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 integrates the valorization of a lignocellulose material into poly(3-hydroxybutyrate), P(3HB), with biopolymer extraction from bacterial cells with the enzyme alcalase. The work focused on Burkholderia thailandensis DSM 13276 as the P(3HB) producer and on eucalyptus bark, a byproduct from the pulp industry, as the sole feedstock for bacterial cultivation. The eucalyptus bark was hydrolyzed by a cellulolytic enzymatic cocktail following steam explosion and further subjected to ultrafiltration for enzyme recovery. The resulting hydrolysate supported good cell growth, achieving a cell dry weight of 7.67 ± 0.16 g/L within 72 h of cultivation, and high P(3HB) content (60.0 ± 2.19 wt %) in the bacterial cells, clearly favoring biopolymer synthesis over cell growth, as demonstrated by the polymer and growth yields (0.190 g P(3HB) /g sugar and 0.026 g X /g sugar , respectively). High extraction efficiency (96%) and biopolymer purity (100 ± 3.38%) were reached by enzymatic treatment, resulting in a sample with properties aligned with those of commercial P(3HB) in terms of molecular mass distribution, crystallinity, and thermal properties. These findings demonstrate the successful use of a sustainable feedstock together with the application of environmentally friendly technologies based on the use of enzymes for both lignocellulosic saccharification and biopolymer recovery to develop high-quality bioplastics, advancing the goals of a circular bioeconomy.

Candidates:
A. The eucalyptus bark was not hydrolyzed by a cellulolytic enzymatic cocktail following steam explosion and further subjected to ultrafiltration for enzyme recovery.
B. The resulting hydrolysate supported good cell growth, achieving a cell dry weight of 8.67 ± 0.16 g/L within 72 h of cultivation, and high P(3HB) content (60.0 ± 2.19 wt %) in the bacterial cells, clearly favoring biopolymer synthesis over cell growth, as demonstrated by the polymer and growth yields (0.190 g P(3HB) /g sugar and 0.026 g X /g sugar , respectively).
C. The work focused on Burkholderia thailandensis DSM 13277 as the P(3HB) producer and on eucalyptus bark, a byproduct from the pulp industry, as the sole feedstock for bacterial cultivation.
D. This study integrates the valorization of a lignocellulose material into poly(4-hydroxybutyrate), P(3HB), with biopolymer extraction from bacterial cells with the enzyme alcalase.
E. This study integrates the valorization of a lignocellulose material into poly(3-hydroxybutyrate), P(3HB), with biopolymer extraction from bacterial cells with the enzyme alcalase.
F. The work focused on Burkholderia thailandensis DSM 13276 as the P(3HB) producer and on eucalyptus bark, a byproduct from the pulp industry, as the sole feedstock for bacterial cultivation.
G. The eucalyptus bark was hydrolyzed by a cellulolytic enzymatic cocktail following steam explosion and further subjected to ultrafiltration for enzyme recovery.
H. The resulting hydrolysate supported good cell growth, achieving a cell dry weight of 7.67 ± 0.16 g/L within 72 h of cultivation, and high P(3HB) content (60.0 ± 2.19 wt %) in the bacterial cells, clearly favoring biopolymer synthesis over cell growth, as demonstrated by the polymer and growth yields (0.190 g P(3HB) /g sugar and 0.026 g X /g sugar , respectively).


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:
Since its discovery as a pivotal enzyme in innate immunity, cyclic GMP‐AMP synthase (cGAS) has been extensively studied for its immunological significance and catalytic mechanism. However, its potential as a biocatalyst for the efficient synthesis of the second messenger 2′3′‐cyclic GMP‐AMP (2′3′‐cGAMP) remains underexplored. This review provides a comprehensive biotechnological perspective on cGAS, highlighting its enzymatic and structural features, substrate promiscuity, homologs, and engineered variants. We examined the expression systems reported in previous studies and assessed their suitability for scalable cGAS production. Furthermore, we explored reaction engineering strategies for 2′3′‐cGAMP synthesis by comparing published production and purification methods. This review aims to bridge the gap between fundamental enzymology and applied bioprocessing by positioning cGAS as a promising biocatalyst for the pharmaceutical industry, with potential applications in immunotherapy, vaccine adjuvants, and beyond. Keywords: biocatalysis, bioprocess, cGAMP, cGAS, cyclic dinucleotides

Candidates:
A. We examined the expression systems reported in previous studies and assessed their suitability for scalable cGAS production.
B. The evidence does not state that since its discovery as a pivotal enzyme in innate immunity, cyclic GMP‐AMP synthase (cGAS) has been extensively studied for its immunological significance and catalytic mechanism.
C. The evidence does not state that we examined the expression systems reported in previous studies and assessed their suitability for scalable cGAS production.
D. The evidence does not state that this review provides a comprehensive biotechnological perspective on cGAS, highlighting its enzymatic and structural features, substrate promiscuity, homologs, and engineered variants.
E. This review provides a comprehensive biotechnological perspective on cGAS, highlighting its enzymatic and structural features, substrate promiscuity, homologs, and engineered variants.
F. However, its potential as a biocatalyst for the efficient synthesis of the second messenger 2′3′‐cyclic GMP‐AMP (2′3′‐cGAMP) remains underexplored.
G. Since its discovery as a pivotal enzyme in innate immunity, cyclic GMP‐AMP synthase (cGAS) has been extensively studied for its immunological significance and catalytic mechanism.
H. However, its potential as a biocatalyst for the efficient synthesis of the second messenger 3′3′‐cyclic GMP‐AMP (2′3′‐cGAMP) remains underexplored.


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: ["24 h", "25 h", "48 h", "73 h", "49 h", "72 h"]

Evidence:
Chinese hamster ovary (CHO) cells are widely utilised in the biopharmaceutical industry to produce therapeutic proteins. Understanding the mechanisms of endoplasmic reticulum (ER) stress and its interplay with protein degradation pathways remains pivotal for improving production efficiency and product quality. In this study, we investigated the proteomic responses of CHO-K1 (non-producer), CHO DP-12 (IgG-producer), and NISTCHO (IgG-producer) cell lines under ER stress induced by a combination of the proteasome inhibitor MG132 and the glycosylation inhibitor tunicamycin. Viability, cell growth, and IgG titre were measured after 24 h, 48 h, and 72 h of treatment and the 48 h timepoint was used for the comparative analysis of the proteomic data across the three cell lines. Proteasome inhibition with MG132 intensified ER stress and altered ER-associated protein degradation (ERAD). Combined tunicamycin + MG132 treatment was associated with cell line-specific proteomic changes: NISTCHO upregulated ER translocation and glycoprotein quality control proteins (SSR4, SEC24C, UGGT1), CHO DP-12 activated redox/disulfide regulators (DNAJC10, CAPN1), while CHO-K1 showed broad proteome shifts, suggesting differences in baseline stress handling. These findings provide mechanistic insights into ER stress and protein quality control in CHO cells, offering a foundation for strategies to enhance cell line robustness and optimise biopharmaceutical production.


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

Evidence:
Microfluidics-based preparation methods for cell-laden hydrogel microspheres are well-suited for large-scale comparative analysis of single or few cells. However, in existing studies, the preparation of cell-laden hydrogel microspheres and the cell culture process are typically separated, requiring the fabricated microspheres to be eluted and transferred from the preparation device to cell culture dishes or plates for cultivation. This transfer process can easily compromise sterility, while conventional cell culture methods consume more reagents and cause microsphere stacking, hindering single-cell observation and analysis. To address these issues, this paper presents an integrated microfluidic chip that sequentially enables droplet generation with cell encapsulation, gel droplet solidification, hydrogel microsphere trapping, and microsphere-based cell culture and analysis, facilitating the cultivation and observation of single or small numbers of cells. Integrating cell-laden microsphere preparation and 3D cell culture within a sealed chip structure reduces contamination risks associated with cell transfer, enables automation of multiple cell analysis workflows, and minimizes reagent and sample consumption. Using polydimethylsiloxane (PDMS) with good gas permeability and processability as the chip material, biocompatible fluorinated oil was selected as the oil phase for microsphere preparation. A mild sodium alginate-calcium ion gelation system was employed, where calcium ions were released under acidic conditions after droplet generation to trigger solidification, yielding uniform hydrogel microspheres. Under optimized conditions, the single-cell encapsulation efficiency for test samples of human myeloid leukemia cells (K562) was 33.8% ± 1.8%, with a size uniformity coefficient of variation (CV) reaching 3.85%. Cells encapsulated within hydrogel microspheres were cultured in 286 on-chip independent cell culture chambers, achieving >95% viability after 24 h.


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:
Fermentation, one of the oldest biotransformation processes, has become a key element of contemporary sustainable biotechnology. In modern food systems, it enables the simultaneous resolution of environmental, nutritional, and economic challenges by converting agricultural and food residues into high-value-added products, such as bioactive compounds, organic acids, biofuels, enzymes, and proteins. Consistent with the concept of a circular bioeconomy, fermentation supports resource recycling, waste minimization, and greenhouse gas reduction, contributing to the achievement of selected United Nations Sustainable Development Goals (SDGs). The importance of fermentation extends beyond its environmental aspects—fermented foods and postbiotics support the modulation of the gut microbiome, strengthen immunity, and can act as a preventative measure against metabolic and inflammatory conditions. Simultaneously, the dynamic development of precision fermentation and synthetic biology enables the design of microorganisms that produce specific food ingredients without the use of animals or traditional agriculture, paving the way for more responsible production and consumption. This review presents the categories of organic residues valorized through fermentation, explains their role in circular food and healthcare systems, and identifies key technological and regulatory barriers limiting the scaling of this approach. Collectively, fermentation emerges as a biotechnology platform with significant transformative potential for future sustainable food systems.

