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:
Gene therapy is based on the introduction of nucleic acids into cells using vectors to enable therapeutic benefits. Hereby, adeno-associated virus (AAV) vectors are preferred for in vivo gene delivery to target cells due to their stability, their ability to transduce a broad range of cell types, and the long-term expression of vector-encoded therapeutic genes in slowly or non-replicating cells. However, large-scale production and quality control during and after manufacturing represent significant challenges. Currently, methods for the analysis of AAVs are rather complex, laborious, expensive, and poorly standardized. Most analyses have to be performed off-line and are not suitable for adaptation toward real-time monitoring during the manufacturing process. Optical analytical methods such as infrared attenuated total reflection (IR-ATR) spectroscopy offer a promising alternative serving as a rapid, cost-efficient, and non-destructive analytical technique with potential on-line monitoring capabilities. The present study fundamentally demonstrates the feasibility of IR-ATR spectroscopy to classify and quantify reference standard materials of full vs. empty AAV type 2 virus capsids. Chemometric methods enable rapid and automated processing of the obtained spectra using specifically partial least squares regression (PLS-R) models. Excellent classification results were achieved, unambiguously identifying samples with predominantly empty capsids vs. those containing predominantly full capsids. The obtained results confirm that IR-ATR spectroscopy in combination with multivariate data evaluation strategies is indeed capable of providing information on the presence of genome-containing virus particles along with the corresponding genomic titer. This analytical strategy may potentially translate into a real-world on-line bioreactor process analytical monitoring technology (PAT) augmenting viral vector production scenarios. The online version contains supplementary material available at 10.1007/s00216-026-06429-x.

Candidates:
A. Hereby, adeno-associated virus (AAV) vectors are preferred for in vivo gene delivery to target cells due to their stability, their ability to transduce a broad range of cell types, and the long-term expression of vector-encoded therapeutic genes in slowly or non-replicating cells.
B. Gene therapy is not based on the introduction of nucleic acids into cells using vectors to enable therapeutic benefits.
C. Currently, methods for the analysis of AAVs are not rather complex, laborious, expensive, and poorly standardized.
D. Currently, methods for the analysis of AAVs are rather complex, laborious, expensive, and poorly standardized.
E. Gene therapy is based on the introduction of nucleic acids into cells using vectors to enable therapeutic benefits.
F. However, large-scale production and quality control during and after manufacturing represent significant challenges.
G. Hereby, adeno-associated virus (AAV) vectors are not preferred for in vivo gene delivery to target cells due to their stability, their ability to transduce a broad range of cell types, and the long-term expression of vector-encoded therapeutic genes in slowly or non-replicating cells.
H. The evidence does not state that however, large-scale production and quality control during and after manufacturing represent significant challenges.


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: ["90%", "68%", "92%", "89%", "1.10 h", "0.10 h", "67%", "93%"]

Evidence:
Efficient large-scale expansion of adherent mammalian cells is essential for the production of recombinant proteins, viral vectors, and vaccines. Conventional multilayer flasks rely on scale-out approaches, require extensive manual handling, and lack monitoring process control compared to bioreactors. Here, we evaluate the CellScrew®, a dynamic 2D culture system available in three sizes ranging from 851 to 10,197 cm 2 , in which cell culture surfaces are continuously rotated, generating controlled hydrodynamic conditions that enhance mass transfer while reducing material consumption. Hydrodynamic conditions and oxygen transfer were characterized, with k L a values up to 6.69 ± 0.10 h −1 . In this context, we assessed cell attachment, proliferation, metabolism, and harvest performance using human embryonic kidney 293 (HEK-293), murine melanoma (B16-F10), and murine colon carcinoma (CT-26 WT) cells, which serve as established model systems for biotherapeutic manufacturing applications such as viral vectors and vaccine production. The system supported cell densities up to 7.7 × 10E5 cells per cm 2 (HEK-293), 8.7 × 10E4 cells per cm 2 (B16-F10), and 1.1 × 10E5 cells per cm 2 (CT-26 WT) with viabilities above 92%, comparable to multilayer flasks. Glucose consumption and lactate accumulation indicated stable metabolic activity. Mechanical harvesting enabled recovery efficiencies exceeding 89% while reducing detachment reagent usage by 67%. Overall, these results suggest that the CellScrew® can support robust adherent cell expansion with potential improvements in process efficiency and reduced resource consumption, indicating that it may represent a scalable and potentially more sustainable alternative to traditional multilayer platforms. • CellScrew® reaches up to 0.77 ± 0.14 × 10E6 cells per cm 2 (cell-line dependent). • Harvest efficiency with 67% less detachment reagent use. • High oxygen transfer (kLa up to 6.69 ± 0.10 h−1) enables efficient culture. The online version contains supplementary material available at 10.1007/s00253-026-13880-4.


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:
Bacillus methanolicus MGA3 is a thermotolerant methylotroph that utilizes methanol, a renewable C₁ substrate, as its sole carbon and energy source. The strain naturally overproduces and secretes L-glutamate, making it a promising platform for engineering pathways toward L-glutamate-derived amino acids such as L-proline, which has applications in nutrition, stress protection, and industry. Heterologous expression of an osmotic stress–responsive L-proline biosynthetic operon from the mesophile Bacillus licheniformis in the thermotolerant B. methanolicus strain MGA3 did not increase L-proline levels but instead led to accumulation of L-citrulline. This was likely due to heat sensitivity of pyrroline-5-carboxylate reductase (ProH), the last enzyme of the osmoregulatory L-proline biosynthetic route, and metabolic crosstalk between L-proline and L-arginine pathways operating in Bacilli. To overcome these limitations, a synthetic operon containing the native anabolic proBA and proI L-proline biosynthetic genes from B. methanolicus MGA3 was engineered to remove transcriptional T-box regulation and biochemical feedback inhibition of ProB enzyme activity. Expression of this engineered operon enhanced L-proline synthesis and triggered its secretion during methanol-based growth of B. methanolicus MGA3 at 50° C. In fed-batch fermentation with methanol as carbon and energy source, extracellular L-proline levels reached 262 ± 20 mg L⁻ 1 after 40 h. During the fermentation process, a stepwise increase in medium osmolarity was observed, likely due to large-scale L-glutamate excretion, which impaired cellular growth. This study links osmolarity dynamics to methanol-based fermentation in B. methanolicus MGA3 and demonstrates its potential as a cell factory for L-proline and L-citrulline production. These findings support further strain optimization for producing value-added amino acids and highlight the relevance of methylotrophic thermophiles in sustainable biotechnology. The online version contains supplementary material available at 10.1186/s12934-026-03032-8.


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:
Disruptions in the gut microbiome are linked to various diseases, but their roles in conditions such as age-related muscle loss (sarcopenia) and drug-induced microbial changes remain poorly understood. To address this gap, MyBioScope , a novel in vitro model using the DASbox ® mini bioreactor system and human stool samples, was developed to simulate the anaerobic environment of the gastrointestinal (GI) tract. Bioreactors containing 120–200 mL of cultivation media were inoculated with stool slurry, stabilized over 24 h, and maintained with a customizable feeding protocol for multi-day experiments. Samples were analyzed using 16S rRNA gene sequencing, quantitative PCR, and metabolomics. Four pilot studies were conducted to validate the platform and model specific disease states, including proton pump inhibitor-induced GI tract oralization and microbiome alterations associated with sarcopenia. The workflow incorporated an anaerobic stool collection kit for user-friendly, room-temperature sample transport and storage. Our results demonstrated consistent microbial community structure and metabolic activity within disease-mimicking conditions. MyBioScope enabled reproducible, controlled studies of gut microbial dynamics and provided a scalable tool for investigating disease-specific microbiome changes. This platform may support translational efforts to integrate microbiome insights into clinical research, therapeutic development, and personalized medicine. In conclusion, this novel bioreactor-based in vitro model, MyBioScope, shows strong potential for in-depth exploration of disease-specific microbiomes and can facilitate new ways for integrating the knowledge of the microbiome’s impact on human health and disease into clinical practice. The online version contains supplementary material available at 10.1186/s40643-026-01044-1.


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: ["80%", "1.24 mg/mL", "81%", "13 g/L", "21 °C", "12 g/L", "20 °C", "0.24 mg/mL"]

Evidence:
Chitin, the second most abundant polysaccharide, is poorly soluble and underutilized, whereas its monomer N-acetylglucosamine (GlcNAc) has broad pharmaceutical applications. Efficient enzymatic conversion of chitin to GlcNAc remains challenging. Chitinibacter mangrovi FCG-7 T , a novel chitinolytic strain isolated from mangrove sediments, exhibited exceptional chitin degradation, achieving a hydrolysis zone ratio (H/C) of 4.75 and maintained genetic stability over ten successive passages. Sequential optimization using One-Factor-at-a-Time (OFAT), Plackett-Burman designs (PBD), and Box–Behnken designs (BBD) enhanced chitinase production by 49.43-fold, yielding a final activity of 1.211 U/mL under optimized conditions (pH 7.4, 20 °C, 12 g/L powdered chitin). Purification yielded a 100 kDa bifunctional chitinase Cm Chi, exhibiting dual chitobiosidase and N-acetylglucosaminidase (NAGase) activities. The purified enzyme showed a specific activity of 7.96 U/mg toward colloidal chitin. Cm Chi demonstrated a high affinity for colloidal chitin with a K m value of 0.24 mg/mL and retained over 80% stability across a broad pH range 4.0–11.0. Furthermore, Cm Chi completely converted colloidal chitin to N-acetylglucosamine (GlcNAc) within 48 h and demonstrated tolerance to various reagents, including 1% methanol, acetonitrile, Tween 20, and Tween 80 (v/v). Notably, in cyclophosphamide (CTX)-induced immunosuppressed mice, GlcNAc (200 mg/kg) significantly restored body weight, thymic/splenic indices, and tissue architecture while elevating serum TNF- α , IL-2, IgG, and IgM ( p < 0.05). This study establishes Cm Chi as an efficient biocatalyst for industrial GlcNAc production and validates the immunomodulatory potential of GlcNAc, highlighting its dual applicability in biotechnology and immunotherapy.


