultures of single cell types (monocultures) that do not capture the complexity of the in vivo environment of muscle and fatty tissues in the body. These systems are simplified for operational practicalities and cost reduction rather than to improve product quality. Irrespective of the manufacturing process, a food product must be of high quality to survive in competitive markets. Co‐culture of multiple cell types is a well‐established practice that enhances tissue qualities by allowing the cells to exchange cytokines and nutrients and to engage in cell‐cell signaling. These synergies promote tissue function and phenotype stability and alter cellular and tissue composition. Co‐culture can improve processes by reducing the need for exogenous media components such as growth factors. Expanding this system to include the products of rumen fermentation, such as B‐vitamins and short‐chain fatty acid (SCFA) has potential to further improve culture conditions and product qualities, particularly for cultivated meat of ruminant species (e.g., cattle, goats, sheep, and lambs). In this review, we cover the role of multiple cell types (notably fibroblasts and liver cells) as well as the role of rumen fermentation products in supporting the development and quality of the desired muscle and fatty tissues. We identify the opportunities and highlight the challenges associated with connecting these cultures as modules in a multi‐organ bioreactor series to further enable synergies and improve product qualities as part of a novel, bio‐inspired, multi‐organ production system.

Candidates:
A. The evidence does not state that irrespective of the manufacturing process, a food product must be of high quality to survive in competitive markets.
B. These systems are not simplified for operational practicalities and cost reduction rather than to improve product quality.
C. Co‐culture of multiple cell types is not a well‐established practice that enhances tissue qualities by allowing the cells to exchange cytokines and nutrients and to engage in cell‐cell signaling.
D. Irrespective of the manufacturing process, a food product must be of high quality to survive in competitive markets.
E. These systems are simplified for operational practicalities and cost reduction rather than to improve product quality.
F. The evidence does not state that current academic research and industrial processes for cultivated meat production primarily utilize cultures of single cell types (monocultures) that do not capture the complexity of the in vivo environment of muscle and fatty tissues in the body.
G. Co‐culture of multiple cell types is a well‐established practice that enhances tissue qualities by allowing the cells to exchange cytokines and nutrients and to engage in cell‐cell signaling.
H. Current academic research and industrial processes for cultivated meat production primarily utilize cultures of single cell types (monocultures) that do not capture the complexity of the in vivo environment of muscle and fatty tissues in the body.