Candidates:
A. Fermentation, one of the oldest biotransformation processes, has become a key element of contemporary sustainable biotechnology.
B. In modern food systems, it enables the simultaneous resolution of environmental, nutritional, and economic challenges by converting agricultural and food residues into high-value-added products, such as bioactive compounds, organic acids, biofuels, enzymes, and proteins.
C. The importance of fermentation extends beyond its environmental aspects—fermented foods and postbiotics support the modulation of the gut microbiome, strengthen immunity, and can act as a preventative measure against metabolic and inflammatory conditions.
D. The evidence does not state that consistent with the concept of a circular bioeconomy, fermentation supports resource recycling, waste minimization, and greenhouse gas reduction, contributing to the achievement of selected United Nations Sustainable Development Goals (SDGs).
E. The importance of fermentation extends beyond its environmental aspects—fermented foods and postbiotics support the modulation of the gut microbiome, strengthen immunity, and cannot act as a preventative measure against metabolic and inflammatory conditions.
F. The evidence does not state that in modern food systems, it enables the simultaneous resolution of environmental, nutritional, and economic challenges by converting agricultural and food residues into high-value-added products, such as bioactive compounds, organic acids, biofuels, enzymes, and proteins.
G. The evidence does not state that fermentation, one of the oldest biotransformation processes, has become a key element of contemporary sustainable biotechnology.
H. Consistent with the concept of a circular bioeconomy, fermentation supports resource recycling, waste minimization, and greenhouse gas reduction, contributing to the achievement of selected United Nations Sustainable Development Goals (SDGs).


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 revalorization of food processing by-products represents a critical strategy for enhancing resource efficiency and advancing circularity within the food system. This review examines the potential of three major plant-based agro-industrial by-products—fruit and vegetable residues, brewer’s spent grain, and spent coffee grounds—as sources of high-value functional ingredients. These by-products contain bioactive compounds, including dietary fibers, polyphenols, proteins, peptides, oils, and antioxidants, that can be recovered using emerging green extraction and bioprocessing technologies. Conventional extraction methods are progressively being replaced or hybridized with enzyme-assisted, ultrasound-assisted, microwave-assisted, and deep eutectic solvent techniques to improve yield, reduce solvent consumption, and preserve bioactivity. The recovered compounds have demonstrated promising applications as gelling agents (pectin), natural colorants and antioxidants, protein-enriched flours, prebiotic fibers, and bioactive extracts for functional food and nutraceutical formulations. However, challenges persist in standardizing feedstock composition, scaling continuous extraction processes, ensuring safety and regulatory compliance, and generating robust techno-economic and life-cycle assessments to validate sustainability claims. This review synthesizes biochemical composition data, processing pathways, food applications, and regulatory considerations, and identifies research priorities for developing integrated, scalable biorefinery models that valorize food by-products into market-ready functional ingredients.

Candidates:
A. Conventional extraction methods are progressively being replaced or hybridized with enzyme-assisted, ultrasound-assisted, microwave-assisted, and deep eutectic solvent techniques to improve yield, reduce solvent consumption, and preserve bioactivity.
B. The revalorization of food processing by-products represents a critical strategy for enhancing resource efficiency and advancing circularity within the food system.
C. The evidence does not state that the revalorization of food processing by-products represents a critical strategy for enhancing resource efficiency and advancing circularity within the food system.
D. The evidence does not state that this review examines the potential of three major plant-based agro-industrial by-products—fruit and vegetable residues, brewer’s spent grain, and spent coffee grounds—as sources of high-value functional ingredients.
E. These by-products contain bioactive compounds, including dietary fibers, polyphenols, proteins, peptides, oils, and antioxidants, that can be recovered using emerging green extraction and bioprocessing technologies.
F. This review examines the potential of three major plant-based agro-industrial by-products—fruit and vegetable residues, brewer’s spent grain, and spent coffee grounds—as sources of high-value functional ingredients.
G. Conventional extraction methods are not progressively being replaced or hybridized with enzyme-assisted, ultrasound-assisted, microwave-assisted, and deep eutectic solvent techniques to improve yield, reduce solvent consumption, and preserve bioactivity.
H. These by-products contain bioactive compounds, including dietary fibers, polyphenols, proteins, peptides, oils, and antioxidants, that cannot be recovered using emerging green extraction and bioprocessing technologies.


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 prevalence of diabetes and its worldwide co-morbidities is escalating. Therefore, the number of users of therapeutic peptides including insulin analogs and glucagon-like peptide 1 receptor agonists (GLP-1RAs), will unavoidably increase in the coming years. However, access to these two antidiabetic classes remains limited in some countries due to their high cost. Even when available, their long-term therapeutic efficiency is often compromised by challenges in sustained treatment adherence, mainly resulting from their mode of administration through repeated subcutaneous injections. This repeated invasive delivery not only affects patient comfort but also complicates long-term disease management and monitoring. Therefore, there is an urgent need to improve the accessibility, affordability, and long-term patient adherence to insulin and GLP-1RAs. In this review, we highlight as promising alternatives the potential of plants and microalgae to serve as host organisms, as well as the use of their polysaccharides as drug carriers, for the production of low-cost and non-invasive antidiabetic drugs.

Candidates:
A. The evidence does not state that however, access to these two antidiabetic classes remains limited in some countries due to their high cost.
B. However, access to these two antidiabetic classes remains limited in some countries due to their high cost.
C. Even when available, their long-term therapeutic efficiency is not often compromised by challenges in sustained treatment adherence, mainly resulting from their mode of administration through repeated subcutaneous injections.
D. The prevalence of diabetes and its worldwide co-morbidities is escalating.
E. The prevalence of diabetes and its worldwide co-morbidities is not escalating.
F. Even when available, their long-term therapeutic efficiency is often compromised by challenges in sustained treatment adherence, mainly resulting from their mode of administration through repeated subcutaneous injections.
G. Therefore, the number of users of therapeutic peptides including insulin analogs and glucagon-like peptide 2 receptor agonists (GLP-1RAs), will unavoidably increase in the coming years.
H. Therefore, the number of users of therapeutic peptides including insulin analogs and glucagon-like peptide 1 receptor agonists (GLP-1RAs), will unavoidably increase in the coming years.


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:
Elastane is ubiquitous in polyester-based textiles and complicates depolymerization-based recycling because it can undergo thermal degradation and chemical bond cleavage, consuming reagents and forming low-molecular by-products that may compromise monomer quality. Here, we investigate alkaline PET depolymerization of PET/elastane blends under an intentional base-competition scenario in a laboratory kneader. Pure PET (100/0) and PET/EL blends (95/5 and 85/15, wt/wt) were processed under quasi-solid-state conditions at 140 °C for 5 min using solid NaOH dosed at 2.1 mol per mol PET repeat unit and pelletized feedstocks to ensure scale-relevant mixing and reproducible chamber filling. Torque and bulk-temperature profiles were similar across compositions, and isolated terephthalic acid yields remained in a narrow corridor (68–71%), indicating that PET depolymerization is not measurably impaired by 5–15 wt% elastane within this reaction window. Differential scanning calorimetry of water-insoluble residues revealed pronounced changes in elastane-related thermal transitions, evidencing elastane modification during treatment. Targeted 1 H NMR screening of recovered TA against a 4,4′-methylenedianiline spiked reference showed no detectable co-isolated aromatic diamines. Overall, the study demonstrates robust monomer recovery from mixed PET/EL textiles under solid-NaOH, short-residence, solvent-lean processing, while identifying residue analytics as the key bottleneck for quantifying elastane fate and closing component balances.

Candidates:
A. Here, we investigate alkaline PET depolymerization of PET/elastane blends under an intentional base-competition scenario in a laboratory kneader.
B. Elastane is ubiquitous in polyester-based textiles and complicates depolymerization-based recycling because it can undergo thermal degradation and chemical bond cleavage, consuming reagents and forming low-molecular by-products that may compromise monomer quality.
C. Pure PET (100/0) and PET/EL blends (95/5 and 85/15, wt/wt) were processed under quasi-solid-state conditions at 140 °C for 5 min using solid NaOH dosed at 2.1 mol per mol PET repeat unit and pelletized feedstocks to ensure scale-relevant mixing and reproducible chamber filling.
D. Torque and bulk-temperature profiles were similar across compositions, and isolated terephthalic acid yields remained in a narrow corridor (68–71%), indicating that PET depolymerization is not measurably impaired by 5–15 wt% elastane within this reaction window.
E. Pure PET (101/0) and PET/EL blends (95/5 and 85/15, wt/wt) were processed under quasi-solid-state conditions at 140 °C for 5 min using solid NaOH dosed at 2.1 mol per mol PET repeat unit and pelletized feedstocks to ensure scale-relevant mixing and reproducible chamber filling.
F. Elastane is not ubiquitous in polyester-based textiles and complicates depolymerization-based recycling because it can undergo thermal degradation and chemical bond cleavage, consuming reagents and forming low-molecular by-products that may compromise monomer quality.
G. The evidence does not state that here, we investigate alkaline PET depolymerization of PET/elastane blends under an intentional base-competition scenario in a laboratory kneader.
H. Torque and bulk-temperature profiles were similar across compositions, and isolated terephthalic acid yields remained in a narrow corridor (69–71%), indicating that PET depolymerization is not measurably impaired by 5–15 wt% elastane within this reaction window.