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:
1,12-Dodecanediol is a high-value chemical widely used in the polymer and fine chemical industries. However, its industrial production is currently based on petroleum-derived processes associated with high energy consumption and environmental concerns. To address these limitations, microbial bioconversion of 1,12-dodecanedioic acid (C12 diacid) to 1,12-dodecanediol (C12 diol) has been investigated as a sustainable production route. In this pathway, carboxylic acid reductase (CAR) requires ATP and NADPH as essential cofactors, making efficient cofactor supply an important factor for achieving high conversion efficiency. In this study, a recombinant Escherichia coli –based biocatalytic system was developed to enhance C12 diol production through ATP regeneration. ATP availability was identified as an important factor for whole-cell bioconversion, and a polyphosphate kinase 2 (PPK2) from Erysipelotrichaceae bacterium was introduced to regenerate ATP from AMP, increasing the conversion yield from 22.0% to 31.8% at 40 mM substrate. Supplementation with sodium hexametaphosphate (SHMP, polyP 6 ) further improved diol production, reaching 19.5 mM C12 diol with a conversion yield of 48.8%. In the cell-lysate-based conversion, NADPH and NADH exhibited comparable conversion efficiencies up to 70 mM MgCl 2 . Diol production was influenced by MgCl 2 concentration, with the highest conversion obtained at 100 mM MgCl 2 in the presence of NADPH, yielding approximately 28 mM C12 diol (~ 70% conversion) within 1 h. Overall, this study highlights the importance of ATP regeneration and cofactor supply for improving C12 diol production. The online version contains supplementary material available at 10.1186/s40643-026-01069-6. Keywords: 1,12-Dodecanediol; Biocatalytic conversion; Carboxylic acid reductase; ATP regeneration; Polyphosphate kinase (PPK2)

Candidates:
A. 2,12-Dodecanediol is a high-value chemical widely used in the polymer and fine chemical industries.
B. However, its industrial production is not currently based on petroleum-derived processes associated with high energy consumption and environmental concerns.
C. The evidence does not state that in this pathway, carboxylic acid reductase (CAR) requires ATP and NADPH as essential cofactors, making efficient cofactor supply an important factor for achieving high conversion efficiency.
D. However, its industrial production is currently based on petroleum-derived processes associated with high energy consumption and environmental concerns.
E. In this pathway, carboxylic acid reductase (CAR) requires ATP and NADPH as essential cofactors, making efficient cofactor supply an important factor for achieving high conversion efficiency.
F. To address these limitations, microbial bioconversion of 1,12-dodecanedioic acid (C12 diacid) to 1,12-dodecanediol (C12 diol) has been investigated as a sustainable production route.
G. To address these limitations, microbial bioconversion of 2,12-dodecanedioic acid (C12 diacid) to 1,12-dodecanediol (C12 diol) has been investigated as a sustainable production route.
H. 1,12-Dodecanediol is a high-value chemical widely used in the polymer and fine chemical industries.


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:
Nanocellulose, derived from renewable cellulose resources, has emerged as a highly promising candidate for biomedical scaffold membrane applications owing to its excellent mechanical properties, tunable surface chemistry, biodegradability, and biocompatibility. The performance of nanocellulose-based membrane materials can be significantly enhanced through the integrated regulation of raw material sources, processing and functionalization. This review provides a comprehensive overview of the advances in nanocellulose scaffold membranes from raw material sources, processing technologies, functionalization strategies and biomedical applications. The review especially focuses on how to synergistically integrate these parameters to achieve a balanced design for customizable membranes. Furthermore, a design-oriented conceptual framework for the fabrication of regenerated nanocellulose composite membranes by electrospinning is discussed, which can provide guidance for future material and process development. Despite the preliminary progress achieved to date, several critical bottlenecks continue to hinder practical implementation, including difficulties in pore-structure regulation, long-term biosafety assessment, standardized large-scale manufacturing, and cost-effective production. Overall, this review not only summarizes the latest advancements in nanocellulose-based scaffold membranes, but also points out a future direction for their rational design and biomedical translation.

Candidates:
A. This review provides a comprehensive overview of the advances in nanocellulose scaffold membranes from raw material sources, processing technologies, functionalization strategies and biomedical applications.
B. The performance of nanocellulose-based membrane materials cannot be significantly enhanced through the integrated regulation of raw material sources, processing and functionalization.
C. The evidence does not state that this review provides a comprehensive overview of the advances in nanocellulose scaffold membranes from raw material sources, processing technologies, functionalization strategies and biomedical applications.
D. Nanocellulose, derived from renewable cellulose resources, has emerged as a highly promising candidate for biomedical scaffold membrane applications owing to its excellent mechanical properties, tunable surface chemistry, biodegradability, and biocompatibility.
E. The evidence does not state that nanocellulose, derived from renewable cellulose resources, has emerged as a highly promising candidate for biomedical scaffold membrane applications owing to its excellent mechanical properties, tunable surface chemistry, biodegradability, and biocompatibility.
F. The review especially focuses on how to synergistically integrate these parameters to achieve a balanced design for customizable membranes.
G. The evidence does not state that the review especially focuses on how to synergistically integrate these parameters to achieve a balanced design for customizable membranes.
H. The performance of nanocellulose-based membrane materials can be significantly enhanced through the integrated regulation of raw material sources, processing and functionalization.


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: ["51 L", "50 L"]

Evidence:
Chicken feathers constitute a major keratin-rich agro-industrial residue whose sustainable valorization remains challenging due to the recalcitrant structure of keratin and the energy-intensive nature of conventional treatments. In this study, we investigated the feasibility of converting raw, mechanically untreated chicken feathers into soluble nitrogen compounds and extracellular keratinases through an inoculum-free microbial process operated at pilot scale. A 50 L horizontal bioreactor was run using only tap water and the indigenous feather-associated microbiota, without sterilization, nutrient supplementation or pretreatment, under alternating aerated and oxygen-limited regimes. Substantial feather degradation was achieved, accompanied by the accumulation of ammonium in the liquid phase up to 5 g L⁻¹ and keratinase activities exceeding 31,000 U mL⁻¹, particularly under oxygen-limited conditions. High-throughput 16 S rRNA gene sequencing revealed a compact but resilient core microbiota whose temporal succession correlated with process performance. Dominance shifts among Comamonas , Wolinella , Tissierella and Pseudoxanthomonas suggested functional complementarity between taxa involved in keratin hydrolysis, peptide fermentation and nitrogen mineralization. The progressive enrichment of anaerobic or facultative anaerobic members during oxygen limitation was associated with intensified ammonium release and sustained enzymatic activity. Overall, this work demonstrates that native feather-associated microbial consortia can self-organize into a functionally stable community capable of driving keratin depolymerization and nitrogen recovery in the absence of an external inoculum. These findings highlight the biotechnological potential of starter-free microbial systems for low-input valorization of keratinous wastes and contribute to a deeper understanding of community-level processes underpinning industrially relevant bioconversions. Conceptual scheme illustrating the proposed sustainable microbial biorefinery strategy for raw chicken feather valorization, highlighting inoculum-free operation, keratin hydrolysis, ammonium recovery and keratinase production in comparison with conventional feather disposal routes such as landfilling, incineration and high-temperature rendering The online version contains supplementary material available at 10.1007/s11274-026-04976-0.