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 research has developed a promising edible vaccine to combat Salmonella infections in poultry, a major source of foodborne illness worldwide. Using advanced computational and immunoinformatic tools, the team designed two multi-epitope vaccine constructs targeting conserved proteins involved in bacterial adhesion and biofilm formation. The vaccines, engineered for expression in the microalga Chlorella vulgaris, incorporate immune-activating adjuvants—β-defensin-3 and lipopolysaccharide (LPS)—to stimulate robust immune responses. Structural modeling and molecular docking revealed that the LPS-based construct (Construct 2) binds strongly to Toll-like receptor 3, suggesting potent innate immune activation. Simulated immune responses showed effective IgM-to-IgG class switching and long-lasting antibody production, indicating strong protection potential. Codon optimization confirmed high expression feasibility in algae, paving the way for scalable, low-cost oral vaccine production. This innovation aligns with One Health principles, aiming to reduce antibiotic use in agriculture, enhance food safety, and mitigate antimicrobial resistance. Experimental trials are underway to validate the vaccine’s efficacy in live poultry.

Candidates:
A. Structural modeling and molecular docking revealed that the LPS-based construct (Construct 2) binds strongly to Toll-like receptor 3, suggesting potent innate immune activation.
B. Structural modeling and molecular docking revealed that the LPS-based construct (Construct 3) binds strongly to Toll-like receptor 3, suggesting potent innate immune activation.
C. The evidence does not state that this research has developed a promising edible vaccine to combat Salmonella infections in poultry, a major source of foodborne illness worldwide.
D. The vaccines, engineered for expression in the microalga Chlorella vulgaris, incorporate immune-activating adjuvants—β-defensin-3 and lipopolysaccharide (LPS)—to stimulate robust immune responses.
E. The evidence does not state that using advanced computational and immunoinformatic tools, the team designed two multi-epitope vaccine constructs targeting conserved proteins involved in bacterial adhesion and biofilm formation.
F. This research has developed a promising edible vaccine to combat Salmonella infections in poultry, a major source of foodborne illness worldwide.
G. The vaccines, engineered for expression in the microalga Chlorella vulgaris, incorporate immune-activating adjuvants—β-defensin-4 and lipopolysaccharide (LPS)—to stimulate robust immune responses.
H. Using advanced computational and immunoinformatic tools, the team designed two multi-epitope vaccine constructs targeting conserved proteins involved in bacterial adhesion and biofilm formation.


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 CO 2 capture coupled with biosurfactant production represents a promising strategy for greenhouse gas mitigation and sustainable biomanufacturing. This review examines the metabolic and engineering aspects of microbial carbon capture, focusing on both anaerobic and CO 2 -enriched systems within the Microbial-CCUS framework. The structural diversity, physicochemical properties, and industrial applications of microbial biosurfactants are discussed, along with emerging evidence of anaerobic biosurfactant synthesis linked to CO 2 metabolism. Advances in genetic and synthetic biology, pathway modularization, and systems-level modeling are reshaping the potential to coordinate CO 2 fixation with biosurfactant biosynthesis. Integrating artificial intelligence with metabolic engineering may further optimize productivity, scalability, and energy efficiency. Despite technical and economic challenges, the convergence of CO 2 utilization, biotechnology, and digital innovation offers a transformative route toward circular carbon systems and climate mitigation. • Microbial CO 2 capture drives biosurfactant synthesis within Microbial-CCUS systems . • Anaerobic and CO 2 -enriched cultures unlock new routes for sustainable biomanufacturing. • Synthetic biology links carbon-fixation modules to biosurfactant pathways . Keywords: Biosurfactants, Microbial CO 2 capture, Anaerobic metabolism, Microbial, CCUS, Circular economy, Bioprocess engineering

Candidates:
A. Advances in genetic and synthetic biology, pathway modularization, and systems-level modeling are reshaping the potential to coordinate CO 3 fixation with biosurfactant biosynthesis.
B. Microbial CO 2 capture coupled with biosurfactant production represents a promising strategy for greenhouse gas mitigation and sustainable biomanufacturing.
C. The structural diversity, physicochemical properties, and industrial applications of microbial biosurfactants are discussed, along with emerging evidence of anaerobic biosurfactant synthesis linked to CO 3 metabolism.
D. Advances in genetic and synthetic biology, pathway modularization, and systems-level modeling are reshaping the potential to coordinate CO 2 fixation with biosurfactant biosynthesis.
E. This review examines the metabolic and engineering aspects of microbial carbon capture, focusing on both anaerobic and CO 2 -enriched systems within the Microbial-CCUS framework.
F. This review examines the metabolic and engineering aspects of microbial carbon capture, focusing on both anaerobic and CO 3 -enriched systems within the Microbial-CCUS framework.
G. The structural diversity, physicochemical properties, and industrial applications of microbial biosurfactants are discussed, along with emerging evidence of anaerobic biosurfactant synthesis linked to CO 2 metabolism.
H. Microbial CO 3 capture coupled with biosurfactant production represents a promising strategy for greenhouse gas mitigation and sustainable biomanufacturing.


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:
Augmentation therapy is a treatment option available in the market that has been approved by the U.S. Food and Drug Administration (FDA) for alpha-1-antitrypsin (A1AT) deficient patients. The treatment requires weekly injections of purified A1AT for the patients and relies on plasma donor. The demand for A1AT is also high due to its functional role in various diseases. However, scaling up production of purified human plasma A1AT remained costly and challenging. It is therefore of great interest to generate A1AT at larger scale in ensuring a consistent supply to the market. In this paper, we evaluated the stability and productivity of ten Chinese Hamster Ovary (CHO) single cell clones over 12 weeks. This was followed by scaling up the fed-batch production of A1AT with the selected cell clone in a 10L single-use surface aerated orbital shaken bioreactor SB10-X. The cell specific productivity of the two bioreactor runs were at 9.6 and 12 pg/cell/day (pcd) respectively, which were comparable to shake flasks. While the paper focuses on the possibility to scale up A1AT production, process conditions such as feeding regime could be investigated to further prolong the culture longevity and increase productivity. The online version contains supplementary material available at 10.1038/s41598-026-37353-w. Keywords: Surface aerated bioreactor, Biologics, Scale up, Bioprocess development, Recombinant protein, Shear sensitive Subject terms: Biotechnology, Biologics

Candidates:
A. The demand for A2AT is also high due to its functional role in various diseases.
B. The demand for A1AT is also high due to its functional role in various diseases.
C. Augmentation therapy is a treatment option available in the market that has been approved by the U.S.
D. The treatment requires weekly injections of purified A1AT for the patients and relies on plasma donor.
E. Food and Drug Administration (FDA) for alpha-1-antitrypsin (A1AT) deficient patients.
F. Food and Drug Administration (FDA) for alpha-2-antitrypsin (A1AT) deficient patients.
G. Augmentation therapy is not a treatment option available in the market that has been approved by the U.S.
H. The treatment requires weekly injections of purified A2AT for the patients and relies on plasma donor.


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:
(+)-Bicyclogermacrene and its derivatives, with promising antimicrobial, anticancer, and insecticidal properties, hold significant potential for applications in pharmaceuticals, agriculture, and industry. However, traditional extraction methods from plant essential oils are unsustainable. In this study, we achieved the de novo biosynthesis of (+)-bicyclogermacrene using a metabolically engineered Escherichia coli strain. The biosynthetic pathway of (+)-bicyclogermacrene was partitioned into upstream and downstream modules to enable precise regulation. This was accomplished through the genome-integrated overexpression of the endogenous methylerythritol phosphate pathway to ensure an adequate supply of terpenoid precursors, which pulled the titer from the initial 11.3 mg/L to 50.1 mg/L. Production was further enhanced to 96.9 mg/L by fusion of downstream key genes to facilitate precursor channeling, along with expression level optimization to improve pathway efficiency. Additionally, NADPH supply was fine-tuned through overexpressing dehydrogenases to improve the overall metabolic balance and this approach achieved a titer of 119 mg/L. Following site-directed of (+)-bicyclogermacrene synthase, the engineered E. coli strain M6-36 produced 565 mg/L of (+)-bicyclogermacrene in a 5-L bioreactor, an approximately 50-fold increase from the initial. To the best of our knowledge, the obtained titer in this study represents the highest level ever reported for the production of (+)-bicyclogermacrene. This study demonstrates an effective approach for the heterologous biosynthesis of sesquiterpenoids in E. coli and provides a scalable platform for the sustainable production of terpenoid-derived valuable chemicals. The online version contains supplementary material available at 10.1186/s40643-026-01017-4.

Candidates:
A. The evidence does not state that (+)-Bicyclogermacrene and its derivatives, with promising antimicrobial, anticancer, and insecticidal properties, hold significant potential for applications in pharmaceuticals, agriculture, and industry.
B. However, traditional extraction methods from plant essential oils are not unsustainable.
C. The evidence does not state that in this study, we achieved the de novo biosynthesis of (+)-bicyclogermacrene using a metabolically engineered Escherichia coli strain.
D. However, traditional extraction methods from plant essential oils are unsustainable.
E. The biosynthetic pathway of (+)-bicyclogermacrene was partitioned into upstream and downstream modules to enable precise regulation.
F. In this study, we achieved the de novo biosynthesis of (+)-bicyclogermacrene using a metabolically engineered Escherichia coli strain.
G. The biosynthetic pathway of (+)-bicyclogermacrene was not partitioned into upstream and downstream modules to enable precise regulation.
H. (+)-Bicyclogermacrene and its derivatives, with promising antimicrobial, anticancer, and insecticidal properties, hold significant potential for applications in pharmaceuticals, agriculture, and industry.


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: ["1.46 g/L", "2.46 g/L"]

Evidence:
In contrast to the extensively researched animal CYP11A1 system, the catalytic mechanism of sterol side-chain cleavage by plant-derived cytochrome P450scc enzymes remains poorly understood. Through the integration of computational structural biology and enzyme channel engineering, this study successfully elucidated the key intermediates in the stepwise hydroxylation-cleavage catalytic process of Digitalis purpurea -derived DlCYP87A enzyme. Building on this foundation, we implemented structure-guided rational design to precisely engineer the substrate channel and catalytic pocket, systematically delineating their structure-activity relationships, which ultimately overcame the critical catalytic bottleneck of low conversion efficiency in heterologous microbial systems expressing plant-derived P450scc. This study established an efficient steroid synthesis system in Saccharomyces cerevisiae through integrated systematic enzyme engineering and transcriptome-guided organelle optimization. In a 5-liter fermentation system, engineered strain P4 achieved a pregnenolone titer of 1.46 g/L. This achievement represents the first gram-scale breakthrough in de novo pregnenolone biosynthesis, laying a crucial technological foundation for scalable bio-manufacturing of steroid precursors and pioneering a new industrial production pathway. Keywords: Cytochrome P450 scc , CYP87A, Steroids, Synthetic biology, Enzyme engineering


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:
Fungi play a dual role as indispensable ecological engineers and as major agents of disease in humans, animals, and plants. Recent estimates highlight their substantial impact, with millions of invasive infections annually and severe agricultural losses threatening food security. At the same time, fungi underpin ecosystem services such as decomposition, soil aggregation, and carbon sequestration, while also serving as prolific sources of enzymes, metabolites, and sustainable biomaterials. Advances in single-cell and spatial omics, cryo-electron microscopy, AlphaFold-based structural predictions, and machine learning applied to biosynthetic gene clusters are transforming the study of fungal pathogenicity, symbiosis, and metabolism. These approaches are shifting fungal research from descriptive biology toward predictive, translational pipelines that connect mechanistic insights to drug discovery, resistance management, and biotechnological innovation. Nevertheless, challenges remain, including antifungal resistance, climate-driven emergence of new pathogens, limited therapeutic options, and bottlenecks in scaling fungal applications for sustainability. Addressing these requires integrated One Health strategies that bridge clinical, agricultural, and environmental perspectives. By uniting structural biology, omics, genome editing, and computational tools within a global framework, fungal biology can be harnessed not only to mitigate emerging risks but also to drive innovations in medicine, agriculture, and green technologies.

Candidates:
A. Fungi play a dual role as indispensable ecological engineers and as major agents of disease in humans, animals, and plants.
B. Recent estimates highlight their substantial impact, with millions of invasive infections annually and severe agricultural losses threatening food security.
C. The evidence does not state that at the same time, fungi underpin ecosystem services such as decomposition, soil aggregation, and carbon sequestration, while also serving as prolific sources of enzymes, metabolites, and sustainable biomaterials.
D. At the same time, fungi underpin ecosystem services such as decomposition, soil aggregation, and carbon sequestration, while also serving as prolific sources of enzymes, metabolites, and sustainable biomaterials.
E. The evidence does not state that recent estimates highlight their substantial impact, with millions of invasive infections annually and severe agricultural losses threatening food security.
F. Advances in single-cell and spatial omics, cryo-electron microscopy, AlphaFold-based structural predictions, and machine learning applied to biosynthetic gene clusters are not transforming the study of fungal pathogenicity, symbiosis, and metabolism.
G. Advances in single-cell and spatial omics, cryo-electron microscopy, AlphaFold-based structural predictions, and machine learning applied to biosynthetic gene clusters are transforming the study of fungal pathogenicity, symbiosis, and metabolism.
H. The evidence does not state that fungi play a dual role as indispensable ecological engineers and as major agents of disease in humans, animals, and plants.


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:
Mesenchymal stem cells (MSCs) are highly valuable for their potential in cell therapy and tissue engineering because of their self-renewal, multilineage differentiation, and immunomodulatory capabilities. Adipose-derived mesenchymal stem cells (AD-MSCs) are advantageous in regenerative medicine because of their accessibility and ease of isolation. However, the clinical application of MSCs faces challenges related to large-scale culture (LSC) expansion, which is required to generate enough cells for transplantation but also decreases their therapeutic properties. This review assesses the impact of LSC on MSC functionality, differentiation potential, and immunomodulatory properties, and identifies key factors, such as metabolic shifts, genetic instability, and altered secretory profiles, that can compromise their therapeutic potential. We explored how prolonged in vitro passaging decreases MSC functionality and increases the risk of genetic alterations. In addition, strategies to preserve the efficacy of MSCs during scaling are discussed. A comprehensive literature review was conducted using PubMed, focusing on in vitro and in vivo studies that evaluated the effects of LSC on MSCs. These findings provide insights into optimizing culture protocols to maintain the clinical efficacy of AD-MSCs in regenerative therapies, addressing the critical need to balance large-scale expansion and functional integrity. Keywords: Adipose-derived stem cells, Large-scale culture expansion, Mesenchymal stem cells, Regenerative medicine, Therapeutic potential, Cell functionality


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:
Demand for recombinant proteins is rapidly growing, driven by their use as biotherapeutics, vaccine components, industrial enzymes, and food ingredients. The growing market requires novel strategies for increasing protein production in cellular hosts. Systems-level frameworks have been used to improve production, but have had difficulty relating complex cellular pathways with protein expression. Here, we demonstrate a method for mapping relationships between gene expression signatures and carbon source-related phenotypes related to recombinant protein production. Our approach induces systematic perturbations in cultures of K. phaffii using varied co-feeds of carbon sources. The different carbon sources significantly impacted cell growth, specific productivity, and transcriptional states. With these data, we identified metagenes for both immunoglobulin G1 monoclonal antibody (IgG1) and Variable domain on a heavy chain (VHH) antibody that explained significant transcriptomic variance. These metagenes strongly associated with two phenotypes: production of recombinant protein-to-biomass ratio, and response to methanol induction. We used these results to identify and knockout 31 novel gene targets for which expression inversely correlated with productivity. Nine of these genes improved productivity of IgG1 by up to 3x and ten genes increased productivity of VHH by up to 1.7x. Many of these genes are involved in the modulation and progression of the cell cycle but interestingly, disruption had little to no impact on cell growth. This study establishes a framework for relating gene signatures to complex cellular phenotypes, providing a robust methodology for assessing production processes and identifying new targets for cellular engineering. While the identified specific metagenes depend on the complexity and structure of the recombinant protein produced, this framework is extensible across diverse proteins and potentially other host organisms. These signatures may serve as scale-independent, cellular-level metrics for traits like efficiency of production of recombinant proteins, facilitating the translation of findings across different scales and cultivation modes. Furthermore, this framework enables the identification of novel targets for genomic modifications that can improve strain performance, offering a predictive tool for the rational design of high-performing microbial cell factories. The online version contains supplementary material available at 10.1186/s12934-026-02948-5. Keywords: Pichia pastoris, Systems biology, Transcriptomics, Monoclonal antibody

Candidates:
A. The evidence does not state that here, we demonstrate a method for mapping relationships between gene expression signatures and carbon source-related phenotypes related to recombinant protein production.
B. The evidence does not state that systems-level frameworks have been used to improve production, but have had difficulty relating complex cellular pathways with protein expression.
C. The evidence does not state that the growing market requires novel strategies for increasing protein production in cellular hosts.
D. Here, we demonstrate a method for mapping relationships between gene expression signatures and carbon source-related phenotypes related to recombinant protein production.
E. Demand for recombinant proteins is rapidly growing, driven by their use as biotherapeutics, vaccine components, industrial enzymes, and food ingredients.
F. Demand for recombinant proteins is not rapidly growing, driven by their use as biotherapeutics, vaccine components, industrial enzymes, and food ingredients.
G. The growing market requires novel strategies for increasing protein production in cellular hosts.
H. Systems-level frameworks have been used to improve production, but have had difficulty relating complex cellular pathways with protein expression.