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:
Human papillomaviruses (HPVs) are a major cause of cervical cancer, which ranks fourth among cancers in women worldwide. A prophylactic vaccine composed of virus-like particles (VLPs) formed by the major capsid protein L1 of human papillomavirus (HPV) effectively prevents HPV infection but provides limited cross-protection against other HPV subtypes, highlighting the importance of multivalent prophylactic vaccines. In this study, cultivation conditions for recombinant HPV58 L1 protein production in the yeast Hansenula polymorpha were optimized to enhance growth and the volumetric yield of L1 protein. The results indicated that induction with methanol (at 1% v/v) was necessary for HPV58 L1 production in SYN6 medium, whereas a cultivation temperature of 37 °C was optimal for growth and production of HPV58 L1 protein. Subsequently, SYN6 medium supplemented with 10 g/L of either Hy-Express™ System II or HySoy was evaluated in batch and fed-batch bioreactor cultivations. Fed-batch cultivation with HySoy supplementation under a constant feeding rate achieved an OD 660 of 117 (26.18 g-CDW/L), a volumetric L1 yield of 312 mg/L, and a productivity of 4.7 mg/L/h, which were significantly higher than those obtained with the control SYN6 medium (92 mg/L; 2.2 mg/L/h). These findings demonstrate that fed-batch cultivation with HySoy supplementation offers a practical and efficient strategy for HPV58 L1 production at the bioreactor scale. The online version contains supplementary material available at 10.1186/s40643-026-01059-8. Keywords: HPV58 L1, Hansenula polymorpha , Virus-like particles, VLP, Bioreactor


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:
Laccase (LAC) was immobilized onto sponge gourd ( Luffa cylindrica ) through a two step method involving physical adsorption followed by glutaraldehyde-mediated crosslinking. The immobilization parameters were systematically optimized, resulting in maximum catalytic activity (52.98 ± 0.85 U/g) and protein loading efficiency (62.09%) under the following conditions: 2.5 mg/L LAC concentration, 15 mg carrier, 90 min adsorption time, 120 min crosslinking time, and 3% glutaraldehyde. Compared to the free LAC, the immobilized LAC showed a 5 °C increase in optimum temperature and a broader pH activity range. It also exhibited significantly improved thermal and pH stability, retaining 33% and 39% more activity after 240 min at 50 °C and 60 °C, respectively. Storage stability was enhanced, with 57% activity retained after 30 days at 4 °C, compared to only 16% for free LAC. Kinetic analysis revealed a moderate increase in K m (from 0.53 to 0.72 mM) and a decrease in V max (from 0.85 to 0.69 mmol/L min), suggesting minor diffusional resistance. In Bisphenol A (BPA) degradation tests, the immobilized enzyme achieved complete removal of 15 mg/L BPA within 240 min and maintained 46% removal efficiency after 10 reuse cycles. These findings demonstrate that sponge gourd is a cost effective, biodegradable support material that can significantly enhance LAC performance for potential applications in environmental remediation.

Candidates:
A. Laccase (LAC) was immobilized onto sponge gourd ( Luffa cylindrica ) through a two step method involving physical adsorption followed by glutaraldehyde-mediated crosslinking.
B. The immobilization parameters were systematically optimized, resulting in maximum catalytic activity (53.98 ± 0.85 U/g) and protein loading efficiency (62.09%) under the following conditions: 2.5 mg/L LAC concentration, 15 mg carrier, 90 min adsorption time, 120 min crosslinking time, and 3% glutaraldehyde.
C. The immobilization parameters were systematically optimized, resulting in maximum catalytic activity (52.98 ± 0.85 U/g) and protein loading efficiency (62.09%) under the following conditions: 2.5 mg/L LAC concentration, 15 mg carrier, 90 min adsorption time, 120 min crosslinking time, and 3% glutaraldehyde.
D. Compared to the free LAC, the immobilized LAC showed a 5 °C increase in optimum temperature and a broader pH activity range.
E. Compared to the free LAC, the immobilized LAC showed a 6 °C increase in optimum temperature and a broader pH activity range.
F. It also exhibited significantly improved thermal and pH stability, retaining 33% and 39% more activity after 240 min at 50 °C and 60 °C, respectively.
G. It also exhibited significantly improved thermal and pH stability, retaining 34% and 39% more activity after 240 min at 50 °C and 60 °C, respectively.
H. Laccase (LAC) was not immobilized onto sponge gourd ( Luffa cylindrica ) through a two step method involving physical adsorption followed by glutaraldehyde-mediated crosslinking.


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:
Fungal exopolysaccharides (EPSs) are increasingly recognized as structurally programmable microbial polymers with applications spanning biomedicine, materials engineering, food systems, and environmental technologies. While previous reviews have often addressed fungal EPS diversity, production variables, or application domains separately, an integrated framework linking biosynthesis, molecular architecture, process control, and translational manufacturing remains underdeveloped. This review positions fungal EPSs as next-generation biomaterials by integrating (i) biochemical and genetic regulation of EPS biosynthesis, (ii) structure–function mapping across major polymer classes, (iii) cultivation and downstream processing workflows that enable reproducible product specifications, and (iv) industrial translation pathways within scalable and sustainability-aligned biomanufacturing systems. Gene-cluster–resolved case studies and process-to-product design principles illustrate how metabolic flux, fermentation parameters, and polymer modification shape functional performance. Current bottlenecks—including strain-dependent variability, purification complexity, quality harmonization, and techno-economic constraints—are critically evaluated to distinguish laboratory potential from scalable feasibility. By shifting from descriptive cataloging toward platform-based engineering logic, this review provides a translational roadmap for rational fungal EPS design within standardized and application-driven manufacturing frameworks.


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: ["49.3%", "46.9%", "50.3%", "86.7%", "49.6%", "45.9%", "48.6%", "87.7%"]

Evidence:
Antibiotic use in food-producing animals can drive antimicrobial resistance (AMR). Communication is a central strategy to promote behavioral change. As such, there is a need to determine from whom and where farmers receive information about antibiotic use. We aim to identify communication pathways that influence farmers’ antibiotic use in food-producing animals and understand the barriers and facilitators for using these information sources. PRISMA Extension for Scoping Reviews guidelines were followed. PubMed, CAB Abstracts, ProQuest Social Science Premium Collection, and PsycINFO were searched for primary peer-reviewed studies published from 2000 to April 10 th , 2025, in any language. We sought studies from any country, that state from whom or where farmers or farm workers get information about antibiotic use. Title and abstract, full text screening and data charting were performed by two reviewers. One researcher inductively coded points of influence, barriers and facilitators, which were thematically combined through discussion. Quantitative and qualitative analyses were then performed. Our search retrieved 5925 records. After abstract review, 140 records met the inclusion criteria. After review of full texts, 75 papers were included for data charting. Overall, we identify a complex network of 25 stakeholders, all of whom can exert influence. The most mentioned actors across eligible studies were veterinarians (86.7%), followed by peers (49.3%). The major barriers to advice were access/availability (48.6%) and cost (45.9%), and the major facilitators were trust (41.7%), followed by access/availability (33.3%). Differences in mechanics and impact on antibiotic use, along with recommendations from the literature, were also summarized Our results suggest that knowledge and awareness about appropriate antibiotic use and AMR must permeate multiple layers of actors with differing interests. We suggest potential strategies to address barriers, including ways to reduce costs, increase access and build trust. The online version contains supplementary material available at 10.1186/s42522-026-00215-6.


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:
Silver nanoparticles (AgNPs) have attracted substantial scientific interest in biomedical research owing to their unique physicochemical characteristics, broad-spectrum antimicrobial activity, plasmonic properties, and therapeutic versatility. Although conventional physicochemical synthesis methods enable controlled NPs fabrication, their dependence on hazardous reagents, elevated energy input, and environmentally detrimental processing conditions has stimulated the development of sustainable biogenic alternatives. Biological synthesis utilizing plants, microorganisms, fungi, algae, and purified biomolecules has emerged as an eco-friendly and bio-compatible strategy for AgNP fabrication, enabling simultaneous reduction, stabilization, and intrinsic biofunctionalization of NPs. However, traditional biogenic synthesis remains constrained by limited mechanistic understanding, poor batch reproducibility, inadequate control over physicochemical properties, and challenges in large-scale manufacturing. Recent advances in bioengineering have transformed this field through the integration of metabolic engineering, synthetic biology, microfluidic-assisted synthesis, artificial intelligence-guided process optimization, and continuous-flow biomanufacturing, collectively enabling precision fabrication of biogenic AgNPs with enhanced uniformity, scalability, and functional tunability. Furthermore, strategic surface engineering and functionalization have expanded the applicability of biogenic AgNPs across targeted anticancer therapy, antimicrobial intervention, wound healing, regenerative medicine, drug delivery, and theranostic imaging. Despite these advancements, critical challenges remain regarding nano–bio interactions, toxicological safety, regulatory compliance, and translational scalability. Unlike conventional reviews focused primarily on green synthesis approaches, this review critically highlights emerging bioengineering paradigms that enable programmable, scalable, and precision-controlled biogenic AgNP fabrication. This review comprehensively examines next-generation paradigms and strategies for AgNPs biosynthesis, elucidates the molecular mechanisms governing their formation, highlights emerging functionalization and biomedical application paradigms, and discusses current translational barriers. Forming biogenic composites of AgNPs and heteroatom doped carbon nanodots needs intense research in near future.

Candidates:
A. The evidence does not state that however, traditional biogenic synthesis remains constrained by limited mechanistic understanding, poor batch reproducibility, inadequate control over physicochemical properties, and challenges in large-scale manufacturing.
B. The evidence does not state that biological synthesis utilizing plants, microorganisms, fungi, algae, and purified biomolecules has emerged as an eco-friendly and bio-compatible strategy for AgNP fabrication, enabling simultaneous reduction, stabilization, and intrinsic biofunctionalization of NPs.
C. Silver nanoparticles (AgNPs) have attracted substantial scientific interest in biomedical research owing to their unique physicochemical characteristics, broad-spectrum antimicrobial activity, plasmonic properties, and therapeutic versatility.
D. The evidence does not state that silver nanoparticles (AgNPs) have attracted substantial scientific interest in biomedical research owing to their unique physicochemical characteristics, broad-spectrum antimicrobial activity, plasmonic properties, and therapeutic versatility.
E. The evidence does not state that although conventional physicochemical synthesis methods enable controlled NPs fabrication, their dependence on hazardous reagents, elevated energy input, and environmentally detrimental processing conditions has stimulated the development of sustainable biogenic alternatives.
F. Although conventional physicochemical synthesis methods enable controlled NPs fabrication, their dependence on hazardous reagents, elevated energy input, and environmentally detrimental processing conditions has stimulated the development of sustainable biogenic alternatives.
G. Biological synthesis utilizing plants, microorganisms, fungi, algae, and purified biomolecules has emerged as an eco-friendly and bio-compatible strategy for AgNP fabrication, enabling simultaneous reduction, stabilization, and intrinsic biofunctionalization of NPs.
H. However, traditional biogenic synthesis remains constrained by limited mechanistic understanding, poor batch reproducibility, inadequate control over physicochemical properties, and challenges in large-scale manufacturing.


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:
Alternative proteins and novel processing technologies are crucial to transforming contemporary food systems into ones with lower environmental impact while meeting the rising global demand for protein. Alternative protein sources from plants, microbes, insects, and cultivated cells offer diverse nutritional and techno-functional attributes that can partially or fully replace conventional animal proteins in meat analogs and related products. This review synthesizes the current knowledge on major categories of alternative protein sources, including plant-based ingredients, microbial- and fermentation-derived proteins, insect and other emerging sources, and cultivated (cell-based) meat, with a specific focus on their suitability for structured meat analog applications. Modern structuring and processing technologies are discussed, including the traditional wet and dry extrusion to modern technologies like high-moisture extrusion, high-pressure processing, shear-cell technology, 3D printing, fermentation-based structuring, and enzymatic protein modification. Furthermore, this review critically evaluates product design and quality attributes of meat analogs, including physicochemical properties, sensory performance, nutritional aspects, and safety considerations. This review highlights technological and scale-up challenges, as well as the necessity of multi-criteria optimization in sensory quality, nutrition, sustainability, and affordability, and presents research priorities focused on combining multiple protein sources and advanced processing pathways for next-generation meat analog. This review provides an integrated framework linking protein sources, processing technologies, antioxidant functionality, and sustainability considerations to support the development of next-generation meat analogs. In addition, this review highlights the intrinsic antioxidant potential of alternative proteins, emphasizing the role of bioactive peptides, polyphenols, and structure–function relationships in enhancing oxidative stability and product quality.

Candidates:
A. Modern structuring and processing technologies are discussed, including the traditional wet and dry extrusion to modern technologies like high-moisture extrusion, high-pressure processing, shear-cell technology, 4D printing, fermentation-based structuring, and enzymatic protein modification.
B. The evidence does not state that this review synthesizes the current knowledge on major categories of alternative protein sources, including plant-based ingredients, microbial- and fermentation-derived proteins, insect and other emerging sources, and cultivated (cell-based) meat, with a specific focus on their suitability for structured meat analog applications.
C. This review synthesizes the current knowledge on major categories of alternative protein sources, including plant-based ingredients, microbial- and fermentation-derived proteins, insect and other emerging sources, and cultivated (cell-based) meat, with a specific focus on their suitability for structured meat analog applications.
D. Modern structuring and processing technologies are discussed, including the traditional wet and dry extrusion to modern technologies like high-moisture extrusion, high-pressure processing, shear-cell technology, 3D printing, fermentation-based structuring, and enzymatic protein modification.
E. Alternative protein sources from plants, microbes, insects, and cultivated cells offer diverse nutritional and techno-functional attributes that cannot partially or fully replace conventional animal proteins in meat analogs and related products.
F. Alternative proteins and novel processing technologies are crucial to transforming contemporary food systems into ones with lower environmental impact while meeting the rising global demand for protein.
G. Alternative proteins and novel processing technologies are not crucial to transforming contemporary food systems into ones with lower environmental impact while meeting the rising global demand for protein.
H. Alternative protein sources from plants, microbes, insects, and cultivated cells offer diverse nutritional and techno-functional attributes that can partially or fully replace conventional animal proteins in meat analogs and related products.


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:
Plant-derived nanovesicles (PDNVs) are a class of nanoscale vesicles derived from plant tissues; they are particles with a lipid bilayer and no ability to replicate autonomously. As a type of bioactive natural nanocarrier, they demonstrate immense potential for application in 21st-century nanomedicine, skincare and nutritional health, owing to their excellent biocompatibility, low immunogenicity and targeted delivery capabilities. However, the clinical translation of PDNVs still faces key bottlenecks, including low extraction efficiency, complex purification processes, and immature engineering modification techniques. Compared to the wealth of systematic reviews in the field of Mammalian Extracellular Vesicles (M-EVs), research on PDNVs still lacks a comprehensive exposition of its multifaceted research progress. This review endeavours to comprehensively summarise the shortcomings over the last 60 years regarding PDNV purification processes, research progress, composition and characterisation, engineering modifications, functional mechanisms, clinical translation and market regulation. It discusses the feasibility of innovative approaches such as AI deep learning technologies, interdisciplinary integration and cross-application, and outlines the latest frontiers in PDNV research. It provides comprehensive and reliable reference material for future research and application strategies regarding PDNVs, offering theoretical support and practical guidance to overcome barriers to their industrialisation. This will facilitate the transition from limited laboratory research to clinical application and drive technological innovation in the next generation of naturally derived nanomedicines.


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:
Synthetic biology is reshaping in vitro diagnostics (IVD) by enabling programmable and modular biosensing elements that can be integrated into point-of-care testing (POCT) platforms. Compared with conventional assays that depend on fixed chemistries and centralized instrumentation, synthetic biology-based systems offer adaptable molecular recognition, tunable signal processing, and flexible readout formats for decentralized diagnostics. In this review, we present synthetic biology-enabled IVD as programmable biosensing platforms organized into four functional layers: molecular recognition, signal transduction and amplification, output generation, and system integration. We discuss four major enabling modules, including cell-free protein synthesis (CFPS) systems, aptamer and riboswitch sensors, CRISPR-Cas diagnostic platforms, and microfluidic integration technologies. We summarize representative clinical applications from 2021 to 2025 in infectious disease detection, cancer biomarker analysis, and drug metabolism/toxicity screening. In addition, we examine practical considerations beyond analytical sensitivity, including matrix tolerance, workflow complexity, manufacturability, quantitative capability, and regulatory readiness. Finally, we highlight future directions for programmable diagnostics, including AI-assisted biosensor design, multimodal readouts, interoperable platform architectures, and real-world clinical validation.

Candidates:
A. Compared with conventional assays that depend on fixed chemistries and centralized instrumentation, synthetic biology-based systems offer adaptable molecular recognition, tunable signal processing, and flexible readout formats for decentralized diagnostics.
B. Synthetic biology is reshaping in vitro diagnostics (IVD) by enabling programmable and modular biosensing elements that can be integrated into point-of-care testing (POCT) platforms.
C. Synthetic biology is not reshaping in vitro diagnostics (IVD) by enabling programmable and modular biosensing elements that can be integrated into point-of-care testing (POCT) platforms.
D. The evidence does not state that compared with conventional assays that depend on fixed chemistries and centralized instrumentation, synthetic biology-based systems offer adaptable molecular recognition, tunable signal processing, and flexible readout formats for decentralized diagnostics.
E. The evidence does not state that we discuss four major enabling modules, including cell-free protein synthesis (CFPS) systems, aptamer and riboswitch sensors, CRISPR-Cas diagnostic platforms, and microfluidic integration technologies.
F. The evidence does not state that in this review, we present synthetic biology-enabled IVD as programmable biosensing platforms organized into four functional layers: molecular recognition, signal transduction and amplification, output generation, and system integration.
G. We discuss four major enabling modules, including cell-free protein synthesis (CFPS) systems, aptamer and riboswitch sensors, CRISPR-Cas diagnostic platforms, and microfluidic integration technologies.
H. In this review, we present synthetic biology-enabled IVD as programmable biosensing platforms organized into four functional layers: molecular recognition, signal transduction and amplification, output generation, and system integration.


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:
The growing global protein demand and environmental concerns from conventional animal agriculture have driven the exploration of sustainable alternative protein sources. Single-cell proteins (SCPs) from microbial fermentation offer a promising solution. This study comprehensively evaluated the nutritional value and safety profile of SCP produced from Ralstonia eutropha H16 through integrated in vitro and in vivo assessments. Nutritional analyses revealed a high crude protein content of 71.87 ± 5.05 g/100 g dry weight, with total amino acids of 53.67 ± 1.05 g/100 g. The essential amino acid content was 24.38 ± 0.51 g/100 g, accounting for 45% of the total amino acids. An essential amino acid index (EAAI) of 1.46 ± 0.04 and an amino acid score (AAS) of 0.83 ± 0.06 confirmed its classification as a high-quality protein source according to FAO/WHO standards. In vivo rat feeding trials demonstrated an adjusted protein efficiency ratio (PER) of 1.81, exceeding common plant proteins such as wheat (0.8–1.1). True digestibility (TD) reached 85.73%, with a biological value (BV) of 49.37%, net protein utilization (NPU) of 42.33%, and protein digestibility-corrected amino acid score (PDCAAS) of 0.71. Comprehensive safety assessments included chemical contaminant screening, acute oral toxicity studies in rats and mice, in vitro chromosome aberration tests, and erythrocyte micronucleus tests. Heavy metals and aflatoxin B 1 levels were below regulatory limits. Acute oral toxicity studies established LD 50 values exceeding 10,000 mg/kg body weight in both rodent species, classifying this protein source as practically non-toxic. The 28-day sub-acute toxicity study showed no significant adverse effects at low doses (6.25% protein replacement). Both genotoxicity assays (mammalian cell chromosome aberration assay and mammalian erythrocyte micronucleus test) returned negative results. These findings establish R. eutropha H16-derived SCP as a safe, nutritious, and sustainable protein source with considerable potential for feed and food applications, contributing to global food security and environmental sustainability.