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 bioconversion of fish by-products has been evidenced as a sustainable process to convert food waste into high-value products. In the present study, protein hydrolysates were produced from fish by-products by different bioprocesses and evaluated as fertilizers in wheat ( Triticum aestivum L. ) on a nitrogen-equivalent basis. Fish by-products were processed through grinding prior to bioconversion. Enzymatic hydrolysis was performed using Alcalase at 55 °C, pH 6.5, and a 3-h reaction, while microbial conversion was assessed using a lactic culture at 40 °C, pH 6.5, and a 10-day culture. Hydrolysates obtained by enzymatic and microbial bioconversion were evaluated as fertilizers by adding 30 mg after 7 and 14 days to wheat seeds sown under controlled conditions. Protease and microbial hydrolysis generated high concentrations of α -amino groups, yielding 100 mM and 170 mM, respectively. The combined process exhibited a synergistic effect, yielding 226 mM of α -amino groups and 33% of protein recovery. Plant growth assays were conducted under controlled conditions using nitrogen-equivalent doses of each hydrolysate. Microbial and combined enzymatic-microbial hydrolysates generated average plant lengths of 52 cm and 54 cm compared to 44 cm in the control, while plant biomass reached 1.7 g and 2.3 g with microbial and combined enzymatic-microbial hydrolysates compared to 0.7 g in the control. Photosynthetic parameters remained within normal physiological ranges from 2.5 to 3.3 for performance index (PI) and from 0.78 to 0.80 for maximum quantum efficiency (Fv/Fm). The integration of enzymatic and microbial catalysis produced the most effective biostimulant activity, highlighting the value of combining enzymatic specificity with microbial metabolic versatility. These findings support fish-derived protein hydrolysates as efficient and eco-friendly fertilizers that are capable of improving plant growth while contributing to sustainable and integral utilization of natural resources.

Candidates:
A. In the present study, protein hydrolysates were produced from fish by-products by different bioprocesses and evaluated as fertilizers in wheat ( Triticum aestivum L.
B. The bioconversion of fish by-products has been evidenced as a sustainable process to convert food waste into high-value products.
C. Enzymatic hydrolysis was performed using Alcalase at 55 °C, pH 6.5, and a 3-h reaction, while microbial conversion was assessed using a lactic culture at 40 °C, pH 6.5, and a 10-day culture.
D. In the present study, protein hydrolysates were not produced from fish by-products by different bioprocesses and evaluated as fertilizers in wheat ( Triticum aestivum L.
E. Enzymatic hydrolysis was performed using Alcalase at 56 °C, pH 6.5, and a 3-h reaction, while microbial conversion was assessed using a lactic culture at 40 °C, pH 6.5, and a 10-day culture.
F. The evidence does not state that the bioconversion of fish by-products has been evidenced as a sustainable process to convert food waste into high-value products.
G. Fish by-products were processed through grinding prior to bioconversion.
H. Fish by-products were not processed through grinding prior to bioconversion.


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:
Keratinous waste, a major by-product of agriculture and animal husbandry, is produced in massive quantities and is notoriously recalcitrant to degradation. With the expansion of the poultry and livestock industries, keratinous waste accumulation (e.g., feathers, hooves, and horns) has become a pressing environmental concern. Keratin’s highly cross-linked disulfide bond structure is resistant to breakdown by common proteases. Keratinase, a specialized protease capable of specifically degrading keratin, has emerges as a pivotal tool for the valorization of keratinous waste, demonstrating significant potential in waste management and resource recovery. This review systematically summarizes the enzymatic properties, mechanisms of action, and microbial sources of keratinases. It elaborates on innovative keratinase applications in waste valorization (including biogas production, the generation of bioactive peptides and amino acid feedstocks, and bioplastic manufacturing) and green industries (including leather and textile processing), as well as in the pharmaceutical, cosmetic, and detergent sectors. This review provides an in-depth discussion of the major challenges hindering industrial-scale keratinase application, including low heterologous expression efficiency and insufficient stability under industrial conditions. Finally, it outlines future research directions, encompassing protein engineering, artificial intelligence (AI)-assisted design, and multi-enzyme synergistic catalysis systems, aiming to offer forward-looking theoretical insights for advanced keratinase development and industrial application.

Candidates:
A. Keratinase, a specialized protease capable of specifically degrading keratin, has emerges as a pivotal tool for the valorization of keratinous waste, demonstrating significant potential in waste management and resource recovery.
B. The evidence does not state that keratinase, a specialized protease capable of specifically degrading keratin, has emerges as a pivotal tool for the valorization of keratinous waste, demonstrating significant potential in waste management and resource recovery.
C. Keratin’s highly cross-linked disulfide bond structure is not resistant to breakdown by common proteases.
D. Keratinous waste, a major by-product of agriculture and animal husbandry, is produced in massive quantities and is notoriously recalcitrant to degradation.
E. The evidence does not state that with the expansion of the poultry and livestock industries, keratinous waste accumulation (e.g., feathers, hooves, and horns) has become a pressing environmental concern.
F. Keratin’s highly cross-linked disulfide bond structure is resistant to breakdown by common proteases.
G. With the expansion of the poultry and livestock industries, keratinous waste accumulation (e.g., feathers, hooves, and horns) has become a pressing environmental concern.
H. Keratinous waste, a major by-product of agriculture and animal husbandry, is not produced in massive quantities and is notoriously recalcitrant to degradation.


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:
Plastics drive twin crises: persistent pollution and greenhouse gas emissions. Bio-based approaches using enzymes and microorganisms to depolymerise plastics and valorise monomers show promise but raise societal, ethical and regulatory questions central to Responsible Research and Innovation (RRI). In this Perspective, we reflect on RRI implications of bio-based plastic degradation, informed by stakeholder discussions across the plastics value chain and public engagement. We identify broad support alongside concerns about scalability, interaction with existing recycling, governance and containment of genetically modified organisms, management of additives and contaminants, and the roles of regulation and economic incentives in enabling adoption. Subject terms: Bioremediation, Environmental biotechnology

Candidates:
A. The evidence does not state that in this Perspective, we reflect on RRI implications of bio-based plastic degradation, informed by stakeholder discussions across the plastics value chain and public engagement.
B. The evidence does not state that bio-based approaches using enzymes and microorganisms to depolymerise plastics and valorise monomers show promise but raise societal, ethical and regulatory questions central to Responsible Research and Innovation (RRI).
C. We identify broad support alongside concerns about scalability, interaction with existing recycling, governance and containment of genetically modified organisms, management of additives and contaminants, and the roles of regulation and economic incentives in enabling adoption.
D. In this Perspective, we reflect on RRI implications of bio-based plastic degradation, informed by stakeholder discussions across the plastics value chain and public engagement.
E. Plastics drive twin crises: persistent pollution and greenhouse gas emissions.
F. The evidence does not state that we identify broad support alongside concerns about scalability, interaction with existing recycling, governance and containment of genetically modified organisms, management of additives and contaminants, and the roles of regulation and economic incentives in enabling adoption.
G. Bio-based approaches using enzymes and microorganisms to depolymerise plastics and valorise monomers show promise but raise societal, ethical and regulatory questions central to Responsible Research and Innovation (RRI).
H. The evidence does not state that plastics drive twin crises: persistent pollution and greenhouse gas emissions.


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:
Listeria monocytogenes is a foodborne pathogen of global concern, particularly for immunocompromised individuals at risk of severe disease. In mice, infection outcomes are strongly influenced by host immunity and gut microbiome composition. The Oligo-MM 12 defined microbiota mouse model, containing a simplified community of 12 bacterial strains, offers a controlled system to study L. monocytogenes pathogenesis and microbiome interactions. Defined or reduced-complexity microbiota models are increasingly used to investigate colonisation resistance and identify protective taxa. In this study, we compared Oligo-MM 12 mice with conventionally raised Specific Pathogen Free (SPF) mice to assess how microbiome complexity shapes infection. This allowed us to explore how microbiome complexity affects resistance to L. monocytogenes . We performed an in vivo infection study to assess host responses and pathogen-related outcomes, alongside an ex vivo fermentation assay that simulated the murine distal colon, to monitor microbial dynamics. Building on our earlier work, we now demonstrate that in vivo, Oligo-MM 12 mice showed significantly higher L. monocytogenes shedding in faeces during infection, whereas SPF mice progressively reduced L. monocytogenes levels. Despite this, L. monocytogenes dissemination to internal organs after three days of infection was similar in both models. Alterations to gut Prevotella , Akkermansia and Blautia species following L. monocytogenes infection were noteworthy. Ex vivo fermentation mirrored in vivo patterns, validating the Oligo-MM 12 system for mechanistic studies. Together, these results highlight the importance of microbiome complexity in modulating infection outcomes and establish a foundation for identifying protective taxa and mechanisms of colonization resistance. The online version contains supplementary material available at 10.1038/s41598-026-37294-4. Keywords: Listeria monocytogenes , SPF, Oligo-MM 12 , micro-Matrix bioreactor, Gut Microbiome Subject terms: Immunology, Microbiology

Candidates:
A. The Oligo-MM 12 defined microbiota mouse model, containing a simplified community of 12 bacterial strains, offers a controlled system to study L.
B. Listeria monocytogenes is not a foodborne pathogen of global concern, particularly for immunocompromised individuals at risk of severe disease.
C. Listeria monocytogenes is a foodborne pathogen of global concern, particularly for immunocompromised individuals at risk of severe disease.
D. In mice, infection outcomes are not strongly influenced by host immunity and gut microbiome composition.
E. In mice, infection outcomes are strongly influenced by host immunity and gut microbiome composition.
F. monocytogenes pathogenesis and microbiome interactions.
G. The Oligo-MM 13 defined microbiota mouse model, containing a simplified community of 12 bacterial strains, offers a controlled system to study L.
H. The evidence does not state that monocytogenes pathogenesis and microbiome interactions.