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: ["2.4%", "41%", "40%", "1.4%"]

Evidence:
Mixtures of seven n‐ carboxylic acids ( n‐ CA), ranging from acetic acid to n‐ octanoic acid, that can be gained from the microbial conversion of waste biomass, were converted into liquid drop‐in fuel via Kolbe electrolysis. We demonstrate a strong, sigmoidal correlation between the total n ‐CA concentration and the Coulombic efficiency for fuel‐like liquid products (CE fuel ). Crucially, only above an apparent threshold concentration of 0.1 mol L −1 n‐ CA, Kolbe electrolysis takes place, whereas at lower concentrations the oxygen evolution reaction (OER) dominates. Optimal performance was achieved at the highest concentration tested, 1.5 mol L −1 , yielding a maximum CE fuel of 63.8 ± 1.4%, whereas for n‐ CA to 0.1 mol L −1 and 0.5 mol L −1 , the chain shortening reaction (CSR) strongly affects CE fuel . Furthermore, the study provides valuable insights into reaction selectivity, demonstrating that longer‐chain n ‐CA react preferentially compared to shorter‐chain n ‐CA, a phenomenon that is attributed to their higher hydrophobicity and hence accumulation in the hydrophobic layer formed on the electrode. Using a one‐chamber cell system at optimized concentration significantly lowered energy demand to 2.92 ± 0.04 kWhL −1 of fuel produced, representing a 40% reduction compared to similar two‐chamber setups.


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: ["101°C", "100°C"]

Evidence:
Contra‐thermodynamic chain‐walking isomerization has emerged as a powerful strategy in green chemistry for the selective transformation of alkenes and functionalized hydrocarbons under relatively mild (typically below 100°C) conditions. Unlike conventional thermodynamically driven isomerizations that favor the most stable olefin products, contra‐thermodynamic processes enable the migration of double bonds toward less stable, yet more synthetically valuable positions. This directional control allows access to otherwise difficult‐to‐obtain isomers without the need for pre‐functionalized substrates or multistep synthesis. By employing tailored, selective transition‐metal catalysts, light‐driven systems, or redox‐mediated pathways, contra‐thermodynamic chain‐walking can proceed with high atom economy and minimal waste generation. Moreover, this methodology enables even the late‐stage functionalization and valorization of biomass‐derived feedstocks. Continued development of more efficient and recyclable catalytic systems is expected to further expand the environmental and industrial relevance of contra‐thermodynamic chain‐walking isomerization. Keywords: contra‐thermodynamic | chain‐walking | isomerization | tandem reactions | terminal alkenes


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: ["1 L", "2 L", "50%", "51%"]

Evidence:
Lignocellulosic biomass represents an abundant and renewable carbon source, and its valorization through microalgal cultivation offers a sustainable route to resource-efficient bioprocessing. This study examined the effects of various carbon and nitrogen sources on the growth and lipid metabolism of Desmodesmus subspicatus , with a focus on ryegrass enzymatic hydrolysates as an alternative carbon source. Cultures were supplied with glucose, xylose, or arabinose at different concentrations, along with sodium nitrate or yeast extract, under different carbon-to-nitrogen ratios. Additionally, the impacts of alkaline- and acid-pretreated enzymatic ryegrass hydrolysates were evaluated. Growth was assessed by optical density and gravimetric analysis, and fatty acid profiles by gas chromatography. Glucose supplementation enhanced lipid accumulation, yielding fatty acid profiles dominated by C16 and C18 fatty acids, which are favorable for the quality of the produced biodiesel. Nitrogen limitation further promoted lipid accumulation; cultures supplied with sodium nitrate achieved higher total lipid content, while yeast extract favored greater proportions of PUFAs. Alkaline-pretreated ryegrass hydrolysate supported dose-dependent biomass formation reaching approximately 12 g L −1 at 50%, whereas the acid-pretreated hydrolysate exhibited inhibitory effects at the same concentration. Scale-up in a 1 L photobioreactor yielded lower biomass but higher lipid content with a fatty acid profile shifted to SFA. These results support ryegrass as a viable alternative carbon source and highlight cultivation parameters that influence growth and lipid quality relevant for biofuel applications.


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:
Morphological control in submerged fermentation is a well-established method for enhancing bioactive metabolite production in filamentous fungi. However, the molecular mechanisms linking morphology to fermentation efficiency remain insufficiently understood. In this study, supplementing 1.5% Tween 80 (P80) at the seed culture stage of Cordyceps militaris consistently induced the formation of compact, uniform mycelial pellets. This morphological induction at the seed stage enhanced fermentation performance, increasing exopolysaccharide (EPS) titer by 71.1% and reducing the production cycle by 24 h. Transcriptomic analysis revealed that pelletized cultures exhibited transcriptional patterns associated with MAPK signaling related to cell wall integrity and upregulation of genes involved in cell wall remodeling. Additionally, pelletized cultures displayed a reduced oxidative burden and were associated with enhanced antioxidant capacity. These findings link morphology induction to cell wall remodeling and oxidative stress defense, offering a potentially scalable strategy for industrial polysaccharide production in medicinal fungi.


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:
Microbial biodegradation represents a promising approach to addressing global plastic pollution, yet the metabolic pathways and environmental origins of polymer-degrading microorganisms remain incompletely characterized. This review synthesizes current knowledge on biodegradation mechanisms across major polymer classes and identifies key environmental reservoirs harboring native plastic-degrading microbiota. Biodegradation pathways differ fundamentally according to polymer chemistry. Polyesters such as PET undergo hydrolytic cleavage by PETases and MHETases, releasing terephthalic acid and ethylene glycol for assimilation via the β-ketoadipate pathway and the TCA cycle. Biodegradable polyesters (PLA, PBAT, PHAs, PCL) are similarly hydrolyzed by cutinases, lipases, and depolymerases. In contrast, polyolefins (PE, PP) and polystyrene lack hydrolyzable bonds and require oxidative attack by laccases, peroxidases, and alkane monooxygenases, followed by β-oxidation to acetyl-CoA. Three principal environmental reservoirs supply plastic-degrading microorganisms: contaminated ecosystems including landfills and the plastisphere; soil microbiota contributing ligninolytic fungi and actinomycetes; and compost environments yielding thermostable enzymes such as leaf-branch compost cutinase. Across all environments, microbial consortia demonstrate superior degradation efficiency compared to single-species cultures, reflecting the enzymatic complexity required for complete polymer mineralization. Understanding these pathways and their environmental origins provides a foundation for biological plastic waste management strategies.

Candidates:
A. Biodegradation pathways differ fundamentally according to polymer chemistry.
B. This review synthesizes current knowledge on biodegradation mechanisms across major polymer classes and identifies key environmental reservoirs harboring native plastic-degrading microbiota.
C. Polyesters such as PET undergo hydrolytic cleavage by PETases and MHETases, releasing terephthalic acid and ethylene glycol for assimilation via the β-ketoadipate pathway and the TCA cycle.
D. The evidence does not state that this review synthesizes current knowledge on biodegradation mechanisms across major polymer classes and identifies key environmental reservoirs harboring native plastic-degrading microbiota.
E. Microbial biodegradation represents a promising approach to addressing global plastic pollution, yet the metabolic pathways and environmental origins of polymer-degrading microorganisms remain incompletely characterized.
F. The evidence does not state that microbial biodegradation represents a promising approach to addressing global plastic pollution, yet the metabolic pathways and environmental origins of polymer-degrading microorganisms remain incompletely characterized.
G. The evidence does not state that biodegradation pathways differ fundamentally according to polymer chemistry.
H. The evidence does not state that polyesters such as PET undergo hydrolytic cleavage by PETases and MHETases, releasing terephthalic acid and ethylene glycol for assimilation via the β-ketoadipate pathway and the TCA cycle.


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:
Background: Live-attenuated Listeria monocytogenes (Lm) vectors are a clinically validated cancer immunotherapy platform, but translation requires reproducible, clinically realistic workflows for dose preparation and infusion. For live bacterial products, in-use stability and device compatibility can drive dose variability through adsorption, settling, and device losses. Methods: We developed and GMP-manufactured an attenuated Lm vaccine expressing human GUCY2C (Lm-GUCY2C) and performed translational characterization, including construct verification and immunogenicity readouts, and defined the administration-focused in-use stability and device compatibility. Post-thaw stability was assessed in primary cryovials and during preparation and delivery from 250 mL saline infusion bags using standard clinical devices (syringes/needles, filter-free IV tubing) and OnGuard2 closed-system components. Samples were collected over 24 h at room temperature, and viable Lm-GUCY2C were quantified by CFU recovery. Results: Lm-GUCY2C remained stable in thawed cryovials for 24 h with no significant CFU loss. High-dose infusion bags (3 × 10 9 CFU/bag) maintained CFU recovery through 6 h, whereas low-dose bags (3 × 10 8 CFU/bag) exhibited significant losses beginning at 3 h, supporting a practical in-use window of up to 2 h for low-dose preparations. OnGuard2 intravenous (i.v.) connectors did not measurably affect CFU recovery, while OnGuard2 vial adapters reduced recovery. Conclusions: This work provides an end-to-end, translationally focused characterization of a GMP-manufactured Lm cancer vaccine, including clinically actionable in-use handling constraints and device compatibility. These data define preparation and administration guardrails (notably, time-to-infusion limits for low-dose bag preparations) that can improve dose accuracy and reproducibility in clinical testing.

Candidates:
A. Methods: We developed and GMP-manufactured an attenuated Lm vaccine expressing human GUCY3C (Lm-GUCY2C) and performed translational characterization, including construct verification and immunogenicity readouts, and defined the administration-focused in-use stability and device compatibility.
B. Background: Live-attenuated Listeria monocytogenes (Lm) vectors are not a clinically validated cancer immunotherapy platform, but translation requires reproducible, clinically realistic workflows for dose preparation and infusion.
C. Background: Live-attenuated Listeria monocytogenes (Lm) vectors are a clinically validated cancer immunotherapy platform, but translation requires reproducible, clinically realistic workflows for dose preparation and infusion.
D. Methods: We developed and GMP-manufactured an attenuated Lm vaccine expressing human GUCY2C (Lm-GUCY2C) and performed translational characterization, including construct verification and immunogenicity readouts, and defined the administration-focused in-use stability and device compatibility.
E. For live bacterial products, in-use stability and device compatibility cannot drive dose variability through adsorption, settling, and device losses.
F. Post-thaw stability was assessed in primary cryovials and during preparation and delivery from 251 mL saline infusion bags using standard clinical devices (syringes/needles, filter-free IV tubing) and OnGuard2 closed-system components.
G. Post-thaw stability was assessed in primary cryovials and during preparation and delivery from 250 mL saline infusion bags using standard clinical devices (syringes/needles, filter-free IV tubing) and OnGuard2 closed-system components.
H. For live bacterial products, in-use stability and device compatibility can drive dose variability through adsorption, settling, and device losses.


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:
Background/Objectives: Vaccination is a critical public health intervention, yet its global implementation is hindered by high production costs and cold-chain requirements. This review aims to evaluate plant-based systems as sustainable, cost-efficient alternatives for vaccine production. Methods : A comprehensive literature search was conducted across major databases (PubMed, Scopus, Web of Science). The peer-reviewed references were critically assessed, focusing on molecular expression strategies, phytochemical immunomodulators, and plant-mediated oral delivery. Results : Plant and microalgae systems effectively support nuclear, chloroplast, and transient expression of diverse antigens. Furthermore, specific plant-derived compounds were found to act as potent adjuvants and immunostimulants, enhancing the immunogenicity of vaccine formulations. Edible plant tissues also provide a viable platform for oral delivery, reducing the need for extensive purification and refrigerated logistics. Conclusions : Integrating recombinant expression technologies with bioactive plant metabolites offers a flexible and scalable foundation for next-generation vaccines. These biological platforms show promise for addressing some immunization challenges, particularly in low-resource settings.


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:
Sustainable biofuels have spurred interest in cyanobacterial ethanol production, yet large-scale application is severely hindered by microbial contamination—a devastating challenge that lacks universally effective, biocompatible mitigation strategies. Traditional methods such as pH manipulation or antibiotic application are often physiologically incompatible, environmentally unsustainable, or ineffective against diverse contaminant consortia. Here, we propose and validate alginate encapsulation as a physical barrier strategy to address this challenge. Alginate, a biodegradable polysaccharide derived from brown algae, forms a porous hydrogel matrix that encapsulates cyanobacterial cells. This matrix blocks direct contact, uptake or ingestion by microbial contaminants while allowing the efficient diffusion of gases (CO 2 , O 2 ), light, and nutrients to maintain photosynthetic and metabolic functions. Using our previously engineered, high-performance ethanol-producing strain (EP), we find that unprotected cultures rapidly collapse and cease ethanol production upon inoculation with a contaminant consortium, with cumulative ethanol yield falling to undetectable levels, whereas encapsulated cells sustain normal growth and photosynthetic activity under identical contamination pressure. After rinsing and re-cultivation, the encapsulated biomass partially restored ethanol productivity, achieving a cumulative titer of 320 mg/L over 4 days post-recovery. This suggests that the protective strategy is non-invasive and enables functional recovery of the production system following contamination exposure. This study demonstrates that alginate encapsulation represents a promising strategy to mitigate microbial contamination, with the potential to enhance the technical resilience and operational stability of cyanobacterial biofuel production. The online version contains supplementary material available at 10.1186/s40643-026-01076-7. Keywords: Cyanobacteria, Ethanol, Biocontamination, Alginate encapsulation, Microbial contaminants

Candidates:
A. Sustainable biofuels have spurred interest in cyanobacterial ethanol production, yet large-scale application is not severely hindered by microbial contamination—a devastating challenge that lacks universally effective, biocompatible mitigation strategies.
B. Sustainable biofuels have spurred interest in cyanobacterial ethanol production, yet large-scale application is severely hindered by microbial contamination—a devastating challenge that lacks universally effective, biocompatible mitigation strategies.
C. The evidence does not state that here, we propose and validate alginate encapsulation as a physical barrier strategy to address this challenge.
D. The evidence does not state that alginate, a biodegradable polysaccharide derived from brown algae, forms a porous hydrogel matrix that encapsulates cyanobacterial cells.
E. Traditional methods such as pH manipulation or antibiotic application are often physiologically incompatible, environmentally unsustainable, or ineffective against diverse contaminant consortia.
F. Here, we propose and validate alginate encapsulation as a physical barrier strategy to address this challenge.
G. Alginate, a biodegradable polysaccharide derived from brown algae, forms a porous hydrogel matrix that encapsulates cyanobacterial cells.
H. Traditional methods such as pH manipulation or antibiotic application are not often physiologically incompatible, environmentally unsustainable, or ineffective against diverse contaminant consortia.


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:
We present MechaSuite , an open-source modular software suite designed to streamline the analysis of quantum-chemical reaction mechanisms. MechaSuite combines an intuitive data manager ( MechaData ), a molecular geometry editor ( MechaEdit ), and a microkinetic modeling engine ( MechaKinetics ). It facilitates the calculation of thermodynamic and kinetic parameters from quantum chemical outputs, the visualization and editing of molecular structures, and the simulation of complex reaction networks. This integration enables chemists to transition seamlessly from ab initio calculations to kinetic predictions in a user-friendly and efficient manner. MechaSuite is primarily implemented in Python with its high-performance 3D visualization engine written in C++ for optimal rendering and interactivity.

Candidates:
A. The evidence does not state that mechaSuite combines an intuitive data manager ( MechaData ), a molecular geometry editor ( MechaEdit ), and a microkinetic modeling engine ( MechaKinetics ).
B. The evidence does not state that we present MechaSuite , an open-source modular software suite designed to streamline the analysis of quantum-chemical reaction mechanisms.
C. We present MechaSuite , an open-source modular software suite designed to streamline the analysis of quantum-chemical reaction mechanisms.
D. The evidence does not state that this integration enables chemists to transition seamlessly from ab initio calculations to kinetic predictions in a user-friendly and efficient manner.
E. The evidence does not state that it facilitates the calculation of thermodynamic and kinetic parameters from quantum chemical outputs, the visualization and editing of molecular structures, and the simulation of complex reaction networks.
F. MechaSuite combines an intuitive data manager ( MechaData ), a molecular geometry editor ( MechaEdit ), and a microkinetic modeling engine ( MechaKinetics ).
G. This integration enables chemists to transition seamlessly from ab initio calculations to kinetic predictions in a user-friendly and efficient manner.
H. It facilitates the calculation of thermodynamic and kinetic parameters from quantum chemical outputs, the visualization and editing of molecular structures, and the simulation of complex reaction networks.


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:
Spectral monitoring combined with chemometrics models resulting from machine learning approaches allows cell culture to be monitored almost in real time. This process analytical technology offers to drastically reduce the amount of hands-on time and laboratory testing needed to monitor this crucial biomanufacturing step. In this article, we propose a method to anticipate future spectra. The method is based on extrapolation of the spectra in a reduced-dimensionality space, followed by retroprojection in the original space. Passed to regular chemometrics models already fitted, these anticipated spectra enable predictive cell culture monitoring up to several dozen hours with satisfactory quality. This anticipation paves the way for course-correction and enhanced operations such as reduced need for night shifts.

Candidates:
A. The method is based on extrapolation of the spectra in a reduced-dimensionality space, followed by retroprojection in the original space.
B. This process analytical technology offers to drastically reduce the amount of hands-on time and laboratory testing needed to monitor this crucial biomanufacturing step.
C. The method is not based on extrapolation of the spectra in a reduced-dimensionality space, followed by retroprojection in the original space.
D. In this article, we propose a method to anticipate future spectra.
E. The evidence does not state that this process analytical technology offers to drastically reduce the amount of hands-on time and laboratory testing needed to monitor this crucial biomanufacturing step.
F. The evidence does not state that in this article, we propose a method to anticipate future spectra.
G. Spectral monitoring combined with chemometrics models resulting from machine learning approaches allows cell culture to be monitored almost in real time.
H. The evidence does not state that spectral monitoring combined with chemometrics models resulting from machine learning approaches allows cell culture to be monitored almost in real time.