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:
Algae-derived bioactive peptides are gaining recognition as functional ingredients offering health benefits and sustainability advantages over conventional proteins. This review aimed to evaluate the current evidence on algal peptides, focusing on their nutritional content, mechanistic actions, health effects, potential for sustainability, and translational challenges. A comprehensive literature search was conducted across PubMed, Scopus, ScienceDirect, Springer, Elsevier, and Google Scholar. Peer-reviewed studies reporting bioactive peptides derived from microalgae, macroalgae, or Cyanophyceae were included. In vitro , animal, and human intervention studies evaluating molecular mechanisms, metabolic outcomes, or clinical relevance were considered. Available evidence shows that algal peptides exert multifunctional bioactivities, including inhibition of angiotensin-converting enzyme and renin, antioxidant and anti-inflammatory effects, modulation of glucose metabolism via α-amylase, α-glucosidase, and DPP-IV inhibition, and regulation of lipid metabolism and adipogenesis. Frequently studied sources included Limnospira , Chlorella , Auxenochlorella , Nannochloropsis , Undaria , Palmaria , Ulva , and Neopyropia . Limited human trials suggest modest but clinically relevant improvements in blood pressure, glycemic control, lipid profiles, and body-weight-related outcomes, primarily using whole algal biomass or extracts. Life-cycle assessments highlight favorable land-use efficiency and carbon sequestration potential, although economic feasibility is constrained by energy-intensive downstream processing. Algal-derived peptides demonstrate promising health-promoting effects and align with sustainable nutrition goals. However, their clinical translation is limited by variability in peptide characterization, uncertain bioavailability, and lack of robust human trials. Standardized production methods, improved delivery strategies, comprehensive safety assessments, and well-designed clinical studies are essential to support their application in functional foods and nutraceuticals.

Candidates:
A. Algae-derived bioactive peptides are not gaining recognition as functional ingredients offering health benefits and sustainability advantages over conventional proteins.
B. Algae-derived bioactive peptides are gaining recognition as functional ingredients offering health benefits and sustainability advantages over conventional proteins.
C. Peer-reviewed studies reporting bioactive peptides derived from microalgae, macroalgae, or Cyanophyceae were not included.
D. This review aimed to evaluate the current evidence on algal peptides, focusing on their nutritional content, mechanistic actions, health effects, potential for sustainability, and translational challenges.
E. The evidence does not state that this review aimed to evaluate the current evidence on algal peptides, focusing on their nutritional content, mechanistic actions, health effects, potential for sustainability, and translational challenges.
F. Peer-reviewed studies reporting bioactive peptides derived from microalgae, macroalgae, or Cyanophyceae were included.
G. A comprehensive literature search was not conducted across PubMed, Scopus, ScienceDirect, Springer, Elsevier, and Google Scholar.
H. A comprehensive literature search was conducted across PubMed, Scopus, ScienceDirect, Springer, Elsevier, and Google Scholar.


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: ["3 h", "2 h"]

Evidence:
Neoscytalidium dimidiatum is a non-dermatophyte mold that commonly causes skin and nail infections in tropical regions and often resists conventional antifungal therapies. Because its clinical and laboratory features often resemble dermatophyte infections, diagnosis is frequently delayed and treatment is sometimes inappropriate. We therefore developed a dot-immunobinding assay (Dot-Iba) to detect N. dimidiatum antigens. We generated a highly specific monoclonal antibody, 3E6F7 (MAb 3E6F7), for antigen capture, and used goat anti-mouse Ig conjugated with alkaline phosphatase (AP) as the signal generator. The test pad comprised a test hole, a nitrocellulose membrane (NC), and water-absorbent pads in a vertical flow-through format to allow a rapid antigen–antibody reaction. The assembled system detected N. dimidiatum antigens in vitro with high specificity and yielded visible results within 2 h; its detection limit was 0.9 µg without cross-reactivity to dermatophyte or non-dermatophyte fungi. This rapid, specific, and easy-to-use assay shows strong potential as a diagnostic tool, particularly in settings with limited access to fungal culture or advanced molecular diagnostics, where early, accurate identification is crucial. Keywords: Dot-immunobinding assay (Dot-Iba), Fungal foot infection, Nail infection, Neoscytalidium dimidiatum , Rapid test


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:
Pork carcass fabrication is a central determinant of value realization within packing and processing systems, translating biological variation in carcass weight and composition into economic outcomes under dynamic market conditions. The objective of this review is to synthesize current knowledge on the economic drivers of pork carcass fabrication, with a specific focus on the interactions among carcass characteristics, fabrication strategies, and value realization. Regional differences in cutting specifications and market orientation are discussed as key factors shaping primal yields, market allocation, and value distribution across domestic and export channels. The economic contributions of primals, subprimals, trim, fat, and by-products are examined in the context of wholesale pricing signals, carcass merit programs, and packer-specific specifications that link production decisions with downstream processing requirements. Fabrication strategies, including depth of fabrication and primal-specific cutting decisions, are evaluated with respect to yield optimization, labor and packaging costs, and market flexibility. The influence of carcass weight and composition on fabrication efficiency, trim generation, and fixed cost allocation is highlighted, illustrating trade-offs between biological performance and processing constraints. Technological advancements, including instrument grading, automation, and data integration, are reviewed for their role in improving yield prediction, carcass sorting, and operational consistency, while emerging tools such as predictive modeling are identified as promising approaches for managing variability and economic risk. Price volatility, biological variability, and supply chain disruptions are identified as persistent challenges to fabrication economics, underscoring the need for resilient and adaptable processing systems. Beyond economic performance, fabrication decisions are discussed in relation to labor welfare and sustainability outcomes. Collectively, this review emphasizes that optimal pork carcass fabrication is achieved through the strategic integration of biological inputs, economic signals, and operational capabilities. Improved data transparency and collaboration between industry professionals are essential to develop integrated biological-economic frameworks that enhance value realization and long-term sustainability across the pork supply chain.

Candidates:
A. The objective of this review is not to synthesize current knowledge on the economic drivers of pork carcass fabrication, with a specific focus on the interactions among carcass characteristics, fabrication strategies, and value realization.
B. The objective of this review is to synthesize current knowledge on the economic drivers of pork carcass fabrication, with a specific focus on the interactions among carcass characteristics, fabrication strategies, and value realization.
C. Pork carcass fabrication is a central determinant of value realization within packing and processing systems, translating biological variation in carcass weight and composition into economic outcomes under dynamic market conditions.
D. The economic contributions of primals, subprimals, trim, fat, and by-products are not examined in the context of wholesale pricing signals, carcass merit programs, and packer-specific specifications that link production decisions with downstream processing requirements.
E. Regional differences in cutting specifications and market orientation are discussed as key factors shaping primal yields, market allocation, and value distribution across domestic and export channels.
F. The economic contributions of primals, subprimals, trim, fat, and by-products are examined in the context of wholesale pricing signals, carcass merit programs, and packer-specific specifications that link production decisions with downstream processing requirements.
G. Regional differences in cutting specifications and market orientation are not discussed as key factors shaping primal yields, market allocation, and value distribution across domestic and export channels.
H. Pork carcass fabrication is not a central determinant of value realization within packing and processing systems, translating biological variation in carcass weight and composition into economic outcomes under dynamic market conditions.


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 integration of artificial intelligence into medicine has led to significant advances, particularly in diagnostics and treatment planning. However, the reliability of AI models is highly dependent on the quality of the training data, especially in medical imaging, where varying patient data and evolving medical knowledge pose a challenge to the accuracy and generalizability of given datasets. The proposed approach focuses on the integration and enhancement of clinical computed tomography (CT) image series for better findability, accessibility, interoperability, and reusability. Through an automated indexing process, CT image series are semantically enhanced using the TotalSegmentator framework for segmentation and resulting SNOMED CT annotations. The metadata is standardized with HL7 FHIR resources to enable efficient data recognition and data exchange between research projects. The study successfully integrates a robust process within the UKSH MeDIC, leading to the semantic enrichment of over 1.7 million CT image series and over 50 million SNOMED CT annotations. The standardized representation using HL7 FHIR resources improves discoverability and facilitates interoperability, providing a foundation for the FAIRness of medical imaging data. However, developing automated annotation methods that can keep pace with growing clinical datasets remains a challenge to ensure continued progress in large-scale integration and indexing of medical imaging for advanced healthcare AI applications. Keywords: Data standardization, Semantic interoperability, Artificial intelligence, Medical image processing, Computed tomography

Candidates:
A. Through an automated indexing process, CT image series are semantically enhanced using the TotalSegmentator framework for segmentation and resulting SNOMED CT annotations.
B. The evidence does not state that the integration of artificial intelligence into medicine has led to significant advances, particularly in diagnostics and treatment planning.
C. However, the reliability of AI models is highly dependent on the quality of the training data, especially in medical imaging, where varying patient data and evolving medical knowledge pose a challenge to the accuracy and generalizability of given datasets.
D. The integration of artificial intelligence into medicine has led to significant advances, particularly in diagnostics and treatment planning.
E. The proposed approach focuses on the integration and enhancement of clinical computed tomography (CT) image series for better findability, accessibility, interoperability, and reusability.
F. Through an automated indexing process, CT image series are not semantically enhanced using the TotalSegmentator framework for segmentation and resulting SNOMED CT annotations.
G. The evidence does not state that the proposed approach focuses on the integration and enhancement of clinical computed tomography (CT) image series for better findability, accessibility, interoperability, and reusability.
H. However, the reliability of AI models is not highly dependent on the quality of the training data, especially in medical imaging, where varying patient data and evolving medical knowledge pose a challenge to the accuracy and generalizability of given datasets.