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 eukaryotic cell-free protein synthesis (CFPS) system, endowed with intrinsic post-translational modification capabilities and a complex molecular chaperone network, efficiently synthesizes functional proteins with correct conformation and biological activity. This effectively compensates for the structural limitations of prokaryotic systems in expressing complex eukaryotic proteins. This paper aims to comprehensively review and analyze the latest advances in the field of eukaryotic CFPS from a systems engineering perspective. The paper delves into the diversification of host chassis, rational design of core reaction components, and the pivotal role of novel biomaterial integration and high-throughput reaction equipment development in system reconfiguration. At the application level, it summarizes the platform's latest achievements, including elucidation of fundamental mechanisms, complex protein engineering, and metabolic synthesis. It particularly highlights its potential in emerging areas such as the construction of artificial cells, the development of bioelectronic interfaces, and the design of microarray chips. Furthermore, addressing the current standardization and cost bottlenecks hindering industrialization, this paper proposes a solution strategy based on artificial intelligence and synthetic biology tools, aligning with the shift from empirical trial-and-error to rational design paradigms. By integrating the current technological landscape with emerging trends, this review aims to provide theoretical references and practical guidance for constructing an economical, high-throughput eukaryotic cell-free biomanufacturing platform. Keywords: Cell-free synthetic biology, Eukaryotic cell-free protein synthesis, Chassis engineering, Protein engineering, Material integration, Biomanufacturing

Candidates:
A. The evidence does not state that this effectively compensates for the structural limitations of prokaryotic systems in expressing complex eukaryotic proteins.
B. This paper aims to comprehensively review and analyze the latest advances in the field of eukaryotic CFPS from a systems engineering perspective.
C. The paper delves into the diversification of host chassis, rational design of core reaction components, and the pivotal role of novel biomaterial integration and high-throughput reaction equipment development in system reconfiguration.
D. The evidence does not state that the paper delves into the diversification of host chassis, rational design of core reaction components, and the pivotal role of novel biomaterial integration and high-throughput reaction equipment development in system reconfiguration.
E. The evidence does not state that this paper aims to comprehensively review and analyze the latest advances in the field of eukaryotic CFPS from a systems engineering perspective.
F. The eukaryotic cell-free protein synthesis (CFPS) system, endowed with intrinsic post-translational modification capabilities and a complex molecular chaperone network, efficiently synthesizes functional proteins with correct conformation and biological activity.
G. This effectively compensates for the structural limitations of prokaryotic systems in expressing complex eukaryotic proteins.
H. The evidence does not state that the eukaryotic cell-free protein synthesis (CFPS) system, endowed with intrinsic post-translational modification capabilities and a complex molecular chaperone network, efficiently synthesizes functional proteins with correct conformation and biological activity.


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 sequencing-by-synthesis technology by Illumina, Inc. enables efficient and scalable readouts of mutations from genomic data. To enhance sequencing speed and efficiency, Illumina has shifted from the four-color base calling chemistry of the HiSeq series to a two-color fluorescent dye chemistry in the NovaSeq series. Benchmarking sequencing artifacts due to biases in the newer chemistry is important to evaluate the quality of identified mutations. We re-analyze a series of whole-genome sequencing experiments in which the same samples were sequenced on the NovaSeq 6000 (two-color) and HiSeq X10 (four-color) platforms by independent groups. In several samples, we observe a higher frequency of T-to-G and A-to-C putative substitutions (“T > G”) at the read level for NovaSeq 6000 versus HiSeq X10. As the per-base error rate is still low, the artifactual substitutions have a negligible effect in identifying germline or high variant allele frequency (VAF) somatic mutations. However, such errors can confound the detection of low-VAF somatic variants in high-depth sequencing samples, particularly in studies of mosaic mutations in normal tissues, where variants have low read support and are called without a matched normal. The artifactual T > G variant calls disproportionately occur at NT[TG] trinucleotides, and we leverage this observation to bioinformatically reduce the T > G excess in somatic mutation callsets. We identified a recurrent artifact specific to the Illumina two-color chemistry platform on the NovaSeq 6000 with the potential to contaminate low-VAF somatic mutation calls. Thus, an unexpected enrichment of T > G mutations in mosaicism studies warrants caution. The online version contains supplementary material available at 10.1186/s13059-026-04081-3.

Candidates:
A. The sequencing-by-synthesis technology by Illumina, Inc.
B. The evidence does not state that enables efficient and scalable readouts of mutations from genomic data.
C. The evidence does not state that the sequencing-by-synthesis technology by Illumina, Inc.
D. To enhance sequencing speed and efficiency, Illumina has shifted from the four-color base calling chemistry of the HiSeq series to a two-color fluorescent dye chemistry in the NovaSeq series.
E. enables efficient and scalable readouts of mutations from genomic data.
F. Benchmarking sequencing artifacts due to biases in the newer chemistry is important to evaluate the quality of identified mutations.
G. The evidence does not state that to enhance sequencing speed and efficiency, Illumina has shifted from the four-color base calling chemistry of the HiSeq series to a two-color fluorescent dye chemistry in the NovaSeq series.
H. Benchmarking sequencing artifacts due to biases in the newer chemistry is not important to evaluate the quality of identified mutations.


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:
Biopesticides, pesticides based on living organisms and/or their bioactive compounds, are increasingly being used as alternatives and replacements to chemical and synthetic pesticides. This is largely due to human and environmental safety concerns, the emergence of pest resistance (collectively insect pests, weeds and diseases), and the move towards holistic pest and disease control approaches. While the number and diversity of agents used in biopesticides slowly increases, the approach to developing these products remains largely ad hoc, resulting in less than 10% success rate for development projects. Here we review the major characteristics of successful biopesticides. Our treatise focuses on the benefits of considering the entire development pathway and requirements of the intended product application before embarking on the costly process of biopesticide development and commercialisation. By a priori consideration of the characteristics of both the target pest and market, and the potential limitations of the candidate microorganism and/or its bioactives (e.g. environmental persistence, ease and cost of mass production), the development pipeline can be streamlined and targeted on projects with the greatest likelihood of success. We provide a detailed consideration of the key factors that underpin successful (or not) biopesticide development and provide decision trees to support the a priori process. Keywords: Microbial biopesticides, Commercialisation, Decision-support framework, Biocontrol agents, Market driven development, Regulatory pathways, Entomopathogens, Mode of action, Product development pipeline, Uptake and adoption barriers, Sustainable pest management

Candidates:
A. Biopesticides, pesticides based on living organisms and/or their bioactive compounds, are not increasingly being used as alternatives and replacements to chemical and synthetic pesticides.
B. The evidence does not state that here we review the major characteristics of successful biopesticides.
C. Biopesticides, pesticides based on living organisms and/or their bioactive compounds, are increasingly being used as alternatives and replacements to chemical and synthetic pesticides.
D. While the number and diversity of agents used in biopesticides slowly increases, the approach to developing these products remains largely ad hoc, resulting in less than 11% success rate for development projects.
E. While the number and diversity of agents used in biopesticides slowly increases, the approach to developing these products remains largely ad hoc, resulting in less than 10% success rate for development projects.
F. This is not largely due to human and environmental safety concerns, the emergence of pest resistance (collectively insect pests, weeds and diseases), and the move towards holistic pest and disease control approaches.
G. Here we review the major characteristics of successful biopesticides.
H. This is largely due to human and environmental safety concerns, the emergence of pest resistance (collectively insect pests, weeds and diseases), and the move towards holistic pest and disease control approaches.


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: ["21 °C", "75–84%", "51%", "74–84%", "20 °C", "50%", "2–30%", "3–30%"]

Evidence:
White rot fungi obtain nutrients from fibrous substrates through the radical secretion of extracellular ligninolytic enzymes (LE) and spent mushroom substrate (SMS), a by-product of commercial cultivation, is a valuable bioresource of LE. This study aims to compare various techniques for dehydrating and preserving the Pleurotus ostreatus SMS extract in terms of enzymatic activity and ability to improve the fiber rumen degradability (NDFD) of extract-treated wheat straw. To convert the extract into stable enzyme powder, it was concentrated before being dried using freeze, spray and vacuum drying processes. The different extracts reconstituted from powders were evaluated for residual LE activity after multiple successive freeze-thaw cycles, as well as their ability to improve wheat straw NDFD. To produce extracts, fresh SMS was suspended in acetate buffer, homogenized and filtered. A portion of the original extract was stored (− 20 °C) and the remainder was concentrated up to 50% of volume by cross-flow filtration. The concentrated extract was divided into four aliquots to be used without processing or spray, vacuum or freeze drying. Extracts were evaluated for total, Mn and lignin peroxidases and laccases (LAs) enzymatic activity and were subjected to four freezing-thawing cycles. Wheat straw was incubated in reconstituted extracts before being tested for in vitro NDFD. Compared to the original extract, reductions of LE were 2–30% in concentrated and 10–36, 4–48 and 74–84% in freeze, vacuum and spray dried, respectively. LE of extracts decreased with the number of freeze-thaw cycles, apart from the spray dried extract. The NDFD of all extract-treated straws resulted higher ( P < 0.05) than the control, apart from the spray-dried extract and had a high linear relationship with LAs of extracts (R 2 0.87, P < 0.01). Results support the potential of extract preparations from fungal SMS to be a promising bioresource useful for improving the NDFD of high-fiber forages, such as straw.