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: ["83.51%", "83.37%", "82.37%", "89.37%", "78.35%", "84.51%", "88.37%", "77.35%"]

Evidence:
Tilapia significantly contributes to global food security and is an affordable protein source for most developing nations. Tilapia tilapinevirus (TiLV) poses a significant economic threat to the global tilapia industry. This study aimed to develop a rapid and accurate detection method for TiLV by synthesizing a monoclonal antibody (MAb) against it. A novel peptide, KLH-CQ, derived from the TiLV sequence, was designed considering physicochemical properties like net cationic charge, amphipathicity, helicity, and hydrophobicity. The KLH-CQ (50 µg) was used to immunize Balb/c mice with Freund's complete adjuvant. The presence of specific antibodies in the mice serum was confirmed by ELISA, which showed a high antibody titre of 2.67:0.12 (mean OD of treated Vs control sera). The mouse with the strongest immune response was used for spleen donor in hybridoma production. Epitope mapping via ELISA screening identified five positive clones (TiLV-MAb 1–5), with the most reactive clone selected for further analysis. Using Classen's method, a cutoff OD value of 1.24 ± 0.45 was determined for virus detection. The selected TiLV-MAb was then used as a probing antibody to develop a latex slide agglutination assay (TiLV-LAT) using passive adsorption method. Validation of the assay with tissue and mucus samples revealed a specificity of 88.37% and a sensitivity of 82.37% for TiLV detection. The overall accuracy of the assay was 83.51%, with positive and negative likelihood ratios of 7.06 and 0.2, respectively. The TiLV-LAT successfully detected TiLV in various tissues, showing variable sensitivity: liver (77.35%), mucus (73.53%), brain (67.92%), and kidney (62.26%). TiLV-LAT developed here has minimized the tedious steps involved in nucleic acid-based detection assays, with the recorded sensitivity and specificity; it can be used as a presumptive diagnosis for testing and point of care/farm site. Moreover, non-lethal sampling and virus testing in mucus samples would be useful for fish health monitoring.


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:
Extremophiles are microorganisms that thrive in environments previously thought to be uninhabitable, including extreme temperature, salinity, pH, pressure, and radiation. These organisms, found in Archaea, Bacteria, and Eukarya, exhibit distinct structural, metabolic, and genetic adaptations, such as enhanced enzyme stability, efficient DNA repair mechanisms, and robust stress-response systems that enable survival under extreme conditions. Understanding these adaptation mechanisms is key to engineering similar traits in mesophilic organisms. This review discusses the diversity of extremophiles and presents phylogenetic and comparative genomic insights which may provide insights into the origins and evolution of early life on Earth We highlight recent advances in CRISPR/Cas-based genome editing, genome-scale metabolic modeling (GEM), and synthetic biology that have expanded the use of extremophiles in sustainable industrial biotechnology. The exceptional stability and catalytic efficiency of extremozymes under harsh conditions underscore their potential in various biotechnological applications. Finally, we discuss the ecological significance of extremophiles in climate change mitigation and outline current challenges and future directions in extremophile research.

Candidates:
A. Extremophiles are microorganisms that thrive in environments previously thought to be uninhabitable, including extreme temperature, salinity, pH, pressure, and radiation.
B. Extremophiles are not microorganisms that thrive in environments previously thought to be uninhabitable, including extreme temperature, salinity, pH, pressure, and radiation.
C. The evidence does not state that these organisms, found in Archaea, Bacteria, and Eukarya, exhibit distinct structural, metabolic, and genetic adaptations, such as enhanced enzyme stability, efficient DNA repair mechanisms, and robust stress-response systems that enable survival under extreme conditions.
D. Understanding these adaptation mechanisms is not key to engineering similar traits in mesophilic organisms.
E. The evidence does not state that this review discusses the diversity of extremophiles and presents phylogenetic and comparative genomic insights which may provide insights into the origins and evolution of early life on Earth We highlight recent advances in CRISPR/Cas-based genome editing, genome-scale metabolic modeling (GEM), and synthetic biology that have expanded the use of extremophiles in sustainable industrial biotechnology.
F. Understanding these adaptation mechanisms is key to engineering similar traits in mesophilic organisms.
G. These organisms, found in Archaea, Bacteria, and Eukarya, exhibit distinct structural, metabolic, and genetic adaptations, such as enhanced enzyme stability, efficient DNA repair mechanisms, and robust stress-response systems that enable survival under extreme conditions.
H. This review discusses the diversity of extremophiles and presents phylogenetic and comparative genomic insights which may provide insights into the origins and evolution of early life on Earth We highlight recent advances in CRISPR/Cas-based genome editing, genome-scale metabolic modeling (GEM), and synthetic biology that have expanded the use of extremophiles in sustainable industrial biotechnology.


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:
Cancer treatment and imaging are still limited since many drugs and imaging agents can neither effectively nor selectively reach tumour tissues. Therefore, new strategies are needed to improve drug and imaging agent delivery and reduce side effects. This review focuses on targeted protein-based nanocarriers as innovative devices for cancer diagnosis and therapy (theranostics), capable of delivering drug(s) and imaging agent(s) simultaneously. We discuss overexpressed protein receptors in cancer cells that differ from normal tissue expression and can be exploited for targeted delivery. This review summarises recent preclinical studies using protein nanocarriers as targeted theranostic platforms to improve cancer treatment, reduce side effects, and enable non-invasive tracking of treatment progress. Overall, protein nanocarriers represent promising devices that combine imaging modalities and targeting strategies for more effective cancer diagnosis and therapy in the future.

Candidates:
A. Cancer treatment and imaging are still limited since many drugs and imaging agents can neither effectively nor selectively reach tumour tissues.
B. Cancer treatment and imaging are not still limited since many drugs and imaging agents can neither effectively nor selectively reach tumour tissues.
C. We discuss overexpressed protein receptors in cancer cells that differ from normal tissue expression and can be exploited for targeted delivery.
D. Therefore, new strategies are needed to improve drug and imaging agent delivery and reduce side effects.
E. We discuss overexpressed protein receptors in cancer cells that differ from normal tissue expression and cannot be exploited for targeted delivery.
F. This review focuses on targeted protein-based nanocarriers as innovative devices for cancer diagnosis and therapy (theranostics), capable of delivering drug(s) and imaging agent(s) simultaneously.
G. The evidence does not state that this review focuses on targeted protein-based nanocarriers as innovative devices for cancer diagnosis and therapy (theranostics), capable of delivering drug(s) and imaging agent(s) simultaneously.
H. Therefore, new strategies are not needed to improve drug and imaging agent delivery and reduce side effects.


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:
Tulathromycin (TULA) is primarily used for treating respiratory diseases in livestock. However, its misuse may lead to bacterial resistance and poses potential health risks such as chronic toxicity and allergic reactions through the food chain. Therefore, it is essential to develop rapid and accurate detection methods. In this study, two quantum dot-based fluorescent immunosorbent assays—direct competitive FLISA (dc-FLISA) and indirect competitive FLISA (ic-FLISA)—were established for detecting TULA residues in milk. The dc-FLISA exhibited a half-maximal inhibitory concentration (IC 50 ) of 1.99 ng·mL −1 , a limit of detection (LOD) of 0.018 ng·mL −1 , and a detection range of 0.058–69.18 ng·mL −1 . The ic-FLISA showed an IC 50 of 0.89 ng·mL −1 , an LOD of 0.005 ng·mL −1 , and a detection range of 0.019–42.65 ng·mL −1 . Spiked recovery tests in milk demonstrated recovery rates ranging from 97.41% to 101.02% for dc-FLISA and from 97.48% to 100.65% for ic-FLISA, with coefficients of variation below 10%. In summary, two simple, effective, rapid, and sensitive methods were successfully developed for detecting TULA residues in milk.

Candidates:
A. However, its misuse may lead to bacterial resistance and poses potential health risks such as chronic toxicity and allergic reactions through the food chain.
B. The evidence does not state that in this study, two quantum dot-based fluorescent immunosorbent assays—direct competitive FLISA (dc-FLISA) and indirect competitive FLISA (ic-FLISA)—were established for detecting TULA residues in milk.
C. Therefore, it is essential to develop rapid and accurate detection methods.
D. In this study, two quantum dot-based fluorescent immunosorbent assays—direct competitive FLISA (dc-FLISA) and indirect competitive FLISA (ic-FLISA)—were established for detecting TULA residues in milk.
E. Tulathromycin (TULA) is primarily used for treating respiratory diseases in livestock.
F. Tulathromycin (TULA) is not primarily used for treating respiratory diseases in livestock.
G. The evidence does not state that however, its misuse may lead to bacterial resistance and poses potential health risks such as chronic toxicity and allergic reactions through the food chain.
H. Therefore, it is not essential to develop rapid and accurate detection methods.


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:
Algae-derived proteins and peptides have gained increasing interest as sustainable bioresources with valuable nutritional and functional properties. This review aims to synthesize current knowledge on their characteristics and applications while highlighting the emerging role of computational tools in peptide research. Key findings show that algae provide diverse proteins and bioactive peptides with advantageous amino acid profiles and notable antioxidant, antihypertensive, antidiabetic, anti-inflammatory, and skin-protective activities. Their applications span food formulation, pharmaceuticals, and cosmetics, although large-scale utilization remains constrained by production, stability, and bioavailability challenges. Computational strategies, including virtual enzymatic hydrolysis, machine-learning prediction, QSAR modeling, molecular docking, molecular dynamics, and toxicity/allergenicity assessment, offer promising avenues for efficient peptide discovery, though their use in algae is still limited. Overall, this review underscores the potential of algae-derived proteins and peptides as multifunctional ingredients and emphasizes the need to integrate in silico pipelines with improved processing and delivery systems to accelerate future translational applications.

Candidates:
A. The evidence does not state that their applications span food formulation, pharmaceuticals, and cosmetics, although large-scale utilization remains constrained by production, stability, and bioavailability challenges.
B. The evidence does not state that algae-derived proteins and peptides have gained increasing interest as sustainable bioresources with valuable nutritional and functional properties.
C. The evidence does not state that key findings show that algae provide diverse proteins and bioactive peptides with advantageous amino acid profiles and notable antioxidant, antihypertensive, antidiabetic, anti-inflammatory, and skin-protective activities.
D. Algae-derived proteins and peptides have gained increasing interest as sustainable bioresources with valuable nutritional and functional properties.
E. Their applications span food formulation, pharmaceuticals, and cosmetics, although large-scale utilization remains constrained by production, stability, and bioavailability challenges.
F. Key findings show that algae provide diverse proteins and bioactive peptides with advantageous amino acid profiles and notable antioxidant, antihypertensive, antidiabetic, anti-inflammatory, and skin-protective activities.
G. The evidence does not state that this review aims to synthesize current knowledge on their characteristics and applications while highlighting the emerging role of computational tools in peptide research.
H. This review aims to synthesize current knowledge on their characteristics and applications while highlighting the emerging role of computational tools in peptide research.


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/Objectives : Multiple sclerosis (MS) imposes a substantial clinical, humanistic, and economic burden, and current disease-modifying therapies require lifelong administration without restoring immune tolerance. IMMUTOL, a tolerogenic gene therapy under development within an EU-funded programme, aims to induce durable remission. Methods : This study assessed the early financial feasibility of IMMUTOL using a structured risk-adjusted net present value (rNPV) model, incorporating development and operating costs, probabilities of clinical and regulatory success, manufacturing expenditure, market dynamics, and revenue projections. Uncertainty was examined through one-way, probabilistic, and scenario analyses. Results : Under base-case assumptions, IMMUTOL generated a deterministic rNPV of −$223.8 million with an internal rate of return of 3.4%. Probabilistic analysis yielded a mean rNPV of −$99.4 million and a mean internal rate of return of 10.5%, with 70.2% of simulations producing negative values. Only scenarios combining higher treatment prices with lower manufacturing costs produced consistently positive rNPVs; a price of $1.5 million with a $200,000 production cost resulted in an rNPV of $711.2 million and an internal rate of return of 20.7%. Neither increased market size, reduced time to approval, nor modest cost reductions altered the conclusion. Conclusions : These findings emphasise a structural gap between value-based pricing and the pricing required for commercial viability. Without external support or reductions in cost structures, commercial development may be economically unattractive.

Candidates:
A. Methods : This study assessed the early financial feasibility of IMMUTOL using a structured risk-adjusted net present value (rNPV) model, incorporating development and operating costs, probabilities of clinical and regulatory success, manufacturing expenditure, market dynamics, and revenue projections.
B. The evidence does not state that methods : This study assessed the early financial feasibility of IMMUTOL using a structured risk-adjusted net present value (rNPV) model, incorporating development and operating costs, probabilities of clinical and regulatory success, manufacturing expenditure, market dynamics, and revenue projections.
C. The evidence does not state that iMMUTOL, a tolerogenic gene therapy under development within an EU-funded programme, aims to induce durable remission.
D. IMMUTOL, a tolerogenic gene therapy under development within an EU-funded programme, aims to induce durable remission.
E. The evidence does not state that background/Objectives : Multiple sclerosis (MS) imposes a substantial clinical, humanistic, and economic burden, and current disease-modifying therapies require lifelong administration without restoring immune tolerance.
F. Background/Objectives : Multiple sclerosis (MS) imposes a substantial clinical, humanistic, and economic burden, and current disease-modifying therapies require lifelong administration without restoring immune tolerance.
G. Uncertainty was examined through one-way, probabilistic, and scenario analyses.
H. Uncertainty was not examined through one-way, probabilistic, and scenario analyses.


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:
Testosterone is a vital steroid hormone with important physiological roles and broad clinical significance, serving as a central molecular precursor in the synthesis of many pharmacologically active steroids. Testosterone is traditionally produced through complex chemical synthesis routes that involve hazardous reagents, harsh conditions, and produce significant toxic waste. In recent decades, growing regulatory requirements and environmental sustainability goals have spurred the development of alternative biotechnological methods that use microbial biotransformation. This review offers a comparative analysis of chemical and biological methods for producing testosterone, focusing on microbial steroid biotransformation pathways and the key enzymatic steps involved in testosterone biosynthesis. It examines key advances in sterol breakdown, pathway engineering, and enzyme driven modifications, including the roles of 17β-hydroxysteroid dehydrogenases and cytochrome P450 monooxygenases. The performance, specificity, and environmental impacts of bacterial and fungal cells as cell factories, especially Mycolicibacterium and Aspergillus species, are critically analyzed within the framework of modern green chemistry principles. Overall, by combining molecular insights with process considerations, this review illustrates how microbial platforms could complement and gradually transform traditional chemical synthesis methods, promoting a shift toward more sustainable steroid hormone production through engineered biocatalysts.

Candidates:
A. This review offers a comparative analysis of chemical and biological methods for producing testosterone, focusing on microbial steroid biotransformation pathways and the key enzymatic steps involved in testosterone biosynthesis.
B. Testosterone is not a vital steroid hormone with important physiological roles and broad clinical significance, serving as a central molecular precursor in the synthesis of many pharmacologically active steroids.
C. Testosterone is a vital steroid hormone with important physiological roles and broad clinical significance, serving as a central molecular precursor in the synthesis of many pharmacologically active steroids.
D. The evidence does not state that this review offers a comparative analysis of chemical and biological methods for producing testosterone, focusing on microbial steroid biotransformation pathways and the key enzymatic steps involved in testosterone biosynthesis.
E. In recent decades, growing regulatory requirements and environmental sustainability goals have spurred the development of alternative biotechnological methods that use microbial biotransformation.
F. Testosterone is not traditionally produced through complex chemical synthesis routes that involve hazardous reagents, harsh conditions, and produce significant toxic waste.
G. The evidence does not state that in recent decades, growing regulatory requirements and environmental sustainability goals have spurred the development of alternative biotechnological methods that use microbial biotransformation.
H. Testosterone is traditionally produced through complex chemical synthesis routes that involve hazardous reagents, harsh conditions, and produce significant toxic waste.


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:
Artificial intelligence (AI), including machine learning (ML) and deep learning (DL), is increasingly transforming the study, manufacturing, and clinical translation of adipose-derived stem/stromal cells (ADSCs). ADSC-based therapies face persistent challenges related to donor variability, heterogeneous cell populations, limited standardization of culture protocols, and the need for robust quality control (QC) and potency assessment under Good Manufacturing Practice (GMP) conditions. This review discusses how AI-driven approaches can support the ADSC pipeline from donor and tissue pre-screening, through isolation and expansion, to differentiation and batch release decisions. We highlight major methodological advances in computer vision and label-free imaging for monitoring morphology, confluency, proliferation, senescence, and contamination, as well as AI-assisted optimization strategies for culture parameters and differentiation protocols. In addition, we examine the growing role of multi-omics integration (transcriptomics, proteomics, metabolomics, and secretomics) combined with ML to predict functional potency, stratify donors, and identify biomarkers associated with therapeutic efficacy. Finally, we address current limitations, including data scarcity, inter-laboratory variability, model interpretability, and regulatory requirements, and outline future perspectives such as closed-loop bioprocess control, foundation models, and federated learning frameworks. Overall, AI offers a powerful toolkit to improve the reproducibility, safety, and scalability of ADSC manufacturing and to accelerate the development of standardized, data-driven regenerative medicine products.

Candidates:
A. This review discusses how AI-driven approaches cannot support the ADSC pipeline from donor and tissue pre-screening, through isolation and expansion, to differentiation and batch release decisions.
B. The evidence does not state that we highlight major methodological advances in computer vision and label-free imaging for monitoring morphology, confluency, proliferation, senescence, and contamination, as well as AI-assisted optimization strategies for culture parameters and differentiation protocols.
C. This review discusses how AI-driven approaches can support the ADSC pipeline from donor and tissue pre-screening, through isolation and expansion, to differentiation and batch release decisions.
D. Artificial intelligence (AI), including machine learning (ML) and deep learning (DL), is not increasingly transforming the study, manufacturing, and clinical translation of adipose-derived stem/stromal cells (ADSCs).
E. The evidence does not state that aDSC-based therapies face persistent challenges related to donor variability, heterogeneous cell populations, limited standardization of culture protocols, and the need for robust quality control (QC) and potency assessment under Good Manufacturing Practice (GMP) conditions.
F. Artificial intelligence (AI), including machine learning (ML) and deep learning (DL), is increasingly transforming the study, manufacturing, and clinical translation of adipose-derived stem/stromal cells (ADSCs).
G. We highlight major methodological advances in computer vision and label-free imaging for monitoring morphology, confluency, proliferation, senescence, and contamination, as well as AI-assisted optimization strategies for culture parameters and differentiation protocols.
H. ADSC-based therapies face persistent challenges related to donor variability, heterogeneous cell populations, limited standardization of culture protocols, and the need for robust quality control (QC) and potency assessment under Good Manufacturing Practice (GMP) conditions.