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:
Neglected tropical diseases and the increasing prevalence of antibiotic resistance in nosocomial infections represent a major challenge in medicine. Intensive research into bioactive compounds has identified the myxobacterial natural product aurachin D as promising drug candidate due to its potent antibiotic and antiprotozoal properties. The biosynthesis of aurachin D from 2-methyl-1 H -quinolin-4-one (MQO) is catalyzed by the membrane-bound farnesyltransferase AuaA. Recently, this enzymatic conversion was reconstructed in the heterologous host Escherichia coli , yielding an aurachin D titer of 17 mg L −1 . In this study, we investigated the influence of the medium on aurachin D biosynthesis in E. coli in order to improve the product titer. Our analyses revealed that the glycerol concentration has a major impact on both the growth of the production host and the biocatalytic formation of aurachin D. After switching to a glycerol-based minimal medium and adjustment of the MQO precursor concentration, an aurachin D titer of 124 mg L −1 was achieved in a microbioreactor system, highlighting the superior performance of minimal over complex medium as well as the beneficial effect of glycerol compared to glucose. Further studies were conducted to elucidate the mechanism underlying glycerol’s effects. While improved growth and substrate solubilization were found to be insignificant for increasing the product titer, it was demonstrated that glycerol supplementation positively influenced the expression of the enzyme AuaA. Using a folding reporter, we were able to show that the availability of correctly folded AuaA increased until growth defects of the host, presumably resulting from osmotic stress, began to appear. • Product toxicity is negligible in the production of aurachin D using E. coli • The media supplement glycerol improves the availability of the enzyme AuaA • Aurachin D titers increase after switching to a minimal medium The online version contains supplementary material available at 10.1007/s00253-026-13890-2.

Candidates:
A. Recently, this enzymatic conversion was reconstructed in the heterologous host Escherichia coli , yielding an aurachin D titer of 17 mg L −1 .
B. Neglected tropical diseases and the increasing prevalence of antibiotic resistance in nosocomial infections represent a major challenge in medicine.
C. The biosynthesis of aurachin D from 2-methyl-1 H -quinolin-4-one (MQO) is catalyzed by the membrane-bound farnesyltransferase AuaA.
D. The evidence does not state that intensive research into bioactive compounds has identified the myxobacterial natural product aurachin D as promising drug candidate due to its potent antibiotic and antiprotozoal properties.
E. The biosynthesis of aurachin D from 3-methyl-1 H -quinolin-4-one (MQO) is catalyzed by the membrane-bound farnesyltransferase AuaA.
F. The evidence does not state that neglected tropical diseases and the increasing prevalence of antibiotic resistance in nosocomial infections represent a major challenge in medicine.
G. Recently, this enzymatic conversion was reconstructed in the heterologous host Escherichia coli , yielding an aurachin D titer of 18 mg L −1 .
H. Intensive research into bioactive compounds has identified the myxobacterial natural product aurachin D as promising drug candidate due to its potent antibiotic and antiprotozoal properties.


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:
Supercritical CO₂ extraction was applied to Heliotropium arbainense to evaluate how extraction pressure influences phytochemical composition and biological activity. Extractions of phenolic and flavonoid were performed at two pressures (200 and 550 bar), by using Supercritical CO₂. Phenolic and flavonoid compounds, were determined by HPLC analysis. Higher pressure extraction (550 bar) resulted in a greater extraction yield and a marked enrichment of phenolic and flavonoid compounds. Gallic acid (1496.01 µg/g) was the most prevalent component under SFE 1, followed by catechin (859.51 µg/g), rutin (824.55 µg/g), and coumaric acid (559.93 µg/g). Both extracts exhibited broad antimicrobial activity against Gram-positive and Gram-negative bacteria as well as Candida albicans , with consistently lower MIC and MBC/MFC values observed for the high-pressure extract. Cytotoxic evaluation against SKOV3 ovarian cancer cells revealed a clear dose-dependent reduction in cell viability. The calculated IC₅₀ values further confirm the greater cytotoxic potency of extract at 550 bar, with a lower IC₅₀ (153.04 ± 0.4 µg/mL) compared to extract at 200 bar (183.18 ± 2.29 µg/mL) against SKOV3 cells. However, flow cytometry revealed that extract at 200 bar produced slightly higher early apoptosis (43.79% vs. 43.39%) and greater necrosis (21.83% vs. 13.29%) in SKOV3 ovarian cells overall. Anti-inflammatory activity, assessed by protein denaturation inhibition, was observed for both extracts, with the lower-pressure extract showing slightly stronger inhibitory efficiency. Pressure-dependent supercritical CO₂ extraction significantly influenced the phytochemical richness and biological performance of H. arbainense . These findings highlight the importance of extraction conditions in maximizing the functional potential of plant-derived bioactive compounds and support further investigation of H. arbainense as a source of biologically active phenolics.

Candidates:
A. Extractions of phenolic and flavonoid were performed at two pressures (201 and 550 bar), by using Supercritical CO₂.
B. Phenolic and flavonoid compounds, were determined by HPLC analysis.
C. Higher pressure extraction (551 bar) resulted in a greater extraction yield and a marked enrichment of phenolic and flavonoid compounds.
D. Phenolic and flavonoid compounds, were not determined by HPLC analysis.
E. Extractions of phenolic and flavonoid were performed at two pressures (200 and 550 bar), by using Supercritical CO₂.
F. Supercritical CO₂ extraction was applied to Heliotropium arbainense to evaluate how extraction pressure influences phytochemical composition and biological activity.
G. Supercritical CO₂ extraction was not applied to Heliotropium arbainense to evaluate how extraction pressure influences phytochemical composition and biological activity.
H. Higher pressure extraction (550 bar) resulted in a greater extraction yield and a marked enrichment of phenolic and flavonoid compounds.


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:
This study evaluated the anti-acne potential of extracellular proteins derived from Lactobacillus gasseri (LG-EPs) by integrating physicochemical characterization, antibacterial assessment, cellular assays, and an in vivo sebaceous gland model. LG-EPs were obtained by ammonium sulfate precipitation and characterized using gel permeation chromatography/light scattering (GPC/LS) and liquid chromatography–tandem mass spectrometry (LC–MS/MS). The molecular weight of LG-EPs was mainly distributed between 1.0 × 10⁴ and 2.0 × 10⁴ g/mol, and peptide analysis identified four peptides with antimicrobial potential and twelve peptides with antioxidant potential. LG-EPs exhibited direct antibacterial activity against acne-associated bacteria, with MIC and MBC values of 600 µg/mL against Cutibacterium acnes and MIC and MBC values of 700 µg/mL and 1 mg/mL, respectively, against Staphylococcus aureus . Growth-curve and biofilm adhesion assays further showed that LG-EPs inhibited bacterial proliferation and adhesion. In LPS-stimulated HaCaT cells, LG-EPs reduced the secretion of pro-inflammatory cytokines, including IL-6, IL-8, IL-1β, and TNF-α, while increasing the expression of barrier-related factors such as AQP3, FLG, and LOR. In a golden hamster model, topical LG-EP treatment decreased sebum production and downregulated the transcription of lipogenesis-related genes, including SREBP-1, FAS, and ACC1. These effects were associated with reduced transcript levels of PI3K, AKT, and mTOR, while the anti-sebum effect was accompanied by increased AMPK transcription. Overall, LG-EPs showed multi-target anti-acne potential through antibacterial, anti-inflammatory, barrier-protective, and sebum-suppressive effects.

Candidates:
A. LG-EPs were not obtained by ammonium sulfate precipitation and characterized using gel permeation chromatography/light scattering (GPC/LS) and liquid chromatography–tandem mass spectrometry (LC–MS/MS).
B. LG-EPs exhibited direct antibacterial activity against acne-associated bacteria, with MIC and MBC values of 600 µg/mL against Cutibacterium acnes and MIC and MBC values of 700 µg/mL and 1 mg/mL, respectively, against Staphylococcus aureus .
C. LG-EPs were obtained by ammonium sulfate precipitation and characterized using gel permeation chromatography/light scattering (GPC/LS) and liquid chromatography–tandem mass spectrometry (LC–MS/MS).
D. This study evaluated the anti-acne potential of extracellular proteins derived from Lactobacillus gasseri (LG-EPs) by integrating physicochemical characterization, antibacterial assessment, cellular assays, and an in vivo sebaceous gland model.
E. The molecular weight of LG-EPs was mainly distributed between 2.0 × 10⁴ and 2.0 × 10⁴ g/mol, and peptide analysis identified four peptides with antimicrobial potential and twelve peptides with antioxidant potential.
F. LG-EPs exhibited direct antibacterial activity against acne-associated bacteria, with MIC and MBC values of 601 µg/mL against Cutibacterium acnes and MIC and MBC values of 700 µg/mL and 1 mg/mL, respectively, against Staphylococcus aureus .
G. The evidence does not state that this study evaluated the anti-acne potential of extracellular proteins derived from Lactobacillus gasseri (LG-EPs) by integrating physicochemical characterization, antibacterial assessment, cellular assays, and an in vivo sebaceous gland model.
H. The molecular weight of LG-EPs was mainly distributed between 1.0 × 10⁴ and 2.0 × 10⁴ g/mol, and peptide analysis identified four peptides with antimicrobial potential and twelve peptides with antioxidant potential.